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Sample records for expression systems yield

  1. Choosing Between Yeast and Bacterial Expression Systems: Yield Dependent

    NASA Technical Reports Server (NTRS)

    Miller, Rebecca S.; Malone, Christine C.; Moore, Blake P.; Burk, Melissa; Crawford, Lisa; Karr, Laurel J.; Curreri, Peter A. (Technical Monitor)

    2002-01-01

    Green fluorescent protein (GFP) is a naturally occurring fluorescent protein isolated from the jellyfish Aequorea victoria. The intrinsic fluorescence of the protein is due to a chromophore located in the center of the molecule. Its usefulness has been established as a marker for gene expression and localization of gene products. GFP has recently been utilized as a model protein for crystallization studies at NASA/MSFC, both in earth-based and in microgravity experiments. Because large quantities of purified protein were needed, the cDNA of GFP was cloned into the Pichia pastoris pPICZ(alpha) C strain, with very little protein secreted into the media. Microscopic analysis prior to harvest showed gigantic green fluorescent yeast, but upon harvesting most protein was degraded. Trial fermentations of GFP cloned into pPICZ A for intracellular expression provided unsatisfactory yield. GFP cloned into E, coli was overexpressed at greater than 150 mg/liter, with purification yields at greater than 100mg/liter.

  2. Choosing Between Yeast and Bacterial Expression Systems: Yield Dependent

    NASA Technical Reports Server (NTRS)

    Miller, Rebecca S.; Malone, Christine C.; Moore, Blake P.; Burk, Melissa; Crawford, Lisa; Karr, Laurel J.; Curreri, Peter A. (Technical Monitor)

    2002-01-01

    Green fluorescent protein (GFP) is a naturally occurring fluorescent protein isolated from the jellyfish Aequorea victoria. The intrinsic fluorescence of the protein is due to a chromophore located in the center of the molecule. Its usefulness has been established as a marker for gene expression and localization of gene products. GFP has recently been utilized as a model protein for crystallization studies at NASA/MSFC, both in earth-based and in microgravity experiments. Because large quantities of purified protein were needed, the cDNA of GFP was cloned into the Pichia pastoris pPICZ(alpha) C strain, with very little protein secreted into the media. Microscopic analysis prior to harvest showed gigantic green fluorescent yeast, but upon harvesting most protein was degraded. Trial fermentations of GFP cloned into pPICZ A for intracellular expression provided unsatisfactory yield. GFP cloned into E, coli was overexpressed at greater than 150 mg/liter, with purification yields at greater than 100mg/liter.

  3. Design and implementation of a high yield production system for recombinant expression of peptides

    PubMed Central

    2014-01-01

    Background Making peptide pharmaceuticals involves challenging processes where many barriers, which include production and manufacture, need to be overcome. A non common but interesting research area is related to peptides with intracellular targets, which opens up new possibilities, allowing the modulation of processes occurring within the cell or interference with signaling pathways. However, if the bioactive sequence requires fusion to a carrier peptide to allow access into the cell, the resulting peptide could be such a length that traditional production could be difficult. The goal of the present study was the development of a flexible recombinant expression and purification system for peptides, as a contribution to the discovery and development of these potentially new drugs. Results In this work, a high throughput recombinant expression and purification system for production of cell penetrating peptides in Escherichia coli has been designed and implemented. The system designed produces target peptides in an insoluble form by fusion to a hexahistidine tagged ketosteroid isomerase which is then separated by a highly efficient thrombin cleavage reaction procedure. The expression system was tested on the anticancer peptides p53pAnt and PNC27. These peptides comprise the C-terminal region and the N-terminal region of the protein p53, respectively, fused by its carboxyl terminal extreme to the cell penetrating peptide Penetratin. High yields of purified recombinant fused peptides were obtained in both cases; nevertheless, thrombin cleavage reaction was successful only for p53pAnt peptide release. The features of the system, together with the procedure developed, allow achievement of high production yields of over 30 mg of highly pure p53pAnt peptide per g of dry cell mass. It is proposed that the system could be used for production of other peptides at a similar yield. Conclusions This study provides a system suitable for recombinant production of peptides for

  4. A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells.

    PubMed

    Dotiwala, Farokh; Fellay, Isabelle; Filgueira, Luis; Martinvalet, Denis; Lieberman, Judy; Walch, Michael

    2015-06-10

    When cytotoxic T lymphocytes (CTL) or natural killer (NK) cells recognize tumor cells or cells infected with intracellular pathogens, they release their cytotoxic granule content to eliminate the target cells and the intracellular pathogen. Death of the host cells and intracellular pathogens is triggered by the granule serine proteases, granzymes (Gzms), delivered into the host cell cytosol by the pore forming protein perforin (PFN) and into bacterial pathogens by the prokaryotic membrane disrupting protein granulysin (GNLY). To investigate the molecular mechanisms of target cell death mediated by the Gzms in experimental in-vitro settings, protein expression and purification systems that produce high amounts of active enzymes are necessary. Mammalian secreted protein expression systems imply the potential to produce correctly folded, fully functional protein that bears posttranslational modification, such as glycosylation. Therefore, we used a cost-efficient calcium precipitation method for transient transfection of HEK293T cells with human Gzms cloned into the expression plasmid pHLsec. Gzm purification from the culture supernatant was achieved by immobilized nickel affinity chromatography using the C-terminal polyhistidine tag provided by the vector. The insertion of an enterokinase site at the N-terminus of the protein allowed the generation of active protease that was finally purified by cation exchange chromatography. The system was tested by producing high levels of cytotoxic human Gzm A, B and M and should be capable to produce virtually every enzyme in the human body in high yields.

  5. Wheat germ cell-free expression system as a pathway to improve protein yield and solubility for the SSGCID pipeline.

    PubMed

    Guild, Katherine; Zhang, Yang; Stacy, Robin; Mundt, Elizabeth; Benbow, Sarah; Green, Amanda; Myler, Peter J

    2011-09-01

    Recombinant expression of proteins of interest in Escherichia coli is an important tool in the determination of protein structure. However, lack of expression and insolubility remain significant challenges to the expression and crystallization of these proteins. The SSGCID program uses a wheat germ cell-free expression system as a rescue pathway for proteins that are either not expressed or insoluble when produced in E. coli. Testing indicates that the system is a valuable tool for these protein targets. Further increases in solubility were obtained by the addition of the NVoy polymer reagent to the reaction mixture. These data indicate that this eukaryotic cell-free expression system has a high success rate and that the addition of specific reagents can increase the yield of soluble protein.

  6. Increasing the yield of middle silk gland expression system through transgenic knock-down of endogenous sericin-1.

    PubMed

    Ma, Sanyuan; Xia, Xiaojuan; Li, Yufeng; Sun, Le; Liu, Yue; Liu, Yuanyuan; Wang, Xiaogang; Shi, Run; Chang, Jiasong; Zhao, Ping; Xia, Qingyou

    2017-08-01

    Various genetically modified bioreactor systems have been developed to meet the increasing demands of recombinant proteins. Silk gland of Bombyx mori holds great potential to be a cost-effective bioreactor for commercial-scale production of recombinant proteins. However, the actual yields of proteins obtained from the current silk gland expression systems are too low for the proteins to be dissolved and purified in a large scale. Here, we proposed a strategy that reducing endogenous sericin proteins would increase the expression yield of foreign proteins. Using transgenic RNA interference, we successfully reduced the expression of BmSer1 to 50%. A total 26 transgenic lines expressing Discosoma sp. red fluorescent protein (DsRed) in the middle silk gland (MSG) under the control of BmSer1 promoter were established to analyze the expression of recombinant. qRT-PCR and western blotting showed that in BmSer1 knock-down lines, the expression of DsRed had significantly increased both at mRNA and protein levels. We did an additional analysis of DsRed/BmSer1 distribution in cocoon and effect of DsRed protein accumulation on the silk fiber formation process. This study describes not only a novel method to enhance recombinant protein expression in MSG bioreactor, but also a strategy to optimize other bioreactor systems.

  7. Yield Optimisation of Hepatitis B Virus Core Particles in E. coli Expression System for Drug Delivery Applications.

    PubMed

    Bin Mohamed Suffian, Izzat Fahimuddin; Garcia-Maya, Mitla; Brown, Paul; Bui, Tam; Nishimura, Yuya; Palermo, Amir Rafiq Bin Mohammad Johari; Ogino, Chiaki; Kondo, Akihiko; Al-Jamal, Khuloud T

    2017-03-03

    An E. coli expression system offers a mean for rapid, high yield and economical production of Hepatitis B Virus core (HBc) particles. However, high-level production of HBc particles in bacteria is demanding and optimisation of HBc particle yield from E. coli is required to improve laboratory-scale productivity for further drug delivery applications. Production steps involve bacterial culture, protein isolation, denaturation, purification and finally protein assembly. In this study, we describe a modified E. coli based method for purifying HBc particles and compare the results with those obtained using a conventional purification method. HBc particle morphology was confirmed by Atomic Force Microscopy (AFM). Protein specificity and secondary structure were confirmed by Western Blot and Circular Dichroism (CD), respectively. The modified method produced ~3-fold higher yield and greater purity of wild type HBc particles than the conventional method. Our results demonstrated that the modified method produce a better yield and purity of HBc particles in an E. coli-expression system, which are fully characterised and suitable to be used for drug delivery applications.

  8. Yield Optimisation of Hepatitis B Virus Core Particles in E. coli Expression System for Drug Delivery Applications

    PubMed Central

    Bin Mohamed Suffian, Izzat Fahimuddin; Garcia-Maya, Mitla; Brown, Paul; Bui, Tam; Nishimura, Yuya; Palermo, Amir Rafiq Bin Mohammad Johari; Ogino, Chiaki; Kondo, Akihiko; Al-Jamal, Khuloud T.

    2017-01-01

    An E. coli expression system offers a mean for rapid, high yield and economical production of Hepatitis B Virus core (HBc) particles. However, high-level production of HBc particles in bacteria is demanding and optimisation of HBc particle yield from E. coli is required to improve laboratory-scale productivity for further drug delivery applications. Production steps involve bacterial culture, protein isolation, denaturation, purification and finally protein assembly. In this study, we describe a modified E. coli based method for purifying HBc particles and compare the results with those obtained using a conventional purification method. HBc particle morphology was confirmed by Atomic Force Microscopy (AFM). Protein specificity and secondary structure were confirmed by Western Blot and Circular Dichroism (CD), respectively. The modified method produced ~3-fold higher yield and greater purity of wild type HBc particles than the conventional method. Our results demonstrated that the modified method produce a better yield and purity of HBc particles in an E. coli-expression system, which are fully characterised and suitable to be used for drug delivery applications. PMID:28256592

  9. [A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].

    PubMed

    Mo, Qianzhen; Mai, Rongjia; Yang, Zhixiao; Chen, Minfang; Yang, Tiezhao; Lai, Huafang; Yang, Peiliang; Chen, Qiang; Zhou, Xiaohong

    2012-06-01

    To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions. To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA. Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration. We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.

  10. High-yield expression of recombinant soybean agglutinin in plants using transient and stable systems.

    PubMed

    Tremblay, Reynald; Feng, Mary; Menassa, Rima; Huner, Norman P A; Jevnikar, Anthony M; Ma, Shengwu

    2011-04-01

    Soybean agglutinin (SBA) is a specific N-acetylgalactosamine-binding plant lectin that can agglutinate a wide variety of cells. SBA has great potential for medical and biotechnology-focused applications, including screening and treatment of breast cancer, isolation of fetal cells from maternal blood for genetic screening, the possibility as a carrier system for oral drug delivery, and utilization as an affinity tag for high-quality purification of tagged proteins. The success of these applications, to a large degree, critically depends on the development of a highly efficient expression system for a source of recombinant SBA (rSBA). Here, we demonstrate the utility of transient and stable expression systems in Nicotiana benthamiana and potato, respectively, for the production of rSBA, with the transgenic protein accumulated to 4% of total soluble protein (TSP) in Nicotiana benthamiana leaves and 0.3% of TSP in potato tubers. Furthermore, we show that both plant-derived rSBAs retain their ability to induce the agglutination of red blood cells, are similarly glycosylated when compared with native SBA, retained their binding specificity for N-acetylgalactosamine, and were highly resistant to degradation in simulated gastric and intestinal fluids. Affinity column purification using N-acetylgalactosamine as a specific ligand resulted in high recovery and purity of rSBA. This work is the first step toward use of rSBA for various new applications, including the development of rSBA as a novel affinity tag for simplified purification of tagged proteins and as a new carrier molecule for delivery of oral drugs.

  11. Laue Crystal Structure of Shewanella oneidensis Cytochrome c Nitrite Reductase from a High-yield Expression System

    PubMed Central

    Youngblut, Matthew; Judd, Evan T.; Srajer, Vukica; Sayyed, Bilal; Goelzer, Tyler; Elliott, Sean J.; Schmidt, Marius; Pacheco, A. Andrew

    2012-01-01

    The high-yield expression and purification of Shewanella oneidensis cytochrome c nitrite reductase (ccNiR), and its characterization by a variety of methods, notably Laue crystallography, is reported. A key component of the expression system is an artificial ccNiR gene in which the N-terminal signal peptide from the highly expressed S. oneidensis protein “Small Tetra-heme c” replaces the wild-type signal peptide. This gene, inserted into the plasmid pHSG298 and expressed in S. oneidensis TSP-1 strain, generated ~20 mg crude ccNiR/L culture, compared with 0.5–1 mg/L for untransformed cells. Purified ccNiR has nitrite and hydroxylamine reductase activities comparable to those previously reported for E. coli ccNiR, and is stable for over two weeks in pH 7 solution at 4° C. UV/Vis spectropotentiometric titrations and protein film voltammetry identified 5 independent 1-electron reduction processes. Global analysis of the spectropotentiometric data also allowed determination of the extinction coefficient spectra for the 5 reduced ccNiR species. The characteristics of the individual extinction coefficient spectra suggest that, within each reduced species, the electrons are distributed amongst the various hemes, rather than being localized on specific heme centers. The purified ccNiR yielded good quality crystals, with which the 2.59 Å resolution structure was solved at room temperature using the Laue diffraction method. The structure is similar to that of E. coli ccNiR, except in the region where the enzyme interacts with its physiological electron donor (CymA in the case of S. oneidensis ccNiR, NrfB in the case of the E. coli protein). PMID:22382353

  12. Laue crystal structure of Shewanella oneidensis cytochrome c nitrite reductase from a high-yield expression system

    SciTech Connect

    Youngblut, Matthew; Judd, Evan T.; Srajer, Vukica; Sayyed, Bilal; Goelzer, Tyler; Elliott, Sean J.; Schmidt, Marius; Pacheco, A. Andrew

    2012-09-11

    The high-yield expression and purification of Shewanella oneidensis cytochrome c nitrite reductase (ccNiR) and its characterization by a variety of methods, notably Laue crystallography, are reported. A key component of the expression system is an artificial ccNiR gene in which the N-terminal signal peptide from the highly expressed S. oneidensis protein 'small tetraheme c' replaces the wild-type signal peptide. This gene, inserted into the plasmid pHSG298 and expressed in S. oneidensis TSP-1 strain, generated approximately 20 mg crude ccNiR per liter of culture, compared with 0.5-1 mg/L for untransformed cells. Purified ccNiR has nitrite and hydroxylamine reductase activities comparable to those previously reported for Escherichia coli ccNiR, and is stable for over 2 weeks in pH 7 solution at 4 C. UV/vis spectropotentiometric titrations and protein film voltammetry identified five independent one-electron reduction processes. Global analysis of the spectropotentiometric data also allowed determination of the extinction coefficient spectra for the five reduced ccNiR species. The characteristics of the individual extinction coefficient spectra suggest that, within each reduced species, the electrons are distributed among the various hemes, rather than being localized on specific heme centers. The purified ccNiR yielded good-quality crystals, with which the 2.59-{angstrom}-resolution structure was solved at room temperature using the Laue diffraction method. The structure is similar to that of E. coli ccNiR, except in the region where the enzyme interacts with its physiological electron donor (CymA in the case of S. oneidensis ccNiR, NrfB in the case of the E. coli protein).

  13. Development of a high yielding E. coli periplasmic expression system for the production of humanized Fab' fragments.

    PubMed

    Ellis, Mark; Patel, Pareshkumar; Edon, Marjory; Ramage, Walter; Dickinson, Robert; Humphreys, David P

    2017-01-01

    Humanized Fab' fragments may be produced in the periplasm of Escherichia coli but can be subject to degradation by host cell proteases. In order to increase Fab' yield and reduce proteolysis we developed periplasmic protease deficient strains of E. coli. These strains lacked the protease activity of Tsp, protease III and DegP. High cell density fermentations indicated Tsp deficient strains increased productivity two fold but this increase was accompanied by premature cell lysis soon after the induction of Fab' expression. To overcome the reduction in cell viability we introduced suppressor mutations into the spr gene. The mutations partially restored the wild type phenotype of the cells. Furthermore, we coexpressed a range of periplasmic chaperone proteins with the Fab', DsbC had the most significant impact, increasing humanized Fab' production during high cell density fermentation. When DsbC coexpression was combined with a Tsp deficient spr strain we observed an increase in yield and essentially restored "wild type" cell viability. We achieved a final periplasmic yield of over 2.4g/L (final cell density OD600 105), 40 h post Fab' induction with minimal cell lysis.The data suggests that proteolysis, periplasm integrity, protein folding and disulphide bond formation are all potential limiting steps in the production of Fab' fragments in the periplasm of E. coli. In this body of work, we have addressed these limiting steps by utilizing stabilized protease deficient strains and chaperone coexpression. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:212-220, 2017.

  14. Removal of bacterial suspension water occupying the intercellular space of detached leaves after agroinfiltration improves the yield of recombinant hemagglutinin in a Nicotiana benthamiana transient gene expression system.

    PubMed

    Fujiuchi, Naomichi; Matsuda, Ryo; Matoba, Nobuyuki; Fujiwara, Kazuhiro

    2016-04-01

    The use of detached leaves instead of whole plants provides an alternative means for recombinant protein production based on Agrobacterium tumefaciens-mediated transient gene overexpression. However, the process for high-level protein production in detached leaves has not yet been established. In this study, we focused on leaf handling and maintenance conditions immediately after infiltration with Agrobacterium suspension (agroinfiltration) to improve recombinant protein expression in detached Nicotiana benthamiana leaves. We demonstrated that the residual water of bacterial suspension in detached leaves had significant impact on the yield of recombinant influenza hemagglutinin (HA). Immediately after agroinfiltration, detached leaves were stored in a dehumidified chamber to allow bacterial suspension water occupying intercellular space to be removed by transpiration. We varied the duration of this water removal treatment from 0.7 to 4.4 h, which resulted in leaf fresh weights ranging from 0.94 to 1.28 g g(-1) relative to weights measured just before agroinfiltration. We used these relative fresh weights (RFWs) as an indicator of the amount of residual water. The detached leaves were then incubated in humidified chambers for 6 days. We found that the presence of residual water significantly decreased HA yield, with a clear inverse correlation observed between HA yield and RFW. We next compared HA yields in detached leaves with those obtained from intact leaves by whole-plant expression performed at the same time. The maximum HA yield obtained from a detached leaf with a RFW of approximately 1.0, namely, 800 μg gFW(-1), was comparable to the mean HA yield of 846 μg gFW(-1) generated in intact leaves. Our results indicate the necessity of removing bacterial suspension water from agroinfiltrated detached leaves in transient overexpression systems and point to a critical factor enabling the detached-leaf system as a viable recombinant protein factory.

  15. High-Yield, Zero-Leakage Expression System with a Translational Switch Using Site-Specific Unnatural Amino Acid Incorporation

    PubMed Central

    Minaba, Masaomi

    2014-01-01

    Synthetic biologists construct complex biological circuits by combinations of various genetic parts. Many genetic parts that are orthogonal to one another and are independent of existing cellular processes would be ideal for use in synthetic biology. However, our toolbox is still limited with respect to the bacterium Escherichia coli, which is important for both research and industrial use. The site-specific incorporation of unnatural amino acids is a technique that incorporates unnatural amino acids into proteins using a modified exogenous aminoacyl-tRNA synthetase/tRNA pair that is orthogonal to any native pairs in a host and is independent from other cellular functions. Focusing on the orthogonality and independency that are suitable for the genetic parts, we designed novel AND gate and translational switches using the unnatural amino acid 3-iodo-l-tyrosine incorporation system in E. coli. A translational switch was turned on after addition of 3-iodo-l-tyrosine in the culture medium within minutes and allowed tuning of switchability and translational efficiency. As an application, we also constructed a gene expression system that produced large amounts of proteins under induction conditions and exhibited zero-leakage expression under repression conditions. Similar translational switches are expected to be applicable also for eukaryotes such as yeasts, nematodes, insects, mammalian cells, and plants. PMID:24375139

  16. Metabolic engineering of the baculovirus-expression system via inverse "shotgun" genomic analysis and RNA interference (dsRNA) increases product yield and cell longevity.

    PubMed

    Kim, Eun Jeong; Kramer, Shannon F; Hebert, Colin G; Valdes, James J; Bentley, William E

    2007-10-15

    RNA interference (RNAi) is as powerful tool for characterizing gene function in eukaryotic organisms and cultured cell lines. Its use in metabolic engineering has been limited and few reports have targeted protein expression systems to increase yield. In this work, we examine the use of in vitro synthesized double stranded RNA (dsRNA) in the baculovirus expression vector system (BEVS), using commercially relevant cultured cells (Spodoptera frugiperda, Sf-9) and larvae (Trichoplusia ni) as hosts. First, we employed an inverse "shotgun" genomic analysis to "find" an array of 16 putative insect gene targets. We then synthesized dsRNA in vitro targeting these genes and investigated the effects of injected dsRNA on larval growth, development, and product yield. Growth and development was at times stunted and in several cases, the effects were lethal. However, dsRNA targeting an acidic juvenile hormone-suppressible protein (AJHSP1), and translational elongation factor 2 (Ef-2) resulted in significantly increased yield of model product, GFP. Next, we targeted known genes, v-cath and apoptosis inducer, sf-caspase 1, in cultured Sf-9 cells. We confirm RNAi-mediated sf-caspase 1 suppression in Sf-9 cells, but not in baculovirus-infected cells, likely due to the overriding effects of inhibitor of apoptosis protein, p35. We also demonstrate suppression of v-cath in infected cells, which leads to a approximately 3-fold increase in product yield. Overall, our results support the application of RNAi in metabolic engineering, specifically for enhancing protein productivity in the baculovirus expression vector system. Copyright 2007 Wiley Periodicals, Inc.

  17. Effects of plant density on recombinant hemagglutinin yields in an Agrobacterium-mediated transient gene expression system using Nicotiana benthamiana plants.

    PubMed

    Fujiuchi, Naomichi; Matsuda, Ryo; Matoba, Nobuyuki; Fujiwara, Kazuhiro

    2017-08-01

    Agrobacterium-mediated transient expression systems enable plants to rapidly produce a wide range of recombinant proteins. To achieve economically feasible upstream production and downstream processing, it is beneficial to obtain high levels of two yield-related quantities of upstream production: recombinant protein content per fresh mass of harvested biomass (g gFM(-1) ) and recombinant protein productivity per unit area-time (g m(-2) /month). Here, we report that the density of Nicotiana benthamiana plants during upstream production had significant impacts on the yield-related quantities of recombinant hemagglutinin (HA). The two quantities were smaller at a high plant density of 400 plants m(-2) than at a low plant density of 100 plants m(-2) . The smaller quantities at the high plant density were attributed to: (i) a lower HA content in young leaves, which usually have high HA accumulation potentials; (ii) a lower biomass allocation to the young leaves; and (iii) a high area-time requirement for plants. Thus, plant density is a key factor for improving upstream production in Agrobacterium-mediated transient expression systems. Biotechnol. Bioeng. 2017;114: 1762-1770. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  18. High yield expression and purification of human endothelin-1.

    PubMed

    Fassina, G; Merli, S; Germani, S; Ciliberto, G; Cassani, G

    1994-12-01

    A DNA construct encoding human big endothelin (Big ET) preceded by the factor Xa protease recognition site (Ile-Glu-Gly-Arg), fused in frame to the maltose binding protein sequence, has been introduced in DH5-alpha cells. The fusion product (MBP-Big ET) was expressed at a concentration close to 100 micrograms/ml of culture broth and constituted approximately 50% of the total protein content. Crude cell extracts containing the fusion product have been directly treated with trypsin under mild denaturing conditions in order to release big endothelin (1-37) from the adduct. Cleavage yield of the MBP-Big ET adduct was close to 70%. Big ET(1-37) was separated from unrelated peptides derived from the tryptic digest of the bacterial extract by affinity chromatography. The affinity column was prepared by immobilizing a protease resistant peptide ligand able to recognize Big ET with sufficient affinity, selectivity, and specificity. From the affinity step (recovery, 90%), recombinant Big ET(1-37) was obtained with a purity close to 80%. The affinity-purified recombinant product was then digested with alpha-chymotrypsin in order to release endothelin (1-21), which was then purified by RP-HPLC. With this two-step purification protocol, 3 micrograms of endothelin was recovered from 1 ml of bacterial broth, with a purity close to 95%.

  19. Yielding Torque-Tube System Reduces Crash Injuries

    NASA Technical Reports Server (NTRS)

    Mcsmith, D. G.

    1982-01-01

    Yielding torque-tube system minimizes injuries by limiting load transferred to occupant in crash. When properly integrated into seat structure, torque tube yields in plastic deformation stage of material and maintains a relatively constant resistance to applied torque for many degrees of rotation. Yielding torque-tube system is expected to find application in aircraft and automobile industries.

  20. Direct laboratory tensile testing of select yielding rock bolt systems

    SciTech Connect

    VandeKraats, J.D.; Watson, S.O.

    1996-08-01

    Yielding rock bolt support systems have been developed to accommodate ground movement in shifting ground such as in coal operations; in creeping ground such as salt, trona, and potash; and in swelling ground associated with some clays. These systems, designed to remain intact despite ground movement, should enhance mine safety and help contain costs in areas where rebolting of rigid non-yielding systems is typically required. Four such systems were tested in straight tensile pulls in the laboratory. They include the Slip Nut System from Dywidag Systems International USA, Inc., Ischebeck`s bolt mounted Titan Load Indicator, Rocky Mountain Bolt Company`s Yielding Cable Bolt, and a rock bolt installed variation of the yielding steel post developed by RE/SPEC Inc. The first two systems are currently marketed products and the latter two are prototype systems. Each system responds to load and displacement by yielding in an unique manner. All are designed to yield at predetermined loads. A description of each system and its yield function is provided. Each system was tested over its prescribed yield range in a test machine. At least five tests were performed on each system. Each system yielded and continued to provide support according to its design. Each shows promise for ground control use in shifting or creeping rock. This work helps to illustrate the comparative differences in performance between these specialized systems and the applications where they may be most useful.

  1. [Prokaryotic expression systems].

    PubMed

    Porowińska, Dorota; Wujak, Magdalena; Roszek, Katarzyna; Komoszyński, Michał

    2013-03-01

    For overproduction of recombinant proteins both eukaryotic and prokaryotic expression systems are used. Choosing the right system depends, among other things, on the growth rate and culture of host cells, level of the target gene expression and posttranslational processing of the synthesized protein. Regardless of the type of expression system, its basic elements are the vector and the expression host. The most widely used system for protein overproduction, both on a laboratory and industrial scale, is the prokaryotic system. This system is based primarily on the bacteria E. coli, although increasingly often Bacillus species are used. The prokaryotic system allows one to obtain large quantities of recombinant proteins in a short time. A simple and inexpensive bacterial cell culture and well-known mechanisms of transcription and translation facilitate the use of these microorganisms. The simplicity of genetic modifications and the availability of many bacterial mutants are additional advantages of the prokaryotic system. In this article we characterize the structural elements of prokaryotic expression vectors. Also strategies for preparation of the target protein gene that increase productivity, facilitate detection and purification of recombinant protein and provide its activity are discussed. Bacterial strains often used as host cells in expression systems as well as the potential location of heterologous proteins are characterized. Knowledge of the basic elements of the prokaryotic expression system allows for production of biologically active proteins in a short time and in satisfactory quantities. 

  2. Surprising yields with no-till cropping systems

    USDA-ARS?s Scientific Manuscript database

    Producers using no-till systems have found that crop yields often exceed their expectation based on nutrient and water supply. For example, corn yields 7% higher in a no-till system in central South Dakota than in a tilled system in eastern South Dakota. This is surprising because rainfall is 5 in...

  3. Vacuum transfer system increases sugar maple sap yield

    Treesearch

    Russell S. Walters

    1978-01-01

    Yields of sugar maple sap collected from three plastic pipeline systems by gravity, vacuum pump, and a vacuum pump with a transfer tank were compared during 2 years in northern Vermont. The transfer system yielded 27 percent more sap one year and 17 percent more the next year. Higher vacuum levels at the tapholes were observed in the transfer system.

  4. Estimating yield gaps at the cropping system level.

    PubMed

    Guilpart, Nicolas; Grassini, Patricio; Sadras, Victor O; Timsina, Jagadish; Cassman, Kenneth G

    2017-05-01

    Yield gap analyses of individual crops have been used to estimate opportunities for increasing crop production at local to global scales, thus providing information crucial to food security. However, increases in crop production can also be achieved by improving cropping system yield through modification of spatial and temporal arrangement of individual crops. In this paper we define the cropping system yield potential as the output from the combination of crops that gives the highest energy yield per unit of land and time, and the cropping system yield gap as the difference between actual energy yield of an existing cropping system and the cropping system yield potential. Then, we provide a framework to identify alternative cropping systems which can be evaluated against the current ones. A proof-of-concept is provided with irrigated rice-maize systems at four locations in Bangladesh that represent a range of climatic conditions in that country. The proposed framework identified (i) realistic alternative cropping systems at each location, and (ii) two locations where expected improvements in crop production from changes in cropping intensity (number of crops per year) were 43% to 64% higher than from improving the management of individual crops within the current cropping systems. The proposed framework provides a tool to help assess food production capacity of new systems (e.g. with increased cropping intensity) arising from climate change, and assess resource requirements (water and N) and associated environmental footprint per unit of land and production of these new systems. By expanding yield gap analysis from individual crops to the cropping system level and applying it to new systems, this framework could also be helpful to bridge the gap between yield gap analysis and cropping/farming system design.

  5. Sex Roles and Yielded/Expressed Self-Control.

    ERIC Educational Resources Information Center

    Ganong, Lawrence H.; Coleman, Marilyn

    1987-01-01

    Results of a study of the impact of sex and sex role orientation on reported self-control behaviors showed that sex did not affect self-control or satisfaction with self-control, but sex role orientation did. Androgynous persons reported using more expressed self-control than others. (PS)

  6. Sex Roles and Yielded/Expressed Self-Control.

    ERIC Educational Resources Information Center

    Ganong, Lawrence H.; Coleman, Marilyn

    1987-01-01

    Results of a study of the impact of sex and sex role orientation on reported self-control behaviors showed that sex did not affect self-control or satisfaction with self-control, but sex role orientation did. Androgynous persons reported using more expressed self-control than others. (PS)

  7. Improved, integrated yield-accounting system saves time and money

    SciTech Connect

    Bayard, P. )

    1993-11-22

    Cit-Con Corp.'s Lake Charles, La., lube plant has improved operating efficiency by employing a computerized yield-accounting process that uses new balancing methods and reconciles plant data. Cit-Con produces high-quality lubricating oils and paraffin waxes using modern refining methods. The yield-accounting task, therefore, is particularly critical because: Cit-Con is jointly owned 65% by Citgo Petroleum Corp. and 35% by Conoco Inc. Accurate yield accounting thus is essential for the owners and for tax purposes. Cit-Con produces 9,000 b/d of lube oil base stocks and 300,00 lb/day of fully refined waxes, and has an estimated replacement value of $356 million. The paper discusses lube production, accounting concerns, tracking feeds, reporting yields, monitoring operations, then describes the yield accounting system, network setup, data integration, data reconciliation, and cost savings.

  8. New Mammalian Expression Systems.

    PubMed

    Zhu, Jie; Hatton, Diane

    2017-06-06

    There are an increasing number of recombinant antibodies and proteins in preclinical and clinical development for therapeutic applications. Mammalian expression systems are key to enabling the production of these molecules, and Chinese hamster ovary (CHO) cell platforms continue to be central to delivery of the stable cell lines required for large-scale production. Increasing pressure on timelines and efficiency, further innovation of molecular formats and the shift to new production systems are driving developments of these CHO cell line platforms. The availability of genome and transcriptome data coupled with advancing gene editing tools are increasing the ability to design and engineer CHO cell lines to meet these challenges. This chapter aims to give an overview of the developments in CHO expression systems and some of the associated technologies over the past few years.

  9. Hyper-inducible expression system for streptomycetes.

    PubMed

    Herai, Sachio; Hashimoto, Yoshiteru; Higashibata, Hiroki; Maseda, Hideaki; Ikeda, Haruo; Omura, Satoshi; Kobayashi, Michihiko

    2004-09-28

    Streptomycetes produce useful enzymes and a wide variety of secondary metabolites with potent biological activities (e.g., antibiotics, immunosuppressors, pesticides, etc.). Despite their importance in the pharmaceutical and agrochemical fields, there have been no reports for practical expression systems in streptomycetes. Here, we developed a "P(nitA)-NitR" system for regulatory gene expression in streptomycetes based on the expression mechanism of Rhodococcus rhodochrous J1 nitrilase, which is highly induced by an inexpensive and safe inducer, epsilon-caprolactam. Heterologous protein expression experiments demonstrated that the system allowed suppressed basal expression and hyper-inducible expression, yielding target protein levels of as high as approximately 40% of all soluble protein. Furthermore, the system functioned in important streptomycete strains. Thus, the P(nitA)-NitR system should be a powerful tool for improving the productivity of various useful products in streptomycetes.

  10. EarthSat spring wheat yield system test 1975

    NASA Technical Reports Server (NTRS)

    1976-01-01

    The results of an operational test of the EarthSat System during the period 1 June - 30 August 1975 over the spring wheat regions of North Dakota, South Dakota, and Minnesota are presented. The errors associated with each sub-element of the system during the operational test and the sensitivity of the complete system and each major functional sub-element of the system to the observed errors were evaluated. Evaluations and recommendations for future operational users of the system include: (1) changes in various system sub-elements, (2) changes in the yield model to affect improved accuracy, (3) changes in the number of geobased cells needed to develop an accurate aggregated yield estimate, (4) changes associated with the implementation of future operational satellites and data processing systems, and (5) detailed system documentation.

  11. Yield and yield gaps in central U.S. corn production systems

    USDA-ARS?s Scientific Manuscript database

    The magnitude of yield gaps (YG) (potential yield – farmer yield) provides some indication of the prospects for increasing crop yield. Quantile regression analysis was applied to county maize (Zea mays L.) yields (1972 – 2011) from Kentucky, Iowa and Nebraska (irrigated) (total of 115 counties) to e...

  12. A High-Yield Co-Expression System for the Purification of an Intact Drs2p-Cdc50p Lipid Flippase Complex, Critically Dependent on and Stabilized by Phosphatidylinositol-4-Phosphate

    PubMed Central

    Azouaoui, Hassina; Montigny, Cédric; Ash, Miriam-Rose; Fijalkowski, Frank; Jacquot, Aurore; Grønberg, Christina; López-Marqués, Rosa L.; Palmgren, Michael G.; Garrigos, Manuel; le Maire, Marc; Decottignies, Paulette; Gourdon, Pontus; Nissen, Poul; Champeil, Philippe; Lenoir, Guillaume

    2014-01-01

    P-type ATPases from the P4 subfamily (P4-ATPases) are energy-dependent transporters, which are thought to establish lipid asymmetry in eukaryotic cell membranes. Together with their Cdc50 accessory subunits, P4-ATPases couple ATP hydrolysis to lipid transport from the exoplasmic to the cytoplasmic leaflet of plasma membranes, late Golgi membranes, and endosomes. To gain insights into the structure and function of these important membrane pumps, robust protocols for expression and purification are required. In this report, we present a procedure for high-yield co-expression of a yeast flippase, the Drs2p-Cdc50p complex. After recovery of yeast membranes expressing both proteins, efficient purification was achieved in a single step by affinity chromatography on streptavidin beads, yielding ∼1–2 mg purified Drs2p-Cdc50p complex per liter of culture. Importantly, the procedure enabled us to recover a fraction that mainly contained a 1∶1 complex, which was assessed by size-exclusion chromatography and mass spectrometry. The functional properties of the purified complex were examined, including the dependence of its catalytic cycle on specific lipids. The dephosphorylation rate was stimulated in the simultaneous presence of the transported substrate, phosphatidylserine (PS), and the regulatory lipid phosphatidylinositol-4-phosphate (PI4P), a phosphoinositide that plays critical roles in membrane trafficking events from the trans-Golgi network (TGN). Likewise, overall ATP hydrolysis by the complex was critically dependent on the simultaneous presence of PI4P and PS. We also identified a prominent role for PI4P in stabilization of the Drs2p-Cdc50p complex towards temperature- or C12E8-induced irreversible inactivation. These results indicate that the Drs2p-Cdc50p complex remains functional after affinity purification and that PI4P as a cofactor tightly controls its stability and catalytic activity. This work offers appealing perspectives for detailed structural and

  13. A high-yield co-expression system for the purification of an intact Drs2p-Cdc50p lipid flippase complex, critically dependent on and stabilized by phosphatidylinositol-4-phosphate.

    PubMed

    Azouaoui, Hassina; Montigny, Cédric; Ash, Miriam-Rose; Fijalkowski, Frank; Jacquot, Aurore; Grønberg, Christina; López-Marqués, Rosa L; Palmgren, Michael G; Garrigos, Manuel; le Maire, Marc; Decottignies, Paulette; Gourdon, Pontus; Nissen, Poul; Champeil, Philippe; Lenoir, Guillaume

    2014-01-01

    P-type ATPases from the P4 subfamily (P4-ATPases) are energy-dependent transporters, which are thought to establish lipid asymmetry in eukaryotic cell membranes. Together with their Cdc50 accessory subunits, P4-ATPases couple ATP hydrolysis to lipid transport from the exoplasmic to the cytoplasmic leaflet of plasma membranes, late Golgi membranes, and endosomes. To gain insights into the structure and function of these important membrane pumps, robust protocols for expression and purification are required. In this report, we present a procedure for high-yield co-expression of a yeast flippase, the Drs2p-Cdc50p complex. After recovery of yeast membranes expressing both proteins, efficient purification was achieved in a single step by affinity chromatography on streptavidin beads, yielding ∼ 1-2 mg purified Drs2p-Cdc50p complex per liter of culture. Importantly, the procedure enabled us to recover a fraction that mainly contained a 1:1 complex, which was assessed by size-exclusion chromatography and mass spectrometry. The functional properties of the purified complex were examined, including the dependence of its catalytic cycle on specific lipids. The dephosphorylation rate was stimulated in the simultaneous presence of the transported substrate, phosphatidylserine (PS), and the regulatory lipid phosphatidylinositol-4-phosphate (PI4P), a phosphoinositide that plays critical roles in membrane trafficking events from the trans-Golgi network (TGN). Likewise, overall ATP hydrolysis by the complex was critically dependent on the simultaneous presence of PI4P and PS. We also identified a prominent role for PI4P in stabilization of the Drs2p-Cdc50p complex towards temperature- or C12E8-induced irreversible inactivation. These results indicate that the Drs2p-Cdc50p complex remains functional after affinity purification and that PI4P as a cofactor tightly controls its stability and catalytic activity. This work offers appealing perspectives for detailed structural and

  14. Ectopic expression of specific GA2 oxidase mutants promotes yield and stress tolerance in rice.

    PubMed

    Lo, Shuen-Fang; Ho, Tuan-Hua David; Liu, Yi-Lun; Jiang, Mirng-Jier; Hsieh, Kun-Ting; Chen, Ku-Ting; Yu, Lin-Chih; Lee, Miin-Huey; Chen, Chi-Yu; Huang, Tzu-Pi; Kojima, Mikiko; Sakakibara, Hitoshi; Chen, Liang-Jwu; Yu, Su-May

    2017-07-01

    A major challenge of modern agricultural biotechnology is the optimization of plant architecture for enhanced productivity, stress tolerance and water use efficiency (WUE). To optimize plant height and tillering that directly link to grain yield in cereals and are known to be tightly regulated by gibberellins (GAs), we attenuated the endogenous levels of GAs in rice via its degradation. GA 2-oxidase (GA2ox) is a key enzyme that inactivates endogenous GAs and their precursors. We identified three conserved domains in a unique class of C20 GA2ox, GA2ox6, which is known to regulate the architecture and function of rice plants. We mutated nine specific amino acids in these conserved domains and observed a gradient of effects on plant height. Ectopic expression of some of these GA2ox6 mutants moderately lowered GA levels and reprogrammed transcriptional networks, leading to reduced plant height, more productive tillers, expanded root system, higher WUE and photosynthesis rate, and elevated abiotic and biotic stress tolerance in transgenic rice. Combinations of these beneficial traits conferred not only drought and disease tolerance but also increased grain yield by 10-30% in field trials. Our studies hold the promise of manipulating GA levels to substantially improve plant architecture, stress tolerance and grain yield in rice and possibly in other major crops. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  15. Applicability and methodology of determining sustainable yield in groundwater systems

    NASA Astrophysics Data System (ADS)

    Kalf, Frans R. P.; Woolley, Donald R.

    2005-03-01

    There is currently a need for a review of the definition and methodology of determining sustainable yield. The reasons are: (1) current definitions and concepts are ambiguous and non-physically based so cannot be used for quantitative application, (2) there is a need to eliminate varying interpretations and misinterpretations and provide a sound basis for application, (3) the notion that all groundwater systems either are or can be made to be sustainable is invalid, (4) often there are an excessive number of factors bound up in the definition that are not easily quantifiable, (5) there is often confusion between production facility optimal yield and basin sustainable yield, (6) in many semi-arid and arid environments groundwater systems cannot be sensibly developed using a sustained yield policy particularly where ecological constraints are applied. Derivation of sustainable yield using conservation of mass principles leads to expressions for basin sustainable, partial (non-sustainable) mining and total (non-sustainable) mining yields that can be readily determined using numerical modelling methods and selected on the basis of applied constraints. For some cases there has to be recognition that the groundwater resource is not renewable and its use cannot therefore be sustainable. In these cases, its destiny should be the best equitable use.

  16. High-yield positron systems for linear colliders

    SciTech Connect

    Clendenin, J.E.

    1989-04-01

    Linear colliders, such as the SLC, are among those accelerators for which a high-yield positron source operating at the repetition rate of the accelerator is desired. The SLC, having electron energies up to 50 GeV, presents the possibility of generating positron bunches with useful charge even exceeding that of the initial electron bunch. The exact positron yield to be obtained depends on the particular capture, transport and damping system employed. Using 31 GeV electrons impinging on a W-type converter phase-space at the target to the acceptance of the capture rf section, the SLC source is capable of producing, for every electron, up to two positrons within the acceptance of the positron damping ring. The design of this source and the performance of the positron system as built are described. Also, future prospects and limitations for high-yield positron systems are discussed. 11 refs., 5 figs., 3 tabs.

  17. High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension

    PubMed Central

    Subedi, Ganesh P.; Johnson, Roy W.; Moniz, Heather A.; Moremen, Kelley W.; Barb, Adam

    2015-01-01

    The art of producing recombinant proteins with complex post-translational modifications represents a major challenge for studies of structure and function. The rapid establishment and high recovery from transiently-transfected mammalian cell lines addresses this barrier and is an effective means of expressing proteins that are naturally channeled through the ER and Golgi-mediated secretory pathway. Here is one protocol for protein expression using the human HEK293F and HEK293S cell lines transfected with a mammalian expression vector designed for high protein yields. The applicability of this system is demonstrated using three representative glycoproteins that expressed with yields between 95-120 mg of purified protein recovered per liter of culture. These proteins are the human FcγRIIIa and the rat α2-6 sialyltransferase, ST6GalI, both expressed with an N-terminal GFP fusion, as well as the unmodified human immunoglobulin G1 Fc. This robust system utilizes a serum-free medium that is adaptable for expression of isotopically enriched proteins and carbohydrates for structural studies using mass spectrometry and nuclear magnetic resonance spectroscopy. Furthermore, the composition of the N-glycan can be tuned by adding a small molecule to prevent certain glycan modifications in a manner that does not reduce yield. PMID:26779721

  18. Yield response and economics of shallow subsurface drip irrigation systems

    USDA-ARS?s Scientific Manuscript database

    Field tests were conducted using shallow subsurface drip irrigation (S3DI) on cotton (Gossypium hirsutum, L.), corn (Zea mays, L.), and peanut (Arachis hypogeae, L.) in rotation to investigate yield potential and economic sustainability of this irrigation system technique over a six year period. Dri...

  19. Genome-wide transcript analysis of maize hybrids: allelic additive gene expression and yield heterosis.

    PubMed

    Guo, Mei; Rupe, Mary A; Yang, Xiaofeng; Crasta, Oswald; Zinselmeier, Christopher; Smith, Oscar S; Bowen, Ben

    2006-09-01

    Heterosis, or hybrid vigor, has been widely exploited in plant breeding for many decades, but the molecular mechanisms underlying the phenomenon remain unknown. In this study, we applied genome-wide transcript profiling to gain a global picture of the ways in which a large proportion of genes are expressed in the immature ear tissues of a series of 16 maize hybrids that vary in their degree of heterosis. Key observations include: (1) the proportion of allelic additively expressed genes is positively associated with hybrid yield and heterosis; (2) the proportion of genes that exhibit a bias towards the expression level of the paternal parent is negatively correlated with hybrid yield and heterosis; and (3) there is no correlation between the over- or under-expression of specific genes in maize hybrids with either yield or heterosis. The relationship of the expression patterns with hybrid performance is substantiated by analysis of a genetically improved modern hybrid (Pioneer hybrid 3394) versus a less improved older hybrid (Pioneer hybrid 3306) grown at different levels of plant density stress. The proportion of allelic additively expressed genes is positively associated with the modern high yielding hybrid, heterosis and high yielding environments, whereas the converse is true for the paternally biased gene expression. The dynamic changes of gene expression in hybrids responding to genotype and environment may result from differential regulation of the two parental alleles. Our findings suggest that differential allele regulation may play an important role in hybrid yield or heterosis, and provide a new insight to the molecular understanding of the underlying mechanisms of heterosis.

  20. Analysis on differential expressed genes of ovarian tissue between high- and low-yield laying hen.

    PubMed

    Chen, Wei; Song, Ling-Jun; Zeng, Yong-Qing; Yang, Yun; Wang, Hui

    2013-01-01

    In order to elucidate molecular genetic mechanism of laying hen reproduction at the transcriptional level and the structure of significantly differential genes, the mRNA differential display and reverse northern dot-blot were used to detect the differential expression of genes in the ovary tissue of low-yield laying hens and high-yield laying hens in the present study. Sixteen 32-week-old CAU-pink laying hens divided into two groups were used and the laying performance was measured. The results showed that only the egg numbers were significantly different between the two groups; and from 15 primer pairs, a total of 336 bands were displayed of which 59 cDNA bands were found to be differentially expressed in both high-yield and low-yield laying hen. The sequence analysis indicated that the expression of such bands as H-AP5, H-P5, and H-P4 was significantly potentiated in high-yield laying hen using primer pairs AP5/HT11G, P5/HT11G and P4/HT11G and these transcripts had high homology (98%) to HoxDb, HoxCa, and HoxBa, respectively. The differentially expressed gene fragments may be relevant to the progression of the high-yield hens to the egg-laying stage. And further study is required to elucidate the molecular function to improve the productivity of laying hens.

  1. Modified harvest system for enhancing Factor VIII yield in alternating tangential flow perfusion culture.

    PubMed

    Kim, Seung-Chul; An, Sora; Kim, Hyun-Ki; Park, Beom-Soo; Na, Kyu-Heum; Kim, Byung-Gee

    2016-05-01

    This study describes the development and experimental verification of a modified harvest system to enhance Factor VIII (FVIII) yield in an alternating tangential flow (ATF) perfusion culture. The main innovation of the modified harvest system is the use of check and pinch valves, eliminating the need of a peristaltic pump for harvest. The system was applied to perfusion cultures of Chinese hamster ovary cells, which co-express both recombinant human FVIII (rhFVIII) and von Willebrand factor (vWF). The modified harvest system showed comparable cell growth with the conventional harvest system using a peristaltic pump. The perfusion rate was successfully controlled using the system. In addition, the modified harvest system achieved an approximately 13.6-fold increase in the final concentration yield of FVIII activity and a 1.47-fold increase in the production yield of FVIII activity compared with a peristaltic pump. Enhancement of the yield of FVIII activity resulted from the reduction of FVIII antigen ( Ag) retention. As a result of transmembrane pressure (TMP) measurement, the reduction of the retained Ag was due to the increased TMP, which was caused by the characteristic function of a check valve, compared with a peristaltic harvest system. The modified harvest system developed in this study could be useful to enhance the production yield of other recombinant proteins in ATF perfusion culture. Copyright © 2015. Published by Elsevier B.V.

  2. Environment Control to Improve Recombinant Protein Yields in Plants Based on Agrobacterium-Mediated Transient Gene Expression

    PubMed Central

    Fujiuchi, Naomichi; Matoba, Nobuyuki; Matsuda, Ryo

    2016-01-01

    Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area–time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant

  3. Environment Control to Improve Recombinant Protein Yields in Plants Based on Agrobacterium-Mediated Transient Gene Expression.

    PubMed

    Fujiuchi, Naomichi; Matoba, Nobuyuki; Matsuda, Ryo

    2016-01-01

    Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area-time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant

  4. A Simplified Protocol for High-Yield Expression and Purification of Bacterial Topoisomerase I

    PubMed Central

    Jones, Jesse A.; Price, Emily; Miller, Donovan; Hevener, Kirk E.

    2016-01-01

    Type IA topoisomerases represent promising antibacterial drug targets. Data exists suggesting that the two bacterial type IA topoisomerase enzymes—topoisomerase I and topoisomerase III—share an overlapping biological role. Furthermore, topoisomerase I has been shown to be essential for the survival of certain organisms lacking topoisomerase III. With this in mind, it is plausible that topoisomerase I may represent a potential target for selective antibacterial drug development. As many reported bacterial topoisomerase I purification protocols have either suffered from relatively low yield, numerous steps, or a simple failure to report target protein yield altogether, a high-yield and high-purity bacterial topoisomerase I expression and purification protocol is highly desirable. The goal of this study was therefore to optimize the expression and purification of topoisomerase I from Streptococcus mutans, a clinically relevant organism that plays a significant role in oral and extra-oral infection, in order to quickly and easily attain the requisite quantities of pure target enzyme suitable for use in assay development, compound library screening, and carrying out further structural and biochemical characterization analyses. Herein we report the systematic implementation and analysis of various expression and purification techniques leading to the development and optimization of a rapid and straightforward protocol for the auto-induced expression and two-step, affinity tag purification of Streptococcus mutans topoisomerase I yielding >20 mg/liter of enzyme at over 95% purity. PMID:27117979

  5. Different universality classes at the yielding transition of amorphous systems

    NASA Astrophysics Data System (ADS)

    Jagla, E. A.

    2017-08-01

    We study the yielding transition of a two-dimensional amorphous system under shear by using a mesoscopic elasto-plastic model. The model combines a full (tensorial) description of the elastic interactions in the system and the possibility of structural reaccommodations that are responsible for the plastic behavior. The possible structural reaccommodations are encoded in the form of a "plastic disorder" potential, which is chosen independently at each position of the sample to account for local heterogeneities. We observe that the stress must exceed a critical value σc in order for the system to yield. In addition, when the system yields a flow curve (relating stress σ and strain rate γ ˙) of the form γ ˙˜(σ-σc) β is obtained. Remarkably, we observe the value of β to depend on some details of the plastic disorder potential. For smooth potentials a value of β ≃2.0 is obtained, whereas for potentials obtained as a concatenation of smooth pieces a value β ≃1.5 is observed in the simulations. This indicates a dependence of critical behavior on details of the plastic behavior. In addition, by integrating out nonessential, harmonic degrees of freedom, we derive a simplified scalar version of the model that represents a collection of interacting Prandtl-Tomlinson particles. A mean-field treatment of this interaction reproduces the difference of β exponents for the two classes of plastic disorder potentials and provides values of β that compare favorably with those found in the full simulations.

  6. Interpretation of quantum yields exceeding unity in photoelectrochemical systems

    SciTech Connect

    Szklarczyk, M.; Allen, R.E.

    1986-10-20

    In photoelectrochemical systems involving light shining on a semiconductor interfaced with an electrolyte, the quantum yield as a function of photon frequency ..nu.. is observed to exhibit a peak at h..nu..roughly-equal2E/sub g/, where E/sub g/ is the band gap of the semiconductor. The maximum in this peak is sometimes found to exceed unity. We provide an interpretation involving surface states and inelastic electron-electron scattering. The theory indicates that the effect should be observable for p-type semiconductors, but not n-type.

  7. Generalized storage-reliability-yield relationships for rainwater harvesting systems

    NASA Astrophysics Data System (ADS)

    Hanson, L. S.; Vogel, R. M.

    2014-07-01

    Sizing storage for rainwater harvesting (RWH) systems is often a difficult design consideration, as the system must be designed specifically for the local rainfall pattern. We introduce a generally applicable method for estimating the required storage by using regional regression equations to account for climatic differences in the behavior of RWH systems across the entire continental United States. A series of simulations for 231 locations with continuous daily precipitation records enable the development of storage-reliability-yield (SRY) relations at four useful reliabilities, 0.8, 0.9, 0.95, and 0.98. Multivariate, log-linear regression results in storage equations that include demand, collection area and local precipitation statistics. The continental regression equations demonstrated excellent goodness-of-fit (R2 0.96-0.99) using only two precipitation parameters, and fits improved when three geographic regions with more homogeneous rainfall characteristics were considered. The SRY models can be used to obtain a preliminary estimate of how large to build a storage tank almost anywhere in the United States based on desired yield and reliability, collection area, and local rainfall statistics. Our methodology could be extended to other regions of world, and the equations presented herein could be used to investigate how RWH systems would respond to changes in climatic variability. The resulting model may also prove useful in regional planning studies to evaluate the net benefits which result from the broad use of RWH to meet water supply requirements. We outline numerous other possible extensions to our work, which when taken together, illustrate the value of our initial generalized SRY model for RWH systems.

  8. Leishmania-based expression systems.

    PubMed

    Taheri, Tahereh; Seyed, Negar; Mizbani, Amir; Rafati, Sima

    2016-09-01

    Production of therapeutic or medical recombinant proteins, such as monoclonal antibodies, proteins, or active enzymes, requires a highly efficient system allowing natural folding and perfect post-translation modifications of the expressed protein. These requirements lead to the generation of a variety of gene expression systems from bacteria to eukaryotes. To achieve the best form of eukaryotic proteins, two factors need to be taken into consideration: choosing a suitable organism to express the protein of interest, and selecting an efficient delivery system. For this reason, the expression of recombinant proteins in eukaryotic nonpathogenic Leishmania parasites is an interesting approach which meets both criteria. Here, new Leishmania-based expression systems are compared with current systems that have long histories in research and industry.

  9. Expression of the Arabidopsis thaliana BBX32 Gene in Soybean Increases Grain Yield

    PubMed Central

    Preuss, Sasha B.; Meister, Robert; Xu, Qingzhang; Urwin, Carl P.; Tripodi, Federico A.; Screen, Steven E.; Anil, Veena S.; Zhu, Shuquan; Morrell, James A.; Liu, Grace; Ratcliffe, Oliver J.; Reuber, T. Lynne; Khanna, Rajnish; Goldman, Barry S.; Bell, Erin; Ziegler, Todd E.; McClerren, Amanda L.; Ruff, Thomas G.; Petracek, Marie E.

    2012-01-01

    Crop yield is a highly complex quantitative trait. Historically, successful breeding for improved grain yield has led to crop plants with improved source capacity, altered plant architecture, and increased resistance to abiotic and biotic stresses. To date, transgenic approaches towards improving crop grain yield have primarily focused on protecting plants from herbicide, insects, or disease. In contrast, we have focused on identifying genes that, when expressed in soybean, improve the intrinsic ability of the plant to yield more. Through the large scale screening of candidate genes in transgenic soybean, we identified an Arabidopsis thaliana B-box domain gene (AtBBX32) that significantly increases soybean grain yield year after year in multiple transgenic events in multi-location field trials. In order to understand the underlying physiological changes that are associated with increased yield in transgenic soybean, we examined phenotypic differences in two AtBBX32-expressing lines and found increases in plant height and node, flower, pod, and seed number. We propose that these phenotypic changes are likely the result of changes in the timing of reproductive development in transgenic soybean that lead to the increased duration of the pod and seed development period. Consistent with the role of BBX32 in A. thaliana in regulating light signaling, we show that the constitutive expression of AtBBX32 in soybean alters the abundance of a subset of gene transcripts in the early morning hours. In particular, AtBBX32 alters transcript levels of the soybean clock genes GmTOC1 and LHY-CCA1-like2 (GmLCL2). We propose that through the expression of AtBBX32 and modulation of the abundance of circadian clock genes during the transition from dark to light, the timing of critical phases of reproductive development are altered. These findings demonstrate a specific role for AtBBX32 in modulating soybean development, and demonstrate the validity of expressing single genes in crops to

  10. Hydrodynamic Tail Vein Injection as a Simple Tool for Yielding Extended Transgene Expression in Solid Tumors.

    PubMed

    Takayama, Takuma; Ukawa, Masami; Kanazawa, Yuki; Ando, Hidenori; Shimizu, Taro; Ishida, Tatsuhiro

    2016-01-01

    Hydrodynamic tail vein injection was considered an in vivo transfection method that yields a higher level of gene expression mainly in the liver. This method has been applied to cancer gene therapy targeting both hepatic and non-hepatic cancers. However, intratumor transgene expression in non-hepatic tumors has not been well studied. In this study, we showed an extended transgene expression of β-galactosidase (LacZ), a nonsecretory protein, in a subcutaneously implanted murine solid tumor following the hydrodynamic injection of plasmid DNA (LacZ pDNA). Our result may indicate that the hydrodynamic injection method is a powerful tool that can be used to gain transgene expression not only in the liver but also in solid tumors.

  11. Constitutive expression of cell wall invertase genes increases grain yield and starch content in maize.

    PubMed

    Li, Bei; Liu, Hua; Zhang, Yue; Kang, Tao; Zhang, Li; Tong, Jianhua; Xiao, Langtao; Zhang, Hongxia

    2013-12-01

    Grain size, number and starch content are important determinants of grain yield and quality. One of the most important biological processes that determine these components is the carbon partitioning during the early grain filling, which requires the function of cell wall invertase. Here, we showed the constitutive expression of cell wall invertase-encoding gene from Arabidopsis, rice (Oryza sativa) or maize (Zea mays), driven by the cauliflower mosaic virus (CaMV) 35S promoter, all increased cell wall invertase activities in different tissues and organs, including leaves and developing seeds, and substantially improved grain yield up to 145.3% in transgenic maize plants as compared to the wild-type plants, an effect that was reproduced in our 2-year field trials at different locations. The dramatically increased grain yield is due to the enlarged ears with both enhanced grain size and grain number. Constitutive expression of the invertase-encoding gene also increased total starch content up to 20% in the transgenic kernels. Our results suggest that cell wall invertase gene can be genetically engineered to improve both grain yield and grain quality in crop plants. © 2013 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  12. A Single Point Mutation within the Coding Sequence of Cholera Toxin B Subunit Will Increase Its Expression Yield

    PubMed Central

    Bakhshi, Bita; Boustanshenas, Mina; Ghorbani, Masoud

    2014-01-01

    Background: Cholera toxin B subunit (CTB) has been extensively considered as an immunogenic and adjuvant protein, but its yield of expression is not satisfactory in many studies. The aim of this study was to compare the expression of native and mutant recombinant CTB (rCTB) in pQE vector. Methods: ctxB fragment from Vibrio cholerae O1 ATCC14035 containing the substitution of mutant ctxB for amino acid S128T was amplified by PCR and cloned in pGETM-T easy vector. It was then transformed to E. coli Top 10F' and cultured on LB agar plate containing ampicillin. Sequence analysis confirmed the mature ctxB gene sequence and the mutant one in both constructs which were further subcloned to pQE-30 vector. Both constructs were subsequently transformed to E. coli M15 (pREP4) for expression of mature and mutant rCTB. Results: SDS-PAGE analysis showed the maximum expression of rCTB in both systems at 5 hours after induction and Western-blot analysis confirmed the presence of rCTB in blotting membranes. The expression of mutant rCTB was much higher than mature rCTB, which may be the result of serine-to-threonine substitution at position 128 of mature rCTB amino acid sequence created by PCR mutagenesis. The mutant rCTB retained pentameric stability and its ability to bind to anti- cholera toxin IgG antibodies. Conclusion: Point mutation in ctxB sequence resulted in over-expression of rCTB, probably due to the increase of solubility of produced rCTB. Consequently, this expression system can be used to produce rCTB in high yield. PMID:24842138

  13. Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli

    PubMed Central

    Ihssen, Julian; Reiss, Renate; Luchsinger, Ronny; Thöny-Meyer, Linda; Richter, Michael

    2015-01-01

    Laccases are multi-copper oxidases that oxidize a broad range of substrates at the expense of molecular oxygen, without any need for co-factor regeneration. These enzymes bear high potential for the sustainable synthesis of fine chemicals and the modification of (bio)polymers. Here we describe cloning and expression of five novel bacterial laccase-like multi copper oxidases (LMCOs) of diverse origin which were identified by homology searches in online databases. Activity yields under different expression conditions and temperature stabilities were compared to three previously described enzymes from Bacillus subtilis, Bacillus pumilus and Bacillus clausii. In almost all cases, a switch to oxygen-limited growth conditions after induction increased volumetric activity considerably. For proteins with predicted signal peptides for secretion, recombinant expression with and without signal sequence was investigated. Bacillus CotA-type LMCOs outperformed enzymes from Streptomyces and Gram-negative bacteria with respect to activity yields in Escherichia coli and application relevant biochemical properties. The novel Bacillus coagulans LMCO combined high activity yields in E. coli with unprecedented activity at strong alkaline pH and high storage stability, making it a promising candidate for further development. PMID:26068013

  14. Biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in Escherichia coli.

    PubMed

    Ihssen, Julian; Reiss, Renate; Luchsinger, Ronny; Thöny-Meyer, Linda; Richter, Michael

    2015-06-12

    Laccases are multi-copper oxidases that oxidize a broad range of substrates at the expense of molecular oxygen, without any need for co-factor regeneration. These enzymes bear high potential for the sustainable synthesis of fine chemicals and the modification of (bio)polymers. Here we describe cloning and expression of five novel bacterial laccase-like multi copper oxidases (LMCOs) of diverse origin which were identified by homology searches in online databases. Activity yields under different expression conditions and temperature stabilities were compared to three previously described enzymes from Bacillus subtilis, Bacillus pumilus and Bacillus clausii. In almost all cases, a switch to oxygen-limited growth conditions after induction increased volumetric activity considerably. For proteins with predicted signal peptides for secretion, recombinant expression with and without signal sequence was investigated. Bacillus CotA-type LMCOs outperformed enzymes from Streptomyces and Gram-negative bacteria with respect to activity yields in Escherichia coli and application relevant biochemical properties. The novel Bacillus coagulans LMCO combined high activity yields in E. coli with unprecedented activity at strong alkaline pH and high storage stability, making it a promising candidate for further development.

  15. Establishing a high yielding streptomyces-based cell-free protein synthesis system.

    PubMed

    Li, Jian; Wang, He; Kwon, Yong-Chan; Jewett, Michael C

    2017-06-01

    Cell-free protein synthesis (CFPS) has emerged as a powerful platform for applied biotechnology and synthetic biology, with a range of applications in synthesizing proteins, evolving proteins, and prototyping genetic circuits. To expand the current CFPS repertoire, we report here the development and optimization of a Streptomyces-based CFPS system for the expression of GC-rich genes. By developing a streamlined crude extract preparation protocol and optimizing reaction conditions, we were able to achieve active enhanced green fluorescent protein (EGFP) yields of greater than 50 μg/mL with batch reactions lasting up to 3 h. By adopting a semi-continuous reaction format, the EGFP yield could be increased to 282 ± 8 μg/mL and the reaction time was extended to 48 h. Notably, our extract preparation procedures were robust to multiple Streptomyces lividans and Streptomyces coelicolor strains, although expression yields varied. We show that our optimized Streptomyces lividans system provides benefits when compared to an Escherichia coli-based CFPS system for increasing percent soluble protein expression for four Streptomyces-originated high GC-content genes that are involved in biosynthesis of the nonribosomal peptides tambromycin and valinomycin. Looking forward, we believe that our Streptomyces-based CFPS system will contribute significantly towards efforts to express complex natural product gene clusters (e.g., nonribosomal peptides and polyketides), providing a new avenue for obtaining and studying natural product biosynthesis pathways. Biotechnol. Bioeng. 2017;114: 1343-1353. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  16. TaER Expression Is Associated with Transpiration Efficiency Traits and Yield in Bread Wheat

    PubMed Central

    Zheng, Jiacheng; Yang, Zhiyuan; Madgwick, Pippa J.; Carmo-Silva, Elizabete; Parry, Martin A. J.; Hu, Yin-Gang

    2015-01-01

    ERECTA encodes a receptor-like kinase and is proposed as a candidate for determining transpiration efficiency of plants. Two genes homologous to ERECTA in Arabidopsis were identified on chromosomes 6 (TaER2) and 7 (TaER1) of bread wheat (Triticum aestivum L.), with copies of each gene on the A, B and D genomes of wheat. Similar expression patterns were observed for TaER1 and TaER2 with relatively higher expression of TaER1 in flag leaves of wheat at heading (Z55) and grain-filling (Z73) stages. Significant variations were found in the expression levels of both TaER1 and TaER2 in the flag leaves at both growth stages among 48 diverse bread wheat varieties. Based on the expression of TaER1 and TaER2, the 48 wheat varieties could be classified into three groups having high (5 varieties), medium (27 varieties) and low (16 varieties) levels of TaER expression. Significant differences were also observed between the three groups varying for TaER expression for several transpiration efficiency (TE)- related traits, including stomatal density (SD), transpiration rate, photosynthetic rate (A), instant water use efficiency (WUEi) and carbon isotope discrimination (CID), and yield traits of biomass production plant-1 (BYPP) and grain yield plant-1 (GYPP). Correlation analysis revealed that the expression of TaER1 and TaER2 at the two growth stages was significantly and negatively associated with SD (P<0.01), transpiration rate (P<0.05) and CID (P<0.01), while significantly and positively correlated with flag leaf area (FLA, P<0.01), A (P<0.05), WUEi (P<0.05), BYPP (P<0.01) and GYPP (P<0.01), with stronger correlations for TaER1 than TaER2 and at grain-filling stage than at heading stage. These combined results suggested that TaER involved in development of transpiration efficiency -related traits and yield in bread wheat, implying a function for TaER in regulating leaf development of bread wheat and contributing to expression of these traits. Moreover, the results indicate

  17. TaER Expression Is Associated with Transpiration Efficiency Traits and Yield in Bread Wheat.

    PubMed

    Zheng, Jiacheng; Yang, Zhiyuan; Madgwick, Pippa J; Carmo-Silva, Elizabete; Parry, Martin A J; Hu, Yin-Gang

    2015-01-01

    ERECTA encodes a receptor-like kinase and is proposed as a candidate for determining transpiration efficiency of plants. Two genes homologous to ERECTA in Arabidopsis were identified on chromosomes 6 (TaER2) and 7 (TaER1) of bread wheat (Triticum aestivum L.), with copies of each gene on the A, B and D genomes of wheat. Similar expression patterns were observed for TaER1 and TaER2 with relatively higher expression of TaER1 in flag leaves of wheat at heading (Z55) and grain-filling (Z73) stages. Significant variations were found in the expression levels of both TaER1 and TaER2 in the flag leaves at both growth stages among 48 diverse bread wheat varieties. Based on the expression of TaER1 and TaER2, the 48 wheat varieties could be classified into three groups having high (5 varieties), medium (27 varieties) and low (16 varieties) levels of TaER expression. Significant differences were also observed between the three groups varying for TaER expression for several transpiration efficiency (TE)- related traits, including stomatal density (SD), transpiration rate, photosynthetic rate (A), instant water use efficiency (WUEi) and carbon isotope discrimination (CID), and yield traits of biomass production plant-1 (BYPP) and grain yield plant-1 (GYPP). Correlation analysis revealed that the expression of TaER1 and TaER2 at the two growth stages was significantly and negatively associated with SD (P<0.01), transpiration rate (P<0.05) and CID (P<0.01), while significantly and positively correlated with flag leaf area (FLA, P<0.01), A (P<0.05), WUEi (P<0.05), BYPP (P<0.01) and GYPP (P<0.01), with stronger correlations for TaER1 than TaER2 and at grain-filling stage than at heading stage. These combined results suggested that TaER involved in development of transpiration efficiency -related traits and yield in bread wheat, implying a function for TaER in regulating leaf development of bread wheat and contributing to expression of these traits. Moreover, the results indicate

  18. Expression of cyanobacterial FBP/SBPase in soybean prevents yield depression under future climate conditions.

    PubMed

    Köhler, Iris H; Ruiz-Vera, Ursula M; VanLoocke, Andy; Thomey, Michell L; Clemente, Tom; Long, Stephen P; Ort, Donald R; Bernacchi, Carl J

    2016-12-12

    Predictions suggest that current crop production needs to double by 2050 to meet global food and energy demands. Based on theory and experimental studies, overexpression of the photosynthetic enzyme sedoheptulose-1,7-bisphosphatase (SBPase) is expected to enhance C3 crop photosynthesis and yields. Here we test how expression of the cyanobacterial, bifunctional fructose-1,6/sedoheptulose-1,7-bisphosphatase (FBP/SBPase) affects carbon assimilation and seed yield (SY) in a major crop (soybean, Glycine max). For three growing seasons, wild-type (WT) and FBP/SBPase-expressing (FS) plants were grown in the field under ambient (400 μmol mol(-1)) and elevated (600 μmol mol(-1)) CO2 concentrations [CO2] and under ambient and elevated temperatures (+2.7 °C during daytime, +3.4 °C at night) at the SoyFACE research site. Across treatments, FS plants had significantly higher carbon assimilation (4-14%), V c,max (5-8%), and J max (4-8%). Under ambient [CO2], elevated temperature led to significant reductions of SY of both genotypes by 19-31%. However, under elevated [CO2] and elevated temperature, FS plants maintained SY levels, while the WT showed significant reductions between 11% and 22% compared with plants under elevated [CO2] alone. These results show that the manipulation of the photosynthetic carbon reduction cycle can mitigate the effects of future high CO2 and high temperature environments on soybean yield.

  19. Expression of cyanobacterial FBP/SBPase in soybean prevents yield depression under future climate conditions

    PubMed Central

    Köhler, Iris H.; Ruiz-Vera, Ursula M.; Thomey, Michell L.; Clemente, Tom; Ort, Donald R.

    2017-01-01

    Abstract Predictions suggest that current crop production needs to double by 2050 to meet global food and energy demands. Based on theory and experimental studies, overexpression of the photosynthetic enzyme sedoheptulose-1,7-bisphosphatase (SBPase) is expected to enhance C3 crop photosynthesis and yields. Here we test how expression of the cyanobacterial, bifunctional fructose-1,6/sedoheptulose-1,7-bisphosphatase (FBP/SBPase) affects carbon assimilation and seed yield (SY) in a major crop (soybean, Glycine max). For three growing seasons, wild-type (WT) and FBP/SBPase-expressing (FS) plants were grown in the field under ambient (400 μmol mol−1) and elevated (600 μmol mol−1) CO2 concentrations [CO2] and under ambient and elevated temperatures (+2.7 °C during daytime, +3.4 °C at night) at the SoyFACE research site. Across treatments, FS plants had significantly higher carbon assimilation (4–14%), Vc,max (5–8%), and Jmax (4–8%). Under ambient [CO2], elevated temperature led to significant reductions of SY of both genotypes by 19–31%. However, under elevated [CO2] and elevated temperature, FS plants maintained SY levels, while the WT showed significant reductions between 11% and 22% compared with plants under elevated [CO2] alone. These results show that the manipulation of the photosynthetic carbon reduction cycle can mitigate the effects of future high CO2 and high temperature environments on soybean yield. PMID:28204603

  20. High-yield expression in Escherichia coli of soluble human MT2A with native functions.

    PubMed

    Yang, Fang; Zhou, Min; He, Zhimin; Liu, Xiaorong; Sun, Lin; Sun, Yu; Chen, Zhuchu

    2007-05-01

    Metallothioneins (MTs) are a family of low molecular weight, cysteine rich heavy metal binding proteins with multifunction, such as metal detoxification and antioxidation, and are involved in a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. However, high yield expression of human MT in Escherichia coli has not been established effectively. To produce large amounts of human MT protein at low cost, recombinant human metallothionein 2A (MT2A) protein with an N-terminal GST tag was successfully expressed at high levels in soluble form in E. coli and high purification of it was established by affinity chromatography under native conditions. The final yield was about 5mg of the recombinant MT2A per liter of bacterial culture with the purity of 97.9%. Chemical and functional characteristics analysis of the recombinant human MT2A exhibited intact metal binding ability, hydroxyl radical scavenging ability and significant protective role against DNA damage caused by UVC radiation. Establishment of highly purified recombinant human MT2A protein with native characteristics at low cost would improve its function study and wide applications in protecting against oxidative damage and UV radiation.

  1. Surprising yields with no-till cropping systems

    USDA-ARS?s Scientific Manuscript database

    Producers using no-till practices have observed that crop yields can greatly exceed expectations based on nutrient and water supply. For example, Ralph Holzwarth, who farms near Gettysburg, SD, has averaged 150 bu/ac of corn on his farm for the past 6 years. We were surprised with this yield, as c...

  2. Zilpaterol hydrochloride improves beef yield, changes palatability traits, and increases calpain-calpastatin gene expression in Nellore heifers.

    PubMed

    Cônsolo, Nara Regina Brandão; Ferrari, Viviane Borba; Mesquita, Ligia Garcia; Goulart, Rodrigo Silva; Silva, Luis Felipe Prada E

    2016-11-01

    This research aimed to evaluate the effects of the beta-agonist zilpaterol hydrochloride (ZH) on carcass traits, subprimal yield, meat quality, palatability traits, and gene expression in Nellore heifers. Zilpaterol increased Longissimus lumborum area and did not change back fat thickness, meat color, and cooking loss. Heifers fed ZH had greater hindquarter weight and carcass percentage. Muscles from hindquarter were heavier for animals fed ZH. Forequarter (% of carcass) decreased and brisket did not change with ZH supplementation. There were no differences between treatments for steak aroma, beef flavor, and off-flavor. However, tenderness and juiciness were reduced by ZH, depending on postmortem aging. Zilpaterol increased Calpain-1, Calpain-2, and calpastatin mRNA expression, with no effect of day of slaughter or ZH×Day interaction. In conclusion, ZH supplementation improved hypertrophy, meat production, and debone yield in Nellore heifers, which led to decreased tenderness and to increased mRNA expression in the calpain-calpastatin system. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Improving starch yield in cereals by over-expression of ADPglucose pyrophosphorylase: expectations and unanticipated outcomes.

    PubMed

    Tuncel, Aytug; Okita, Thomas W

    2013-10-01

    Significant improvements in crop productivity are required to meet the nutritional requirements of a growing world population. This challenge is magnified by an increased demand for bioenergy as a means to mitigate carbon inputs into the environment. Starch is a major component of the harvestable organs of many crop plants, and various endeavors have been taken to improve the yields of starchy organs through the manipulation of starch synthesis. Substantial efforts have centered on the starch regulatory enzyme ADPglucose pyrophosphorylase (AGPase) due to its pivotal role in starch biosynthesis. These efforts include over-expression of this enzyme in cereal plants such as maize, rice and wheat as well as potato and cassava, as they supply the bulk of the staple food worldwide. In this perspective, we describe efforts to increase starch yields in cereal grains by first providing an introduction about the importance of source-sink relationship and the motives behind the efforts to alter starch biosynthesis and turnover in leaves. We then discuss the catalytic and regulatory properties of AGPase and the molecular approaches used to enhance starch synthesis by manipulation of this process during grain filling using seed-specific promoters. Several studies have demonstrated increases in starch content per seed using endosperm-specific promoters, but other studies have demonstrated an increase in seed number with only marginal impact on seed weight. Potential mechanisms that may be responsible for this paradoxical increase in seed number will also be discussed. Finally, we describe current efforts and future prospects to improve starch yield in cereals. These efforts include further enhancement of starch yield in rice by augmenting the process of ADPglucose transport into amyloplast as well as other enzymes involved in photoassimilate partitioning in seeds. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  4. Inducible gene expression systems and plant biotechnology.

    PubMed

    Corrado, Giandomenico; Karali, Marianthi

    2009-01-01

    Plant biotechnology relies heavily on the genetic manipulation of crops. Almost invariantly, the gene of interest is expressed in a constitutive fashion, although this may not be strictly necessary for several applications. Currently, there are several regulatable expression systems for the temporal, spatial and quantitative control of transgene activity. These molecular switches are based on components derived from different organisms, which range from viruses to higher eukaryotes. Many inducible systems have been designed for fundamental and applied research and since their initial development, they have become increasingly popular in plant molecular biology. This review covers a broad number of inducible expression systems examining their properties and relevance for plant biotechnology in its various guises, from molecular breeding to pharmaceutical and industrial applications. For each system, we examine some advantages and limitations, also in relation to the strategy on which they rely. Besides being necessary to control useful genes that may negatively affect crop yield and quality, we discuss that inducible systems can be also used to increase public acceptance of GMOs, reducing some of the most common concerns. Finally, we suggest some directions and future developments for their further diffusion in agriculture and biotechnology.

  5. Pluto Express power system architecture

    SciTech Connect

    Carr, G.A.

    1996-12-31

    The Pluto Express power system must answer the challenge of the next generation spacecraft by reducing its power, mass and volume envelopes. Technology developed by the New Millennium Program will enable the power system to meet the stringent requirements for the Pluto Express mission without exceeding the spacecraft mass and volume budgets. Traditionally, there has been an increasing trend of the percentage of mass of the power system electronics with respect to the total spacecraft mass. With all of the previous technology focus on high density digital packaging, the power system electronics have not been keeping pace forcing the spacecraft to absorb a relative increase in the power system mass. The increasing trend can be reversed by using mixed signal ASICs and high density multi-chip-module (MCM) packaging techniques validated by the New Millennium Program. As the size of the spacecraft shrinks, the power system electronics must become tightly integrated with the spacecraft loads. The power system architecture needs the flexibility to accommodate the specific load requirements without sacrificing the capability for growth or reduction as the spacecraft requirements change throughout the development. Modularity is a key requirement that will reduce the overall power system cost. Although the focus has been on shrinking the power system volume and mass, the efficiency and functionality cannot be ignored. Increased efficiency and functionality will only enhance the power systems capability to reduce spacecraft power requirements. The combination of the New Millennium packaging technologies with the Pluto Express power system architecture will produce a product with the capability to meet a wide range of mission profiles while reducing system development costs.

  6. Seed-specific expression of the wheat puroindoline genes improves maize wet milling yields.

    PubMed

    Zhang, Jinrui; Martin, John M; Beecher, Brian; Morris, Craig F; Curtis Hannah, L; Giroux, Michael J

    2009-10-01

    The texture of maize (Zea mays L.) seeds is important to seed processing properties, and soft dent maize is preferred for both wet-milling and livestock feed applications. The puroindoline genes (Pina and Pinb) are the functional components of the wheat (Triticum aestivum L.) Hardness locus and together function to create soft grain texture in wheat. The PINs (PINA and PINB) are believed to act by binding to lipids on the surface of starch granules, preventing tight adhesion between starch granules and the surrounding protein matrix during seed maturation. Here, maize kernel structure and wet milling properties were successfully modified by the endosperm-specific expression of wheat Pins (Pina and Pinb). Pins were introduced into maize under the control of a maize gamma-Zein promoter. Three Pina/Pinb expression positive transgenic lines were evaluated over two growing seasons. Textural analysis of the maize seeds indicated that the expression of PINs decreased adhesion between starch and protein matrix and reduced maize grain hardness significantly. Reduction in pressure required to fracture kernels ranged from 15.65% to 36.86% compared with control seeds. Further, the PINs transgenic maize seeds had increased levels of extractable starch as characterized by a small scale wet milling method. Starch yield was increased by 4.86% on average without negatively impacting starch purity. The development of softer maize hybrids with higher starch extractability would be of value to maize processors.

  7. Transgenic Arabidopsis Gene Expression System

    NASA Technical Reports Server (NTRS)

    Ferl, Robert; Paul, Anna-Lisa

    2009-01-01

    The Transgenic Arabidopsis Gene Expression System (TAGES) investigation is one in a pair of investigations that use the Advanced Biological Research System (ABRS) facility. TAGES uses Arabidopsis thaliana, thale cress, with sensor promoter-reporter gene constructs that render the plants as biomonitors (an organism used to determine the quality of the surrounding environment) of their environment using real-time nondestructive Green Fluorescent Protein (GFP) imagery and traditional postflight analyses.

  8. Site-specific integration and constitutive expression of key genes into Escherichia coli chromosome increases shikimic acid yields.

    PubMed

    Liu, Xianglei; Lin, Jun; Hu, Haifeng; Zhou, Bin; Zhu, Baoquan

    2016-01-01

    As the key starting material for the chemical synthesis of Oseltamivir, shikimic acid (SA) has captured worldwide attention. Many researchers have tried to improve SA production by metabolic engineering, yet expression plasmids were used generally. In recent years, site-specific integration of key genes into chromosome to increase the yield of metabolites showed considerable advantages. The genes could maintain stably and express constitutively without induction. Herein, crucial genes aroG, aroB, tktA, aroE (encoding 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase, dehydroquinate synthase, transketolase and shikimate dehydrogenase, respectively) of SA pathway and glk, galP (encoding glucokinase and galactose permease) were integrated into the locus of ptsHIcrr (phosphoenolpyruvate: carbohydrate phosphotransferase system operon) in a shikimate kinase genetic defect strain Escherichia coli BW25113 (ΔaroL/aroK, DE3). Furthermore, another key gene ppsA (encoding phosphoenolpyruvate synthase) was integrated into tyrR (encoding Tyr regulator protein). As a result, SA production of the recombinant (SA5/pGBAE) reached to 4.14 g/L in shake flask and 27.41 g/L in a 5-L bioreactor. These data suggested that integration of key genes increased SA yields effectively. This strategy is environmentally friendly for no antibiotic is added, simple to handle without induction, and suitable for industrial production.

  9. Positively regulated bacterial expression systems

    PubMed Central

    Brautaset, Trygve; Lale, Rahmi; Valla, Svein

    2009-01-01

    Summary Regulated promoters are useful tools for many aspects related to recombinant gene expression in bacteria, including for high‐level expression of heterologous proteins and for expression at physiological levels in metabolic engineering applications. In general, it is common to express the genes of interest from an inducible promoter controlled either by a positive regulator or by a repressor protein. In this review, we discuss established and potentially useful positively regulated bacterial promoter systems, with a particular emphasis on those that are controlled by the AraC‐XylS family of transcriptional activators. The systems function in a wide range of microorganisms, including enterobacteria, soil bacteria, lactic bacteria and streptomycetes. The available systems that have been applied to express heterologous genes are regulated either by sugars (l‐arabinose, l‐rhamnose, xylose and sucrose), substituted benzenes, cyclohexanone‐related compounds, ε‐caprolactam, propionate, thiostrepton, alkanes or peptides. It is of applied interest that some of the inducers require the presence of transport systems, some are more prone than others to become metabolized by the host and some have been applied mainly in one or a limited number of species. Based on bioinformatics analyses, the AraC‐XylS family of regulators contains a large number of different members (currently over 300), but only a small fraction of these, the XylS/Pm, AraC/PBAD, RhaR‐RhaS/rhaBAD, NitR/PnitA and ChnR/Pb regulator/promoter systems, have so far been explored for biotechnological applications. PMID:21261879

  10. Gastrointestinal Spatiotemporal mRNA Expression of Ghrelin vs Growth Hormone Receptor and New Growth Yield Machine Learning Model Based on Perturbation Theory.

    PubMed

    Ran, Tao; Liu, Yong; Li, Hengzhi; Tang, Shaoxun; He, Zhixiong; Munteanu, Cristian R; González-Díaz, Humberto; Tan, Zhiliang; Zhou, Chuanshe

    2016-07-27

    The management of ruminant growth yield has economic importance. The current work presents a study of the spatiotemporal dynamic expression of Ghrelin and GHR at mRNA levels throughout the gastrointestinal tract (GIT) of kid goats under housing and grazing systems. The experiments show that the feeding system and age affected the expression of either Ghrelin or GHR with different mechanisms. Furthermore, the experimental data are used to build new Machine Learning models based on the Perturbation Theory, which can predict the effects of perturbations of Ghrelin and GHR mRNA expression on the growth yield. The models consider eight longitudinal GIT segments (rumen, abomasum, duodenum, jejunum, ileum, cecum, colon and rectum), seven time points (0, 7, 14, 28, 42, 56 and 70 d) and two feeding systems (Supplemental and Grazing feeding) as perturbations from the expected values of the growth yield. The best regression model was obtained using Random Forest, with the coefficient of determination R(2) of 0.781 for the test subset. The current results indicate that the non-linear regression model can accurately predict the growth yield and the key nodes during gastrointestinal development, which is helpful to optimize the feeding management strategies in ruminant production system.

  11. Neuroglobin expression in the mammalian auditory system

    PubMed Central

    Reuss, Stefan; Banica, Ovidiu; Elgurt, Mirra; Mitz, Stephanie; Disque-Kaiser, Ursula; Riemann, Randolf; Hill, Marco; Jaquish, Dawn V.; Koehrn, Fred J.; Burmester, Thorsten; Hankeln, Thomas; Woolf, Nigel K.

    2015-01-01

    The energy-yielding pathways that provide the large amounts of metabolic energy required by inner ear sensorineural cells are poorly understood. Neuroglobin (Ngb) is a neuron-specific hemoprotein of the globin family, which is suggested to be involved in oxidative energy metabolism. Here we present quantitative real-time reverse transcription PCR, in situ hybridization, immunohistochemical and Western blot evidence that neuroglobin is highly expressed in the mouse and rat cochlea. For primary cochlea neurons, Ngb expression is limited to the subpopulation of type I spiral ganglion cells, those which innervate inner hair cells, while the subpopulation of type II spiral ganglion cells which innervate the outer hair cells do not express Ngb. We further investigated Ngb distribution in rat, mouse and human auditory brainstem centers, and found that the cochlear nuclei and superior olivary complex (SOC) also express considerable amounts of Ngb. Notably, the majority of olivocochlear neurons, those which provide efferent innervation of outer hair cells as identified by neuronal tract tracing, were Ngb-immunoreactive. We also observed that neuroglobin in the SOC frequently co-localized with neuronal nitric oxide synthase, the enzyme responsible for nitric oxide production. Our findings suggest that neuroglobin is well positioned to play an important physiologic role in the oxygen homeostasis of the peripheral and central auditory nervous system, and provides the first evidence that Ngb signal differentiates the central projections of the inner and outer hair cells. PMID:25636685

  12. Neuroglobin Expression in the Mammalian Auditory System.

    PubMed

    Reuss, Stefan; Banica, Ovidiu; Elgurt, Mirra; Mitz, Stephanie; Disque-Kaiser, Ursula; Riemann, Randolf; Hill, Marco; Jaquish, Dawn V; Koehrn, Fred J; Burmester, Thorsten; Hankeln, Thomas; Woolf, Nigel K

    2016-04-01

    The energy-yielding pathways that provide the large amounts of metabolic energy required by inner ear sensorineural cells are poorly understood. Neuroglobin (Ngb) is a neuron-specific hemoprotein of the globin family, which is suggested to be involved in oxidative energy metabolism. Here, we present quantitative real-time reverse transcription PCR, in situ hybridization, immunohistochemical, and Western blot evidence that neuroglobin is highly expressed in the mouse and rat cochlea. For primary cochlea neurons, Ngb expression is limited to the subpopulation of type I spiral ganglion cells, those which innervate inner hair cells, while the subpopulation of type II spiral ganglion cells which innervate the outer hair cells do not express Ngb. We further investigated Ngb distribution in rat, mouse, and human auditory brainstem centers, and found that the cochlear nuclei and superior olivary complex (SOC) also express considerable amounts of Ngb. Notably, the majority of olivocochlear neurons, those which provide efferent innervation of outer hair cells as identified by neuronal tract tracing, were Ngb-immunoreactive. We also observed that neuroglobin in the SOC frequently co-localized with neuronal nitric oxide synthase, the enzyme responsible for nitric oxide production. Our findings suggest that neuroglobin is well positioned to play an important physiologic role in the oxygen homeostasis of the peripheral and central auditory nervous system, and provides the first evidence that Ngb signal differentiates the central projections of the inner and outer hair cells.

  13. Upper Limits for Power Yield in Thermal, Chemical, and Electrochemical Systems

    NASA Astrophysics Data System (ADS)

    Sieniutycz, Stanislaw

    2010-03-01

    We consider modeling and power optimization of energy converters, such as thermal, solar and chemical engines and fuel cells. Thermodynamic principles lead to expressions for converter's efficiency and generated power. Efficiency equations serve to solve the problems of upgrading or downgrading a resource. Power yield is a cumulative effect in a system consisting of a resource, engines, and an infinite bath. While optimization of steady state systems requires using the differential calculus and Lagrange multipliers, dynamic optimization involves variational calculus and dynamic programming. The primary result of static optimization is the upper limit of power, whereas that of dynamic optimization is a finite-rate counterpart of classical reversible work (exergy). The latter quantity depends on the end state coordinates and a dissipation index, h, which is the Hamiltonian of the problem of minimum entropy production. In reacting systems, an active part of chemical affinity constitutes a major component of the overall efficiency. The theory is also applied to fuel cells regarded as electrochemical flow engines. Enhanced bounds on power yield follow, which are stronger than those predicted by the reversible work potential.

  14. Corn and soybean grain yields in a long-term tillage and cropping systems study

    USDA-ARS?s Scientific Manuscript database

    Reports on the long-term effects of tillage and cropping systems on corn and soybean yields are limited. Yields have been measured in a long-term experiment (30+ years) with three cropping systems [continuous corn (CC), continuous soybean (CSB), and soybean-corn (SB-C)] in six primary tillage system...

  15. Differential gene expression at different stages of mesocarp development in high- and low-yielding oil palm.

    PubMed

    Wong, Yick Ching; Teh, Huey Fang; Mebus, Katharina; Ooi, Tony Eng Keong; Kwong, Qi Bin; Koo, Ka Loo; Ong, Chuang Kee; Mayes, Sean; Chew, Fook Tim; Appleton, David R; Kulaveerasingam, Harikrishna

    2017-06-21

    The oil yield trait of oil palm is expected to involve multiple genes, environmental influences and interactions. Many of the underlying mechanisms that contribute to oil yield are still poorly understood. In this study, we used a microarray approach to study the gene expression profiles of mesocarp tissue at different developmental stages, comparing genetically related high- and low- oil yielding palms to identify genes that contributed to the higher oil-yielding palm and might contribute to the wider genetic improvement of oil palm breeding populations. A total of 3412 (2001 annotated) gene candidates were found to be significantly differentially expressed between high- and low-yielding palms at at least one of the different stages of mesocarp development evaluated. Gene Ontologies (GO) enrichment analysis identified 28 significantly enriched GO terms, including regulation of transcription, fatty acid biosynthesis and metabolic processes. These differentially expressed genes comprise several transcription factors, such as, bHLH, Dof zinc finger proteins and MADS box proteins. Several genes involved in glycolysis, TCA, and fatty acid biosynthesis pathways were also found up-regulated in high-yielding oil palm, among them; pyruvate dehydrogenase E1 component Subunit Beta (PDH), ATP-citrate lyase, β- ketoacyl-ACP synthases I (KAS I), β- ketoacyl-ACP synthases III (KAS III) and ketoacyl-ACP reductase (KAR). Sucrose metabolism-related genes such as Invertase, Sucrose Synthase 2 and Sucrose Phosphatase 2 were found to be down-regulated in high-yielding oil palms, compared to the lower yield palms. Our findings indicate that a higher carbon flux (channeled through down-regulation of the Sucrose Synthase 2 pathway) was being utilized by up-regulated genes involved in glycolysis, TCA and fatty acid biosynthesis leading to enhanced oil production in the high-yielding oil palm. These findings are an important stepping stone to understand the processes that lead to

  16. Enhanced artemisinin yield by expression of rol genes in Artemisia annua.

    PubMed

    Dilshad, Erum; Cusido, Rosa Maria; Palazon, Javier; Estrada, Karla Ramirez; Bonfill, Mercedes; Mirza, Bushra

    2015-10-29

    Despite of many advances in the treatment of malaria, it is still the fifth most prevalent disease worldwide and is one of the major causes of death in the developing countries which accounted for 584,000 deaths in 2013, as estimated by World Health Organization. Artemisinin from Artemisia annua is still one of the most effective treatments for malaria. Increasing the artemisinin content of A. annua plants by genetic engineering would improve the availability of this much-needed drug. In this regard, a high artemisinin-yielding hybrid of A. annua produced by the centre for novel agricultural products of the University of York, UK, was selected (artemisinin maximally 1.4 %). As rol genes are potential candidates of biochemical engineering, genetic transformation of A. annua with Agrobacterium tumefaciens GV3101 harbouring vectors with rol B and rol C genes was carried out with the objective of enhancement of artemisinin content. Transgenic lines produced were analysed by the LC-MS for quantitative analysis of artemisinin and analogues. These high artemisinin yielding transgenics were also analysed by real time quantitative PCR to find the molecular dynamics of artemisinin enhancement. Genes of artemisinin biosynthetic pathway were studied including amorphadiene synthase (ADS), cytochrome P450, (CYP71AV1) and aldehyde dehydrogenase 1 (ALDH1). Trichome-specific fatty acyl-CoA reductase 1(TAFR1) is an enzyme involved in both trichome development and sesquiterpenoid biosynthesis and both processes are important for artemisinin biosynthesis. Thus, real time qPCR analysis of the TAFR1 gene was carried out, and trichome density was determined. Transgenics of rol B gene showed two- to ninefold (the decimal adds nothing in the abstract, please simplify to two- to ninefold) increase in artemisinin, 4-12-fold increase in artesunate and 1.2-3-fold increase in dihydroartemisinin. Whereas in the case of rol C gene transformants, a fourfold increase in artemisinin, four to

  17. Cloning, expression, purification and characterization of his-tagged human glucose-6-phosphate dehydrogenase: a simplified method for protein yield.

    PubMed

    Gómez-Manzo, Saúl; Terrón-Hernández, Jessica; de la Mora-de la Mora, Ignacio; García-Torres, Itzhel; López-Velázquez, Gabriel; Reyes-Vivas, Horacio; Oria-Hernández, Jesús

    2013-10-01

    Glucose-6-phosphate dehydrogenase (G6PD) catalyzes the first step of the pentose phosphate pathway. In erythrocytes, the functionality of the pathway is crucial to protect these cells against oxidative damage. G6PD deficiency is the most frequent enzymopathy in humans with a global prevalence of 4.9 %. The clinical picture is characterized by chronic or acute hemolysis in response to oxidative stress, which is related to the low cellular activity of G6PD in red blood cells. The disease is heterogeneous at genetic level with around 160 mutations described, mostly point mutations causing single amino acid substitutions. The biochemical studies aimed to describe the detrimental effects of mutations on the functional and structural properties of human G6PD are indispensable to understand the molecular physiopathology of this disease. Therefore, reliable systems for efficient expression and purification of the protein are highly desirable. In this work, human G6PD was heterologously expressed in Escherichia coli and purified by immobilized metal affinity chromatography in a single chromatographic step. The structural and functional characterization indicates that His-tagged G6PD resembles previous preparations of recombinant G6PD. In contrast with previous protein yield systems, our method is based on commonly available resources and fully accessible laboratory equipment; therefore, it can be readily implemented.

  18. Heterologous high yield expression and purification of neurotensin and its functional fragment in Escherichia coli.

    PubMed

    Tapaneeyakorn, Satita; Ross, Simon; Attrill, Helen; Watts, Anthony

    2010-11-01

    Peptide synthesis is widely used for the production of small proteins and peptides, but producing uniformly isotopically labelled peptides for NMR and other biophysical studies could be limited for economic reasons. Here, we propose a use of a modified pGEV-1 plasmid to express neurotensin (NT(1-13)), pGlu(1)-Leu(2)-Tyr(3)-Glu(4)-Asn(5)-Lys(6)-Pro(7)-Arg(8)-Arg(9)-Pro(10)-Tyr(11)-Ile(12)-Leu(13)-OH, as a C-terminal fusion protein with the GB1 domain of streptococcal protein G. The free carboxyl-terminus is important for the function of several peptide hormones, including neurotensin. Therefore, for the pGEV-NT(1-13) construct, the C-terminal pGEV-encoded 6xHis tag was removed and an N-terminal 8xHis tag was introduced for affinity purification. To facilitate removal of tags using CNBr cleavage, a methionine was introduced at the N-terminal of the peptide. Furthermore, this pGEV-NT(1-13) plasmid was used as a template to include a Pro-7 to Met mutation for CNBr cleavage, giving NT(8-13), the sub-fragment crucial for the biological activity of this peptide. These two constructs are being used to produce uniformly labelled NT(1-13) and NT(8-13) in high yield and in a cost effective way, using cheap (15)N and/or (13)C source. The modification proposed here using the pGEV-1 plasmid could be an alternative option for the high expression of other isotopically labelled and unlabelled short peptides, including hormones and hydrophobic membrane peptides.

  19. Identification of expressed genes in the mapped QTLs for yield related traits in rice

    USDA-ARS?s Scientific Manuscript database

    Improvement of grain yield is a perpetual goal in rice breeding. Yield and its component traits are quantitatively inherited and controlled by many genes. To identify the Quantitative Trait Loci (QTL) involved in yield, a recombinant inbred line (RIL) population consisting of 259 progeny was devel...

  20. Overexpression of wheat gene TaMOR improves root system architecture and grain yield in Oryza sativa

    PubMed Central

    Li, Bo; Liu, Dan; Li, Qiaoru; Mao, Xinguo; Li, Ang; Wang, Jingyi; Chang, Xiaoping; Jing, Ruilian

    2016-01-01

    Improved root architecture is an effective strategy to increase crop yield. We demonstrate that overexpression of transcription factor gene MORE ROOT (TaMOR) from wheat (Triticum aestivum L.) results in more roots and higher grain yield in rice (Oryza sativa). TaMOR, encoding a plant-specific transcription factor belonging to the ASYMMETRIC LEAVES2/LATERAL ORGAN BOUNDARIES (AS2/LOB) protein family, is highly conserved in wheat and its wild relatives. In this study, tissue expression patterns indicated that TaMOR mainly localizes to root initiation sites. The consistent gene expression pattern suggests that TaMOR is involved in root initiation. Exogenous auxin treatment induced TaMOR expression without de novo protein biosynthesis. Both in vivo and in vitro experiments demonstrated that TaMOR interacts with TaMOR-related protein TaMRRP, which contains a four-tandem-pentatricopeptide repeat motif. Overexpression of TaMOR led to more lateral roots in Arabidopsis thaliana, and TaMOR-overexpressing rice plants had more crown roots, a longer main panicle, a higher number of primary branches on the main panicle, a higher grain number per plant, and higher yield per plant than the plants of wild type. In general, TaMOR-D-overexpressing lines had larger root systems in Arabidopsis and rice, and produce a higher grain yield per plant. TaMOR therefore offers an opportunity to improve root architecture and increase yield in crop plants. PMID:27229732

  1. High-yield expression of human vascular endothelial growth factor VEGF(165) in Escherichia coli and purification for therapeutic applications.

    PubMed

    Pizarro, Shelly A; Gunson, Jane; Field, Matthew J; Dinges, Rachel; Khoo, Stefanie; Dalal, Milind; Lee, Michael; Kaleas, Kimberly A; Moiseff, Kathryn; Garnick, Susan; Reilly, Dorothea E; Laird, Michael W; Schmelzer, Charles H

    2010-08-01

    Vascular endothelial growth factor (VEGF(165)) is a potent mitogen that induces angiogenesis and vascular permeability in vivo and has demonstrated potential in therapeutic applications for accelerating wound healing. An industrial production method that provides high yield as well as high purity, quality, and potency is needed. The process described in this report involves a bacterial expression system capable of producing approximately 9g of rhVEGF per liter of broth and a downstream purification process consisting of protein refolding and three chromatography steps prior to formulation of the drug substance. A high cell density (HCD) fed-batch fermentation process was used to produce rhVEGF in periplasmic inclusion bodies. The inclusion bodies are harvested from the cell lysate and subjected to a single-step protein solubilization and refolding operation to extract the rhVEGF for purification. Overall recovery yields observed during development, including refolding and chromatography, were 30+/-6%. Host cell impurities are consistently cleared below target levels at both laboratory and large-scale demonstrating process robustness. The structure of the refolded and purified rhVEGF was confirmed by mass spectrometry, N-terminal sequencing, and tryptic peptide mapping while product variants were analyzed by multiple HPLC assays. Biological activity was verified by the proliferation of human umbilical vein derived endothelial cells. Copyright 2010 Elsevier Inc. All rights reserved.

  2. Pentopyranosyl Oligonucleotide Systems. Part 11: Systems with Shortened Backbones: D)-beta-Ribopyranosyl-(4 yields 3 )- and (L)-alpha - Lyxopyranosyl-(4 yields 3 )-oligonucleotides

    NASA Technical Reports Server (NTRS)

    Wippo, Harald; Reck, Folkert; Kudick, Rene; Ramaseshan, Mahesh; Ceulemans, Griet; Bolli, Martin; Krishnamurthy, Ramanarayanan; Eschenmoser, Albert

    2001-01-01

    The (L)-a-lyxopyranosyl-(4'yields 3')-oligonucleotide system-a member of a pentopyranosyl oligonucleotide family containing a shortened backbone-is capable of cooperative base-pairing and of cross-pairing with DNA and RNA. In contrast, corresponding (D)-beta-ribopyransoyl-(4' yields 3')-oligonucleotides do not show base-pairing under similar conditions. We conclude that oligonucleotide systems can violate the six-bonds-per-backbone-unit rule by having five bonds instead, if their vicinally bound phosphodiester bridges can assume an antiperiplanar conformation. An additional structural feature that seems relevant to the cross-pairing capability of the (L)-a-lyxopyranosyl-(4' yields 3')-oligonucleotide system is its (small) backbone/basepair axes inclination. An inclination which is similar to that in B-DNA seems to be a prerequisite for an oligonucleotide system s capability to cross-pair with DNA.

  3. Effect of transmembrane pressure on Factor VIII yield in ATF perfusion culture for the production of recombinant human Factor VIII co-expressed with von Willebrand factor.

    PubMed

    Kim, Seung-Chul; An, Sora; Kim, Hyun-Ki; Park, Beom-Soo; Na, Kyu-Heum; Kim, Byung-Gee

    2016-10-01

    In this study, we evaluated three cell retention devices, an alternating tangential flow (ATF) system, a spin-filter, and a Centritech Lab III centrifuge, for the production of recombinant human Factor VIII co-expressed with von Willebrand factor. From the results, it was found that the FVIII activity in bioreactor was significantly higher in the ATF perfusion culture than two other perfusion cultures. Moreover, the FVIII activity yield was unexpectedly low in the ATF perfusion culture. We have, therefore, studied the reasons for this low FVIII activity yield. It was revealed that the inactivation and the surface adsorption of FVIII onto the harvest bag were not the main reasons for the low yield in the ATF perfusion culture. The FVIII activity yield was not increased by the use of a hollow fiber filter with 0.5 μm pore size instead of 0.2 μm pore size. Additionally, the retention of FVIII molecules by the hollow fiber filter was a dominant factor in the low FVIII activity yield in the ATF perfusion culture. We demonstrated that FVIII yield was significantly improved by controlling transmembrane pressure (TMP) across the hollow fiber filter membrane. Taken together, these results suggest that TMP control could be an efficient method for the enhancement of FVIII yield in an ATF perfusion culture.

  4. High-yield expression and purification of the Hsp90-associated p23, FKBP52, HOP and SGTα proteins.

    PubMed

    Hildenbrand, Zacariah L; Molugu, Sudheer K; Paul, Atanu; Avila, Gustavo A; Herrera, Nadia; Xiao, Chuan; Cox, Marc B; Bernal, Ricardo A

    2010-10-15

    Hsp90 is a ubiquitous molecular chaperone that plays a key role in the malignant development of hormone-dependent pathologies such as cancer. An important role for Hsp90 is to facilitate the stable binding of steroid hormones to their respective receptors enabling the ligand-based signal to be carried to the nucleus and ultimately resulting in the up-regulation of gene expression. Along with Hsp90, this dynamic and transient process also involves the recruitment of additional proteins and co-chaperones that add further stability to the mature receptor-chaperone complex. In the work presented here, we describe four new protocols for the bacterial over-expression and column chromatographic purification of the human p23, FKBP52, HOP and SGTα proteins. Each of these proteins plays a distinct role in the steroid hormone receptor regulatory cycle. Affinity, ion-exchange and size-exclusion techniques were used to produce target yields greater than 50mg/L of cultured media, with each purified sample reaching near absolute sample homogeneity. These results reveal a reliable system for the production of p23, FKBP52, HOP and SGTα substrate proteins for use in the investigation of the Hsp90-associated protein interactions of the steroid hormone receptor cycle. Copyright © 2010 Elsevier B.V. All rights reserved.

  5. High-yield recombinant expression of the chicken antimicrobial peptide fowlicidin-2 in Escherichia coli.

    PubMed

    Feng, Xingjun; Xu, Wenshan; Qu, Pei; Li, Xiaochong; Xing, Liwei; Liu, Di; Jiao, Jian; Wang, Jue; Li, Zhongqiu; Liu, Chunlong

    2015-01-01

    The antimicrobial peptide fowlicidin-2 identified in chicken is a member of the cathelicidins family. The mature fowlicidin-2 possesses high antibacterial efficacy and lipopolysaccharide (LPS) neutralizing activity, and also represents an excellent candidate as an antimicrobial agent. In the present study, the recombinant fowlicidin-2 was successfully produced by Escherichia coli (E. coli) recombinant expression system. The gene encoding fowlicidin-2 with the codon preference of E. coli was designed through codon optimization and synthesized in vitro. The gene was then ligated into the plasmid pET-32a(+), which features fusion protein thioredoxin at the N-terminal. The recombinant plasmid was transformed into E. coli BL21(DE3) and cultured in Luria-Bertani (LB) medium. After isopropyl-β-D-thiogalactopyranoside (IPTG) induction, the fowlicidin-2 fusion protein was successfully expressed as inclusion bodies. The inclusion bodies were dissolved and successfully released the peptide in 70% formic acid solution containing cyanogen bromide (CNBr) in a single step. After purification by reverse-phase high-performance liquid chromatography (RP-HPLC), ∼6.0 mg of fowlicidin-2 with purity more than 97% was obtained from 1 litre of bacteria culture. The recombinant peptide exhibited high antibacterial activity against the Gram-positive and Gram-negative bacteria, and even drug-resistant strains. This system could be used to rapidly and efficiently produce milligram quantities of a battery of recombinant antimicrobial peptides as well as for large-scale production. © 2015 American Institute of Chemical Engineers.

  6. Efficient expression systems for cysteine proteases of malaria parasites

    PubMed Central

    Sarduy, Emir Salas; de los A. Chávez Planes, María

    2013-01-01

    Papain-like cysteine proteases of malaria parasites are considered important chemotherapeutic targets or valuable models for the evaluation of drug candidates. Consequently, many of these enzymes have been cloned and expressed in Escherichia coli for their biochemical characterization. However, their expression has been problematic, showing low yield and leading to the formation of insoluble aggregates. Given that highly-productive expression systems are required for the high-throughput evaluation of inhibitors, we analyzed the existing expression systems to identify the causes of such apparent issues. We found that significant divergences in codon and nucleotide composition from host genes are the most probable cause of expression failure, and propose several strategies to overcome these limitations. Finally we predict that yeast hosts Saccharomyces cerevisiae and Pichia pastoris may be better suited than E. coli for the efficient expression of plasmodial genes, presumably leading to soluble and active products reproducing structural and functional characteristics of the natural enzymes. PMID:23018863

  7. Topical imiquimod yields systemic effects due to unintended oral uptake.

    PubMed

    Grine, Lynda; Steeland, Sophie; Van Ryckeghem, Sara; Ballegeer, Marlies; Lienenklaus, Stefan; Weiss, Siegfried; Sanders, Niek N; Vandenbroucke, Roosmarijn E; Libert, Claude

    2016-01-28

    Repetitive application of topical imiquimod is used as an experimental model for the induction of psoriasiform skin lesions in mice. The model is characterized by several inflammatory processes, including cytokine production both locally and systemically, cellular infiltration, and splenomegaly. To investigate the production of type I interferons in response to imiquimod-containing Aldara cream, IFNβ-luciferase reporter mice were imaged in vivo and ex vivo. Type I interferons were found to be produced in the skin, but also in the intestinal system caused by unintended ingestion of imiquimod by the mice. Through the use of Elizabethan collars to prevent ingestion, these effects, including psoriasiform lesions were nearly completely prevented. Our findings reveal that topical treatment with Aldara induces a psoriasiform skin inflammation, but that its mode of action depends on ingestion of the chemical, which leads to systemic responses and affects local inflammation. Therefore, potential ingestion of topical treatments during experimental procedures should be taken into account during assessment of cutaneous inflammatory parameters in skin disease models.

  8. Topical imiquimod yields systemic effects due to unintended oral uptake

    PubMed Central

    Grine, Lynda; Steeland, Sophie; Van Ryckeghem, Sara; Ballegeer, Marlies; Lienenklaus, Stefan; Weiss, Siegfried; Sanders, Niek N.; Vandenbroucke, Roosmarijn E.; Libert, Claude

    2016-01-01

    Repetitive application of topical imiquimod is used as an experimental model for the induction of psoriasiform skin lesions in mice. The model is characterized by several inflammatory processes, including cytokine production both locally and systemically, cellular infiltration, and splenomegaly. To investigate the production of type I interferons in response to imiquimod-containing Aldara cream, IFNβ-luciferase reporter mice were imaged in vivo and ex vivo. Type I interferons were found to be produced in the skin, but also in the intestinal system caused by unintended ingestion of imiquimod by the mice. Through the use of Elizabethan collars to prevent ingestion, these effects, including psoriasiform lesions were nearly completely prevented. Our findings reveal that topical treatment with Aldara induces a psoriasiform skin inflammation, but that its mode of action depends on ingestion of the chemical, which leads to systemic responses and affects local inflammation. Therefore, potential ingestion of topical treatments during experimental procedures should be taken into account during assessment of cutaneous inflammatory parameters in skin disease models. PMID:26818707

  9. EarthSat spring wheat yield system test 1975, appendix 4

    NASA Technical Reports Server (NTRS)

    1976-01-01

    A computer system is presented which processes meteorological data from both ground observations and meteorologic satellites to define plant weather aspects on a four time per day basis. Plant growth stages are calculated and soil moisture profiles are defined by the system. The EarthSat system assesses plant stress and prepares forecasts of end-of-year yields. The system was used to forecast spring wheat yields in the upper Great Plains states. Hardware and software documentation is provided.

  10. Expressing the Geobacter metallireducens PilA in Geobacter sulfurreducens Yields Pili with Exceptional Conductivity

    PubMed Central

    Tan, Yang; Adhikari, Ramesh Y.; Malvankar, Nikhil S.; Ward, Joy E.; Woodard, Trevor L.; Nevin, Kelly P.

    2017-01-01

    ABSTRACT The electrically conductive pili (e-pili) of Geobacter sulfurreducens serve as a model for a novel strategy for long-range extracellular electron transfer. e-pili are also a new class of bioelectronic materials. However, the only other Geobacter pili previously studied, which were from G. uraniireducens, were poorly conductive. In order to obtain more information on the range of pili conductivities in Geobacter species, the pili of G. metallireducens were investigated. Heterologously expressing the PilA gene of G. metallireducens in G. sulfurreducens yielded a G. sulfurreducens strain, designated strain MP, that produced abundant pili. Strain MP exhibited phenotypes consistent with the presence of e-pili, such as high rates of Fe(III) oxide reduction and high current densities on graphite anodes. Individual pili prepared at physiologically relevant pH 7 had conductivities of 277 ± 18.9 S/cm (mean ± standard deviation), which is 5,000-fold higher than the conductivity of G. sulfurreducens pili at pH 7 and nearly 1 million-fold higher than the conductivity of G. uraniireducens pili at the same pH. A potential explanation for the higher conductivity of the G. metallireducens pili is their greater density of aromatic amino acids, which are known to be important components in electron transport along the length of the pilus. The G. metallireducens pili represent the most highly conductive pili found to date and suggest strategies for designing synthetic pili with even higher conductivities. PMID:28096491

  11. Expression, Purification, and Characteristic of Tibetan Sheep Breast Lysozyme Using Pichia pastoris Expression System

    PubMed Central

    Li, Jianbo; Jiang, Mingfeng; Wang, Yong

    2014-01-01

    A lysozyme gene from breast of Tibetan sheep was successfully expressed by secretion using a-factor signal sequence in the methylotrophic yeast, Pichia pastoris GS115. An expression yield and specific activity greater than 500 mg/L and 4,000 U/mg was obtained. Results at optimal pH and temperature showed recombinant lysozyme has higher lytic activity at pH 6.5 and 45°C. This study demonstrates the successful expression of recombinant lysozyme using the eukaryotic host organism P. pastoris paving the way for protein engineering. Additionally, this study shows the feasibility of subsequent industrial manufacture of the enzyme with this expression system together with a high purity scheme for easy high-yield purification. PMID:25049990

  12. Transient co-expression for fast and high-yield production of antibodies with human-like N-glycans in plants.

    PubMed

    Vézina, Louis-P; Faye, Loïc; Lerouge, Patrice; D'Aoust, Marc-André; Marquet-Blouin, Estelle; Burel, Carole; Lavoie, Pierre-Olivier; Bardor, Muriel; Gomord, Véronique

    2009-06-01

    Plant-based transient expression is potentially the most rapid and cost-efficient system for the production of recombinant pharmaceutical proteins, but safety concerns associated with plant-specific N-glycosylation have hampered its adoption as a commercial production system. In this article, we describe an approach based on the simultaneous transient co-expression of an antibody, a suppressor of silencing and a chimaeric human beta1,4-galactosyltransferase targeted for optimal activity to the early secretory pathway in agroinfiltrated Nicotiana benthamiana leaves. This strategy allows fast and high-yield production of antibodies with human-like N-glycans and, more generally, provides solutions to many critical problems posed by the large-scale production of therapeutic and vaccinal proteins, specifically yield, volume and quality.

  13. Production of different glycosylation variants of the tumour-targeting mAb H10 in Nicotiana benthamiana: influence on expression yield and antibody degradation.

    PubMed

    Lombardi, Raffaele; Donini, Marcello; Villani, Maria Elena; Brunetti, Patrizia; Fujiyama, Kazuhito; Kajiura, Hiroyuki; Paul, Matthew; Ma, Julian K-C; Benvenuto, Eugenio

    2012-10-01

    We previously described the expression of a tumour-targeting antibody (mAb H10) in Nicotiana benthamiana by vacuum-agro-infiltration and the remarkable yields of highly pure protein achieved. The objective of the present work was to investigate different strategies for transient overexpression of the mAb H10 in which glycan configuration was modulated and assess how these strategies affect the accumulation yield and stability of the antibody. To this aim, three procedures have been assayed: (1) Site-directed mutagenesis to abolish the glycosylation site; (2) endoplasmic reticulum retention (C-terminal SEKDEL fusion) to ensure predominantly high-mannose type glycans; and (3) expression in a N. benthamiana RNAi down-regulated line in which β1,2-xylosyltransferase and α1,3-fucosyltransferase gene expression is silenced. The three antibody variants (H10-Mut) (H10-SEKDEL) (H10(XylT/FucT)) were transiently expressed, purified and characterised for their glycosylation profile, expression/purification yield and antibody degradation pattern. Glycosylation analysis of H10(XylT/FucT) demonstrated the absence of plant complex-type sugars, while H10-SEKDEL, although substantially retained in the ER, revealed the presence of β1,2-xylose and α1,3-fucose residues, indicating a partial escape from the ER retrieval system. Antibody accumulation and purification yields were not enhanced by ER retention. All H10 antibody glyco-forms revealed greater degradation compared to the original, resulting mostly in the formation of Fab fragments. In the case of aglycosylated H10-Mut, more than 95% of the heavy chain was cleaved, confirming the pivotal role of the sugar moiety in protein stability. Identification of possible 'fragile' sites in the H10 antibody hinge region could be of general interest for the development of new strategies to reduce antibody degradation and increase the yield of intact IgGs in plants.

  14. Knowledge-based software system for fast yield loss detection in a semiconductor fab

    NASA Astrophysics Data System (ADS)

    Martin Santamaria, Victorino; Recio, Miguel; Merino, Miguel A.; Moreno, Julian; Fernandez, Almudena; Gonzalez, Gerardo; Sanchez, Guillermo; Barrios, Luis J.; del Castillo, Maria D.; Lemus, Lissette; Gonzalez, Angel L.

    1997-09-01

    The comparative analysis of process machines in terms of yield related metrics (such as probe and E-Test data, process and particle data,. ..) is a source of a great deal of information for yield improvement. With this aim we published on SPIE's Microelectronic Manufacturing an Advanced Software System to detect machine-related yield limitors using a comparative analysis. This paper presents the natural expansion of that Software System by converting it into a more knowledge-based tool for fast yield loss detection on a semiconductor fab. The new System performs, in an automatic mode, the comparison among machines for every single step selected in the fabrication routing. The detection of statistically significative differences among machines at every step is performed using algorithms that incorporate the overall analysts experience on our fab. The output of the System allows a fast detection and reaction to yield issues, mainly to those that are still on the initial or baseline stages.

  15. Performance benchmarking of four cell-free protein expression systems.

    PubMed

    Gagoski, Dejan; Polinkovsky, Mark E; Mureev, Sergey; Kunert, Anne; Johnston, Wayne; Gambin, Yann; Alexandrov, Kirill

    2016-02-01

    Over the last half century, a range of cell-free protein expression systems based on pro- and eukaryotic organisms have been developed and have found a range of applications, from structural biology to directed protein evolution. While it is generally accepted that significant differences in performance among systems exist, there is a paucity of systematic experimental studies supporting this notion. Here, we took advantage of the species-independent translation initiation sequence to express and characterize 87 N-terminally GFP-tagged human cytosolic proteins of different sizes in E. coli, wheat germ (WGE), HeLa, and Leishmania-based (LTE) cell-free systems. Using a combination of single-molecule fluorescence spectroscopy, SDS-PAGE, and Western blot analysis, we assessed the expression yields, the fraction of full-length translation product, and aggregation propensity for each of these systems. Our results demonstrate that the E. coli system has the highest expression yields. However, we observe that high expression levels are accompanied by production of truncated species-particularly pronounced in the case of proteins larger than 70 kDa. Furthermore, proteins produced in the E. coli system display high aggregation propensity, with only 10% of tested proteins being produced in predominantly monodispersed form. The WGE system was the most productive among eukaryotic systems tested. Finally, HeLa and LTE show comparable protein yields that are considerably lower than the ones achieved in the E. coli and WGE systems. The protein products produced in the HeLa system display slightly higher integrity, whereas the LTE-produced proteins have the lowest aggregation propensity among the systems analyzed. The high quality of HeLa- and LTE-produced proteins enable their analysis without purification and make them suitable for analysis of multi-domain eukaryotic proteins.

  16. Using FACE systems to screen wheat cultivars for yield increases at elevated CO2

    USDA-ARS?s Scientific Manuscript database

    Because of continuing increases in atmospheric CO2, identifying cultivars of crops with larger yield increases at elevated CO2 may provide an avenue to increase crop yield potential in future climates. Free-air CO2 enrichment (FACE) systems have most often been used with multiple replications of ea...

  17. Heterologous expression of chloroplast-localized geranylgeranyl pyrophosphate synthase confers fast plant growth, early flowering and increased seed yield.

    PubMed

    Tata, Sandeep Kumar; Jung, Jihye; Kim, Yoon-Ha; Choi, Jun Young; Jung, Ji-Yul; Lee, In-Jung; Shin, Jeong Sheop; Ryu, Stephen Beungtae

    2016-01-01

    Geranylgeranyl pyrophosphate synthase (GGPS) is a key enzyme for a structurally diverse class of isoprenoid biosynthetic metabolites including gibberellins, carotenoids, chlorophylls and rubber. We expressed a chloroplast-targeted GGPS isolated from sunflower (Helianthus annuus) under control of the cauliflower mosaic virus 35S promoter in tobacco (Nicotiana tabacum). The resulting transgenic tobacco plants expressing heterologous GGPS showed remarkably enhanced growth (an increase in shoot and root biomass and height), early flowering, increased number of seed pods and greater seed yield compared with that of GUS-transgenic lines (control) or wild-type plants. The gibberellin levels in HaGGPS-transgenic plants were higher than those in control plants, indicating that the observed phenotype may result from increased gibberellin content. However, in HaGGPS-transformant tobacco plants, we did not observe the phenotypic defects such as reduced chlorophyll content and greater petiole and stalk length, which were previously reported for transgenic plants expressing gibberellin biosynthetic genes. Fast plant growth was also observed in HaGGPS-expressing Arabidopsis and dandelion plants. The results of this study suggest that GGPS expression in crop plants may yield desirable agronomic traits, including enhanced growth of shoots and roots, early flowering, greater numbers of seed pods and/or higher seed yield. This research has potential applications for fast production of plant biomass that provides commercially valuable biomaterials or bioenergy. © 2015 Korea Research Institute of Bioscience & Biotechnology. Plant Biotechnology Journal published by John Wiley & Sons Ltd and Society for Experimental Biology, Association of Applied Biologists.

  18. Field-scale experiments reveal persistent yield gaps in low-input and organic cropping systems

    PubMed Central

    Kravchenko, Alexandra N.; Snapp, Sieglinde S.; Robertson, G. Philip

    2017-01-01

    Knowledge of production-system performance is largely based on observations at the experimental plot scale. Although yield gaps between plot-scale and field-scale research are widely acknowledged, their extent and persistence have not been experimentally examined in a systematic manner. At a site in southwest Michigan, we conducted a 6-y experiment to test the accuracy with which plot-scale crop-yield results can inform field-scale conclusions. We compared conventional versus alternative, that is, reduced-input and biologically based–organic, management practices for a corn–soybean–wheat rotation in a randomized complete block-design experiment, using 27 commercial-size agricultural fields. Nearby plot-scale experiments (0.02-ha to 1.0-ha plots) provided a comparison of plot versus field performance. We found that plot-scale yields well matched field-scale yields for conventional management but not for alternative systems. For all three crops, at the plot scale, reduced-input and conventional managements produced similar yields; at the field scale, reduced-input yields were lower than conventional. For soybeans at the plot scale, biological and conventional managements produced similar yields; at the field scale, biological yielded less than conventional. For corn, biological management produced lower yields than conventional in both plot- and field-scale experiments. Wheat yields appeared to be less affected by the experimental scale than corn and soybean. Conventional management was more resilient to field-scale challenges than alternative practices, which were more dependent on timely management interventions; in particular, mechanical weed control. Results underscore the need for much wider adoption of field-scale experimentation when assessing new technologies and production-system performance, especially as related to closing yield gaps in organic farming and in low-resourced systems typical of much of the developing world. PMID:28096409

  19. Field-scale experiments reveal persistent yield gaps in low-input and organic cropping systems.

    PubMed

    Kravchenko, Alexandra N; Snapp, Sieglinde S; Robertson, G Philip

    2017-01-31

    Knowledge of production-system performance is largely based on observations at the experimental plot scale. Although yield gaps between plot-scale and field-scale research are widely acknowledged, their extent and persistence have not been experimentally examined in a systematic manner. At a site in southwest Michigan, we conducted a 6-y experiment to test the accuracy with which plot-scale crop-yield results can inform field-scale conclusions. We compared conventional versus alternative, that is, reduced-input and biologically based-organic, management practices for a corn-soybean-wheat rotation in a randomized complete block-design experiment, using 27 commercial-size agricultural fields. Nearby plot-scale experiments (0.02-ha to 1.0-ha plots) provided a comparison of plot versus field performance. We found that plot-scale yields well matched field-scale yields for conventional management but not for alternative systems. For all three crops, at the plot scale, reduced-input and conventional managements produced similar yields; at the field scale, reduced-input yields were lower than conventional. For soybeans at the plot scale, biological and conventional managements produced similar yields; at the field scale, biological yielded less than conventional. For corn, biological management produced lower yields than conventional in both plot- and field-scale experiments. Wheat yields appeared to be less affected by the experimental scale than corn and soybean. Conventional management was more resilient to field-scale challenges than alternative practices, which were more dependent on timely management interventions; in particular, mechanical weed control. Results underscore the need for much wider adoption of field-scale experimentation when assessing new technologies and production-system performance, especially as related to closing yield gaps in organic farming and in low-resourced systems typical of much of the developing world.

  20. Methane yield phenotypes linked to differential gene expression in the sheep rumen microbiome

    PubMed Central

    Moon, Christina D.; Leahy, Sinead C.; Kang, Dongwan; Froula, Jeff; Kittelmann, Sandra; Fan, Christina; Deutsch, Samuel; Gagic, Dragana; Seedorf, Henning; Kelly, William J.; Atua, Renee; Sang, Carrie; Soni, Priya; Li, Dong; Pinares-Patiño, Cesar S.; McEwan, John C.; Janssen, Peter H.; Chen, Feng; Visel, Axel; Wang, Zhong; Attwood, Graeme T.

    2014-01-01

    Ruminant livestock represent the single largest anthropogenic source of the potent greenhouse gas methane, which is generated by methanogenic archaea residing in ruminant digestive tracts. While differences between individual animals of the same breed in the amount of methane produced have been observed, the basis for this variation remains to be elucidated. To explore the mechanistic basis of this methane production, we measured methane yields from 22 sheep, which revealed that methane yields are a reproducible, quantitative trait. Deep metagenomic and metatranscriptomic sequencing demonstrated a similar abundance of methanogens and methanogenesis pathway genes in high and low methane emitters. However, transcription of methanogenesis pathway genes was substantially increased in sheep with high methane yields. These results identify a discrete set of rumen methanogens whose methanogenesis pathway transcription profiles correlate with methane yields and provide new targets for CH4 mitigation at the levels of microbiota composition and transcriptional regulation. PMID:24907284

  1. Methane yield phenotypes linked to differential gene expression in the sheep rumen microbiome.

    PubMed

    Shi, Weibing; Moon, Christina D; Leahy, Sinead C; Kang, Dongwan; Froula, Jeff; Kittelmann, Sandra; Fan, Christina; Deutsch, Samuel; Gagic, Dragana; Seedorf, Henning; Kelly, William J; Atua, Renee; Sang, Carrie; Soni, Priya; Li, Dong; Pinares-Patiño, Cesar S; McEwan, John C; Janssen, Peter H; Chen, Feng; Visel, Axel; Wang, Zhong; Attwood, Graeme T; Rubin, Edward M

    2014-09-01

    Ruminant livestock represent the single largest anthropogenic source of the potent greenhouse gas methane, which is generated by methanogenic archaea residing in ruminant digestive tracts. While differences between individual animals of the same breed in the amount of methane produced have been observed, the basis for this variation remains to be elucidated. To explore the mechanistic basis of this methane production, we measured methane yields from 22 sheep, which revealed that methane yields are a reproducible, quantitative trait. Deep metagenomic and metatranscriptomic sequencing demonstrated a similar abundance of methanogens and methanogenesis pathway genes in high and low methane emitters. However, transcription of methanogenesis pathway genes was substantially increased in sheep with high methane yields. These results identify a discrete set of rumen methanogens whose methanogenesis pathway transcription profiles correlate with methane yields and provide new targets for CH4 mitigation at the levels of microbiota composition and transcriptional regulation. © 2014 Shi et al.; Published by Cold Spring Harbor Laboratory Press.

  2. The Effect of High Concentrations of Glufosinate Ammonium on the Yield Components of Transgenic Spring Wheat (Triticum aestivum L.) Constitutively Expressing the bar Gene

    PubMed Central

    Áy, Zoltán; Mihály, Róbert; Cserháti, Mátyás; Kótai, Éva; Pauk, János

    2012-01-01

    We present an experiment done on a bar + wheat line treated with 14 different concentrations of glufosinate ammonium—an effective component of nonselective herbicides—during seed germination in a closed experimental system. Yield components as number of spikes per plant, number of grains per spike, thousand kernel weight, and yield per plant were thoroughly analysed and statistically evaluated after harvesting. We found that a concentration of glufosinate ammonium 5000 times the lethal dose was not enough to inhibit the germination of transgenic plants expressing the bar gene. Extremely high concentrations of glufosinate ammonium caused a bushy phenotype, significantly lower numbers of grains per spike, and thousand kernel weights. Concerning the productivity, we observed that concentrations of glufosinate ammonium 64 times the lethal dose did not lead to yield depression. Our results draw attention to the possibilities implied in the transgenic approaches. PMID:22649303

  3. Direct and efficient ethanol production from high-yielding rice using a Saccharomyces cerevisiae strain that express amylases.

    PubMed

    Yamada, Ryosuke; Yamakawa, Syun-Ichi; Tanaka, Tsutomu; Ogino, Chiaki; Fukuda, Hideki; Kondo, Akihiko

    2011-04-07

    Efficient ethanol producing yeast Saccharomyces cerevisiae cannot produce ethanol from raw starch directly. Thus the conventional ethanol production required expensive and complex process. In this study, we developed a direct and efficient ethanol production process from high-yielding rice harvested in Japan by using amylase expressing yeast without any pretreatment or addition of enzymes or nutrients. Ethanol productivity from high-yielding brown rice (1.1g/L/h) was about 5-fold higher than that obtained from purified raw corn starch (0.2g/L/h) when nutrients were added. Using an inoculum volume equivalent to 10% of the fermentation volume without any nutrient supplementation resulted in ethanol productivity and yield reaching 1.2g/L/h and 101%, respectively, in a 24-h period. High-yielding rice was demonstrated to be a suitable feedstock for bioethanol production. In addition, our polyploid amylase-expressing yeast was sufficiently robust to produce ethanol efficiently from real biomass. This is first report of direct ethanol production on real biomass using an amylase-expressing yeast strain without any pretreatment or commercial enzyme addition.

  4. An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation.

    PubMed

    Bursey, Evan H; Kim, Chang-Yub; Yu, Minmin; Terwilliger, Thomas C; Hung, Li-Wei

    2006-12-01

    A highly automated method for rapidly identifying soluble protein variants with good expression yields has been developed. This method is based on a commercially available in vitro protein expression system. It consists of two polymerase chain reactions (PCR) followed by in vitro protein expression and protein quantification by dot blot. The PCR protocols have been improved and optimized to allow automation using commercial fluid handling devices. A PCR primer design program has also been implemented to streamline protein variant design. This automated protocol is highly reliable and has tremendously improved the throughput of expression screening as compared to conventional cell-based methods and manual in vitro methods. We have applied this method to 32 problematic targets from the TB Structural Genomics Consortium. Experimental results of these studies are reported.

  5. Transient expression systems for plant-derived biopharmaceuticals.

    PubMed

    Komarova, Tatiana V; Baschieri, Selene; Donini, Marcello; Marusic, Carla; Benvenuto, Eugenio; Dorokhov, Yuri L

    2010-08-01

    In the molecular farming area, transient expression approaches for pharmaceutical proteins production, mainly recombinant monoclonal antibodies and vaccines, were developed almost two decades ago and, to date, these systems basically depend on Agrobacterium-mediated delivery and virus expression machinery. We survey here the current state-of-the-art of this research field. Several vectors have been designed on the basis of DNA- and RNA-based plant virus genomes and viral vectors are used both as single- and multicomponent expression systems in different combinations depending on the protein of interest. The obvious advantages of these systems are ease of manipulation, speed, low cost and high yield of proteins. In addition, Agrobacterium-mediated expression also allows the production in plants of complex proteins assembled from subunits. Currently, the transient expression methods are preferential over any other transgenic system for the exploitation of large and unrestricted numbers of plants in a contained environment. By designing optimal constructs and related means of delivery into plant cells, the overall technology plan considers scenarios that envisage high yield of bioproducts and ease in monitoring the whole spectrum of upstream production, before entering good manufacturing practice facilities. In this way, plant-derived bioproducts show promise of high competitiveness towards classical eukaryotic cell factory systems.

  6. Co-Expression Network and Pathway Analyses Reveal Important Modules of miRNAs Regulating Milk Yield and Component Traits.

    PubMed

    Do, Duy N; Dudemaine, Pier-Luc; Li, Ran; Ibeagha-Awemu, Eveline M

    2017-07-18

    Co-expression network analyses provide insights into the molecular interactions underlying complex traits and diseases. In this study, co-expression network analysis was performed to detect expression patterns (modules or clusters) of microRNAs (miRNAs) during lactation, and to identify miRNA regulatory mechanisms for milk yield and component traits (fat, protein, somatic cell count (SCC), lactose, and milk urea nitrogen (MUN)) via miRNA target gene enrichment analysis. miRNA expression (713 miRNAs), and milk yield and components (Fat%, Protein%, lactose, SCC, MUN) data of nine cows at each of six different time points (day 30 (D30), D70, D130, D170, D230 and D290) of an entire lactation curve were used. Four modules or clusters (GREEN, BLUE, RED and TURQUOISE) of miRNAs were identified as important for milk yield and component traits. The GREEN and BLUE modules were significantly correlated (|r| > 0.5) with milk yield and lactose, respectively. The RED and TURQUOISE modules were significantly correlated (|r| > 0.5) with both SCC and lactose. In the GREEN module, three abundantly expressed miRNAs (miR-148a, miR-186 and miR-200a) were most significantly correlated to milk yield, and are probably the most important miRNAs for this trait. DDR1 and DDHX1 are hub genes for miRNA regulatory networks controlling milk yield, while HHEX is an important transcription regulator for these networks. miR-18a, miR-221/222 cluster, and transcription factors HOXA7, and NOTCH 3 and 4, are important for the regulation of lactose. miR-142, miR-146a, and miR-EIA17-14144 (a novel miRNA), and transcription factors in the SMAD family and MYB, are important for the regulation of SCC. Important signaling pathways enriched for target genes of miRNAs of significant modules, included protein kinase A and PTEN signaling for milk yield, eNOS and Noth signaling for lactose, and TGF β, HIPPO, Wnt/β-catenin and cell cycle signaling for SCC. Relevant enriched gene ontology (GO)-terms related to

  7. Co-Expression Network and Pathway Analyses Reveal Important Modules of miRNAs Regulating Milk Yield and Component Traits

    PubMed Central

    Do, Duy N.; Dudemaine, Pier-Luc; Li, Ran; Ibeagha-Awemu, Eveline M.

    2017-01-01

    Co-expression network analyses provide insights into the molecular interactions underlying complex traits and diseases. In this study, co-expression network analysis was performed to detect expression patterns (modules or clusters) of microRNAs (miRNAs) during lactation, and to identify miRNA regulatory mechanisms for milk yield and component traits (fat, protein, somatic cell count (SCC), lactose, and milk urea nitrogen (MUN)) via miRNA target gene enrichment analysis. miRNA expression (713 miRNAs), and milk yield and components (Fat%, Protein%, lactose, SCC, MUN) data of nine cows at each of six different time points (day 30 (D30), D70, D130, D170, D230 and D290) of an entire lactation curve were used. Four modules or clusters (GREEN, BLUE, RED and TURQUOISE) of miRNAs were identified as important for milk yield and component traits. The GREEN and BLUE modules were significantly correlated (|r| > 0.5) with milk yield and lactose, respectively. The RED and TURQUOISE modules were significantly correlated (|r| > 0.5) with both SCC and lactose. In the GREEN module, three abundantly expressed miRNAs (miR-148a, miR-186 and miR-200a) were most significantly correlated to milk yield, and are probably the most important miRNAs for this trait. DDR1 and DDHX1 are hub genes for miRNA regulatory networks controlling milk yield, while HHEX is an important transcription regulator for these networks. miR-18a, miR-221/222 cluster, and transcription factors HOXA7, and NOTCH 3 and 4, are important for the regulation of lactose. miR-142, miR-146a, and miR-EIA17-14144 (a novel miRNA), and transcription factors in the SMAD family and MYB, are important for the regulation of SCC. Important signaling pathways enriched for target genes of miRNAs of significant modules, included protein kinase A and PTEN signaling for milk yield, eNOS and Noth signaling for lactose, and TGF β, HIPPO, Wnt/β-catenin and cell cycle signaling for SCC. Relevant enriched gene ontology (GO)-terms related to

  8. Predictive models for transient protein expression in tobacco (Nicotiana tabacum L.) can optimize process time, yield, and downstream costs.

    PubMed

    Buyel, J F; Fischer, R

    2012-10-01

    The transient expression of recombinant biopharmaceutical proteins in plants can suffer inter-batch variation, which is considered a major drawback under the strict regulatory demands imposed by current good manufacturing practice (cGMP). However, we have achieved transient expression of the monoclonal antibody 2G12 and the fluorescent marker protein DsRed in tobacco leaves with ∼ 15% intra-batch coefficients of variation, which is within the range reported for transgenic plants. We developed models for the transient expression of both proteins that predicted quantitative expression levels based on five parameters: The OD(600 nm) of Agrobacterium tumefaciens (from 0.13 to 2.00), post-inoculation incubation temperature (15-30°C), plant age (harvest at 40 or 47 days after seeding), leaf age, and position within the leaf. The expression models were combined with a model of plant biomass distribution and extraction, generating a yield model for each target protein that could predict the amount of protein in specific leaf parts, individual leaves, groups of leaves, and whole plants. When the yield model was combined with a cost function for the production process, we were able to perform calculations to optimize process time, yield, or downstream costs. We illustrate this procedure by transferring the cost function from a production process using transgenic plants to a hypothetical process for the transient expression of 2G12. Our models allow the economic evaluation of new plant-based production processes and provide greater insight into the parameters that affect transient protein expression in plants. Copyright © 2012 Wiley Periodicals, Inc.

  9. High-yield expression in Escherichia coli, purification and application of budding yeast K2 killer protein.

    PubMed

    Podoliankaitė, Monika; Lukša, Juliana; Vyšniauskis, Gintautas; Sereikaitė, Jolanta; Melvydas, Vytautas; Serva, Saulius; Servienė, Elena

    2014-07-01

    Saccharomyces cerevisiae K2 toxin is a highly active extracellular protein, important as a biocontrol agent for biotechnological applications in the wine industry. This protein is produced at negligible levels in yeast, making difficult to isolate it in amounts sufficient for investigation and generation of analysis tools. In this work, we demonstrate the use of a bacterial system for expression of the recombinant K2 protein, suitable for generation of antibodies specific for toxin of the yeast origin. Synthesis of the full-length S. cerevisiae K2 preprotoxin in Escherichia coli was found to be toxic to the host cell, resulting in diminished growth. Such effect was abolished by the introduction of the C-terminal truncation into K2 protein, directing it into non-toxic inclusion body fraction. The obtained protein is of limited solubility thus, facilitating the purification by simple and efficient chromatography-free procedure. The protein aggregates were successfully refolded into a soluble form yielding sufficient amounts of a tag-less truncated K2 protein suitable for polyclonal antibody production. Antibodies were raised in rabbit and found to be specific for detection of both antigen and native S. cerevisiae K2 toxin.

  10. Meteorological fluctuations define long-term crop yield patterns in conventional and organic production systems.

    PubMed

    Teasdale, John R; Cavigelli, Michel A

    2017-04-06

    Variability in meteorological patterns presents significant challenges to crop production consistency and yield stability. Meteorological influences on corn and soybean grain yields were analyzed over an 18-year period at a long-term experiment in Beltsville, Maryland, U.S.A., comparing conventional and organic management systems. Precipitation and temperature variables explained much of the yield variability, with precipitation and heat stress during the late vegetative and early reproductive phases of crop growth accounting for the majority of yield variability in all crops and management systems. Crop yields under conventional and organic management followed similar periodic patterns, but yields were 31% and 20% lower in organic than conventional corn and soybean, respectively. The efficiency of grain yield per unit precipitation was higher under conventional than organic management, highlighting the importance of crop management for optimizing production in response to meteorological variability. Periodic yield and precipitation patterns did not consistently align with global meteorological cycles such as the El Niño Southern Oscillation.

  11. Venus Express Chemical Propulsion System - The Mars Express Legacy

    NASA Astrophysics Data System (ADS)

    Hunter, C. J.

    2004-10-01

    ESA's ambition of inter-planetary exploration using a fast-track low cost industrial programme was well achieved with Mars Express. Reusing the platform architecture for the service module and specifically the Propulsion system enabled Venus Express to benefit from several lessons learnt from the Mars Express experience. Using all existing components qualified for previous programmes, many of them commercial telecommunication spacecraft programmes with components available from stock, an industrial organisation familiar from Mars Express was able to compress the schedule to make the November 2005 launch window a realistic target. While initial inspection of the CPS schematic indicates a modified Eurostar type architecture, - a similar system using some Eurostar components - would be a fairer description. The use of many parts of the system on arrival at the destination (Mars or Venus in this case) is a departure from the usual mode of operation, where many components are used during the initial few weeks of GTO or GEO. The system modifications over the basic Eurostar system have catered for this in terms of reliability contingencies by replacing components, or providing different levels of test capability or isolation in flight. This paper aims to provide an introduction to the system, address the evolution from Eurostar, and provide an initial assessment of the success of these modifications using the Mars Express experience, and how measures have been adopted specifically for Venus Express.

  12. Influence of an oxic settling anoxic system on biomass yield, protozoa and filamentous bacteria.

    PubMed

    Rodriguez-Perez, Santiago; Fermoso, Fernando G

    2016-01-01

    An oxic settling anoxic system coupled with an activated sludge process has been studied to reduce sewage sludge production. The reduction of sludge yield, excess sludge production and active biomass yield were 51.7%, 52.9% and 67.1%, respectively, compared with the control system. The oxic reactor of the oxic settling anoxic system, even with a lower active biomass concentration than the oxic reactor of control system, showed a higher metabolic activity in their active biomass. Diversity and crawling ciliates group have been shown as promising bioindicators of active biomass yield reduction. The identification of floc-forming bacteria in the control system suggested that oxic settling anoxic system will improve settling properties compared to a Conventional Activated Sludge process.

  13. Develop a field grid system for yield mapping and machine control. Final report, Invention 544

    SciTech Connect

    1995-12-15

    The objective of this project was to build and test the Field Grid Sense system for yield mapping and machine control during harvesting. Secondly, to use Field Grid Sense system with chemical application equipment to demonstrate a workable in-field system. This document contains summarized quarterly reports.

  14. Greenhouse tomato limited cluster production systems: crop management practices affect yield

    NASA Technical Reports Server (NTRS)

    Logendra, L. S.; Gianfagna, T. J.; Specca, D. R.; Janes, H. W.

    2001-01-01

    Limited-cluster production systems may be a useful strategy to increase crop production and profitability for the greenhouse tomato (Lycopersicon esculentum Mill). In this study, using an ebb-and-flood hydroponics system, we modified plant architecture and spacing and determined the effects on fruit yield and harvest index at two light levels. Single-cluster plants pruned to allow two leaves above the cluster had 25% higher fruit yields than did plants pruned directly above the cluster; this was due to an increase in fruit weight, not fruit number. Both fruit yield and harvest index were greater for all single-cluster plants at the higher light level because of increases in both fruit weight and fruit number. Fruit yield for two-cluster plants was 30% to 40% higher than for single-cluster plants, and there was little difference in the dates or length of the harvest period. Fruit yield for three-cluster plants was not significantly different from that of two-cluster plants; moreover, the harvest period was delayed by 5 days. Plant density (5.5, 7.4, 9.2 plants/m2) affected fruit yield/plant, but not fruit yield/unit area. Given the higher costs for materials and labor associated with higher plant densities, a two-cluster crop at 5.5 plants/m2 with two leaves above the cluster was the best of the production system strategies tested.

  15. Greenhouse tomato limited cluster production systems: crop management practices affect yield.

    PubMed

    Logendra, L S; Gianfagna, T J; Specca, D R; Janes, H W

    2001-08-01

    Limited-cluster production systems may be a useful strategy to increase crop production and profitability for the greenhouse tomato (Lycopersicon esculentum Mill). In this study, using an ebb-and-flood hydroponics system, we modified plant architecture and spacing and determined the effects on fruit yield and harvest index at two light levels. Single-cluster plants pruned to allow two leaves above the cluster had 25% higher fruit yields than did plants pruned directly above the cluster; this was due to an increase in fruit weight, not fruit number. Both fruit yield and harvest index were greater for all single-cluster plants at the higher light level because of increases in both fruit weight and fruit number. Fruit yield for two-cluster plants was 30% to 40% higher than for single-cluster plants, and there was little difference in the dates or length of the harvest period. Fruit yield for three-cluster plants was not significantly different from that of two-cluster plants; moreover, the harvest period was delayed by 5 days. Plant density (5.5, 7.4, 9.2 plants/m2) affected fruit yield/plant, but not fruit yield/unit area. Given the higher costs for materials and labor associated with higher plant densities, a two-cluster crop at 5.5 plants/m2 with two leaves above the cluster was the best of the production system strategies tested.

  16. Greenhouse tomato limited cluster production systems: crop management practices affect yield

    NASA Technical Reports Server (NTRS)

    Logendra, L. S.; Gianfagna, T. J.; Specca, D. R.; Janes, H. W.

    2001-01-01

    Limited-cluster production systems may be a useful strategy to increase crop production and profitability for the greenhouse tomato (Lycopersicon esculentum Mill). In this study, using an ebb-and-flood hydroponics system, we modified plant architecture and spacing and determined the effects on fruit yield and harvest index at two light levels. Single-cluster plants pruned to allow two leaves above the cluster had 25% higher fruit yields than did plants pruned directly above the cluster; this was due to an increase in fruit weight, not fruit number. Both fruit yield and harvest index were greater for all single-cluster plants at the higher light level because of increases in both fruit weight and fruit number. Fruit yield for two-cluster plants was 30% to 40% higher than for single-cluster plants, and there was little difference in the dates or length of the harvest period. Fruit yield for three-cluster plants was not significantly different from that of two-cluster plants; moreover, the harvest period was delayed by 5 days. Plant density (5.5, 7.4, 9.2 plants/m2) affected fruit yield/plant, but not fruit yield/unit area. Given the higher costs for materials and labor associated with higher plant densities, a two-cluster crop at 5.5 plants/m2 with two leaves above the cluster was the best of the production system strategies tested.

  17. Expression of cyanobacterial FBP/SBPase in soybean prevents yield depression under future climate conditions

    USDA-ARS?s Scientific Manuscript database

    Predictions suggest that current crop production needs to double by 2050 to meet global food and energy demands. Based on theory and experimental studies, overexpression of the photosynthetic enzyme sedoheptulose-1,7-bisphosphatase (SBPase) is expected to enhance C3 crop photosynthesis and yields. H...

  18. Identification of crowding stress tolerance co-expression networks involved in sweet corn yield

    USDA-ARS?s Scientific Manuscript database

    Tolerance to crowding stress has played a crucial role in improving agronomic productivity in field corn; however, commercial sweet corn hybrids vary greatly in crowding stress tolerance. The objectives were to 1) explore transcriptional changes among sweet corn hybrids with differential yield under...

  19. Improvement of enzymatic saccharification yield in Arabidopsis thaliana by ectopic expression of the rice SUB1A-1 transcription factor

    PubMed Central

    Núñez-López, Lizeth; Aguirre-Cruz, Andrés

    2015-01-01

    Saccharification of polysaccharides releases monosaccharides that can be used by ethanol-producing microorganisms in biofuel production. To improve plant biomass as a raw material for saccharification, factors controlling the accumulation and structure of carbohydrates must be identified. Rice SUB1A-1 is a transcription factor that represses the turnover of starch and postpones energy-consuming growth processes under submergence stress. Arabidopsis was employed to test if heterologous expression of SUB1A-1 or SUB1C-1 (a related gene) can be used to improve saccharification. Cellulolytic and amylolytic enzymatic treatments confirmed that SUB1A-1 transgenics had better saccharification yield than wild-type (Col-0), mainly from accumulated starch. This improved saccharification yield was developmentally controlled; when compared to Col-0, young transgenic vegetative plants yielded 200–300% more glucose, adult vegetative plants yielded 40–90% more glucose and plants in reproductive stage had no difference in yield. We measured photosynthetic parameters, starch granule microstructure, and transcript abundance of genes involved in starch degradation (SEX4, GWD1), juvenile transition (SPL3-5) and meristematic identity (FUL, SOC1) but found no differences to Col-0, indicating that starch accumulation may be controlled by down-regulation of CONSTANS and FLOWERING LOCUS T by SUB1A-1 as previously reported. SUB1A-1 transgenics also offered less resistance to deformation than wild-type concomitant to up-regulation of AtEXP2 expansin and BGL2 glucan-1,3,-beta-glucosidase. We conclude that heterologous SUB1A-1 expression can improve saccharification yield and softness, two traits needed in bioethanol production. PMID:25780769

  20. Regulated expression of an isopentenyltransferase gene (IPT) in peanut significantly improves drought tolerance and increases yield under field conditions.

    PubMed

    Qin, Hua; Gu, Qiang; Zhang, Junling; Sun, Li; Kuppu, Sundaram; Zhang, Yizheng; Burow, Mark; Payton, Paxton; Blumwald, Eduardo; Zhang, Hong

    2011-11-01

    Isopentenyltransferase (IPT) is a critical enzyme in the cytokinin biosynthetic pathway. The expression of IPT under the control of a maturation- and stress-induced promoter was shown to delay stress-induced plant senescence that resulted in an enhanced drought tolerance in both monocot and dicot plants. This report extends the earlier findings in tobacco and rice to peanut (Arachis hypogaea L.), an important oil crop and protein source. Regulated expression of IPT in peanut significantly improved drought tolerance in both laboratory and field conditions. Transgenic peanut plants maintained higher photosynthetic rates, higher stomatal conductance and higher transpiration than wild-type control plants under reduced irrigation conditions. More importantly, transgenic peanut plants produced significantly higher yields than wild-type control plants in the field, indicating a great potential for the development of crops with improved performance and yield in water-limited areas of the world.

  1. Animal component-free Agrobacterium tumefaciens cultivation media for better GMP-compliance increases biomass yield and pharmaceutical protein expression in Nicotiana benthamiana.

    PubMed

    Houdelet, Marcel; Galinski, Anna; Holland, Tanja; Wenzel, Kathrin; Schillberg, Stefan; Buyel, Johannes Felix

    2017-02-21

    Transient expression systems allow the rapid production of recombinant proteins in plants. Such systems can be scaled up to several hundred kilograms of biomass, making them suitable for the production of pharmaceutical proteins required at short notice, such as emergency vaccines. However, large-scale transient expression requires the production of recombinant Agrobacterium tumefaciens strains with the capacity for efficient gene transfer to plant cells. The complex media often used for the cultivation of this species typically include animal-derived ingredients that can contain human pathogens, thus conflicting with the requirements of good manufacturing practice (GMP). We replaced all the animal-derived components in yeast extract broth (YEB) cultivation medium with soybean peptone, and then used a design-of-experiments approach to optimize the medium composition, increasing the biomass yield while maintaining high levels of transient expression in subsequent infiltration experiments. The resulting plant peptone Agrobacterium medium (PAM) achieved a two-fold increase in OD600 compared to YEB medium during a 4-L batch fermentation lasting 18 h. Furthermore, the yields of the monoclonal antibody 2G12 and the fluorescent protein DsRed were maintained when the cells were cultivated in PAM rather than YEB. We have thus demonstrated a simple, efficient and scalable method for medium optimization that reduces process time and costs. The final optimized medium for the cultivation of A. tumefaciens completely lacks animal-derived components, thus facilitating the GMP-compliant large-scale transient expression of recombinant proteins in plants.

  2. Ecological intensification of cereal production systems: Yield potential, soil quality, and precision agriculture

    PubMed Central

    Cassman, Kenneth G.

    1999-01-01

    Wheat (Triticum aestivum L.), rice (Oryza sativa L.), and maize (Zea mays L.) provide about two-thirds of all energy in human diets, and four major cropping systems in which these cereals are grown represent the foundation of human food supply. Yield per unit time and land has increased markedly during the past 30 years in these systems, a result of intensified crop management involving improved germplasm, greater inputs of fertilizer, production of two or more crops per year on the same piece of land, and irrigation. Meeting future food demand while minimizing expansion of cultivated area primarily will depend on continued intensification of these same four systems. The manner in which further intensification is achieved, however, will differ markedly from the past because the exploitable gap between average farm yields and genetic yield potential is closing. At present, the rate of increase in yield potential is much less than the expected increase in demand. Hence, average farm yields must reach 70–80% of the yield potential ceiling within 30 years in each of these major cereal systems. Achieving consistent production at these high levels without causing environmental damage requires improvements in soil quality and precise management of all production factors in time and space. The scope of the scientific challenge related to these objectives is discussed. It is concluded that major scientific breakthroughs must occur in basic plant physiology, ecophysiology, agroecology, and soil science to achieve the ecological intensification that is needed to meet the expected increase in food demand. PMID:10339523

  3. Ecological intensification of cereal production systems: yield potential, soil quality, and precision agriculture.

    PubMed

    Cassman, K G

    1999-05-25

    Wheat (Triticum aestivum L.), rice (Oryza sativa L.), and maize (Zea mays L.) provide about two-thirds of all energy in human diets, and four major cropping systems in which these cereals are grown represent the foundation of human food supply. Yield per unit time and land has increased markedly during the past 30 years in these systems, a result of intensified crop management involving improved germplasm, greater inputs of fertilizer, production of two or more crops per year on the same piece of land, and irrigation. Meeting future food demand while minimizing expansion of cultivated area primarily will depend on continued intensification of these same four systems. The manner in which further intensification is achieved, however, will differ markedly from the past because the exploitable gap between average farm yields and genetic yield potential is closing. At present, the rate of increase in yield potential is much less than the expected increase in demand. Hence, average farm yields must reach 70-80% of the yield potential ceiling within 30 years in each of these major cereal systems. Achieving consistent production at these high levels without causing environmental damage requires improvements in soil quality and precise management of all production factors in time and space. The scope of the scientific challenge related to these objectives is discussed. It is concluded that major scientific breakthroughs must occur in basic plant physiology, ecophysiology, agroecology, and soil science to achieve the ecological intensification that is needed to meet the expected increase in food demand.

  4. Expression of varied GFPs in Saccharomyces cerevisiae: codon optimization yields stronger than expected expression and fluorescence intensity

    PubMed Central

    Kaishima, Misato; Ishii, Jun; Matsuno, Toshihide; Fukuda, Nobuo; Kondo, Akihiko

    2016-01-01

    Green fluorescent protein (GFP), which was originally isolated from jellyfish, is a widely used tool in biological research, and homologs from other organisms are available. However, researchers must determine which GFP is the most suitable for a specific host. Here, we expressed GFPs from several sources in codon-optimized and non-codon-optimized forms in the yeast Saccharomyces cerevisiae, which represents an ideal eukaryotic model. Surprisingly, codon-optimized mWasabi and mNeonGreen, which are typically the brightest GFPs, emitted less green fluorescence than did the other five codon-optimized GFPs tested in S. cerevisiae. Further, commercially available GFPs that have been optimized for mammalian codon usage (e.g., EGFP, AcGFP1 and TagGFP2) unexpectedly exhibited extremely low expression levels in S. cerevisiae. In contrast, codon-optimization of the GFPs for S. cerevisiae markedly increased their expression levels, and the fluorescence intensity of the cells increased by a maximum of 101-fold. Among the tested GFPs, the codon-optimized monomeric mUkG1 from soft coral showed the highest levels of both expression and fluorescence. Finally, the expression of this protein as a fusion-tagged protein successfully improved the reporting system’s ability to sense signal transduction and protein–protein interactions in S. cerevisiae and increased the detection rates of target cells using flow cytometry. PMID:27782154

  5. Evaluating yield quality and quantity of garlic as affected by different farming systems and garlic clones.

    PubMed

    Mirzaei, R; Liaghati, H; Damghani, A Mahdavi

    2007-07-01

    In order to study the effects of different farming systems and garlic (Allium sativum L.) clones on yield quality and quantity of garlic, an experiment was conducted with split plot arrangement with three completely randomized blockes in the 2005 growing season at the experimental research station of Shahid Beheshti University at Zirab, north of Iran. Two factors were involved in the experiment: farming systems in three levels (intensive, conventional and organic farming), as main plots and garlic clones in three levels (Atoo, Hamedani and Khorassani) as sub-plots. The studied factors in this experiment consisted of leaf number, LAI, stem height and diameter, bulb yield, weight of bulbs, number of cloves, weight of cloves and level of allicin. Results showed that the farming systems had significant effect (p<0.05) on LAI, number of plant and bulb yield, but the effect on the other factors was not significant. The highest and lowest bulb yields were obtained in intensive (9.5 ton ha(-1)) and organic (7.4 ton ha(-1)) systems, respectively. All of the top factors were significantly (p< or =0.01) affected by garlic clones. Maximum and minimum yields were obtained from Hamedani, Atoo (9.2 ton ha(-1)) and Virani (7.1 ton ha(-1)) clones, respectively. Level of allicin was not significantly affected by farming systems but, differences among garlic clones were significant. Maximum and minimum allicin yields were obtained from Hamedan (5.96 mg g(-1)) and Virani (4.52 mg g(-1)) clones, respectively. As a result, however, organic farming systems can not influence the yield in short term, but can increase it by applying crop rotation, use of organic fertilizer and cover crops in the long term.

  6. Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectors.

    PubMed

    Giritch, Anatoli; Marillonnet, Sylvestre; Engler, Carola; van Eldik, Gerben; Botterman, Johan; Klimyuk, Victor; Gleba, Yuri

    2006-10-03

    Plant viral vectors allow expression of heterologous proteins at high yields, but so far, they have been unable to express heterooligomeric proteins efficiently. We describe here a rapid and indefinitely scalable process for high-level expression of functional full-size mAbs of the IgG class in plants. The process relies on synchronous coinfection and coreplication of two viral vectors, each expressing a separate antibody chain. The two vectors are derived from two different plant viruses that were found to be noncompeting. Unlike vectors derived from the same virus, noncompeting vectors effectively coexpress the heavy and light chains in the same cell throughout the plant body, resulting in yields of up to 0.5 g of assembled mAbs per kg of fresh-leaf biomass. This technology allows production of gram quantities of mAbs for research purposes in just several days, and the same protocol can be used on an industrial scale in situations requiring rapid response, such as pandemic or terrorism events.

  7. Fasciclin-like arabinogalactan protein gene expression is associated with yield of flour in the milling of wheat.

    PubMed

    Nirmal, Ravi C; Furtado, Agnelo; Rangan, Parimalan; Henry, Robert J

    2017-10-02

    A large portion of the global wheat crop is milled to produce flour for use in the production of foods such as bread. Pressure to increase food supplies sustainably can be address directly by reducing post-harvest losses during processes such as flour milling. The recovery of flour in the milling of wheat is genetically determined but difficult to assess in wheat breeding due to the requirement for a large sample. Here we report the discovery that human selection for altered expression of putative cell adhesion proteins is associated with wheats that give high yields of flour on milling. Genes encoding fasciclin-like arabinogalactan proteins are expressed at low levels in high milling wheat genotypes at mid grain development. Thirty worldwide wheat genotypes were grouped into good and poor millers based flour yield obtained from laboratory scale milling of mature seeds. Differentially expressed genes were identified by comparing transcript profiles at 14 and 30 days post anthesis obtained from RNA-seq data of all the genotypes. Direct selection for genotypes with appropriate expression of these genes will greatly accelerate wheat breeding and ensure high recoveries of flour from wheat by resulting in grains that break up more easily on milling.

  8. Ectopic expression of SlAGO7 alters leaf pattern and inflorescence architecture and increases fruit yield in tomato.

    PubMed

    Lin, Dongbo; Xiang, Ya; Xian, Zhiqiang; Li, Zhengguo

    2016-08-01

    ARGONAUTE7 (AGO7), a key regulator of the trans-acting small interfering RNAs (ta-siRNA) pathway, plays a conserved role in controlling leaf pattern among species. However, little is known about the ta-siRNA pathway in regulating inflorescence architecture and fruit yield. In this study, we characterized the expression pattern, subcellular localization and developmental functions of SlAGO7 in tomato (Solanum lycopersicum). Overexpressing SlAGO7 in tomato exhibited pleiotropic phenotypes, including improved axillary bud formation, altered leaf morphology and inflorescence architecture, and increased fruit yield. Cross-sectioning of leaves showed that the number of vascular bundles was significantly increased in 35:SlAGO7 lines. Overexpression of SlAGO7 increased the production of ta-siRNA, and repressed the expression ta-siRNA-targeted genes (SlARF2a, SlARF2b, SlARF3 and SlARF4). Further analysis showed that overexpression of SlAGO7 alters the expression of key genes implicated in leaf morphology, inflorescence architecture, auxin transport and signaling. In addition, the altered auxin response of 35:SlAGO7 lines were also investigated. These results suggested that SlAGO7 plays a positive role in determining inflorescence architecture and fruit yield though the ta-siRNA pathway. Therefore, SlAGO7 represents a useful gene that can be incorporated in tomato breeding programs for developing cultivars with yield potential. © 2016 Scandinavian Plant Physiology Society.

  9. Vermont Finance Reform: The Implementation of a Block Grant and Guaranteed Yield System.

    ERIC Educational Resources Information Center

    Mathis, William J.

    This report examines tax reform in Vermont. A unanimous state supreme court decision in 1997 ruled that the state's educational finance system was unconstitutional. In response to this ruling, the legislature passed a new statewide property-tax system with two tiers: a block grant of $5,010 per pupil and a guaranteed yield for spending above the…

  10. Grain yield response to poultry litter application under a wheat-soybean double cropping system

    USDA-ARS?s Scientific Manuscript database

    Poultry litter application and double cropping are management practices that could be used with conservation tillage systems to increase yields compared to conventional monocropping systems. The objective of this study was to evaluate wheat (Triticum aestivum L.) and soybean [Glycine max (L.) Merr.]...

  11. Longer-term potato cropping system effects on soilborne diseases and tuber yield

    USDA-ARS?s Scientific Manuscript database

    In field trials established in 2004, different 3-yr potato cropping systems focused on specific crop management goals of (SC) soil conservation, (SI) soil improvement, and (DS) disease-suppression were evaluated for their effects on soilborne diseases and tuber yield. These systems were compared to ...

  12. Statistical modeling of yield and variance instability in conventional and organic cropping systems

    USDA-ARS?s Scientific Manuscript database

    Cropping systems research was undertaken to address declining crop diversity and verify competitiveness of alternatives to the predominant conventional cropping system in the northern Corn Belt. To understand and capitalize on temporal yield variability within corn and soybean fields, we quantified ...

  13. Expression of salinarum halorhodopsin in Escherichia coli cells: solubilization in the presence of retinal yields the natural state.

    PubMed

    Yamashita, Yasutaka; Kikukawa, Takashi; Tsukamoto, Takashi; Kamiya, Masakatsu; Aizawa, Tomoyasu; Kawano, Keiichi; Miyauchi, Seiji; Kamo, Naoki; Demura, Makoto

    2011-12-01

    Salinarum halorhodopsin (HsHR), a light-driven chloride ion pump of haloarchaeon Halobacterium salinarum, was heterologously expressed in Escherichia coli. The expressed HsHR had no color in the E. coli membrane, but turned purple after solubilization in the presence of all-trans retinal. This colored HsHR was purified by Ni-chelate chromatography in a yield of 3-4 mg per liter culture. The purified HsHR showed a distinct chloride pumping activity by incorporation into the liposomes, and showed even in the detergent-solubilized state, its typical behaviors in both the unphotolyzed and photolyzed states. Upon solubilization, HsHR expressed in the E. coli membrane attains the proper folding and a trimeric assembly comparable to those in the native membranes. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Fertilizer source and tillage effects on yield-scaled nitrous oxide emissions in a corn cropping system.

    PubMed

    Venterea, Rodney T; Bijesh, Maharjan; Dolan, Michael S

    2011-01-01

    Management practices such as fertilizer or tillage regime may affect nitrous oxide (N₂O) emissions and crop yields, each of which is commonly expressed with respect to area (e.g., kg N ha or Mg grain ha). Expressing N₂O emissions per unit of yield can account for both of these management impacts and might provide a useful metric for greenhouse gas inventories by relating N₂O emissions to grain production rates. The objective of this study was to examine the effects of long-term (>17 yr) tillage treatments and N fertilizer source on area- and yield-scaled N₂O emissions, soil N intensity, and nitrogen use efficiency for rainfed corn ( L.) in Minnesota over three growing seasons. Two different controlled-release fertilizers (CRFs) and conventional urea (CU) were surface-applied at 146 kg N ha(-1) several weeks after planting to conventional tillage (CT) and no-till (NT) treatments. Yield-scaled emissions across all treatments represented 0.4 to 1.1% of the N harvested in the grain. Both CRFs reduced soil nitrate intensity, but not N₂O emissions, compared with CU. One CRF, consisting of nitrification and urease inhibitors added to urea, decreased N₂O emissions compared with a polymer-coated urea (PCU). The PCU tended to have lower yields during the drier years of the study, which increased its yield-scaled N₂O emissions. The overall effectiveness of CRFs compared with CU in this study may have been reduced because they were applied several weeks after corn was planted. Across all N treatments, area-scaled N₂O emissions were not significantly affected by tillage. However, when expressed per unit yield of grain, grain N, or total aboveground N, N₂O emissions with NT were 52, 66, and 69% greater, respectively, compared with CT. Thus, in this cropping system and climate regime, production of an equivalent amount of grain using NT would generate substantially more N₂O compared with CT.

  15. The expression of a recombinant glycolate dehydrogenase polyprotein in potato (Solanum tuberosum) plastids strongly enhances photosynthesis and tuber yield.

    PubMed

    Nölke, Greta; Houdelet, Marcel; Kreuzaler, Fritz; Peterhänsel, Christoph; Schillberg, Stefan

    2014-08-01

    We have increased the productivity and yield of potato (Solanum tuberosum) by developing a novel method to enhance photosynthetic carbon fixation based on expression of a polyprotein (DEFp) comprising all three subunits (D, E and F) of Escherichia coli glycolate dehydrogenase (GlcDH). The engineered polyprotein retained the functionality of the native GlcDH complex when expressed in E. coli and was able to complement mutants deficient for the D, E and F subunits. Transgenic plants accumulated DEFp in the plastids, and the recombinant protein was active in planta, reducing photorespiration and improving CO2 uptake with a significant impact on carbon metabolism. Transgenic lines with the highest DEFp levels and GlcDH activity produced significantly higher levels of glucose (5.8-fold), fructose (3.8-fold), sucrose (1.6-fold) and transitory starch (threefold), resulting in a substantial increase in shoot and leaf biomass. The higher carbohydrate levels produced in potato leaves were utilized by the sink capacity of the tubers, increasing the tuber yield by 2.3-fold. This novel approach therefore has the potential to increase the biomass and yield of diverse crops. © 2014 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  16. Auto-induction for high yield expression of recombinant novel isoallergen tropomyosin from King prawn (Melicertus latisulcatus) for improved diagnostics and immunotherapeutics.

    PubMed

    Koeberl, Martina; Kamath, Sandip D; Saptarshi, Shruti R; Smout, Michael J; Rolland, Jennifer M; O'Hehir, Robyn E; Lopata, Andreas L

    2014-12-15

    Food allergies are increasing worldwide, demonstrating a considerable public health concern. Shellfish allergy is one of the major food groups causing allergic sensitization among adults and children, affecting up to 2% of the general world population. Tropomyosin (TM) is the major allergen in shellfish and frequently used in the diagnosis of allergic sensitization and the detection of cross-contaminated food. To improve and establish better and more sensitive diagnostics for allergies and immunotherapeutics, large quantities of pure allergens are required. To establish a reproducible method for the generation of pure recombinant tropomyosin we utilized in this study different Escherichia coli strains (NM522, TOP10 and BL21(DE3)RIPL). In addition, isopropyl-β-D-thiogalactoside (IPTG) induction was compared with a novel auto-induction system to allow the generation of larger quantities of recombinant allergen. We demonstrated that the B-strain of E. coli is better for the expression of TM compared to the K-strain. Moreover, a higher yield could be achieved when using the auto-induction system, with up to 62 mg/l. High yield expressed recombinant TM from King prawn (KP) was compared to recombinant TM from Black tiger prawn (Pen m 1). We demonstrated that recombinant TM from KP and known isoallergen Pen m 1 have very similar molecular and immunological characteristics. Overall, we demonstrate that auto-induction can be used to express larger quantities of recombinant allergens for the development of diagnostic, to quantify allergens as well as immunotherapeutics employing isoallergens.

  17. Expression of Vitreoscilla hemoglobin in Bacillus thuringiensis improve the cell density and insecticidal crystal proteins yield.

    PubMed

    Liang, Feng; Shouwen, Chen; Ming, Sun; Ziniu, Yu

    2007-02-01

    The Vitreoscilla hemoglobin (VHb) gene (vgb) was integrated into the chromosome of Bacillus thuringiensis BMB171 using integrative vector pEG491. The production of VHb was confirmed by CO-difference spectra analysis. Fermentation experiments results showed that with the production of VHb, the critical oxygen concentration (COC) of the host strain was reduced from 18 to 12%. The maximum viable cell counts of the VHb+ strain in high, middle, and low aeration/agitation fermentations were 0.94-, 1.23-, and 1.59-fold of those of the VHb- strain, respectively. Under the same conditions, the yields of insecticidal crystal proteins (ICP) by VHb+ strain were 1.22-, 1.63-, and 3.13-fold of those of the VHb- strain. The production of VHb also accelerated the formation of ICP and spores. These results indicated that the production of VHb could improve the cell density and ICP yield of B. thuringiensis, especially under low aeration/agitation condition.

  18. Construction and development of a novel expression system of Streptomyces.

    PubMed

    Guan, Chengran; Cui, Wenjing; He, Xiaotian; Hu, Xu; Xu, Jun; Du, Guocheng; Chen, Jian; Zhou, Zhemin

    2015-09-01

    Streptomyces is well known to be an attractive host for producing large amounts of proteins with potent biological activities into the culture supernatant. To expand its expression system, we constructed a novel expression plasmid for gene expression in Streptomyces by inserting the promoter (P(tg)) and the signal peptide (SP(tg)) of transglutaminase (TGase) from Streptomyces hygroscopicus WSH03-13 into vector pIJ86, followed by multiple cloning sites and a transcriptional terminator fd (fd-ter). The secretion capacity of the vector was further enhanced by optimizing the signal peptidase cleavage site and a rare codon of SP(tg), yielding expression vector pSG02. Using this vector, TGase was actively and greatly expressed in the supernatant in several Streptomyces strains. In addition, the heterologous proteins aminopeptidase from Bacillus subtilis Zj016 (BSAP) and phenylalanine ammonia-lyase from Rhodotorula glutinis (PAL) were also expressed in various Streptomyces strains by this vector. This expression system should be useful for the expression of other proteins. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Mesoderm-specific Stat3 deletion affects expression of Sox9 yielding Sox9-dependent phenotypes

    PubMed Central

    Hall, Michael D.; Murray, Caroline A.; Perantoni, Alan O.

    2017-01-01

    To date, mutations within the coding region and translocations around the SOX9 gene both constitute the majority of genetic lesions underpinning human campomelic dysplasia (CD). While pathological coding-region mutations typically result in a non-functional SOX9 protein, little is known about what mechanism(s) controls normal SOX9 expression, and subsequently, which signaling pathways may be interrupted by alterations occurring around the SOX9 gene. Here, we report the identification of Stat3 as a key modulator of Sox9 expression in nascent cartilage and developing chondrocytes. Stat3 expression is predominant in tissues of mesodermal origin, and its conditional ablation using mesoderm-specific TCre, in vivo, causes dwarfism and skeletal defects characteristic of CD. Specifically, Stat3 loss results in the expansion of growth plate hypertrophic chondrocytes and deregulation of normal endochondral ossification in all bones examined. Conditional deletion of Stat3 with a Sox9Cre driver produces palate and tracheal irregularities similar to those described in Sox9+/- mice. Furthermore, mesodermal deletion of Stat3 causes global embryonic down regulation of Sox9 expression and function in vivo. Mechanistic experiments ex vivo suggest Stat3 can directly activate the expression of Sox9 by binding to its proximal promoter following activation. These findings illuminate a novel role for Stat3 in chondrocytes during skeletal development through modulation of a critical factor, Sox9. Importantly, they further provide the first evidence for the modulation of a gene product other than Sox9 itself which is capable of modeling pathological aspects of CD and underscore a potentially valuable therapeutic target for patients with the disorder. PMID:28166224

  20. High-yield secretion of recombinant proteins expressed in tobacco cell culture with a designer glycopeptide tag: Process development.

    PubMed

    Zhang, Ningning; Gonzalez, Maria; Savary, Brett; Xu, Jianfeng

    2016-03-01

    Low-yield protein production remains the most significant economic hurdle with plant cell culture technology. Fusions of recombinant proteins with hydroxyproline-O-glycosylated designer glycopeptide tags have consistently boosted secreted protein yields. This prompted us to study the process development of this technology aiming to achieve productivity levels necessary for commercial viability. We used a tobacco BY-2 cell culture expressing EGFP as fusion with a glycopeptide tag comprised of 32 repeat of "Ser-Pro" dipeptide, or (SP)32 , to study cell growth and protein secretion, culture scale-up, and establishment of perfusion cultures for continuous production. The BY-2 cells accumulated low levels of cell biomass (~7.5 g DW/L) in Schenk & Hildebrandt medium, but secreted high yields of (SP)32 -tagged EGFP (125 mg/L). Protein productivity of the cell culture has been stable for 6.0 years. The BY-2 cells cultured in a 5-L bioreactor similarly produced high secreted protein yield at 131 mg/L. Successful operation of a cell perfusion culture for 30 days was achieved under the perfusion rate of 0.25 and 0.5 day(-1) , generating a protein volumetric productivity of 17.6 and 28.9 mg/day/L, respectively. This research demonstrates the great potential of the designer glycopeptide technology for use in commercial production of valuable proteins with plant cell cultures. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Development of a telemetry and yield-mapping system of olive harvester.

    PubMed

    Castillo-Ruiz, Francisco J; Pérez-Ruiz, Manuel; Blanco-Roldán, Gregorio L; Gil-Ribes, Jesús A; Agüera, Juan

    2015-02-10

    Sensors, communication systems and geo-reference units are required to achieve an optimized management of agricultural inputs with respect to the economic and environmental aspects of olive groves. In this study, three commercial olive harvesters were tracked during two harvesting seasons in Spain and Chile using remote and autonomous equipment that was developed to determine their time efficiency and effective based on canopy shaking for fruit detachment. These harvesters work in intensive/high-density (HD) and super-high-density (SHD) olive orchards. A GNSS (Global Navigation Satellite System) and GSM (Global System for Mobile Communications) device was installed to track these harvesters. The GNSS receiver did not affect the driver's work schedule. Time elements methodology was adapted to the remote data acquisition system. The effective field capacity and field efficiency were investigated. In addition, the field shape, row length, angle between headland alley and row, and row alley width were measured to determinate the optimum orchard design parameters value. The SHD olive harvester showed significant lower effective field capacity values when alley width was less than 4 m. In addition, a yield monitor was developed and installed on a traditional olive harvester to obtain a yield map from the harvested area. The hedge straddle harvester stood out for its highly effective field capacity; nevertheless, a higher field efficiency was provided by a non-integral lateral canopy shaker. All of the measured orchard parameters have influenced machinery yields, whether effective field capacity or field efficiency. A saving of 40% in effective field capacity was achieved with a reduction from 4 m or higher to 3.5 m in alley width for SHD olive harvester. A yield map was plotted using data that were acquired by a yield monitor, reflecting the yield gradient in spite of the larger differences between tree yields.

  2. Development of a Telemetry and Yield-Mapping System of Olive Harvester

    PubMed Central

    Castillo-Ruiz, Francisco J.; Pérez-Ruiz, Manuel; Blanco-Roldán, Gregorio L.; Gil-Ribes, Jesús A.; Agüera, Juan

    2015-01-01

    Sensors, communication systems and geo-reference units are required to achieve an optimized management of agricultural inputs with respect to the economic and environmental aspects of olive groves. In this study, three commercial olive harvesters were tracked during two harvesting seasons in Spain and Chile using remote and autonomous equipment that was developed to determine their time efficiency and effective based on canopy shaking for fruit detachment. These harvesters work in intensive/high-density (HD) and super-high-density (SHD) olive orchards. A GNSS (Global Navigation Satellite System) and GSM (Global System for Mobile Communications) device was installed to track these harvesters. The GNSS receiver did not affect the driver’s work schedule. Time elements methodology was adapted to the remote data acquisition system. The effective field capacity and field efficiency were investigated. In addition, the field shape, row length, angle between headland alley and row, and row alley width were measured to determinate the optimum orchard design parameters value. The SHD olive harvester showed significant lower effective field capacity values when alley width was less than 4 m. In addition, a yield monitor was developed and installed on a traditional olive harvester to obtain a yield map from the harvested area. The hedge straddle harvester stood out for its highly effective field capacity; nevertheless, a higher field efficiency was provided by a non-integral lateral canopy shaker. All of the measured orchard parameters have influenced machinery yields, whether effective field capacity or field efficiency. A saving of 40% in effective field capacity was achieved with a reduction from 4 m or higher to 3.5 m in alley width for SHD olive harvester. A yield map was plotted using data that were acquired by a yield monitor, reflecting the yield gradient in spite of the larger differences between tree yields. PMID:25675283

  3. Modular Protein Expression Toolbox (MoPET), a standardized assembly system for defined expression constructs and expression optimization libraries

    PubMed Central

    Birkenfeld, Jörg; Franz, Jürgen; Gritzan, Uwe; Linden, Lars; Trautwein, Mark

    2017-01-01

    The design and generation of an optimal expression construct is the first and essential step in in the characterization of a protein of interest. Besides evaluation and optimization of process parameters (e.g. selection of the best expression host or cell line and optimal induction conditions and time points), the design of the expression construct itself has a major impact. However, the path to this final expression construct is often not straight forward and includes multiple learning cycles accompanied by design variations and retesting of construct variants, since multiple, functional DNA sequences of the expression vector backbone, either coding or non-coding, can have a major impact on expression yields. To streamline the generation of defined expression constructs of otherwise difficult to express proteins, the Modular Protein Expression Toolbox (MoPET) has been developed. This cloning platform allows highly efficient DNA assembly of pre-defined, standardized functional DNA modules with a minimal cloning burden. Combining these features with a standardized cloning strategy facilitates the identification of optimized DNA expression constructs in shorter time. The MoPET system currently consists of 53 defined DNA modules divided into eight functional classes and can be flexibly expanded. However, already with the initial set of modules, 792,000 different constructs can be rationally designed and assembled. Furthermore, this starting set was used to generate small and mid-sized combinatorial expression optimization libraries. Applying this screening approach, variants with up to 60-fold expression improvement have been identified by MoPET variant library screening. PMID:28520717

  4. Histology-Based Expression Profiling Yields Novel Prognostic Markers in Human Glioblastoma

    PubMed Central

    Dong, Shumin; Nutt, Catherine L.; Betensky, Rebecca A.; Stemmer-Rachamimov, Anat O.; Denko, Nicholas C.; Ligon, Keith L.; Rowitch, David H.; Louis, David N.

    2006-01-01

    Although the prognosis for patients with glioblastoma is poor, survival is variable, with some patients surviving longer than others. For this reason, there has been longstanding interest in the identi-fication of prognostic markers for glioblastoma. We hypothesized that specific histologic features known to correlate with malignancy most likely express molecules that are directly related to the aggressive behavior of these tumors. We further hypothesized that such molecules could be used as biomarkers to predict behavior in a manner that might add prognostic power to sole histologic observation of the feature. We reasoned that perinecrotic tumor cell palisading, which denotes the most aggressive forms of malignant gliomas, would be a striking histologic feature on which to test this hypothesis. We therefore used laser capture microdissection and oligonucleotide arrays to detect molecules differentially expressed in perinecrotic palisades. A set of RNAs (including POFUT2, PTDSR, PLOD2, ATF5, and HK2) that were differentially expressed in 3 initially studied, micro-dissected glioblastomas also provided prognostic information in an independent set of 28 glioblastomas that did not all have perinecrotic palisades. On validation in a second, larger independent series, this approach could be applied to other human glioma types to derive tissue biomarkers that could offer ancillary prognostic and predictive information alongside standard histopathologic examination. PMID:16254489

  5. Expression of the Aeluropus littoralis AlSAP Gene Enhances Rice Yield under Field Drought at the Reproductive Stage.

    PubMed

    Ghneim-Herrera, Thaura; Selvaraj, Michael G; Meynard, Donaldo; Fabre, Denis; Peña, Alexandra; Ben Romdhane, Walid; Ben Saad, Rania; Ogawa, Satoshi; Rebolledo, Maria C; Ishitani, Manabu; Tohme, Joe; Al-Doss, Abdullah; Guiderdoni, Emmanuel; Hassairi, Afif

    2017-01-01

    We evaluated the yields of Oryza sativa L. 'Nipponbare' rice lines expressing a gene encoding an A20/AN1 domain stress-associated protein, AlSAP, from the halophyte grass Aeluropus littoralis under the control of different promoters. Three independent field trials were conducted, with drought imposed at the reproductive stage. In all trials, the two transgenic lines, RN5 and RN6, consistently out-performed non-transgenic (NT) and wild-type (WT) controls, providing 50-90% increases in grain yield (GY). Enhancement of tillering and panicle fertility contributed to this improved GY under drought. In contrast with physiological records collected during previous greenhouse dry-down experiments, where drought was imposed at the early tillering stage, we did not observe significant differences in photosynthetic parameters, leaf water potential, or accumulation of antioxidants in flag leaves of AlSAP-lines subjected to drought at flowering. However, AlSAP expression alleviated leaf rolling and leaf drying induced by drought, resulting in increased accumulation of green biomass. Therefore, the observed enhanced performance of the AlSAP-lines subjected to drought at the reproductive stage can be tentatively ascribed to a primed status of the transgenic plants, resulting from a higher accumulation of biomass during vegetative growth, allowing reserve remobilization and maintenance of productive tillering and grain filling. Under irrigated conditions, the overall performance of AlSAP-lines was comparable with, or even significantly better than, the NT and WT controls. Thus, AlSAP expression inflicted no penalty on rice yields under optimal growth conditions. Our results support the use of AlSAP transgenics to reduce rice GY losses under drought conditions.

  6. Expression of the Aeluropus littoralis AlSAP Gene Enhances Rice Yield under Field Drought at the Reproductive Stage

    PubMed Central

    Ghneim-Herrera, Thaura; Selvaraj, Michael G.; Meynard, Donaldo; Fabre, Denis; Peña, Alexandra; Ben Romdhane, Walid; Ben Saad, Rania; Ogawa, Satoshi; Rebolledo, Maria C.; Ishitani, Manabu; Tohme, Joe; Al-Doss, Abdullah; Guiderdoni, Emmanuel; Hassairi, Afif

    2017-01-01

    We evaluated the yields of Oryza sativa L. ‘Nipponbare’ rice lines expressing a gene encoding an A20/AN1 domain stress-associated protein, AlSAP, from the halophyte grass Aeluropus littoralis under the control of different promoters. Three independent field trials were conducted, with drought imposed at the reproductive stage. In all trials, the two transgenic lines, RN5 and RN6, consistently out-performed non-transgenic (NT) and wild-type (WT) controls, providing 50–90% increases in grain yield (GY). Enhancement of tillering and panicle fertility contributed to this improved GY under drought. In contrast with physiological records collected during previous greenhouse dry-down experiments, where drought was imposed at the early tillering stage, we did not observe significant differences in photosynthetic parameters, leaf water potential, or accumulation of antioxidants in flag leaves of AlSAP-lines subjected to drought at flowering. However, AlSAP expression alleviated leaf rolling and leaf drying induced by drought, resulting in increased accumulation of green biomass. Therefore, the observed enhanced performance of the AlSAP-lines subjected to drought at the reproductive stage can be tentatively ascribed to a primed status of the transgenic plants, resulting from a higher accumulation of biomass during vegetative growth, allowing reserve remobilization and maintenance of productive tillering and grain filling. Under irrigated conditions, the overall performance of AlSAP-lines was comparable with, or even significantly better than, the NT and WT controls. Thus, AlSAP expression inflicted no penalty on rice yields under optimal growth conditions. Our results support the use of AlSAP transgenics to reduce rice GY losses under drought conditions. PMID:28659945

  7. Ectopic expression of Arabidopsis Target of Rapamycin (AtTOR) improves water-use efficiency and yield potential in rice

    PubMed Central

    Bakshi, Achala; Moin, Mazahar; Kumar, M. Udaya; Reddy, Aramati Bindu Madhava; Ren, Maozhi; Datla, Raju; Siddiq, E. A.; Kirti, P. B.

    2017-01-01

    The target of Rapamycin (TOR) present in all eukaryotes is a multifunctional protein, regulating growth, development, protein translation, ribosome biogenesis, nutrient, and energy signaling. In the present study, ectopic expression of TOR gene of Arabidopsis thaliana in a widely cultivated indica rice resulted in enhanced plant growth under water-limiting conditions conferring agronomically important water-use efficiency (WUE) trait. The AtTOR high expression lines of rice exhibited profuse tillering, increased panicle length, increased plant height, high photosynthetic efficiency, chlorophyll content and low ∆13C. Δ13C, which is inversely related to high WUE, was as low as 17‰ in two AtTOR high expression lines. These lines were also insensitive to the ABA-mediated inhibition of seed germination. The significant upregulation of 15 stress-specific genes in high expression lines indicates their contribution to abiotic stress tolerance. The constitutive expression of AtTOR is also associated with significant transcriptional upregulation of putative TOR complex-1 components, OsRaptor and OsLST8. Glucose-mediated transcriptional activation of AtTOR gene enhanced lateral root formation. Taken together, our findings indicate that TOR, in addition to its multiple cellular functions, also plays an important role in response to abiotic stress and potentially enhances WUE and yield related attributes. PMID:28230163

  8. Ectopic expression of Arabidopsis Target of Rapamycin (AtTOR) improves water-use efficiency and yield potential in rice

    NASA Astrophysics Data System (ADS)

    Bakshi, Achala; Moin, Mazahar; Kumar, M. Udaya; Reddy, Aramati Bindu Madhava; Ren, Maozhi; Datla, Raju; Siddiq, E. A.; Kirti, P. B.

    2017-02-01

    The target of Rapamycin (TOR) present in all eukaryotes is a multifunctional protein, regulating growth, development, protein translation, ribosome biogenesis, nutrient, and energy signaling. In the present study, ectopic expression of TOR gene of Arabidopsis thaliana in a widely cultivated indica rice resulted in enhanced plant growth under water-limiting conditions conferring agronomically important water-use efficiency (WUE) trait. The AtTOR high expression lines of rice exhibited profuse tillering, increased panicle length, increased plant height, high photosynthetic efficiency, chlorophyll content and low ∆13C. Δ13C, which is inversely related to high WUE, was as low as 17‰ in two AtTOR high expression lines. These lines were also insensitive to the ABA-mediated inhibition of seed germination. The significant upregulation of 15 stress-specific genes in high expression lines indicates their contribution to abiotic stress tolerance. The constitutive expression of AtTOR is also associated with significant transcriptional upregulation of putative TOR complex-1 components, OsRaptor and OsLST8. Glucose-mediated transcriptional activation of AtTOR gene enhanced lateral root formation. Taken together, our findings indicate that TOR, in addition to its multiple cellular functions, also plays an important role in response to abiotic stress and potentially enhances WUE and yield related attributes.

  9. A quality assessment of the MARS crop yield forecasting system for the European Union

    NASA Astrophysics Data System (ADS)

    van der Velde, Marijn; Bareuth, Bettina

    2015-04-01

    Timely information on crop production forecasts can become of increasing importance as commodity markets are more and more interconnected. Impacts across large crop production areas due to (e.g.) extreme weather and pest outbreaks can create ripple effects that may affect food prices and availability elsewhere. The MARS Unit (Monitoring Agricultural ResourceS), DG Joint Research Centre, European Commission, has been providing forecasts of European crop production levels since 1993. The operational crop production forecasting is carried out with the MARS Crop Yield Forecasting System (M-CYFS). The M-CYFS is used to monitor crop growth development, evaluate short-term effects of anomalous meteorological events, and provide monthly forecasts of crop yield at national and European Union level. The crop production forecasts are published in the so-called MARS bulletins. Forecasting crop yield over large areas in the operational context requires quality benchmarks. Here we present an analysis of the accuracy and skill of past crop yield forecasts of the main crops (e.g. soft wheat, grain maize), throughout the growing season, and specifically for the final forecast before harvest. Two simple benchmarks to assess the skill of the forecasts were defined as comparing the forecasts to 1) a forecast equal to the average yield and 2) a forecast using a linear trend established through the crop yield time-series. These reveal a variability in performance as a function of crop and Member State. In terms of production, the yield forecasts of 67% of the EU-28 soft wheat production and 80% of the EU-28 maize production have been forecast superior to both benchmarks during the 1993-2013 period. In a changing and increasingly variable climate crop yield forecasts can become increasingly valuable - provided they are used wisely. We end our presentation by discussing research activities that could contribute to this goal.

  10. Cell surface engineering of Bacillus subtilis improves production yields of heterologously expressed α-amylases.

    PubMed

    Cao, Haojie; van Heel, Auke J; Ahmed, Hifza; Mols, Maarten; Kuipers, Oscar P

    2017-04-04

    Bacillus subtilis is widely used as a cell factory for numerous heterologous proteins of commercial value and medical interest. To explore the possibility of further enhancing the secretion potential of this model bacterium, a library of engineered strains with modified cell surface components was constructed, and the corresponding influences on protein secretion were investigated by analyzing the secretion of α-amylase variants with either low-, neutral- or high- isoelectric points (pI). Relative to the wild-type strain, the presence of overall anionic membrane phospholipids (phosphatidylglycerol and cardiolipin) increased dramatically in the PssA-, ClsA- and double KO mutants, which resulted in an up to 47% higher secretion of α-amylase. Additionally, we demonstrated that the appropriate net charge of secreted targets (AmyTS-23, AmyBs and AmyBm) was beneficial for secretion efficiency as well. In B. subtilis, the characteristics of cell membrane phospholipid bilayer and the pIs of heterologous α-amylases appear to be important for their secretion efficiency. These two factors can be engineered to reduce the electrostatic interaction between each other during the secretion process, which finally leads to a better secretion yield of α-amylases.

  11. Impacts of temperature on rice yields of different rice cultivation systems in southern China over the past 40 years

    NASA Astrophysics Data System (ADS)

    Zhang, Jiahua; Yao, Fengmei; Hao, Cui; Boken, Vijendra

    The impact of climate change on rice yield in China remains highly uncertain. We examined the impact of the change of maximum temperature (Tmax) and minimum temperature (Tmin) on rice yields in southern China from 1967 to 2007. The rice yields were simulated by using the DSSAT3.5 (Decision Support System for Agro-technology Transfer)-Rice model. The change of Tmax and Tmin in rice growing seasons and simulated rice yields as well as their correlations were analyzed. The simulated yields of middle rice and early rice had a decreasing trend, but late rice yields showed a weak rise trend. There was significant negative correlation between Tmax and the early rice yields, as well as the late rice yields in most stations, but non-significant negative correlation for the middle rice yields. An obviously negative relationship was found between Tmin and the early and middle rice yields, and a significant positive relationship was found between Tmin and the late rice yields. It indicated that under the recent climate warming, the increased Tmax brought strong negative impacts on early rice yields and late rice yields, but a weak negative impact on the middle rice yields; the increased Tmin had a strong negative impact on the middle rice yields and the early rice yields, but a significant positive impact on the late rice yields. It suggested that it is necessary to adjust rice planting date and adapt to higher Tmin.

  12. Crop yield network and its response to changes in climate system

    NASA Astrophysics Data System (ADS)

    Yokozawa, M.

    2013-12-01

    Crop failure (reduction in crop yield) due to extreme weather and climate change could lead to unstable food supply, reflecting the recent globalization in world agricultural production. Specifically, in several major production countries producing large amount of main cereal crops, wheat, maize, soybean and rice, abrupt crop failures in wide area are significantly serious for world food supply system. We examined the simultaneous changes in crop yield in USA, China and Brazil, in terms of the changes in climate system such as El Nino, La nina and so on. In this study, we defined a crop yield networks, which represent the correlation between yearly changes in crop yields and climate resources during the crop growing season in two regions. The climate resources during the crop growing season represents here the average temperature and the accumulated precipitation during the crop growing season of a target crop. As climate data, we used a reanalysis climate data JRA-25 (Japan Meteorological Agency). The yearly changes in crop yields are based on a gridded crop productivity database with a resolution of 1.125 degree in latitude/longitude (Iizumi et al. 2013). It is constructed from the agriculture statistics issued by local administrative bureau in each country, which covers the period during 1982 to 2006 (25 years). For the regions being lack of data, the data was interpolated referring to NPP values estimated by satellite data. Crop yield network is constructed as follows: (1) let DY(i,y) be negative difference in crop yield of year y from the trend yield at grid i; (2) define the correlation of the differences Cij(y) = DY(i, y) DY(j, y); (3) if Cij(y) > Q, then grids i and j are mutually linked for a threshold value Q. Links between grids make a crop yield network. It is here noted that only negative differences are taken into account because we focused on the lean year cases (i.e. yields of both grids were lower than those in the long-term trend). The arrays of

  13. Sheep Grazing Effect on Dryland Soil Properties and Wheat Yield in the Wheat-Fallow System

    USDA-ARS?s Scientific Manuscript database

    Sheep (Ovis aries L.) grazing during fallow is an effective method of controlling weeds and pests in the wheat (Triticum aestivum L.)-fallow system. Little is known about the effect of sheep grazing on dryland soil properties and wheat yield. We evaluated the effects of fallow management for weed co...

  14. Sheep Grazing in the Wheat-Fallow System Affects Dryland Soil Properties and Grain Yield

    USDA-ARS?s Scientific Manuscript database

    Sheep (Ovis aries L.) grazing during fallow is an effective method of controlling weeds and pests in the wheat (Triticum aestivum L.)-fallow system. Little is known about the effect of sheep grazing on dryland soil properties and wheat yield. We evaluated the effects of fallow management for weed co...

  15. A whole stand growth and yield system for young longleaf pine plantations in Southwest Georgia

    Treesearch

    John R. Brooks; Steven B. Jack

    2006-01-01

    A whole stand growth and yield system for planted longleaf pine (Pinus palustris Mill.) was developed from permanent plot data collected annually over an 8 year period. The dataset consists of 12 intensively-managed longleaf pine plantations that are located in Lee, Worth, Mitchell, and Baker counties in southwest Georgia. Stand survival, dominant...

  16. Destiny-yield relationship for channel catfish reared in a biofloc technology production system

    USDA-ARS?s Scientific Manuscript database

    The effect of stocking density on yield of stocker channel catfish and water quality in a biofloc technology production system was studied in this completely randomized design experiment. Fingerling channel catfish (Ictalurus punctatus; 48.0 g/fish, 17.8 cm/fish) were stocked into nine continuously ...

  17. Does increasing milk yield per cow reduce greenhouse gas emissions? A system approach.

    PubMed

    Zehetmeier, M; Baudracco, J; Hoffmann, H; Heißenhuber, A

    2012-01-01

    Milk yield per cow has continuously increased in many countries over the last few decades. In addition to potential economic advantages, this is often considered an important strategy to decrease greenhouse gas (GHG) emissions per kg of milk produced. However, it should be considered that milk and beef production systems are closely interlinked, as fattening of surplus calves from dairy farming and culled dairy cows play an important role in beef production in many countries. The main objective of this study was to quantify the effect of increasing milk yield per cow on GHG emissions and on other side effects. Two scenarios were modelled: constant milk production at the farm level and decreasing beef production (as co-product; Scenario 1); and both milk and beef production kept constant by compensating the decline in beef production with beef from suckler cow production (Scenario 2). Model calculations considered two types of production unit (PU): dairy cow PU and suckler cow PU. A dairy cow PU comprises not only milk output from the dairy cow, but also beef output from culled cows and the fattening system for surplus calves. The modelled dairy cow PU differed in milk yield per cow per year (6000, 8000 and 10 000 kg) and breed. Scenario 1 resulted in lower GHG emissions with increasing milk yield per cow. However, when milk and beef outputs were kept constant (Scenario 2), GHG emissions remained approximately constant with increasing milk yield from 6000 to 8000 kg/cow per year, whereas further increases in milk yield (10 000 kg milk/cow per year) resulted in slightly higher (8%) total GHG emissions. Within Scenario 2, two different allocation methods to handle co-products (surplus calves and beef from culled cows) from dairy cow production were evaluated. Results showed that using the 'economic allocation method', GHG emissions per kg milk decreased with increasing milk yield per cow per year, from 1.06 kg CO2 equivalents (CO2eq) to 0.89 kg CO2eq for the 6000 and

  18. Algevir: An Expression System for Microalgae Based on Viral Vectors.

    PubMed

    Bañuelos-Hernández, Bernardo; Monreal-Escalante, Elizabeth; González-Ortega, Omar; Angulo, Carlos; Rosales-Mendoza, Sergio

    2017-01-01

    The use of recombinant algae for the production of valuable compounds is opening promising biotechnological applications. However, the development of efficient expression approaches is still needed to expand the exploitation of microalgae in biotechnology. Herein, the concept of using viral expression vectors in microalgae was explored for the first time. An inducible geminiviral vector leading to Rep-mediated replication of the expression cassette allowed the production of antigenic proteins at high levels. This system, called Algevir, allows the production of complex viral proteins (GP1 from Zaire ebolavirus) and bacterial toxin subunits (B subunit of the heat-labile Escherichia coli enterotoxin), which retained their antigenic activity. The highest achieved yield was 1.25 mg/g fresh biomass (6 mg/L of culture), which was attained 3 days after transformation. The Algevir system allows for a fast and efficient production of recombinant proteins, overcoming the difficulties imposed by the low yields and unstable expression patterns frequently observed in stably transformed microalgae at the nuclear level; as well as the toxicity of some target proteins.

  19. Codon Optimization to Enhance Expression Yields Insights into Chloroplast Translation1[OPEN

    PubMed Central

    Chan, Hui-Ting; Williams-Carrier, Rosalind; Barkan, Alice

    2016-01-01

    Codon optimization based on psbA genes from 133 plant species eliminated 105 (human clotting factor VIII heavy chain [FVIII HC]) and 59 (polio VIRAL CAPSID PROTEIN1 [VP1]) rare codons; replacement with only the most highly preferred codons decreased transgene expression (77- to 111-fold) when compared with the codon usage hierarchy of the psbA genes. Targeted proteomic quantification by parallel reaction monitoring analysis showed 4.9- to 7.1-fold or 22.5- to 28.1-fold increase in FVIII or VP1 codon-optimized genes when normalized with stable isotope-labeled standard peptides (or housekeeping protein peptides), but quantitation using western blots showed 6.3- to 8-fold or 91- to 125-fold increase of transgene expression from the same batch of materials, due to limitations in quantitative protein transfer, denaturation, solubility, or stability. Parallel reaction monitoring, to our knowledge validated here for the first time for in planta quantitation of biopharmaceuticals, is especially useful for insoluble or multimeric proteins required for oral drug delivery. Northern blots confirmed that the increase of codon-optimized protein synthesis is at the translational level rather than any impact on transcript abundance. Ribosome footprints did not increase proportionately with VP1 translation or even decreased after FVIII codon optimization but is useful in diagnosing additional rate-limiting steps. A major ribosome pause at CTC leucine codons in the native gene of FVIII HC was eliminated upon codon optimization. Ribosome stalls observed at clusters of serine codons in the codon-optimized VP1 gene provide an opportunity for further optimization. In addition to increasing our understanding of chloroplast translation, these new tools should help to advance this concept toward human clinical studies. PMID:27465114

  20. The yield and decay coefficients of exoelectrogenic bacteria in bioelectrochemical systems.

    PubMed

    Wilson, Erica L; Kim, Younggy

    2016-05-01

    In conventional wastewater treatment, waste sludge management and disposal contribute the major cost for wastewater treatment. Bioelectrochemical systems, as a potential alternative for future wastewater treatment and resources recovery, are expected to produce small amounts of waste sludge because exoelectrogenic bacteria grow on anaerobic respiration and form highly populated biofilms on bioanode surfaces. While waste sludge production is governed by the yield and decay coefficient, none of previous studies have quantified these kinetic constants for exoelectrogenic bacteria. For yield coefficient estimation, we modified McCarty's free energy-based model by using the bioanode potential for the free energy of the electron acceptor reaction. The estimated true yield coefficient ranged 0.1 to 0.3 g-VSS (volatile suspended solids) g-COD(-1) (chemical oxygen demand), which is similar to that of most anaerobic microorganisms. The yield coefficient was sensitively affected by the bioanode potential and pH while the substrate and bicarbonate concentrations had relatively minor effects on the yield coefficient. In lab-scale experiments using microbial electrolysis cells, the observed yield coefficient (including the effect of cell decay) was found to be 0.020 ± 0.008 g-VSS g-COD(-1), which is an order of magnitude smaller than the theoretical estimation. Based on the difference between the theoretical and experimental results, the decay coefficient was approximated to be 0.013 ± 0.002 d(-1). These findings indicate that bioelectrochemical systems have potential for future wastewater treatment with reduced waste sludge as well as for resources recovery. Also, the found kinetic information will allow accurate estimation of wastewater treatment performance in bioelectrochemical systems.

  1. Heterologous expression of VHb can improve the yield and quality of biocontrol fungus Paecilomyces lilacinus, during submerged fermentation.

    PubMed

    Zhang, Shumeng; Wang, Jieping; Wei, Yale; Tang, Qing; Ali, Maria Kanwal; He, Jin

    2014-10-10

    Paecilomyces lilacinus is an egg-parasitic fungus which is effective against plant-parasitic nematodes and it has been successfully commercialized for the control of many plant-parasitic nematodes. However, during the large-scale industrial fermentation process of the filamentous fungus, the dissolved oxygen supply is a limiting factor, which influences yield, product quality and production cost. To solve this problem, we intended to heterologously express VHb in P. lilacinus ACSS. After optimizing the vgb gene, we fused it with a selection marker gene nptII, a promoter PgpdA and a terminator TtrpC. The complete expression cassette PgpdA-nptII-vgb-TtrpC was transferred into P. lilacinus ACSS by Agrobacterium tumefaciens-mediated transformation. Consequently, we successfully screened an applicable fungus strain PNVT8 which efficiently expressed VHb. The submerged fermentation experiments demonstrated that the expression of VHb not only increased the production traits of P. lilacinus such as biomass and spore production, but also improved the beneficial product quality and application value, due to the secretion of more protease and chitinase. It can be speculated that the recombinant strain harboring vgb gene will have a growth advantage over the original strain under anaerobic conditions in soil and therefore will possess higher biocontrol efficiency against plant-parasitic nematodes.

  2. Transgenic Cotton Plants Expressing the HaHR3 Gene Conferred Enhanced Resistance to Helicoverpa armigera and Improved Cotton Yield.

    PubMed

    Han, Qiang; Wang, Zhenzhen; He, Yunxin; Xiong, Yehui; Lv, Shun; Li, Shupeng; Zhang, Zhigang; Qiu, Dewen; Zeng, Hongmei

    2017-08-30

    RNA interference (RNAi) has been developed as an efficient technology. RNAi insect-resistant transgenic plants expressing double-stranded RNA (dsRNA) that is ingested into insects to silence target genes can affect the viability of these pests or even lead to their death. HaHR3, a molt-regulating transcription factor gene, was previously selected as a target expressed in bacteria and tobacco plants to control Helicoverpa armigera by RNAi technology. In this work, we selected the dsRNA-HaHR3 fragment to silence HaHR3 in cotton bollworm for plant mediated-RNAi research. A total of 19 transgenic cotton lines expressing HaHR3 were successfully cultivated, and seven generated lines were used to perform feeding bioassays. Transgenic cotton plants expressing dsHaHR3 were shown to induce high larval mortality and deformities of pupation and adult eclosion when used to feed the newly hatched larvae, and 3rd and 5th instar larvae of H. armigera. Moreover, HaHR3 transgenic cotton also demonstrated an improved cotton yield when compared with controls.

  3. High-Yield Expression of M2e Peptide of Avian Influenza Virus H5N1 in Transgenic Duckweed Plants.

    PubMed

    Firsov, Aleksey; Tarasenko, Irina; Mitiouchkina, Tatiana; Ismailova, Natalya; Shaloiko, Lyubov; Vainstein, Alexander; Dolgov, Sergey

    2015-07-01

    Avian influenza is a major viral disease in poultry. Antigenic variation of this virus hinders vaccine development. However, the extracellular domain of the virus-encoded M2 protein (peptide M2e) is nearly invariant in all influenza A strains, enabling the development of a broad-range vaccine against them. Antigen expression in transgenic plants is becoming a popular alternative to classical expression methods. Here we expressed M2e from avian influenza virus A/chicken/Kurgan/5/2005(H5N1) in nuclear-transformed duckweed plants for further development of avian influenza vaccine. The N-terminal fragment of M2, including M2e, was selected for expression. The M2e DNA sequence fused in-frame to the 5' end of β-glucuronidase was cloned into pBI121 under the control of CaMV 35S promoter. The resulting plasmid was successfully used for duckweed transformation, and western analysis with anti-β-glucuronidase and anti-M2e antibodies confirmed accumulation of the target protein (M130) in 17 independent transgenic lines. Quantitative ELISA of crude protein extracts from these lines showed M130-β-glucuronidase accumulation ranging from 0.09-0.97 mg/g FW (0.12-1.96 % of total soluble protein), equivalent to yields of up to 40 μg M2e/g plant FW. This relatively high yield holds promise for the development of a duckweed-based expression system to produce an edible vaccine against avian influenza.

  4. Expression of an Arabidopsis vacuolar H+-pyrophosphatase gene (AVP1) in cotton improves drought- and salt tolerance and increases fibre yield in the field conditions.

    PubMed

    Pasapula, Vijaya; Shen, Guoxin; Kuppu, Sundaram; Paez-Valencia, Julio; Mendoza, Marisol; Hou, Pei; Chen, Jian; Qiu, Xiaoyun; Zhu, Longfu; Zhang, Xianlong; Auld, Dick; Blumwald, Eduardo; Zhang, Hong; Gaxiola, Roberto; Payton, Paxton

    2011-01-01

    The Arabidopsis gene AVP1 encodes a vacuolar pyrophosphatase that functions as a proton pump on the vacuolar membrane. Overexpression of AVP1 in Arabidopsis, tomato and rice enhances plant performance under salt and drought stress conditions, because up-regulation of the type I H+-PPase from Arabidopsis may result in a higher proton electrochemical gradient, which facilitates enhanced sequestering of ions and sugars into the vacuole, reducing water potential and resulting in increased drought- and salt tolerance when compared to wild-type plants. Furthermore, overexpression of AVP1 stimulates auxin transport in the root system and leads to larger root systems, which helps transgenic plants absorb water more efficiently under drought conditions. Using the same approach, AVP1-expressing cotton plants were created and tested for their performance under high-salt and reduced irrigation conditions. The AVP1-expressing cotton plants showed more vigorous growth than wild-type plants in the presence of 200 mM NaCl under hydroponic growth conditions. The soil-grown AVP1-expressing cotton plants also displayed significantly improved tolerance to both drought and salt stresses in greenhouse conditions. Furthermore, the fibre yield of AVP1-expressing cotton plants is at least 20% higher than that of wild-type plants under dry-land conditions in the field. This research indicates that AVP1 has the potential to be used for improving crop's drought- and salt tolerance in areas where water and salinity are limiting factors for agricultural productivity.

  5. Heterologous Expression of ATG8c from Soybean Confers Tolerance to Nitrogen Deficiency and Increases Yield in Arabidopsis

    PubMed Central

    Liu, Dong; Chai, Wenting; Gong, Qingqiu; Wang, Ning Ning

    2012-01-01

    Nitrogen is an essential element for plant growth and yield. Improving Nitrogen Use Efficiency (NUE) of crops could potentially reduce the application of chemical fertilizer and alleviate environmental damage. To identify new NUE genes is therefore an important task in molecular breeding. Macroautophagy (autophagy) is an intracellular process in which damaged or obsolete cytoplasmic components are encapsulated in double membraned vesicles termed autophagosomes, then delivered to the vacuole for degradation and nutrient recycling. One of the core components of autophagosome formation, ATG8, has been shown to directly mediate autophagosome expansion, and the transcript of which is highly inducible upon starvation. Therefore, we postulated that certain homologs of Saccharomyces cerevisiae ATG8 (ScATG8) from crop species could have potential for NUE crop breeding. A soybean (Glycine max, cv. Zhonghuang-13) ATG8, GmATG8c, was selected from the 11 family members based on transcript analysis upon nitrogen deprivation. GmATG8c could partially complement the yeast atg8 mutant. Constitutive expression of GmATG8c in soybean callus cells not only enhanced nitrogen starvation tolerance of the cells but accelerated the growth of the calli. Transgenic Arabidopsis over-expressing GmATG8c performed better under extended nitrogen and carbon starvation conditions. Meanwhile, under optimum growth conditions, the transgenic plants grew faster, bolted earlier, produced larger primary and axillary inflorescences, eventually produced more seeds than the wild-type. In average, the yield was improved by 12.9%. We conclude that GmATG8c may serve as an excellent candidate for breeding crops with enhanced NUE and better yield. PMID:22629371

  6. Expression, purification and characterization of inactive and active forms of ERK2 from insect expression system.

    PubMed

    Yan, Kelly; Merritt, Hanne; Crawford, Kenneth; Pardee, Gwynn; Cheng, Jan Marie; Widger, Stephania; Hekmat-Nejad, Mohammad; Zaror, Isabel; Sim, Janet

    2015-06-01

    Extracellular signal-regulated kinase 2 (ERK2) is a serine/threonine protein kinase involved in many cellular programs, such as cell proliferation, differentiation, motility and programed cell-death. It is therefore considered an important target in the treatment of cancer. In an effort to support biochemical screening and small molecule drug discovery, we established a robust system to generate both inactive and active forms of ERK2 using insect expression system. We report here, for the first time, that inactive ERK2 can be expressed and purified with 100% homogeneity in the unphosphorylated form using insect system. This resulted in a significant 20-fold yield improvement compared to that previously reported using bacterial expression system. We also report a newly developed system to generate active ERK2 in insect cells through in vivo co-expression with a constitutively active MEK1 (S218D S222D). Isolated active ERK2 was confirmed to be doubly phosphorylated at the correct sites, T185 and Y187, in the activation loop of ERK2. Both ERK2 forms, inactive and active, were well characterized by biochemical activity assay for their kinase function. Inactive and active ERK2 were the two key reagents that enabled successful high through-put biochemical assay screen and structural drug discovery studies.

  7. Effect of tillage system on yield and weed populations of soybean ( Glycin Max L.).

    PubMed

    Hosseini, Seyed Z; Firouzi, Saeed; Aminpanah, Hashem; Sadeghnejhad, Hamid R

    2016-03-01

    Field experiment was conducted at Agricultural and Natural Resources Research Center of Golestan Province, Iran, to determine the effects of tillage system and weed management regime on yield and weed populations in soybean ( Glycin max L.). The experimental design was a split plot where the whole plot portion was a randomized complete block with three replicates. Main plots were tillage system: 1- No-till row crop seeding, 2- No-till seed drilling, 3- Tillage with disc harrow and drill planting, 4- Tillage with chisel packer and drill planting. The subplots were weed management regimes: 1-Weed control with herbicide application, 2- Hand weeding, 3- Herbicide application plus hand weeding, and 4- Non-weeding. Results indicated that the main effects of tillage system and weed management regime were significant for seed yield, pod number per plant, seed number per pod, weed density and biomass, while their interaction were significant only for weed density, weed biomass, and seed number per pod. The highest grain yields (3838 kg ha-1) were recorded for No-till row crop seeding. The highest seed yield (3877 kg ha-1) also was recorded for weed control with herbicide and hand weeding treatment, followed by hand weeding (3379 kg ha-1).

  8. Heterologous and cell free protein expression systems.

    PubMed

    Farrokhi, Naser; Hrmova, Maria; Burton, Rachel A; Fincher, Geoffrey B

    2009-01-01

    In recognition of the fact that a relatively small percentage of 'named' genes in databases have any experimental proof for their annotation, attention is shifting towards the more accurate assignment of functions to individual genes in a genome. The central objective will be to reduce our reliance on nucleotide or amino acid sequence similarities as a means to define the functions of genes and to annotate genome sequences. There are many unsolved technical difficulties associated with the purification of specific proteins from extracts of biological material, especially where the protein is present in low abundance, has multiple isoforms or is found in multiple post-translationally modified forms. The relative ease with which cDNAs can be cloned has led to the development of methods through which cDNAs from essentially any source can be expressed in a limited range of suitable host organisms, so that sufficient levels of the encoded proteins can be generated for functional analysis. Recently, these heterologous expression systems have been supplemented by more robust prokaryotic and eukaryotic cell-free protein synthesis systems. In this chapter, common host systems for heterologous expression are reviewed and the current status of cell-free expression systems will be presented. New approaches to overcoming the special problems encountered during the expression of membrane-associated proteins will also be addressed. Methodological considerations, including the characteristics of codon usage in the expressed DNA, peptide tags that facilitate subsequent purification of the expressed proteins and the role of post-translational modifications, are examined.

  9. Development and validation of equations utilizing lamb vision system output to predict lamb carcass fabrication yields.

    PubMed

    Cunha, B C N; Belk, K E; Scanga, J A; LeValley, S B; Tatum, J D; Smith, G C

    2004-07-01

    This study was performed to validate previous equations and to develop and evaluate new regression equations for predicting lamb carcass fabrication yields using outputs from a lamb vision system-hot carcass component (LVS-HCC) and the lamb vision system-chilled carcass LM imaging component (LVS-CCC). Lamb carcasses (n = 149) were selected after slaughter, imaged hot using the LVS-HCC, and chilled for 24 to 48 h at -3 to 1 degrees C. Chilled carcasses yield grades (YG) were assigned on-line by USDA graders and by expert USDA grading supervisors with unlimited time and access to the carcasses. Before fabrication, carcasses were ribbed between the 12th and 13th ribs and imaged using the LVS-CCC. Carcasses were fabricated into bone-in subprimal/primal cuts. Yields calculated included 1) saleable meat yield (SMY); 2) subprimal yield (SPY); and 3) fat yield (FY). On-line (whole-number) USDA YG accounted for 59, 58, and 64%; expert (whole-number) USDA YG explained 59, 59, and 65%; and expert (nearest-tenth) USDA YG accounted for 60, 60, and 67% of the observed variation in SMY, SPY, and FY, respectively. The best prediction equation developed in this trial using LVS-HCC output and hot carcass weight as independent variables explained 68, 62, and 74% of the variation in SMY, SPY, and FY, respectively. Addition of output from LVS-CCC improved predictive accuracy of the equations; the combined output equations explained 72 and 66% of the variability in SMY and SPY, respectively. Accuracy and repeatability of measurement of LM area made with the LVS-CCC also was assessed, and results suggested that use of LVS-CCC provided reasonably accurate (R2 = 0.59) and highly repeatable (repeatability = 0.98) measurements of LM area. Compared with USDA YG, use of the dual-component lamb vision system to predict cut yields of lamb carcasses improved accuracy and precision, suggesting that this system could have an application as an objective means for pricing carcasses in a value

  10. Expression systems for cloned xenobiotic transporters

    SciTech Connect

    Pritchard, John B.

    2005-05-01

    One challenge of modern biology is to be able to match genes and their encoded proteins with events at the molecular, cellular, tissue, and organism levels, and thus, provide a multi-level understanding of gene function and dysfunction. How well this can be done for xenobiotic transporters depends on a knowledge of the genes expressed in the tissue, the cellular locations of the gene products (do they function for uptake or efflux?), and our ability to match substrates with transporters using information obtained from cloned transporters functioning in heterologous expression systems. Clearly, making a rational choice of expression system to use for the characterization and study of cloned xenobiotic transporters is a critical part of study design. This choice requires well-defined goals, as well as an understanding of the strengths and weaknesses of candidate expression systems.

  11. Baculovirus-insect cell expression systems.

    PubMed

    Jarvis, Donald L

    2009-01-01

    In the early 1980s, the first-published reports of baculovirus-mediated foreign gene expression stimulated great interest in the use of baculovirus-insect cell systems for recombinant protein production. Initially, this system appeared to be the first that would be able to provide the high production levels associated with bacterial systems and the eukaryotic protein processing capabilities associated with mammalian systems. Experience and an increased understanding of basic insect cell biology have shown that these early expectations were not completely realistic. Nevertheless, baculovirus-insect cell expression systems have the capacity to produce many recombinant proteins at high levels and they also provide significant eukaryotic protein processing capabilities. Furthermore, important technological advances over the past 20 years have improved upon the original methods developed for the isolation of baculovirus expression vectors, which were inefficient, required at least some specialized expertise and, therefore, induced some frustration among those who used the original baculovirus-insect cell expression system. Today, virtually any investigator with basic molecular biology training can relatively quickly and efficiently isolate a recombinant baculovirus vector and use it to produce their favorite protein in an insect cell culture. This chapter will begin with background information on the basic baculovirus-insect cell expression system and will then focus on recent developments that have greatly facilitated the ability of an average investigator to take advantage of its attributes.

  12. Estimating the potential intensification of global grazing systems based on climate adjusted yield gap analysis

    NASA Astrophysics Data System (ADS)

    Sheehan, J. J.

    2016-12-01

    We report here a first-of-its-kind analysis of the potential for intensification of global grazing systems. Intensification is calculated using the statistical yield gap methodology developed previously by others (Mueller et al 2012 and Licker et al 2010) for global crop systems. Yield gaps are estimated by binning global pasture land area into 100 equal area sized bins of similar climate (defined by ranges of rainfall and growing degree days). Within each bin, grid cells of pastureland are ranked from lowest to highest productivity. The global intensification potential is defined as the sum of global production across all bins at a given percentile ranking (e.g. performance at the 90th percentile) divided by the total current global production. The previous yield gap studies focused on crop systems because productivity data on these systems is readily available. Nevertheless, global crop land represents only one-third of total global agricultural land, while pasture systems account for the remaining two-thirds. Thus, it is critical to conduct the same kind of analysis on what is the largest human use of land on the planet—pasture systems. In 2013, Herrero et al announced the completion of a geospatial data set that augmented the animal census data with data and modeling about production systems and overall food productivity (Herrero et al, PNAS 2013). With this data set, it is now possible to apply yield gap analysis to global pasture systems. We used the Herrero et al data set to evaluate yield gaps for meat and milk production from pasture based systems for cattle, sheep and goats. The figure included with this abstract shows the intensification potential for kcal per hectare per year of meat and milk from global cattle, sheep and goats as a function of increasing levels of performance. Performance is measured as the productivity achieved at a given ranked percentile within each bin.We find that if all pasture land were raised to their 90th percentile of

  13. Genetic modification of alphaGal expression in xenogeneic endothelial cells yields a complex immunological response.

    PubMed

    Fischbeck, J A; Baier, J M; Akella, R; Hern-Anderson, D; Schmidt, C E

    2001-12-01

    The source of cells for tissue engineering applications remains a hurdle, predominantly for procedures in which there is insufficient time to harvest a patient's own cells. Animal cells are readily available, but undergo immune rejection. Rejection of animal (i.e., xenogeneic) tissue involves practically every component of the immune system. The initial phase, hyperacute rejection (HAR), involves natural xenoreactive antibodies and the complement system, and leads to endothelial cell lysis and rapid tissue destruction. The cell-surface epitope, galactose-alpha(1,3)-galactose (alphaGal), is presumed to play a key role in HAR. The later stage of immune response (delayed xenograft rejection or DXR), is mediated by immune cells such as monocytes. Carbohydrates are likely also involved in DXR, but their role in this phase of the immune response is less clear. A better understanding of all stages of xenogeneic immune rejection may make it feasible to create cell lines that are immune tolerant. In these studies, we have genetically modified bovine endothelial cells to study the roles of carbohydrates in immune rejection. Our studies suggest that one or more epitopes other than alphaGal may influence complement-mediated lysis. Furthermore, antibodies, as instigators in the complement response, and monocytes appear to recognize different cell surface epitopes.

  14. Transgene expression systems in the Triticeae cereals.

    PubMed

    Hensel, Götz; Himmelbach, Axel; Chen, Wanxin; Douchkov, Dimitar K; Kumlehn, Jochen

    2011-01-01

    The control of transgene expression is vital both for the elucidation of gene function and for the engineering of transgenic crops. Given the dominance of the Triticeae cereals in the agricultural economy of the temperate world, the development of well-performing transgene expression systems of known functionality is of primary importance. Transgenes can be expressed either transiently or stably. Transient expression systems based on direct or virus-mediated gene transfer are particularly useful in situations where the need is to rapidly screen large numbers of genes. However, an unequivocal understanding of gene function generally requires that a transgene functions throughout the plant's life and is transmitted through the sexual cycle, since this alone allows its effect to be decoupled from the plant's response to the generally stressful gene transfer event. Temporal, spatial and quantitative control of a transgene's expression depends on its regulatory environment, which includes both its promoter and certain associated untranslated region sequences. While many transgenic approaches aim to manipulate plant phenotype via ectopic gene expression, a transgene sequence can be also configured to down-regulate the expression of its endogenous counterpart, a strategy which exploits the natural gene silencing machinery of plants. In this review, current technical opportunities for controlling transgene expression in the Triticeae species are described. Apart from protocols for transient and stable gene transfer, the choice of promoters and other untranslated regulatory elements, we also consider signal peptides, as they too govern the abundance and particularly the sub-cellular localization of transgene products.

  15. Satellite-based assessment of yield variation and its determinants in smallholder African systems

    PubMed Central

    Lobell, David B.

    2017-01-01

    The emergence of satellite sensors that can routinely observe millions of individual smallholder farms raises possibilities for monitoring and understanding agricultural productivity in many regions of the world. Here we demonstrate the potential to track smallholder maize yield variation in western Kenya, using a combination of 1-m Terra Bella imagery and intensive field sampling on thousands of fields over 2 y. We find that agreement between satellite-based and traditional field survey-based yield estimates depends significantly on the quality of the field-based measures, with agreement highest (R2 up to 0.4) when using precise field measures of plot area and when using larger fields for which rounding errors are smaller. We further show that satellite-based measures are able to detect positive yield responses to fertilizer and hybrid seed inputs and that the inferred responses are statistically indistinguishable from estimates based on survey-based yields. These results suggest that high-resolution satellite imagery can be used to make predictions of smallholder agricultural productivity that are roughly as accurate as the survey-based measures traditionally used in research and policy applications, and they indicate a substantial near-term potential to quickly generate useful datasets on productivity in smallholder systems, even with minimal or no field training data. Such datasets could rapidly accelerate learning about which interventions in smallholder systems have the most positive impact, thus enabling more rapid transformation of rural livelihoods. PMID:28202728

  16. Effect of Pulse Width on Ozone Yield using Inductive Energy Storage System Pulsed Power Generator

    NASA Astrophysics Data System (ADS)

    Yagi, Ippei; Mukaigawa, Seiji; Takaki, Koichi; Fujiwara, Tamiya; Go, Tomio

    Nanosecond pulse voltages of several pulse widths were applied to a cylindrical plasma reactor for ozone synthesis with high energy yield. Nanoseconds pulse voltages were produced by inductive energy storage system pulsed power generators using semiconductor opening switch (SOS) diodes. First recovery diodes were used as SOS diodes in the inductive energy storage system to produce short-pulsed high voltage with high-repetition rate. The short pulse voltage of 9.5 ns width and 33 kV peak voltage was produced at charging voltage of 15 kV and was applied to a 1 mm diameter center wire electrode in the plasma reactor. The copper cylinder of 19 mm inner diameter was used as outer electrode and was connected to a ground. The ozone yield of 271 g/kWh was obtained using the 9.5 ns width pulse voltage at synthesized 412 ppm of ozone concentration. The yield 271 g/kWh was more than twice as much as the yield 114 g/kWh at 401 ppm using a 60 ns pulse voltage.

  17. Ectopic expression of Pokkali phosphoglycerate kinase-2 (OsPGK2-P) improves yield in tobacco plants under salinity stress.

    PubMed

    Joshi, Rohit; Karan, Ratna; Singla-Pareek, Sneh L; Pareek, Ashwani

    2016-01-01

    Our results indicate that OsPGK2a-P gene is differentially regulated in contrasting rice cultivars under stress and its overexpression confers salt stress tolerance in transgenic tobacco. Phosphoglycerate kinase (PGK; EC = 2.7.2.3) plays a major role for ATP production during glycolysis and 1, 3-bisphosphoglycerate production to participate in the Calvin cycle for carbon fixation in plants. Whole genome analysis of rice reveals the presence of four PGK genes (OsPgks) on different chromosomes. Comparative expression analysis of OsPgks in rice revealed highest level of transcripts for OsPgk2 at most of its developmental stages. Detailed characterization of OsPgk2 transcript and protein showed that it is strongly induced by salinity stress in two contrasting genotypes of rice, i.e., cv IR64 (salt sensitive) and landrace Pokkali (salt tolerant). Ectopic expression of OsPgk2a-P (isolated from Pokkali) in transgenic tobacco improved its salinity stress tolerance by higher chlorophyll retention and enhanced proline accumulation, besides maintaining better ion homeostasis. Ectopically expressing OsPgk2a-P transgenic tobacco plants showed tall phenotype with more number of pods than wild-type plants. Therefore, OsPgk2a-P appears to be a potential candidate for increasing salinity stress tolerance and enhanced yield in crop plants.

  18. Kinetics of lipogenic genes expression in milk purified mammary epithelial cells (MEC) across lactation and their correlation with milk and fat yield in buffalo.

    PubMed

    Yadav, Poonam; Kumar, Parveen; Mukesh, Manishi; Kataria, R S; Yadav, Anita; Mohanty, A K; Mishra, B P

    2015-04-01

    Expression patterns of lipogenic genes (LPL, ABCG2, ACSS2, ACACA, SCD, BDH, LIPIN1, SREBF1, PPARα and PPARγ) were studied in milk purified MEC across different stages of lactation (15, 30, 45, 60, 90, 120 and 240 days relative to parturition) in buffalo. PPARα was the most abundant gene while ABCG2 and ACSS2 had moderate level of expression; whereas expression of SREBF and PPARγ was very low. The expression patterns of some genes (BDH1, ACSS2, and LIPIN1) across lactation were positively correlated with milk yield while negatively correlated with fat yield. SCD also showed weak correlation with milk yield (p, 0.53) and fat yield (p, -0.47). On the other hand, expression pattern of ACACA was negatively correlated with milk yield (p, -0.88) and positively correlated with fat yield (p, 0.62). Strong correlation was observed between genes involved in de novo milk fat synthesis (BDH1, ACSS2, LIPIN2 and SCD) and milk yield.

  19. PERUN system and its application for assessing the crop yield potential of the Czech Republic

    NASA Astrophysics Data System (ADS)

    Dubrovsky, M.; Zalud, Z.; Eitzinger, J.; Trnka, M.; Semeradova, D.

    2003-04-01

    The main purpose of the first version of the computer system PERUN, which has been developed in 2001-2002 (presented in EGS 2002), is the probabilistic seasonal crop yield forecasting for a given site. The system is based on the crop growth model WOFOST (version 7, slightly modified) and the six-variate version of the stochastic weather generator Met&Roll. The system is now being enhanced to allow assessment of the crop yield potential of a larger area. As this assessment requires a great amount of meteorological, pedological and crop data to be gathered, but these data are not yet all available to the authors, the presentation will rather focus on the methodological aspects and the results of the sensitivity analysis. The presentation will consist of the following points: (i) Overview of the PERUN system. The results of the validation experiments (spring barley and winter wheat at selected Czech locations) will be presented, too. (ii) Methodology used for a spatial assessment. The assessment is based on integrating model crop yields simulated at multiple sub-regions with region-specific climatic and pedological conditions. The input daily weather series are produced by the stochastic generator. The multi-year crop model simulation is performed for each sub-region to assess the mean and variability of the model yields. (iii) Sensitivity of the regional crop production potential to uncertainties in selected input characteristics: crop cultivar, soil type, hydrological characteristics (e.g. amount of available water at the beginning of the simulation), and climatic conditions (e.g temperature, precipitation). In assessing sensitivity to climate, the climatic characteristics will be varied within the range of values typical for the territory of the Czech Republic. The crops applied in the analysis are spring barley and winter wheat. Acknowledgement: The system PERUN has been developed within the frame of project QC1316 sponsored by the Czech National Agency for

  20. Maximum sustainable yield and species extinction in a prey-predator system: some new results.

    PubMed

    Ghosh, Bapan; Kar, T K

    2013-06-01

    Though the maximum sustainable yield (MSY) approach has been legally adopted for the management of world fisheries, it does not provide any guarantee against from species extinction in multispecies communities. In the present article, we describe the appropriateness of the MSY policy in a Holling-Tanner prey-predator system with different types of functional responses. It is observed that for both type I and type II functional responses, harvesting of either prey or predator species at the MSY level is a sustainable fishing policy. In the case of combined harvesting, both the species coexist at the maximum sustainable total yield (MSTY) level if the biotic potential of the prey species is greater than a threshold value. Further, increase of the biotic potential beyond the threshold value affects the persistence of the system.

  1. Detailed analysis of the energy yield of systems with covered sheet-and-tube PVT collectors

    SciTech Connect

    Santbergen, R.; Rindt, C.C.M.; van Zolingen, R.J.Ch.; Zondag, H.A.

    2010-05-15

    Solar cells have a typical efficiency in the range of 5-20%, implying that 80% or more of the incident solar energy can be harvested in the form of heat and applied for low-temperature heating. In a PVT collector one tries to collect this heat. In this work, the electrical and thermal yield of solar domestic hot water systems with one-cover sheet-and-tube PVT collectors were considered. Objectives of the work were to understand the mechanisms determining these yields, to investigate measures to improve these yields and to investigate the yield consequences if various solar cell technologies are being used. The work was carried out using numerical simulations. A detailed quantitative understanding of all loss mechanisms was obtained, especially of those being inherent to the use of PVT collectors instead of PV modules and conventional thermal collectors. The annual electrical efficiencies of the PVT systems investigated were up to 14% (relative) lower compared to pure PV systems and the annual thermal efficiencies up to 19% (relative) lower compared to pure thermal collector systems. The loss of electrical efficiency is mainly caused by the relatively high fluid temperature. The loss of thermal efficiency is caused both by the high emissivity of the absorber and the withdrawal of electrical energy. However, both the loss of electrical and thermal efficiency can be reduced further by the application of anti-reflective coatings. The thermal efficiency can be improved by the application of a low-emissivity coating on the absorber, however at the cost of a reduced electrical efficiency. (author)

  2. Visual tree grading systems for estimating lumber yields in young and mature southern pine

    Treesearch

    Alexander Clark; Robert H. McAlister

    1998-01-01

    New visual tree grading systems for mature southern pine ? 35 years old and young pine ? 35 years old based on number and size of branches in the lower bole are described. A series of lumber grade yield studies was conducted to test the new grading rules. A total of 214 natural loblolly pine (Pinus taeda L.) and shortleaf pine (P. echinata Mill) trees 9 to 20 inches...

  3. An updated whole stand growth and yield system for planted longleaf pine in southwest Georgia

    Treesearch

    John R. Brooks; Steven B. Jack

    2016-01-01

    An updated whole stand growth and yield system for planted longleaf pine (Pinus palustris) was developed from permanent plot data collected annually over a 13 to 16 year period. The data set consists of 15 intensively managed longleaf pine plantations that are located in Lee, Worth, Mitchell, and Baker counties in southwest Georgia. Stand survival, dominant height,...

  4. Enhancing digestibility and ethanol yield of Populus wood via expression of an engineered monolignol 4-O-methyltransferase

    DOE PAGES

    Cai, Yuanheng; Zhang, Kewei; Kim, Hoon; ...

    2016-06-28

    Producing cellulosic biofuels and bio-based chemicals from woody biomass is impeded by the presence of lignin polymer in the plant cell wall. Manipulating the monolignol biosynthetic pathway offers a promising approach to improved processability, but often impairs plant growth and development. Here, we show that expressing an engineered 4-O-methyltransferase that chemically modifies the phenolic moiety of lignin monomeric precursors, thus preventing their incorporation into the lignin polymer, substantially alters hybrid aspens’ lignin content and structure. Woody biomass derived from the transgenic aspens shows a 62% increase in the release of simple sugars and up to a 49% increase in themore » yield of ethanol when the woody biomass is subjected to enzymatic digestion and yeast-mediated fermentation. Furthermore, the cell wall structural changes do not affect growth and biomass production of the trees. Our study provides a useful strategy for tailoring woody biomass for bio-based applications.« less

  5. Enhancing digestibility and ethanol yield of Populus wood via expression of an engineered monolignol 4-O-methyltransferase.

    PubMed

    Cai, Yuanheng; Zhang, Kewei; Kim, Hoon; Hou, Guichuan; Zhang, Xuebin; Yang, Huijun; Feng, Huan; Miller, Lisa; Ralph, John; Liu, Chang-Jun

    2016-06-28

    Producing cellulosic biofuels and bio-based chemicals from woody biomass is impeded by the presence of lignin polymer in the plant cell wall. Manipulating the monolignol biosynthetic pathway offers a promising approach to improved processability, but often impairs plant growth and development. Here, we show that expressing an engineered 4-O-methyltransferase that chemically modifies the phenolic moiety of lignin monomeric precursors, thus preventing their incorporation into the lignin polymer, substantially alters hybrid aspens' lignin content and structure. Woody biomass derived from the transgenic aspens shows a 62% increase in the release of simple sugars and up to a 49% increase in the yield of ethanol when the woody biomass is subjected to enzymatic digestion and yeast-mediated fermentation. Moreover, the cell wall structural changes do not affect growth and biomass production of the trees. Our study provides a useful strategy for tailoring woody biomass for bio-based applications.

  6. Enhancing digestibility and ethanol yield of Populus wood via expression of an engineered monolignol 4-O-methyltransferase

    PubMed Central

    Cai, Yuanheng; Zhang, Kewei; Kim, Hoon; Hou, Guichuan; Zhang, Xuebin; Yang, Huijun; Feng, Huan; Miller, Lisa; Ralph, John; Liu, Chang-Jun

    2016-01-01

    Producing cellulosic biofuels and bio-based chemicals from woody biomass is impeded by the presence of lignin polymer in the plant cell wall. Manipulating the monolignol biosynthetic pathway offers a promising approach to improved processability, but often impairs plant growth and development. Here, we show that expressing an engineered 4-O-methyltransferase that chemically modifies the phenolic moiety of lignin monomeric precursors, thus preventing their incorporation into the lignin polymer, substantially alters hybrid aspens' lignin content and structure. Woody biomass derived from the transgenic aspens shows a 62% increase in the release of simple sugars and up to a 49% increase in the yield of ethanol when the woody biomass is subjected to enzymatic digestion and yeast-mediated fermentation. Moreover, the cell wall structural changes do not affect growth and biomass production of the trees. Our study provides a useful strategy for tailoring woody biomass for bio-based applications. PMID:27349324

  7. Enhanced expression of OsSPL14 gene and its association with yield components in rice (Oryza sativa) under low nitrogen conditions.

    PubMed

    Srikanth, B; Subhakara Rao, I; Surekha, K; Subrahmanyam, D; Voleti, S R; Neeraja, C N

    2016-01-15

    Nitrogen use efficiency (NUE) in rice crop is the need of the hour for reduction of nitrous oxide emission resulting from excess nitrogen (N) fertilizer application and also in reduction of cost of cultivation. Ten rice genotypes were grown under low and recommended dose of N application and characterized in terms of parameters related to yield, yield related components and NUE indicators. Wide genetic variability under low N conditions was observed with significant variation for 15 yield related parameters in interactions of genotypes and treatment. Limitation of N has led to the decrease of all yield and yield related parameters, but for grain filling % and 1000 grain weight. Two genotypes, Rasi and Varadhan have shown minimum differences between low and recommended N conditions. Correlation analysis of various yield components showed the importance of the secondary branches for the total grains under low N. Expression analysis of OsSPL14 (LOC_Os08g39890) gene reported to be associated with increased panicle branching and higher grain yield through real time PCR in leaf and three stages of panicle has shown differential temporal expression and its association with yield and yield related components across the genotypes. The expression of OsSPL14 at panicle stage 3, has shown correlation (P<0.05) with N% in grain. Since OsSPL14 is a functional transcription activator, its association of expression in leaf and three panicle stages with yield components as observed in the present study suggests the role of nitrogen metabolism related genes in plant growth and development and its conversion into yield components in rice.

  8. Constitutive expression of CaPLA1 conferred enhanced growth and grain yield in transgenic rice plants.

    PubMed

    Park, Ki Youl; Kim, Eun Yu; Seo, Young Sam; Kim, Woo Taek

    2016-03-01

    Phospholipids are not only important components of cell membranes, but participate in diverse processes in higher plants. In this study, we generated Capsicum annuum phospholipiase A1 (CaPLA1) overexpressing transgenic rice (Oryza sativa L.) plants under the control of the maize ubiquitin promoter. The T4 CaPLA1-overexpressing rice plants (Ubi:CaPLA1) had a higher root:shoot mass ratio than the wild-type plants in the vegetative stage. Leaf epidermal cells from transgenic plants had more cells than wild-type plants. Genes that code for cyclin and lipid metabolic enzymes were up-regulated in the transgenic lines. When grown under typical paddy field conditions, the transgenic plants produced more tillers, longer panicles and more branches per panicle than the wild-type plants, all of which resulted in greater grain yield. Microarray analysis suggests that gene expressions that are related with cell proliferation, lipid metabolism, and redox state were widely altered in CaPLA1-overexpressing transgenic rice plants. Ubi:CaPLA1 plants had a reduced membrane peroxidation state, as determined by malondialdehyde and conjugated diene levels and higher peroxidase activity than wild-type rice plants. Furthermore, three isoprenoid synthetic genes encoding terpenoid synthase, hydroxysteroid dehydrogenase and 3-hydroxy-3-methyl-glutaryl-CoA reductase were up-regulated in CaPLA1-overexpressing plants. We suggest that constitutive expression of CaPLA1 conferred increased grain yield with enhanced growth in transgenic rice plants by alteration of gene activities related with cell proliferation, lipid metabolism, membrane peroxidation state and isoprenoid biosynthesis.

  9. Dependence of Initial Oxygen Concentration on Ozone Yield Using Inductive Energy Storage System Pulsed Power Generator

    NASA Astrophysics Data System (ADS)

    Go, Tomio; Tanaka, Yasushi; Yamazaki, Nobuyuki; Mukaigawa, Seiji; Takaki, Koichi; Fujiwara, Tamiya

    Dependence of initial oxygen concentration on ozone yield using streamer discharge reactor driven by an inductive energy storage system pulsed power generator is described in this paper. Fast recovery type diodes were employed as semiconductor opening switch to interrupt a circuit current within 100 ns. This rapid current change produced high-voltage short pulse between a secondary energy storage inductor. The repetitive high-voltage short pulse was applied to a 1 mm diameter center wire electrode placed in a cylindrical pulse corona reactor. The streamer discharge successfully occurred between the center wire electrode and an outer cylinder ground electrode of 2 cm inner diameter. The ozone was produced with the streamer discharge and increased with increasing pulse repetition rate. The ozone yield changed in proportion to initial oxygen concentration contained in the injected gas mixture at 800 ns forward pumping time of the current. However, the decrease of the ozone yield by decreasing oxygen concentration in the gas mixture at 180 ns forward pumping time of the current was lower than the decrease at 800 ns forward pumping time of the current. This dependence of the initial oxygen concentration on ozone yield at 180 ns forward pumping time is similar to that of dielectric barrier discharge reactor.

  10. The Potato Systems Planner: Integrating Cropping System Impacts on Crop Yield and Quality, Soil Biology, Nutrient Cycling, Diseases, and Economics

    USDA-ARS?s Scientific Manuscript database

    Finding and developing profitable cropping systems is a high priority for the potato industry. Consequently, an interdisciplinary team of ARS scientists from the New England Plant, Soil, & Water Laboratory evaluated 14 different rotations for their impacts on crop yield and quality, nutrient availa...

  11. Holographic lens spectrum splitting photovoltaic system for increased diffuse collection and annual energy yield

    NASA Astrophysics Data System (ADS)

    Vorndran, Shelby D.; Wu, Yuechen; Ayala, Silvana; Kostuk, Raymond K.

    2015-09-01

    Concentrating and spectrum splitting photovoltaic (PV) modules have a limited acceptance angle and thus suffer from optical loss under off-axis illumination. This loss manifests itself as a substantial reduction in energy yield in locations where a significant portion of insulation is diffuse. In this work, a spectrum splitting PV system is designed to efficiently collect and convert light in a range of illumination conditions. The system uses a holographic lens to concentrate shortwavelength light onto a smaller, more expensive indium gallium phosphide (InGaP) PV cell. The high efficiency PV cell near the axis is surrounded with silicon (Si), a less expensive material that collects a broader portion of the solar spectrum. Under direct illumination, the device achieves increased conversion efficiency from spectrum splitting. Under diffuse illumination, the device collects light with efficiency comparable to a flat-panel Si module. Design of the holographic lens is discussed. Optical efficiency and power output of the module under a range of illumination conditions from direct to diffuse are simulated with non-sequential raytracing software. Using direct and diffuse Typical Metrological Year (TMY3) irradiance measurements, annual energy yield of the module is calculated for several installation sites. Energy yield of the spectrum splitting module is compared to that of a full flat-panel Si reference module.

  12. Wild pollinators enhance oilseed rape yield in small-holder farming systems in China.

    PubMed

    Zou, Yi; Xiao, Haijun; Bianchi, Felix J J A; Jauker, Frank; Luo, Shudong; van der Werf, Wopke

    2017-02-21

    Insect pollinators play an important role in crop pollination, but the relative contribution of wild pollinators and honey bees to pollination is currently under debate. There is virtually no information available on the strength of pollination services and the identity of pollination service providers from Asian smallholder farming systems, where fields are small, and variation among fields is high. We established 18 winter oilseed rape (Brassica napus L.) fields along a large geographical gradient in Jiangxi province in China. In each field, oilseed rape plants were grown in closed cages that excluded pollinators and open cages that allowed pollinator access. The pollinator community was sampled by pan traps for the entire oilseed rape blooming period. Oilseed rape plants from which insect pollinators were excluded had on average 38% lower seed set, 17% lower fruit set and 12% lower yield per plant, but the seeds were 17% heavier, and the caged plants had 28% more flowers and 18% higher aboveground vegetative biomass than plants with pollinator access. Oilseed rape plants thus compensate for pollination deficit by producing heavier seeds and more flowers. Regression analysis indicated that local abundance and diversity of wild pollinators were positively associated with seed set and yield/straw ratio, while honey bee abundance was not related to yield parameters. Wild pollinator abundance and diversity contribute to oilseed rape yield by enhancing plant resource allocation to seeds rather than to above-ground biomass. This study highlights the importance of the conservation of wild pollinators to support oilseed rape production in small-holder farming systems in China.

  13. Impact of land management system on crop yields and soil fertility in Cameroon

    NASA Astrophysics Data System (ADS)

    Tsozué, D.; Nghonda, J. P.; Mekem, D. L.

    2015-06-01

    The impact of direct-seeding mulch-based cropping systems (DMC), direct seeding (DS) and tillage seeding (TS) on Sorghum yields, soil fertility and the rehabilitation of degraded soils was evaluated in northern Cameroon. Field work consisted of visual examination, soil sampling, yield and rainfall data collection. Three fertilization rates (F1: 100 kg ha-1 NPK + 25 kg ha-1 of urea in DMC, F2: 200 kg ha-1 NPK + 50 kg ha-1 of urea in DMC and F3: 300 kg ha-1 NPK + 100 kg ha-1 of urea in DMC) were applied to each cropping system (DS, TS and DMC), resulting in nine experimental plots. Two types of chemical fertilizer were used (NPK 22.10.15 and urea) and applied each year from 2002 to 2012. Average Sorghum yields were 1239, 863 and 960 kg ha-1 respectively in DMC, DS and TS at F1, 1658, 1139 and 1192 kg ha-1 respectively in DMC, DS and TS at F2, and 2270, 2138 and 1780 kg ha-1 respectively in DMC, DS and TS at F3. pH values were 5.2 to 5.7 under DMC, 4.9 to 5.3 under DS and TS, and 5.6 in the control sample. High values of cation exchange capacity were recorded in the control sample, TS system and F1 of DMC. Base saturation rates, total nitrogen and organic matter contents were high in the control sample and the DMC than in the others systems. All studied soils were permanently not suitable for Sorghum due to the high percentage of nodules. F1 and F2 of the DS were currently not suitable, while F1 and F3 of DMC, F3 of DS and F1, F2 and F3 of TS were marginally suitable for Sorghum due to low soil pH values.

  14. Impact of land management system on crop yields and soil fertility in Cameroon

    NASA Astrophysics Data System (ADS)

    Tsozué, D.; Nghonda, J. P.; Mekem, D. L.

    2015-09-01

    The impact of direct-seeding mulch-based cropping systems (DMC), direct seeding (DS) and tillage seeding (TS) on Sorghum yields, soil fertility and the rehabilitation of degraded soils was evaluated in northern Cameroon. Field work consisted of visual examination, soil sampling, yield and rainfall data collection. Three fertilization rates (F1: 100 kg ha-1 NPK + 25 kg ha-1 of urea in DMC, F2: 200 kg ha-1 NPK + 50 kg ha-1 of urea in DMC and F3: 300 kg ha-1 NPK + 100 kg ha-1 of urea in DMC) were applied to each cropping system (DS, TS and DMC), resulting in nine experimental plots. Two types of chemical fertilizer were used (NPK 22.10.15 and urea) and applied each year from 2002 to 2012. Average Sorghum yields were 1239, 863 and 960 kg ha-1 in DMC, DS and TS, respectively, at F1, 1658, 1139 and 1192 kg ha-1 in DMC, DS and TS, respectively, at F2, and 2270, 2138 and 1780 kg ha-1 in DMC, DS and TS, respectively, at F3. pH values were 5.2-5.7 under DMC, 4.9-5.3 under DS and TS and 5.6 in the control sample. High values of cation exchange capacity were recorded in the control sample, TS system and F1 of DMC. Base saturation rates, total nitrogen and organic matter contents were higher in the control sample and DMC than in the other systems. All studied soils were permanently not suitable for Sorghum due to the high percentage of nodules. F1 and F2 of the DS were currently not suitable, while F1 and F3 of DMC, F3 of DS and F1, F2 and F3 of TS were marginally suitable for Sorghum due to low pH values.

  15. Yield and gas exchange ability of sweetpotato plants cultured in a hydroponic system

    NASA Astrophysics Data System (ADS)

    Kitaya, Y.; Hirai, H.; Saiful Islam, A. F. M.; Yamamoto, M.

    Life support of crews in space is greatly dependent on the amounts of food atmospheric O 2 and clean water produced by plants Therefore the space farming systems with scheduling of crop production obtaining high yields with a rapid turnover rate converting atmospheric CO 2 to O 2 and purifying water should be established with employing suitable plant species and varieties and precisely controlling environmental variables around plants grown at a high density in a limited space In this study three sweetpotato varieties were cultured in a newly developed hydroponic system and the yield the photosynthetic rate and the transpiration rate were compared on the earth as a fundamental study for establishing the space farming systems The varieties were Elegant summer Koukei 14 and Beniazuma The hydroponic system mainly consisted of water channels and rockwool boards A growing space for roots was made between the rockwool board and nutrient solution in the water channel Storage roots were developed on the lower surface of the rockwool plates Fresh weights of the storage roots were 1 6 1 2 and 0 6 kg plant for Koukei 14 Elegant summer and Beniazuma respectively grown for five months from June to October under the sun light in Osaka Japan Koukei 14 and Elegant summer produced greater total phytomass than Beniazuma There were positive correlations among the total phytomass the net photosynthetic rate and the transpiration rate Young stems and leaves as well as storage roots of Elegant summer are edible Therefore Elegant-summer

  16. Online evaluation of a commercial video image analysis system (Computer Vision System) to predict beef carcass red meat yield and for augmenting the assignment of USDA yield grades. United States Department of Agriculture.

    PubMed

    Cannell, R C; Belk, K E; Tatum, J D; Wise, J W; Chapman, P L; Scanga, J A; Smith, G C

    2002-05-01

    Objective quantification of differences in wholesale cut yields of beef carcasses at plant chain speeds is important for the application of value-based marketing. This study was conducted to evaluate the ability of a commercial video image analysis system, the Computer Vision System (CVS) to 1) predict commercially fabricated beef subprimal yield and 2) augment USDA yield grading, in order to improve accuracy of grade assessment. The CVS was evaluated as a fully installed production system, operating on a full-time basis at chain speeds. Steer and heifer carcasses (n = 296) were evaluated using CVS, as well as by USDA expert and online graders, before the fabrication of carcasses into industry-standard subprimal cuts. Expert yield grade (YG), online YG, CVS estimated carcass yield, and CVS measured ribeye area in conjunction with expert grader estimates of the remaining YG factors (adjusted fat thickness, percentage of kidney-pelvic-heart fat, hot carcass weight) accounted for 67, 39, 64, and 65% of the observed variation in fabricated yields of closely trimmed subprimals. The dual component CVS predicted wholesale cut yields more accurately than current online yield grading, and, in an augmentation system, CVS ribeye measurement replaced estimated ribeye area in determination of USDA yield grade, and the accuracy of cutability prediction was improved, under packing plant conditions and speeds, to a level close to that of expert graders applying grades at a comfortable rate of speed offline.

  17. Metabolic engineering of essential oil yield and composition in mint by altering expression of deoxyxylulose phosphate reductoisomerase and menthofuran synthase

    PubMed Central

    Mahmoud, Soheil S.; Croteau, Rodney B.

    2001-01-01

    Peppermint (Mentha × piperita L.) was independently transformed with a homologous sense version of the 1-deoxy-d-xylulose-5-phosphate reductoisomerase cDNA and with a homologous antisense version of the menthofuran synthase cDNA, both driven by the CaMV 35S promoter. Two groups of transgenic plants were regenerated in the reductoisomerase experiments, one of which remained normal in appearance and development; another was deficient in chlorophyll production and grew slowly. Transgenic plants of normal appearance and growth habit expressed the reductoisomerase transgene strongly and constitutively, as determined by RNA blot analysis and direct enzyme assay, and these plants accumulated substantially more essential oil (about 50% yield increase) without change in monoterpene composition compared with wild-type. Chlorophyll-deficient plants did not afford detectable reductoisomerase mRNA or enzyme activity and yielded less essential oil than did wild-type plants, indicating cosuppression of the reductoisomerase gene. Plants transformed with the antisense version of the menthofuran synthase cDNA were normal in appearance but produced less than half of this undesirable monoterpene oil component than did wild-type mint grown under unstressed or stressed conditions. These experiments demonstrate that essential oil quantity and quality can be regulated by metabolic engineering. Thus, alteration of the committed step of the mevalonate-independent pathway for supply of terpenoid precursors improves flux through the pathway that leads to increased monoterpene production, and antisense manipulation of a selected downstream monoterpene biosynthetic step leads to improved oil composition. PMID:11427737

  18. Efficient expression of acetylcholine-binding protein from Aplysia californica in Bac-to-Bac system.

    PubMed

    Lin, Bo; Meng, Hailing; Bing, Hui; Zhangsun, Dongting; Luo, Sulan

    2014-01-01

    The Bac-to-Bac baculovirus expression system can efficiently produce recombinant proteins, but the system may have to be optimized to achieve high-level expression for different candidate proteins. We reported here the efficient expression of acetylcholine-binding proteins from sea hares Aplysia californica (Ac-AChBP) and a convenient method to monitor protein expression level in this expression system. Three key factors affecting expression of Ac-AChBP were optimized for maximizing the yield, which included the cell density, volume of the infecting baculovirus inoculums, and the culturing time of postinfection. We have found it to reach a high yield of ∼5 mg/L, which needs 55 h incubation after infection at the cell density of 2 × 10(6) cells/mL with an inoculum volume ratio of 1 : 100. The optimized expression system in this study was also applied for expressing another protein Ls-AChBP from Lymnaea stagnalis successfully. Therefore, this established method is helpful to produce high yields of AChBP proteins for X-ray crystallographic structural and functional studies.

  19. Economics of fertility in high-yielding dairy cows on confined TMR systems.

    PubMed

    Cabrera, V E

    2014-05-01

    The objective of this review paper was to summarise the latest findings in dairy cattle reproductive economics with an emphasis on high yielding, confined total mixed ration systems. The economic gain increases as the reproductive efficiency improves. These increments follow the law of diminishing returns, but are still positive even at high reproductive performance. Reproductive improvement results in higher milk productivity and, therefore, higher milk income over feed cost, more calf sales and lower culling and breeding expenses. Most high-yielding herds in the United States use a combination of timed artificial insemination (TAI) and oestrous detection (OD) reproductive programme. The ratio of achievable pregnancies between OD and TAI determines the economic value difference between both and their combinations. Nonetheless, complex interactions between reproductive programme, herd relative milk yield, and type of reproductive programme are reported. For example, higher herd relative milk yield would favour programme relying more on TAI. In addition, improved reproductive efficiency produces extra replacements. The availability of additional replacements could allow more aggressive culling policies (e.g. less services for non-pregnant cows) to balance on-farm supply and demand of replacements. Balancing heifer replacement availability in an efficient reproductive programme brings additional economic benefits. New technologies such as the use of earlier chemical tests for pregnancy diagnosis could be economically effective depending on the goals and characteristics of the farm. Opportunities for individual cow reproductive management within defined reproductive programme exist. These decisions would be based on economic metrics derived from the value of a cow such as the value of a new pregnancy, the cost of a pregnancy loss, or the cost of an extra day open.

  20. The relationship between yield and the antioxidant defense system in tomatoes grown under heat stress.

    PubMed

    Rainwater, D T; Gossett, D R; Millhollon, E P; Hanna, H Y; Banks, S W; Lucas, M C

    1996-11-01

    Four putative heat-tolerant tomato (Lycopersicum esculentum) cultivars (Tamasabro, Heat Wave, LHT-24, and Solar Set) and one putative heat-sensitive tomato cultivar (Floradade) were grown in the field under non-stress (average daily temperature of 26 degrees C) and heat-stress (average daily temperature of 34 degrees C) conditions. At anthesis, approximately five weeks after being transplanted to the field, leaf samples were collected for antioxidant analyses. Yield was determined by harvesting ripe fruit seven weeks after the collection of leaf samples. Heat stress resulted in a 79.1% decrease in yield for the heat-sensitive Floradade, while the fruit yield in the heat-tolerant cultivars Heat Wave, LHT-24, Solar Set, and Tamasabro was reduced 51.5%, 22.1%, 43.8%, and 34.8% respectively. When grown under heat stress, antioxidant activities were also greater in the heat-tolerant cultivars. Superoxide dismutase (SOD) activity increased up to 9-fold in the heat-tolerant cultivars but decreased 83.1% in the heat-sensitive Floradade. Catalase, peroxidase, and ascorbate peroxidase activity increased significantly in all cultivars. Only Heat Wave showed a significant increase in glutathione reductase in response to heat stress but all heat-tolerant cultivars exhibited significantly lower oxidized ascorbate/reduced ascorbate ratios, greater reduced glutathione/oxidized glutathione rations, and greater alpha-tocopherol concentrations compared to the heat-sensitive cultivar Floridade. These data indicate that the more heat-tolerant cultivars had an enhanced capacity for scavenging active oxygen species and a more active ascorbate-glutathione cycle and suggest a strong correlation between the ability to up-regulate the antioxidant defense system and the ability of tomatoes to produce greater yields when grown under heat stress.

  1. Efficient cyclic system to yield ectoine using Brevibacterium sp. JCM 6894 subjected to osmotic downshock.

    PubMed

    Nagata, Shinichi; Wang, Yaoqiang; Oshima, Akinobu; Zhang, Linghua; Miyake, Hideyoshi; Sasaki, Hideaki; Ishida, Akio

    2008-03-01

    Brevibacterium sp. JCM 6894 cells grown in the presence of 1.5-2.5 M NaCl for 24 h at 30 degrees C were subjected to the osmotic downshock. Downshocked cells after ectoine release were grown for further 24 h in the fresh medium with same salinity as before shock. When this cyclic system was applied to the strain JCM 6894, the amount of ectoine in the cells increased with an increase of incubation time, which indicates that the cells manipulated by the present conditions were enough active to survive and synthesize ectoine after several times of osmotic downshock. In the presence of 2 M NaCl, the highest yield of ectoine released was achieved in this cyclic system, more than 2.4 g/L during 7 days of incubation. (1)H and (13)C-NMR analyses of solutes released from the cells by the osmotic downshock showed the presence of only ectoine with high purity. Release of ectoine from the cells was carried out within 5 min and its rates were increased by the dilution in the downshock treatment. For the convenience of operations, non-sterilized medium containing 2 M NaCl was examined for the cell growth in the present system, in which almost same level of ectoine yield, release rates, and cell viability were observed as those of sterilized medium.

  2. Method for improving product yields in an anionic metalloporphyrin-based artificial photosynthesis system

    DOEpatents

    Shelnutt, J.A.

    1984-11-29

    A method is disclosed improving product yields in an anionic metalloporphyrin-based artificial photosynthesis system for hydrogen generation. The method comprises forming an aqueous solution comprising an electron donor, methylviologen, and certain metalloporphyrins and metallochlorins, and irradiating said aqueous solution with light in the presence of a catalyst. In the photosynthesis process, solar energy is collected and stored in the form of a hydrogen. Ligands attached above and below the metalloporphyrin and metallochlorin plane are capable of sterically blocking photochemically inactive electrostatically bound ..pi..-..pi.. complexes which can develop.

  3. Method for improving product yields in an anionic metalloporphyrin-based artificial photosynthesis system

    DOEpatents

    Shelnutt, John A.

    1986-01-01

    A method for improving product yields in an anionic metalloporphyrin-based artificial photosynthesis system for hydrogen generation which comprises forming an aqueous solution comprising an electron donor, methylviologen, and certain metalloporphyrins and metallochlorins, and irradiating said aqueous solution with light in the presence of a catalyst. In the photosynthesis process, solar energy is collected and stored in the form of a gas hydrogen. Ligands attached above and below the metalloporphyrin and metallochlorin plane are capable of sterically blocking photochemically inactive electrostatically bound .pi.--.pi. complexes which can develop.

  4. CHO-S antibody titers >1 gram/liter using flow electroporation-mediated transient gene expression followed by rapid migration to high-yield stable cell lines.

    PubMed

    Steger, Krista; Brady, James; Wang, Weili; Duskin, Meg; Donato, Karen; Peshwa, Madhusudan

    2015-04-01

    In recent years, researchers have turned to transient gene expression (TGE) as an alternative to CHO stable cell line generation for early-stage antibody development. Despite advances in transfection methods and culture optimization, the majority of CHO-based TGE systems produce insufficient antibody titers for extensive use within biotherapeutic development pipelines. Flow electroporation using the MaxCyte STX Scalable Transfection System is a highly efficient, scalable means of CHO-based TGE for gram-level production of antibodies without the need for specialized expression vectors or genetically engineered CHO cell lines. CHO cell flow electroporation is easily scaled from milligram to multigram quantities without protocol reoptimization while maintaining transfection performance and antibody productivity. In this article, data are presented that demonstrate the reproducibility, scalability, and antibody production capabilities of CHO-based TGE using the MaxCyte STX. Data show optimization of posttransfection parameters such as cell density, media composition, and feed strategy that result in secreted antibody titers >1 g/L and production of multiple grams of antibody within 2 weeks of a single CHO-S cell transfection. In addition, data are presented to demonstrate the application of scalable electroporation for the rapid generation of high-yield stable CHO cell lines to bridge the gap between early- and late-stage antibody development activities.

  5. Expression of soluble and active interferon consensus in SUMO fusion expression system in E. coli.

    PubMed

    Peciak, Karolina; Tommasi, Rita; Choi, Ji-won; Brocchini, Steve; Laurine, Emmanuelle

    2014-07-01

    Protein production can be improved if methods for soluble protein expression are developed. Interferon consensus (IFN-con) is used to treat hepatitis C. IFN-con has superior activity compared to other clinically used interferon α subtypes. However IFN-con is a challenging protein to produce in a soluble form using an Escherichia coli expression system. Here we describe the expression of soluble and active recombinant IFN-con in E. coli. The IFN-con gene sequence was optimised for expression in E. coli, which was then cloned into the Champion™ pET SUMO expression vector downstream of the SUMO fusion protein and under strong T7lac promoter. The SUMO-IFN-con fusion protein was efficiently expressed using the SHuffle™ E. coli strain and existed in soluble form as 86-88% of the total IFN-con. After removal of the SUMO fusion partner, approximately 50mg of recombinant IFN-con of at least 98% purity (by RP-HPLC) was obtained from a 1L fermentation culture. Using an A549/EMCV antiviral assay, the specific activity of the recombinant IFN-con was determined to be 960×10(6) IU/mg as calculated to NIBSC standard for IFN-con (3×10(5)pfu/mL virus titre). Comparison of the antiviral activity of the produced IFN-con to IFN α-2a showed that IFN-con displays 2.8 times greater activity, which is in good agreement with what has been reported in the literature for pure protein. IFN-con expression in a soluble form from E. coli allowed us to use a simple, two-step purification process to yield highly pure and active IFN-con which is more efficient than obtaining IFN-con from inclusion bodies.

  6. Ectopic expression of a hot pepper bZIP-like transcription factor in potato enhances drought tolerance without decreasing tuber yield.

    PubMed

    Moon, Seok-Jun; Han, Se-Youn; Kim, Dool-Yi; Yoon, In Sun; Shin, Dongjin; Byun, Myung-Ok; Kwon, Hawk-Bin; Kim, Beom-Gi

    2015-11-01

    Over-expression of group A bZIP transcription factor genes in plants improves abiotic stress tolerance but usually reduces yields. Thus, there have been several efforts to overcome yield penalty in transgenic plants. In this study, we characterized that expression of the hot pepper (Capsicum annuum) gene CaBZ1, which encodes a group S bZIP transcription factor, was induced by salt and osmotic stress as well as abscisic acid (ABA). Transgenic potato (Solanum tuberosum) plants over-expressing CaBZ1 exhibited reduced rates of water loss and faster stomatal closure than non transgenic potato plants under drought and ABA treatment conditions. CaBZ1 over-expression in transgenic potato increased the expression of ABA- and stress-related genes (such as CYP707A1, CBF and NAC-like genes) and improved drought stress tolerance. Interestingly, over-expression of CaBZ1 in potato did not produce undesirable growth phenotypes in major agricultural traits such as plant height, leaf size and tuber formation under normal growth conditions. The transgenic potato plants also had higher tuber yields than non transgenic potato plants under drought stress conditions. Thus, CaBZ1 may be useful for improving drought tolerance in tuber crops. This might be the first report of the production of transgenic potato with improved tuber yields under drought conditions.

  7. Recombinant Expression of Trichoderma reesei Cel61A in Pichia pastoris: Optimizing Yield and N-terminal Processing.

    PubMed

    Tanghe, Magali; Danneels, Barbara; Camattari, Andrea; Glieder, Anton; Vandenberghe, Isabel; Devreese, Bart; Stals, Ingeborg; Desmet, Tom

    2015-12-01

    The auxiliary activity family 9 (AA9, formerly GH61) harbors a recently discovered group of oxidative enzymes that boost cellulose degradation. Indeed, these lytic polysaccharide monooxygenases (LPMOs) are able to disrupt the crystalline structure of cellulose, thereby facilitating the work of hydrolytic enzymes involved in biomass degradation. Since these enzymes require an N-terminal histidine residue for activity, their recombinant production as secreted protein is not straightforward. We here report the expression optimization of Trichoderma reesei Cel61A (TrCel61A) in the host Pichia pastoris. The use of the native TrCel61A secretion signal instead of the alpha-mating factor from Saccharomyces cerevisiae was found to be crucial, not only to obtain high protein yields (>400 mg/L during fermentation) but also to enable the correct processing of the N-terminus. Furthermore, the LPMO activity of the enzyme is demonstrated here for the first time, based on its degradation profile of a cellulosic substrate.

  8. High-Yield Expression in E. coli and Refolding of the bZIP Domain of Activating Transcription Factor 5

    PubMed Central

    Ciaccio, Natalie A.; Moreno, Matthew L.; Bauer, Rachel L.; Laurence, Jennifer S.

    2008-01-01

    Activating Transcription Factor 5 (ATF5) recently has been demonstrated to play a critical role in promoting the survival of human glioblastoma cells. Interference with the function of ATF5 in an in vivo rat model caused glioma cell death in primary tumors but did not affect the status of normal cells surrounding the tumor, suggesting ATF5 may prove an ideal target for anti-cancer therapy. In order to examine ATF5 as a pharmaceutical target, the protein must be produced and purified to sufficient quantity to begin analyses. Here, a procedure for expressing and refolding the bZIP domain of ATF5 in sufficient yield and final concentration to permit assay development and structural characterization of this target using solution NMR is reported. Two-dimensional NMR and circular dichrosim analyses indicate the protein exists in the partially α-helical, monomeric x-form conformation with only a small fraction of ATF5 participating in formation of higher-order structure, presumably coiled-coil homodimerization. Despite the persistence of monomers in solution even at high concentration, an electrophoretic mobility shift assay showed that ATF5 is able to bind to the cAMP response element (CRE) DNA motif. Polyacrylamide gel electrophoresis and mass spectrometry were used to confirm that ATF5 can participate in homodimer formation and that this dimerization is mediated by disulfide bond formation. PMID:18718539

  9. Genome-Wide Identification of Differentially Expressed Genes Associated with the High Yielding of Oleoresin in Secondary Xylem of Masson Pine (Pinus massoniana Lamb) by Transcriptomic Analysis

    PubMed Central

    Liu, Qinghua; Zhou, Zhichun; Wei, Yongcheng; Shen, Danyu; Feng, Zhongping; Hong, Shanping

    2015-01-01

    Masson pine is an important timber and resource for oleoresin in South China. Increasing yield of oleoresin in stems can raise economic benefits and enhance the resistance to bark beetles. However, the genetic mechanisms for regulating the yield of oleoresin were still unknown. Here, high-throughput sequencing technology was used to investigate the transcriptome and compare the gene expression profiles of high and low oleoresin-yielding genotypes. A total of 40,690,540 reads were obtained and assembled into 137,499 transcripts from the secondary xylem tissues. We identified 84,842 candidate unigenes based on sequence annotation using various databases and 96 unigenes were candidates for terpenoid backbone biosynthesis in pine. By comparing the expression profiles of high and low oleoresin-yielding genotypes, 649 differentially expressed genes (DEGs) were identified. GO enrichment analysis of DEGs revealed that multiple pathways were related to high yield of oleoresin. Nine candidate genes were validated by QPCR analysis. Among them, the candidate genes encoding geranylgeranyl diphosphate synthase (GGPS) and (-)-alpha/beta-pinene synthase were up-regulated in the high oleoresin-yielding genotype, while tricyclene synthase revealed lower expression level, which was in good agreement with the GC/MS result. In addition, DEG encoding ABC transporters, pathogenesis-related proteins (PR5 and PR9), phosphomethylpyrimidine synthase, non-specific lipid-transfer protein-like protein and ethylene responsive transcription factors (ERFs) were also confirmed to be critical for the biosynthesis of oleoresin. The next-generation sequencing strategy used in this study has proven to be a powerful means for analyzing transcriptome variation related to the yield of oleoresin in masson pine. The candidate genes encoding GGPS, (-)-alpha/beta-pinene, tricyclene synthase, ABC transporters, non-specific lipid-transfer protein-like protein, phosphomethylpyrimidine synthase, ERFs and pathogen

  10. Validation of a land data assimilation system using river discharge and agricultural yield observations

    NASA Astrophysics Data System (ADS)

    Calvet, Jean-Christophe; Barbu, Alina; Fairbairn, David; Gelati, Emiliano

    2015-04-01

    Meteo-France develops the ISBA-A-gs generic Land Surface Model (LSM) able to represent the diurnal cycle of the surface fluxes together with the seasonal, interannual and decadal variability of the vegetation biomass. The LSM is embedded in the SURFEX modeling platform together with a simplified extended Kalman filter. These tools form a Land Data Assimilation System (LDAS). The current version of the LDAS assimilates SPOT-VGT LAI and ASCAT surface soil moisture (SSM) products over France (8km x 8km), and a passive monitoring of albedo, FAPAR and Land Surface temperature (LST) is performed (i.e., the simulated values are compared with the satellite products). The vegetation biomass is analysed together with the root-zone soil moisture. The LDAS was coupled to the MODCOU hydrological model, and this allowed the use of in situ river discharge observations for the validation of the whole system. Moreover, open-loop (i.e. without integrationg satellite observations into the model) simulations of the above-ground biomass of straw cereals were compared with the analyzed values (i.e. after integration of satellite observations into the model), and with agricultural yield observations. It is shown that the assimilation of satellite observations sharply enhances the overall correlation of the simulated above-ground biomass with the agricultural yield observations.

  11. A nonlinear dynamical system approach for the yielding behaviour of a viscoplastic material.

    PubMed

    Burghelea, Teodor; Moyers-Gonzalez, Miguel; Sainudiin, Raazesh

    2017-02-15

    A nonlinear dynamical system model that approximates a microscopic Gibbs field model for the yielding of a viscoplastic material subjected to varying external stresses recently reported in R. Sainudiin, M. Moyers-Gonzalez and T. Burghelea, Soft Matter, 2015, 11(27), 5531-5545 is presented. The predictions of the model are in fair agreement with microscopic simulations and are in very good agreement with the micro-structural semi-empirical model reported in A. M. V. Putz and T. I. Burghelea, Rheol. Acta, 2009, 48, 673-689. With only two internal parameters, the nonlinear dynamical system model captures several key features of the solid-fluid transition observed in experiments: the effect of the interactions between microscopic constituents on the yield point, the abruptness of solid-fluid transition and the emergence of a hysteresis of the micro-structural states upon increasing/decreasing external forces. The scaling behaviour of the magnitude of the hysteresis with the degree of the steadiness of the flow is consistent with previous experimental observations. Finally, the practical usefulness of the approach is demonstrated by fitting a rheological data set measured with an elasto-viscoplastic material.

  12. Antigenic assessment of a recombinant human CD90 protein expressed in prokaryotic expression system.

    PubMed

    Yousefi-Rad, Narges; Shokrgozar, Mohammad Ali; Behdani, Mahdi; Moradi-Kalbolandi, Shima; Motamedi-Rad, Mahdieh; Habibi-Anbouhi, Mahdi

    2015-12-01

    Cluster of Differentiation 90 (CD90, Thy-1) has been proposed as one of the most important biomarkers in several cancer cells including cancer stem cells (CSCs). CD90 is considered as a potential normal stem cell and CSCs biomarker and also has been identified in lung cancer stem cells, hepatocellular carcinoma cells and high-grade gliomas. Using eukaryotic host systems involves complex procedures and frequently results in low protein yields. The expression of recombinant proteins in Escherichia coli is comparatively easier than eukaryotic host cells. The potential of large scale production of recombinant protein has made this system an economic production platform. In this study we expressed the extra-membrane domain of human CD90 (exCD90) antigen (Gln15-Cys130) in E. coli expression host cells. The epitope integrity of purified recombinant antigen was confirmed by antibody-antigen interaction using 5E10 anti-CD90 monoclonal antibody and binding study through ELISA and florescent staining of CD90(+) cells in a flow cytometry experiment. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. An efficient protocol towards site-specifically clickable nanobodies in high yield: cytoplasmic expression in Escherichia coli combined with intein-mediated protein ligation.

    PubMed

    Ta, Duy Tien; Redeker, Erik Steen; Billen, Brecht; Reekmans, Gunter; Sikulu, Josephine; Noben, Jean-Paul; Guedens, Wanda; Adriaensens, Peter

    2015-10-01

    In this study, several expression strategies were investigated in order to develop a generic, highly productive and efficient protocol to produce nanobodies modified with a clickable alkyne function at their C-terminus via the intein-mediated protein ligation (IPL) technique. Hereto, the nanobody targeting the vascular cell adhesion molecule 1 (NbVCAM1) was used as a workhorse. The highlights of the protocol can be ascribed to a cytoplasmic expression of the nanobody-intein-chitin-binding domain fusion protein in the Escherichia coli SHuffle(®) T7 cells with a C-terminal extension, i.e. LEY, EFLEY or His6 spacer peptide, in the commonly used Luria-Bertani medium. The combination of these factors led to a high yield (up to 22 mg/l of culture) and nearly complete alkynation efficiency of the C-terminally modified nanobody via IPL. This yield can even be improved to ∼45 mg/l in the EnPresso(®) growth system but this method is more expensive and time-consuming. The resulting alkynated nanobodies retained excellent binding capacity towards the recombinant human VCAM1. The presented protocol benefits from time- and cost-effectiveness, which allows a feasible production up-scaling of generic alkynated nanobodies. The production of high quantities of site-specifically modified nanobodies paves the way to new biosurface applications that demand for a homogeneously oriented nanobody coupling. Prospectively, the alkynated nanobodies can be covalently coupled to a multitude of azide-containing counterparts, e.g. contrast labeling agents, particles or surfaces for numerous innovative applications.

  14. Effect of zinc application on rice yield under wheat rice system.

    PubMed

    Khan, Rahmatullah; Gurmani, Ali Raza; Khan, Muhammad Sohail; Gurmani, Akber Hussain

    2007-01-15

    A field experiment was conducted during 2004-05 on wheat and rice to study the response of Zinc application in wheat-rice system. Two levels of zinc 5 and 10 kg ha(-1) with control were studied with the basal dose of N, P2O5, K2O as 120-90-60 kg ha(-1) in the form of urea, TSP, SOP and zinc sulphate during both the crops. Wheat variety Naseer 2000 and rice variety IRRI 6, both were planted in R.C.B design with three replications. Zn application, significantly affected wheat grain yield, ranged from 2.7 to 3.5 t ha(-1), giving highest increase of 31.6% over control from 5 kg Zn ha(-1). The number of tillers, spike m(-2), spike length, plant height and 1000 grain weight of wheat were also significantly affected over control with the same treatment. Paddy yield was also significantly affected by Zn levels ranged from 3.9 to 5.9 t ha(-1). The highest yield was obtained from 10 kg Zn ha(-1) each applied to both crops. Similarly Zn application also affected significantly to the yield parameters of rice like the number of spike m(-2), number of spike/plant, spike length, plant height and 1000 grain weight over control from the above said treatment of 10 kg Zn ha(-1). The concentration of zinc in soil and leaves was significantly affected by the application of zinc in wheat and rice, ranged from 0.47-1.37, 22.6-367.37 mg kg(-1) in wheat and 0.45-1.18; 29.32-40.67 mg kg(-1) in rice, respectively (soil and leaves). The highest concentration in soil and leaves was recorded by the cumulative application of 10 kg Zn ha(-1) while lowest from control. The direct application of 5 and 10 kg Zn ha(-1) gave an increase of 39 and 45% while residual effect 30.0 and 43.0%, cumulative effect of 38 and 50% over control, respectively. The residual application of 10 kg Zn ha(-1) can be recommended for economical production in wheat rice system.

  15. High-yield expression in Escherichia coli and purification of mouse ubiquitin-activating enzyme E1.

    PubMed

    Carvalho, Andreia F; Pinto, Manuel P; Grou, Cláudia P; Vitorino, Rui; Domingues, Pedro; Yamao, Fumiaki; Sá-Miranda, Clara; Azevedo, Jorge E

    2012-07-01

    Research in the ubiquitin field requires large amounts of ubiquitin-activating enzyme (E1) for in vitro ubiquitination assays. Typically, the mammalian enzyme is either isolated from natural sources or produced recombinantly using baculovirus/insect cell protein expression systems. Escherichia coli is seldom used to produce mammalian E1 probably due to the instability and insolubility of this high-molecular mass protein. In this report, we show that 5-10 mg of histidine-tagged mouse E1 can be easily obtained from a 1 l E. coli culture. A low temperature during the protein induction step was found to be critical to obtain an active enzyme.

  16. Root-knot Nematode Management and Yield of Soybean as Affected by Winter Cover Crops, Tillage Systems, and Nematicides.

    PubMed

    Minton, N A; Parker, M B

    1987-01-01

    Management of Meloidogyne incognita on soybean as affected by winter small grain crops or fallow, two tillage systems, and nematicides was studied. Numbers of M. incognita did not differ in plots planted to wheat and rye. Yields of soybean planted after these crops also did not differ. Numbers of M. incognita were greater in fallow than in rye plots, but soybean yield was not affected by the two treatments. Soybean yields were greater in subsoil-plant than in moldboard plowed plots. Ethylene dibromide reduced nematode population densities more consistently than aldicarb and phenamiphos. Also, ethylene dibromide increased yields the most and phenamiphos the least. There was a positive correlation (P = 0.001) of seed size (weight of 100 seeds) with yield (r = 0.79), indicating that factors affecting yield also affected seed size.

  17. Recombinant production of mecasermin in E. coli expression system

    PubMed Central

    Jafari, S.; Babaeipour, V.; Seyedi, H.A. Eslampanah; Rahaie, M.; Mofid, M.R.; Haddad, L.; Namvaran, M.M.; Fallah, J.

    2014-01-01

    Human Insulin-like growth factor 1 (hIGF-1) consists of 70 amino acids in a single chain with three intermolecular disulfide bridges possessing valuable therapeutic effects. To date, numerous variants of specifically engineered hIGF-1 have been produced so as to improve hIGF-1 biological activity, stability and stronger binding to IGF-1 receptor. Mecasermin is one of the modified variants with one amino acid substitution near the N-terminal (T4I) approved for the treatment of growth failure diabetes, wound healing, amyotrophic lateral sclerosis and severe primary IGF-1 deficiency. No scientific report for recombinant production of mecasermin in Escherichia coli (E. coli) expression system has been sofar reported. In the present study, we therefore investigated the overexpression of mecasermin in two different E. coli strains in order to obtain higher yield of recombinant protein. To achieve this goal, mecasermin DNA encoding sequence was designed based on polypeptide sequence, optimized according to E. coli codon preference, and cloned in pET15b. Recombinant vector, pET15-mecasermin, transferred into two E. coli strains rigami B (DE3) and BL21 (DE3) and induced for expression in a small scale. Results revealed the E. coli Origami B (DE3) expression system was a preferable host for mecasermin production due to its high expression level being around twice as much as BL21 (DE3). Large scale mecasermin production was performed in batch culture and produced recombinant protein specifically confirmed by western blotting and mass spectroscopy. Since major part of recombinant mecasermin was expressed as inclusion body, isolation and refolding was accomplished through developed purification procedure, and finally recombinant protein was successfully purified by gel filtration chromatography. PMID:26339260

  18. A meta-analysis of maize and wheat yields in low-input vs. conventional and organic systems

    USDA-ARS?s Scientific Manuscript database

    Organic and low-input systems are proposed as ways to reduce the environmental impacts of agriculture. Previous studies have shown that yields of organic systems can be ~19-25% lower than conventional systems. An intermediary, low-input system could be less damaging for the environment than conventi...

  19. ROI on yield data analysis systems through a business process management strategy

    NASA Astrophysics Data System (ADS)

    Rehani, Manu; Strader, Nathan; Hanson, Jeff

    2005-05-01

    The overriding motivation for yield engineering is profitability. This is achieved through application of yield management. The first application is to continually reduce waste in the form of yield loss. New products, new technologies and the dynamic state of the process and equipment keep introducing new ways to cause yield loss. In response, the yield management efforts have to continually come up with new solutions to minimize it. The second application of yield engineering is to aid in accurate product pricing. This is achieved through predicting future results of the yield engineering effort. The more accurate the yield prediction, the more accurate the wafer start volume, the more accurate the wafer pricing. Another aspect of yield prediction pertains to gauging the impact of a yield problem and predicting how long that will last. The ability to predict such impacts again feeds into wafer start calculations and wafer pricing. The question then is that if the stakes on yield management are so high why is it that most yield management efforts are run like science and engineering projects and less like manufacturing? In the eighties manufacturing put the theory of constraints1 into practice and put a premium on stability and predictability in manufacturing activities, why can't the same be done for yield management activities? This line of introspection led us to define and implement a business process to manage the yield engineering activities. We analyzed the best known methods (BKM) and deployed a workflow tool to make them the standard operating procedure (SOP) for yield managment. We present a case study in deploying a Business Process Management solution for Semiconductor Yield Engineering in a high-mix ASIC environment. We will present a description of the situation prior to deployment, a window into the development process and a valuation of the benefits.

  20. Increasing Hematopoietic Stem Cell Yield to Develop Mice with Human Immune Systems

    PubMed Central

    Biancotti, Juan-Carlos; Town, Terrence

    2013-01-01

    Hematopoietic stem cells (HSCs) are unique in their capacity to give rise to all mature cells of the immune system. For years, HSC transplantation has been used for treatment of genetic and neoplastic diseases of the hematopoietic and immune systems. The sourcing of HSCs from human umbilical cord blood has salient advantages over isolation from mobilized peripheral blood. However, poor sample yield has prompted development of methodologies to expand HSCs ex vivo. Cytokines, trophic factors, and small molecules have been variously used to promote survival and proliferation of HSCs in culture, whilst strategies to lower the concentration of inhibitors in the culture media have recently been applied to promote HSC expansion. In this paper, we outline strategies to expand HSCs in vitro, and to improve engraftment and reconstitution of human immune systems in immunocompromised mice. To the extent that these “humanized” mice are representative of the endogenous human immune system, they will be invaluable tools for both basic science and translational medicine. PMID:23509770

  1. Improvement in the yield and quality of kalmegh (Andrographis paniculata Nees) under the sustainable production system.

    PubMed

    Verma, Rajesh Kumar; Verma, Sanjeet K; Pankaj, Umesh; Gupta, Anand K; Khan, Khushboo; Shankar, Karuna

    2015-02-01

    Andrographis paniculata Nees is an annual erect herb with wide medicinal and pharmacological applications due to the presence of andrographolide and other active chemical constituents. The large-scale cultivation of the kalmegh is not in practice. The aim of this study was to establish sustainable production systems of A. paniculata cv CIM-Megha with the application of different bioinoculants and chemical fertilisers. A. paniculata herb and andrographolide yield in the dried leaves was found to be highest (218% and 61.3%, respectively) in treatment T3 (NPK+Bacillus sp.) compared with T1 (control). The soil organic carbon, soil microbial respiration, soil enzymes activity and available nutrients improved significantly with combined application of bioinoculants and chemical fertilisers.

  2. Engineering Camelina sativa (L.) Crantz for Enhanced Oil and Seed Yields by Combining Diacylglycerol Acyltransferase1 and Glycerol-3-Phosphate Dehydrogenase Expression.

    PubMed

    Chhikara, Sudesh; Abdullah, Hesham M; Akbari, Parisa; Schnell, Danny; Dhankher, Om Parkash

    2017-10-04

    Plant seed oils based liquid transportation fuels (i.e., biodiesel and green diesel) have tremendous potential as environmentally, economically and technologically feasible alternatives to petroleum-derived fuels. Due to their nutritional and industrial importance, one of the major objectives is to increase the seed yield and oil production of oilseed crops via biotechnological approaches. Camelina sativa, an emerging oilseed crop, has been proposed as an ideal crop for biodiesel and bioproducts applications. Further increase in seed oil yield by increasing the flux of carbon from increased photosynthesis into triacylglycerol (TAG) synthesis will make this crop more profitable. To increase the oil yield, we engineered Camelina by co-expressing the Arabidopsis thaliana (L.) Heynh. diacylglycerol acyltransferase1 (DGAT1) and a yeast cytosolic glycerol-3-phosphate dehydrogenase (GPD1) genes under the control of seed-specific promoters. Plants co-expressing DGAT1 and GPD1 exhibited up to 13% higher seed oil content and up to 52% increase in seed mass compared to wild type plants. Further, DGAT1 and GDP1 co-expressing lines showed significantly higher seed and oil yields on a dry-weight basis than the wild type controls or plants expressing DGAT1 and GPD1 alone. The oil harvest index (g oil per g total dry matter) for DGTA1 and GPD1 co-expressing lines was almost two-folds higher as compared to wild type and the lines expressing DGAT1 and GPD1 alone. Therefore, combining the overexpression of TAG biosynthetic genes, DGAT1 and GPD1, appears to be a positive strategy to achieve a synergistic effect on the flux through the TAG synthesis pathway, and thereby further increase the oil yield. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  3. Baculovirus expression system and method for high throughput expression of genetic material

    SciTech Connect

    Clark, Robin; Davies, Anthony

    2001-01-01

    The present invention provides novel recombinant baculovirus expression systems for expressing foreign genetic material in a host cell. Such expression systems are readily adapted to an automated method for expression foreign genetic material in a high throughput manner. In other aspects, the present invention features a novel automated method for determining the function of foreign genetic material by transfecting the same into a host by way of the recombinant baculovirus expression systems according to the present invention.

  4. Developing a Hybrid Solar/Wind Powered Drip Irrigation System for Dragon Fruit Yield

    NASA Astrophysics Data System (ADS)

    Widiastuti, I.; Wijayanto, D. S.

    2017-03-01

    Irrigation operations take a large amount of water and energy which impact to total costs of crop production. Development of an efficient irrigation supplying precise amount of water and conserving the use of energy can have benefits not only by reducing the operating costs but also by enhancing the farmland productivity. This article presents an irrigation method that promotes sustainable use of water and energy appropriate for a developing tropical country. It proposes a drip irrigation system supported by a combined solar-wind electric power generation system for efficient use of water in dragon fruit cultivation. The electric power generated is used to drive a water pump filling a storage tank for irrigating a 3000 m2 dragon fruit yield in Nguntoronadi, Wonogiri, Indonesia. In designing the irrigation system, the plant’s water requirement was identified based on the value of reference evapotranspiration of the area. A cost/benefit analysis was performed to evaluate the economic feasibility of the proposed scheme. The installation of this solar and wind drip irrigation helps provide sufficient quantity of water to each plant using renewable energy sources which reduce dependence on fossil fuel.

  5. The TRANSFAC system on gene expression regulation.

    PubMed

    Wingender, E; Chen, X; Fricke, E; Geffers, R; Hehl, R; Liebich, I; Krull, M; Matys, V; Michael, H; Ohnhäuser, R; Prüss, M; Schacherer, F; Thiele, S; Urbach, S

    2001-01-01

    The TRANSFAC database on transcription factors and their DNA-binding sites and profiles (http://www.gene-regulation.de/) has been quantitatively extended and supplemented by a number of modules. These modules give information about pathologically relevant mutations in regulatory regions and transcription factor genes (PathoDB), scaffold/matrix attached regions (S/MARt DB), signal transduction (TRANSPATH) and gene expression sources (CYTOMER). Altogether, these distinct database modules constitute the TRANSFAC system. They are accompanied by a number of program routines for identifying potential transcription factor binding sites or for localizing individual components in the regulatory network of a cell.

  6. Silkworm expression system as a platform technology in life science.

    PubMed

    Kato, Tatsuya; Kajikawa, Mizuho; Maenaka, Katsumi; Park, Enoch Y

    2010-01-01

    Many recombinant proteins have been successfully produced in silkworm larvae or pupae and used for academic and industrial purposes. Several recombinant proteins produced by silkworms have already been commercialized. However, construction of a recombinant baculovirus containing a gene of interest requires tedious and troublesome steps and takes a long time (3-6 months). The recent development of a bacmid, Escherichia coli and Bombyx mori shuttle vector, has eliminated the conventional tedious procedures required to identify and isolate recombinant viruses. Several technical improvements, including a cysteine protease or chitinase deletion bacmid and chaperone-assisted expression and coexpression, have led to significantly increased protein yields and reduced costs for large-scale production. Terminal N-acetyl glucosamine and galactose residues were found in the N-glycan structures produced by silkworms, which are different from those generated by insect cells. Genomic elucidation of silkworm has opened a new chapter in utilization of silkworm. Transgenic silkworm technology provides a stable production of recombinant protein. Baculovirus surface display expression is one of the low-cost approaches toward silkworm larvae-derived recombinant subunit vaccines. The expression of pharmaceutically relevant proteins, including cell/viral surface proteins, membrane proteins, and guanine nucleotide-binding protein (G protein) coupled receptors, using silkworm larvae or cocoons has become very attractive. Silkworm biotechnology is an innovative and easy approach to achieve high protein expression levels and is a very promising platform technology in the field of life science. Like the "Silkroad," we expect that the "Bioroad" from Asia to Europe will be established by the silkworm expression system.

  7. A high loading overland flow system: Impacts on soil characteristics, grass constituents, yields and nutrient removal.

    PubMed

    Wen, C G; Chen, T H; Hsu, F H; Lu, C H; Lin, J B; Chang, C H; Chang, S P; Lee, C S

    2007-04-01

    The objectives of this paper are to determine effects of different grass species and their harvests on pollutant removal, elucidate impacts on soil characteristics and grass constituents, observe grass yield and quantify nutrient uptake by vegetation in an overland flow system (OLFS). Polluted creek water was applied to eight channels in the OLFS, which were planted with Paragrass, Nilegrass, Cattail, and Vetiver, with each two channels being randomly planted with a given grass species. The grass in one channel was harvested while that in the other channel was not. At a high rate of 27.8 m d(-1) hydraulic loading, the removal efficiencies of conventional pollutants such as BOD, COD, suspended solids (SS), and total coliforms in wastewater are not affected by the type of the grasses species, but those of nitrogen and phosphorus are affected by different species. Overall average removal efficiencies of BOD, COD, SS, ammonia, total nitrogen, total phosphorus and total coliforms through the OLFS are 42%, 48%, 78%, 47%, 40%, 33% and 89%, respectively. The concentration of nitrate, however, increases due to nitrification. Soil characteristics in OLFS have been changed significantly; specific conductivity, organic matter, exchangeable magnesium, extractable copper and zinc in soils all increase with time while pHs decrease. During the winter season, there is a significant accumulation of nitrate in grass with the subsequent reduction during the active growing season (Spring). The contents of nitrate and phosphorus in grass tissue are higher than those of grass in general pastureland, probably due to nutrient luxury uptake by grass. The overall grass yield, growth rate and nutrient uptake are quantified and implication of such high rate OLFS discussed.

  8. Subplots facilitate assessment of corn yield losses from weed competition in a long-term systems experiment

    USDA-ARS?s Scientific Manuscript database

    Weeds can potentially limit crop yield, particularly in organic systems where herbicide technologies are unavailable. Weedy and weed-free subplots were established within full plots of a long-term cropping systems experiment, the Farming Systems Project, at Beltsville, Maryland, USA, to determine t...

  9. Thick target neutron yield from 145 MeV 19F+27Al system

    NASA Astrophysics Data System (ADS)

    Sunil, C.; Bandyopadhyay, T.; Nandy, M.; Suman, Vitisha; Paul, S.; Nanal, V.; Pillay, R. G.; Sarkar, P. K.

    2013-09-01

    The double differential neutron energy distribution has been measured for the 19F+27Al system at 145 MeV projectile energy. The time of flight technique was used to measure the energy while pulse shape discrimination has been used to separate the neutrons from photons. The results are compared with the statistical nuclear reaction model codes PACE and EMPIRE. The PACE code appears to predict the slope and the end point energy of the experimental spectra fairly well but over predicts the values. The slope obtained from the EMPIRE calculations appears to be harder while the values being closer to the experimental results. The yield from the Hauser-Feshbach based compound nucleus model calculations agree reasonably well with the experimental results at the backward angles but not in the forward directions. The energy integrated angular distribution from 145 MeV projectiles show an enhanced emission in the forward angles compared to the similar results from 110 MeV projectiles. This analysis suggests some contribution from the pre-equilibrium emissions from the system at the higher projectile energy.

  10. Efficient expression systems for cysteine proteases of malaria parasites: too good to be true?

    PubMed

    Sarduy, Emir Salas; Chávez Planes, María de los A

    2013-01-01

    Papain-like cysteine proteases of malaria parasites are considered important chemotherapeutic targets or valuable models for the evaluation of drug candidates. Consequently, many of these enzymes have been cloned and expressed in Escherichia coli for their biochemical characterization. However, their expression has been problematic, showing low yield and leading to the formation of insoluble aggregates. Given that highly-productive expression systems are required for the high-throughput evaluation of inhibitors, we analyzed the existing expression systems to identify the causes of such apparent issues. We found that significant divergences in codon and nucleotide composition from host genes are the most probable cause of expression failure, and propose several strategies to overcome these limitations. Finally we predict that yeast hosts Saccharomyces cerevisiae and Pichia pastoris may be better suited than E. coli for the efficient expression of plasmodial genes, presumably leading to soluble and active products reproducing structural and functional characteristics of the natural enzymes.

  11. Straw return reduces yield-scaled N2O plus NO emissions from annual winter wheat-based cropping systems in the North China Plain.

    PubMed

    Yao, Zhisheng; Yan, Guangxuan; Zheng, Xunhua; Wang, Rui; Liu, Chunyan; Butterbach-Bahl, Klaus

    2017-07-15

    Straw return in combination with synthetic N fertilizer is considered to be beneficial to soil fertility and crop yield. Such practice, however, can considerably modify soil microbial activity and relative C and N availability, both of which are known to regulate soil nitrous oxide (N2O) and nitric oxide (NO) emissions. Minimizing these emissions per unit of crop yield is a prerequisite to minimize the environmental footprint of agricultural production and thus, a policy objective. In our study, we quantified N2O and NO emissions and determined fertilizer-N use efficiencies (NUE) and crop yields of two double-cropping (summer maize/Welsh onion-winter wheat) systems with and without straw incorporation in the North China Plain. Relative to the fertilized treatment without straw amendments, straw incorporation showed a significant inhibitory effect on annual N2O emissions from the maize-wheat system (-31%), but no significant effect was observed for the Welsh onion-wheat system. However, straw return significantly reduced annual NO emissions by >30% for both systems. Meanwhile, straw return in both systems significantly increased the NUE and crop yields by 34-47% and 7-16%, respectively, as compared to the treatment without straw additions. Across the double-cropping systems, annual direct emission factors of N2O, NO and N2O+NO were 0.37-0.57%, 0.08-0.78% and 0.57-1.36%, respectively. Furthermore, a negative relationship between direct emission factors of N2O+NO and crop NUE was observed, highlighting the importance of optimizing NUE for reducing environmental risks of a cropping system. When expressing emissions on a yield basis, straw return significantly reduced annual yield-scaled N2O+NO emissions by 15-42% for both systems. Overall, our results show that the combined application of crop straw and synthetic N fertilizer is a promising N management strategy for maximizing crop yields while mitigating N-trace gas emissions. Copyright © 2017 Elsevier B.V. All

  12. Crop yield and light / energy efficiency in a closed ecological system: two laboratory biosphere experiments

    NASA Astrophysics Data System (ADS)

    Nelson, M.; Dempster, W. F.; Silverstone, S.; Alling, A.; Allen, J. P.; van Thillo, M.

    field results but somewhat below greenhouse trials at comparable light levels, and the best portion of the crop at 0.22g/mole was inbetween those values. Sweet potato production was overall close to 50% higher than trials using hydroponic methods with TU-82-155 at NASA JSC. Compared to projected yields for the Mars on Earth life support system, these wheat yields were about 15% higher, and the sweet potato yields averaged over 80% higher

  13. Genotype by feeding system interaction in the genetic evaluation of Jersey cattle for milk yield.

    PubMed

    Ramírez-Valverde, R; Peralta-Aban, J A; Núñez-Domínguez, R; Ruíz-Flores, A; García-Muñiz, J G; García-Peniche, T B

    2010-12-01

    Results of studies in dairy cattle about the magnitude of the genotype-environment interaction (GEI) are variable, depending on the definitions of genotype and environment. Therefore, the objective of this study was to determine the magnitude of the interaction of genotype and feeding system (confinement and grazing) in the Mexican genetic evaluation of Jersey cattle for milk yield. The number of lactations and animals in the pedigree used were 5122 and 18 432. An animal model and the MTDFREML program were used to estimate genetic parameters and predict genetic values of the animals. Bivariate analysis was carried out considering the performance of confined and grazing cows as two different traits. Three indicator variables were used to assess GEI: (i) magnitude of the genetic correlation coefficients, (ii) correlation between predicted breeding values and (iii) frequency of coincidence in the ranking of top sires. The magnitude of GEI depended on the choice of the indicator variable. The estimate of genetic correlation coefficient less than unity (0.76; P < 0.05) suggested the presence of biologically important GEI. The differences in phenotypic averages and variances between confinement and grazing systems seem to be the main causes for the genotype by environment interaction detected. However, the correlation coefficient between breeding values from confined and grazing animals (0.96) and the frequency of coincidence between breeding values of common sires within the top 100 in confinement and grazing (0.86) indicated low-to-moderate re-ranking of animals or top sires. In addition, the high correlations between predicted breeding values of Mexican genetic evaluation and the two environments (0.99 and 0.93 for confinement and grazing) indicated that for the two feeding systems, breeding values from national analyses could be safely used.

  14. Application of a "black body" like reactor for measurements of quantum yields of photochemical reactions in heterogeneous systems.

    PubMed

    Emeline, A V; Zhang, X; Jin, M; Murakami, T; Fujishima, A

    2006-04-13

    We report for the first time an experimental application of the concept of a "black body" like reactor to measure quantum yields (Phi) of photochemical reactions in liquid-solid heterogeneous systems. A major advantage of this new method is its simplicity since the fractions of reflected and transmitted light are negligible due to reactor geometry and high optical density of the heterogeneous systems. The average quantum yield of a test reaction (phenol photodegradation) over TiO(2) (Degussa P25) as determined by this method was 0.14, identical to the quantum yield measured earlier for this same reaction under similar conditions by Salinaro and Serpone. We also report the quantum yield of phenol photodegradation over N-doped TiO(2) during photoexcitation at the fundamental absorption band (lambda = 365 nm; Phi = 0.12) and at the N-doping induced extrinsic absorption band (lambda = 436 nm; Phi = 0.08) of the photocatalyst.

  15. Orbital express capture system: concept to reality

    NASA Astrophysics Data System (ADS)

    Stamm, Shane; Motaghedi, Pejmun

    2004-08-01

    The development of autonomous servicing of on-orbit spacecraft has been a sought after objective for many years. A critical component of on-orbit servicing involves the ability to successfully capture, institute mate, and perform electrical and fluid transfers autonomously. As part of a Small Business Innovation Research (SBIR) grant, Starsys Research Corporation (SRC) began developing such a system. Phase I of the grant started in 1999, with initial work focusing on simultaneously defining the parameters associated with successful docking while designing to those parameters. Despite the challenge of working without specific requirements, SRC completed development of a prototype design in 2000. Throughout the following year, testing was conducted on the prototype to characterize its performance. Having successfully completed work on the prototype, SRC began a Phase II SBIR effort in mid-2001. The focus of the second phase was a commercialization effort designed to augment the prototype model into a more flight-like design. The technical requirements, however, still needed clear definition for the design to progress. The advent of the Orbital Express (OE) program provided much of that definition. While still in the proposal stages of the OE program, SRC began tailoring prototype redesign efforts to the OE program requirements. A primary challenge involved striking a balance between addressing the technical requirements of OE while designing within the scope of the SBIR. Upon award of the OE contract, the Phase II SBIR design has been fully developed. This new design, designated the Mechanical Docking System (MDS), successfully incorporated many of the requirements of the OE program. SRC is now completing dynamic testing on the MDS hardware, with a parallel effort of developing a flight design for OE. As testing on the MDS progresses, the design path that was once common to both SBIR effort and the OE program begins to diverge. The MDS will complete the scope of the

  16. Soil and rainfall factors influencing yields of a dryland cropping system in Colorado

    USDA-ARS?s Scientific Manuscript database

    The semi-arid Great Plains of the United States experience a large variation in crop yields due to variability in rainfall, soil, and other factors. We analyzed crop yields (24-year period) from a no-till rotation of wheat(Triticum aestivum)-corn (Zea mays L.) or sorghum[Sorghum bicolor (L.) Moench]...

  17. Meteorological fluctuations define long-term crop yield patterns in conventional and organic production systems

    USDA-ARS?s Scientific Manuscript database

    Periodic variability in meteorological patterns presents significant challenges to crop production consistency and yield stability. Meteorological influences on corn and soybean grain yields were analyzed over an 18-year period at a long-term experiment in Beltsville, Maryland, U.S.A., comparing c...

  18. The sweet potato sporamin promoter confers high-level phytase expression and improves organic phosphorus acquisition and tuber yield of transgenic potato.

    PubMed

    Hong, Ya-Fang; Liu, Chang-Yeu; Cheng, Kuo-Joan; Hour, Ai-Ling; Chan, Min-Tsair; Tseng, Tung-Hai; Chen, Kai-Yi; Shaw, Jei-Fu; Yu, Su-May

    2008-07-01

    The sweet potato sporamin promoter was used to control the expression in transgenic potato of the E. coli appA gene, which encodes a bifunctional enzyme exhibiting both acid phosphatase and phytase activities. The sporamin promoter was highly active in leaves, stems and different size tubers of transgenic potato, with levels of phytase expression ranging from 3.8 to 7.4% of total soluble proteins. Phytase expression levels in transgenic potato tubers were stable over several cycles of propagation. Field tests showed that tuber size, number and yield increased in transgenic potato. Improved phosphorus (P) acquisition when phytate was provided as a sole P source and enhanced microtuber formation in cultured transgenic potato seedlings when phytate was provided as an additional P source were observed, which may account for the increase in leaf chloroplast accumulation (important for photosynthesis) and tuber yield of field-grown transgenic potato supplemented with organic fertilizers. Animal feeding tests indicated that the potato-produced phytase supplement was as effective as a commercially available microbial phytase in increasing the availability of phytate-P to weanling pigs. This study demonstrates that the sporamin promoter can effectively direct high-level recombinant protein expression in potato tubers. Moreover, overexpression of phytase in transgenic potato not only offers an ideal feed additive for improving phytate-P digestibility in monogastric animals but also improves tuber yield, enhances P acquisition from organic fertilizers, and has a potential for phytoremediation.

  19. A mercury-catalyzed, high-yield system for the oxidation of methane to methanol.

    PubMed

    Periana, R A; Taube, D J; Evitt, E R; Löffler, D G; Wentrcek, P R; Voss, G; Masuda, T

    1993-01-15

    A homogeneous system for the selective, catalytic oxidation of methane to methanol via methyl bisulfate is reported. The net reaction catalyzed by mercuric ions, Hg(II), is the oxidation of methane by concentrated sulfuric acid to produce methyl bisulfate, water, and sulfur dioxide. The reaction is efficient. At a methane conversion of 50 percent, 85 percent selectivity to methyl bisulfate ( approximately 43 percent yield; the major side product is carbon dioxide) was achieved at a molar productivity of 10(-7) mole per cubic centimeter per second and Hg(II) turnover frequency of 10(-3) per second. Separate hydrolysis of methyl bisulfate and reoxidation of the sulfur dioxide with air provides a potentially practical scheme for the oxidation of methane to methanol with molecular oxygen. The primary steps of the Hg(II)-catalyzed reaction were individually examined and the essential elements of the mechanism were identified. The Hg(II) ion reacts with methane by an electrophilic displacement mechanism to produce an observable species, CH(3)HgOSO(3)H, 1. Under the reaction conditions, 1 readily decomposes to CH(3)OSO(3)H and the reduced mercurous species, Hg(2)(2+) The catalytic cycle is completed by the reoxidation of Hg(2)(2+) with H(2)SO(4) to regenerate Hg(II) and byproducts SO(2) and H(2)O. Thallium(III), palladium(II), and the cations of platinum and gold also oxidize methane to methyl bisulfate in sulfuric acid.

  20. A Mercury-Catalyzed, High-Yield System for the Oxidation of Methane to Methanol

    NASA Astrophysics Data System (ADS)

    Periana, Roy A.; Taube, Douglas J.; Evitt, Eric R.; Loffler, Daniel G.; Wentrcek, Paul R.; Voss, George; Masuda, Toshihiko

    1993-01-01

    A homogeneous system for the selective, catalytic oxidation of methane to methanol via methyl bisulfate is reported. The net reaction catalyzed by mercuric ions, Hg(II), is the oxidation of methane by concentrated sulfuric acid to produce methyl bisulfate, water, and sulfur dioxide. The reaction is efficient. At a methane conversion of 50 percent, 85 percent selectivity to methyl bisulfate (~43 percent yield; the major side product is carbon dioxide) was achieved at a molar productivity of 10-7 mole per cubic centimeter per second and Hg(II) turnover frequency of 10-3 per second. Separate hydrolysis of methyl bisulfate and reoxidation of the sulfur dioxide with air provides a potentially practical scheme for the oxidation of methane to methanol with molecular oxygen. The primary steps of the Hg(II)-catalyzed reaction were individually examined and the essential elements of the mechanism were identified. The Hg(II) ion reacts with methane by an electrophilic displacement mechanism to produce an observable species, CH_3HgOSO_3H, 1. Under the reaction conditions, 1 readily decomposes to CH_3OSO_3H and the reduced mercurous species, Hg_22+. The catalytic cycle is completed by the reoxidation of Hg_22+ with H_2SO_4 to regenerate Hg(II) and byproducts SO_2 and H_2O. Thallium(III), palladium(II), and the cations of platinum and gold also oxidize methane to methyl bisulfate in sulfuric acid.

  1. Expression of the hemagglutinin HA1 subunit of the equine influenza virus using a baculovirus expression system.

    PubMed

    Sguazza, Guillermo H; Fuentealba, Nadia A; Tizzano, Marco A; Galosi, Cecilia M; Pecoraro, Marcelo R

    2013-01-01

    Equine influenza virus is a leading cause of respiratory disease in horses worldwide. Disease prevention is by vaccination with inactivated whole virus vaccines. Most current influenza vaccines are generated in embryonated hens' eggs. Virions are harvested from allantoic fluid and chemically inactivated. Although this system has served well over the years, the use of eggs as the substrate for vaccine production has several well-recognized disadvantages (cost, egg supply, waste disposal and yield in eggs). The aim of this study was to evaluate a baculovirus system as a potential method for producing recombinant equine influenza hemagglutinin to be used as a vaccine. The hemagglutinin ectodomain (HA1 subunit) was cloned and expressed using a baculovirus expression vector. The expression was determined by SDS-PAGE and immunoblotting. A high yield, 20μg/ml of viral protein, was obtained from recombinant baculovirus-infected cells. The immune response in BALB/c mice was examined following rHA1 inoculation. Preliminary results show that recombinant hemagglutinin expressed from baculovirus elicits a strong antibody response in mice; therefore it could be used as an antigen for subunit vaccines and diagnostic tests. Copyright © 2013 Asociación Argentina de Microbiología. Publicado por Elsevier España. All rights reserved.

  2. ESPRESSO: a system for estimating protein expression and solubility in protein expression systems.

    PubMed

    Hirose, Shuichi; Noguchi, Tamotsu

    2013-05-01

    Recombinant protein technology is essential for conducting protein science and using proteins as materials in pharmaceutical or industrial applications. Although obtaining soluble proteins is still a major experimental obstacle, knowledge about protein expression/solubility under standard conditions may increase the efficiency and reduce the cost of proteomics studies. In this study, we present a computational approach to estimate the probability of protein expression and solubility for two different protein expression systems: in vivo Escherichia coli and wheat germ cell-free, from only the sequence information. It implements two kinds of methods: a sequence/predicted structural property-based method that uses both the sequence and predicted structural features, and a sequence pattern-based method that utilizes the occurrence frequencies of sequence patterns. In the benchmark test, the proposed methods obtained F-scores of around 70%, and outperformed publicly available servers. Applying the proposed methods to genomic data revealed that proteins associated with translation or transcription have a strong tendency to be expressed as soluble proteins by the in vivo E. coli expression system. The sequence pattern-based method also has the potential to indicate a candidate region for modification, to increase protein solubility. All methods are available for free at the ESPRESSO server (http://mbs.cbrc.jp/ESPRESSO). © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Effect of initial biomass on channel catfish yield and water quality in a biofloc technology production system

    USDA-ARS?s Scientific Manuscript database

    Biofloc technology (BFT) production systems are being used more commonly to produce high yields of fish or shrimp because very high feed rates are possible. In an outdoor BFT production system, a complex of living organisms is closely associated with particulate organic matter and is maintained in ...

  4. Efficient Agrobacterium-based transient expression system for the production of biopharmaceuticals in plants

    PubMed Central

    Circelli, Patrizia; Donini, Marcello; Villani, Maria Elena; Benvenuto, Eugenio

    2010-01-01

    We have recently described an efficient transient expression system mediated by Agrobacterium tumefaciens for the production of HIV-1 Nef protein in Nicotiana benthamiana plants. In order to enhance the yield of recombinant protein we assayed the effect of three gene-silencing viral suppressor proteins (P25 of Potato Virus X, P19 of Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus) on Nef expression levels. Results demonstrated that AMCV-P19 gave the highest Nef yield (1.3% of total soluble protein) and that this effect was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs) indicating an effective modulation of RNA silencing mechanisms. Here we report additional data on the production of different heterologous proteins including human immunoglobulin heavy and light chains and a virus coat protein that demonstrate the robustness of this co-agroinfiltration expression system boosted by the AMCV-P19 gene-silencing suppressor. PMID:21326930

  5. An evaluation of the lamb vision system as a predictor of lamb carcass red meat yield percentage.

    PubMed

    Brady, A S; Belk, K E; LeValley, S B; Dalsted, N L; Scanga, J A; Tatum, J D; Smith, G C

    2003-06-01

    An objective method for predicting red meat yield in lamb carcasses is needed to accurately assess true carcass value. This study was performed to evaluate the ability of the lamb vision system (LVS; Research Management Systems USA, Fort Collins, CO) to predict fabrication yields of lamb carcasses. Lamb carcasses (n = 246) were evaluated using LVS and hot carcass weight (HCW), as well as by USDA expert and on-line graders, before fabrication of carcass sides to either bone-in or boneless cuts. On-line whole number, expert whole-number, and expert nearest-tenth USDA yield grades and LVS + HCW estimates accounted for 53, 52, 58, and 60%, respectively, of the observed variability in boneless, saleable meat yields, and accounted for 56, 57, 62, and 62%, respectively, of the variation in bone-in, saleable meat yields. The LVS + HCW system predicted 77, 65, 70, and 87% of the variation in weights of boneless shoulders, racks, loins, and legs, respectively, and 85, 72, 75, and 86% of the variation in weights of bone-in shoulders, racks, loins, and legs, respectively. Addition of longissimus muscle area (REA), adjusted fat thickness (AFT), or both REA and AFT to LVS + HCW models resulted in improved prediction of boneless saleable meat yields by 5, 3, and 5 percentage points, respectively. Bone-in, saleable meat yield estimations were improved in predictive accuracy by 7.7, 6.6, and 10.1 percentage points, and in precision, when REA alone, AFT alone, or both REA and AFT, respectively, were added to the LVS + HCW output models. Use of LVS + HCW to predict boneless red meat yields of lamb carcasses was more accurate than use of current on-line whole-number, expert whole-number, or expert nearest-tenth USDA yield grades. Thus, LVS + HCW output, when used alone or in combination with AFT and/or REA, improved on-line estimation of boneless cut yields from lamb carcasses. The ability of LVS + HCW to predict yields of wholesale cuts suggests that LVS could be used as an objective

  6. Fluorescence quantum yield measurement in nanoparticle-fluorophore systems by thermal lens spectroscopy

    NASA Astrophysics Data System (ADS)

    Ferreira, M.; Piscitelli, V.

    2016-04-01

    Metallic nanoparticles have been used as a way to tailor the fluorescence properties like quantum yield, but regular fluorescence quantum yield measurements have to counter the reflection and dispersion of a sample for an accurate result. Thermal lens spectroscopy is a good alternative to resolve this problem because doesn't measure the fluorescence intensity but the heat generated by absorption. We studied the changes induced by silver nanoparticles, generated by laser ablation, in the fluorescence peak and quantum yield of Rhodamine B. We fund that the silver nanoparticles lowered the fluorescence peak and quenched the fluorescence of the Rhodamine B and how much is quenched also depends on its concentration.

  7. Expression of the Arabidopsis vacuolar H⁺-pyrophosphatase gene (AVP1) improves the shoot biomass of transgenic barley and increases grain yield in a saline field.

    PubMed

    Schilling, Rhiannon K; Marschner, Petra; Shavrukov, Yuri; Berger, Bettina; Tester, Mark; Roy, Stuart J; Plett, Darren C

    2014-04-01

    Cereal varieties with improved salinity tolerance are needed to achieve profitable grain yields in saline soils. The expression of AVP1, an Arabidopsis gene encoding a vacuolar proton pumping pyrophosphatase (H⁺-PPase), has been shown to improve the salinity tolerance of transgenic plants in greenhouse conditions. However, the potential for this gene to improve the grain yield of cereal crops in a saline field has yet to be evaluated. Recent advances in high-throughput nondestructive phenotyping technologies also offer an opportunity to quantitatively evaluate the growth of transgenic plants under abiotic stress through time. In this study, the growth of transgenic barley expressing AVP1 was evaluated under saline conditions in a pot experiment using nondestructive plant imaging and in a saline field trial. Greenhouse-grown transgenic barley expressing AVP1 produced a larger shoot biomass compared to null segregants, as determined by an increase in projected shoot area, when grown in soil with 150 mM NaCl. This increase in shoot biomass of transgenic AVP1 barley occurred from an early growth stage and also in nonsaline conditions. In a saline field, the transgenic barley expressing AVP1 also showed an increase in shoot biomass and, importantly, produced a greater grain yield per plant compared to wild-type plants. Interestingly, the expression of AVP1 did not alter barley leaf sodium concentrations in either greenhouse- or field-grown plants. This study validates our greenhouse-based experiments and indicates that transgenic barley expressing AVP1 is a promising option for increasing cereal crop productivity in saline fields.

  8. Tradeoffs between vigor and yield for crops grown under different management systems

    NASA Astrophysics Data System (ADS)

    Simic Milas, Anita; Keller Vincent, Robert; Romanko, Matthew; Feitl, Melina; Rupasinghe, Prabha

    2016-04-01

    Remote sensing can provide an effective means for rapid and non-destructive monitoring of crop status and biochemistry. Monitoring pattern of traditional vigor algorithms generated from Landsat 8 OLI satellite data represents a robust method that can be widely used to differentiate the status of crops, as well as to monitor nutrient uptake functionality of differently treated seeds grown under different managements. This study considers 24 factorial parcels of winter wheat in 2013, corn in 2014, and soybeans in 2015, grown under four different types of agricultural management. The parcels are located at the Kellogg Biological Station, Long-Term Ecological Research site in the State of Michigan USA. At maturity, the organic crops exhibit significantly higher vigor and significantly lower yield than conventionally managed crops under different treatments. While organic crops invest in their metabolism at the expense of their yield, the conventional crops manage to increase their yield at the expense of their vigor. Landsat 8 OLI is capable of 1) differentiating the biochemical status of crops under different treatments at maturity, and 2) monitoring the tradeoff between crop yield and vigor that can be controlled by the seed treatments and proper conventional applications, with the ultimate goal of increasing food yield and food availability, and 3) distinguishing between organic and conventionally treated crops. Timing, quantity and types of herbicide applications have a great impact on early and pre-harvest vigor, maturity and yield of conventionally treated crops. Satellite monitoring using Landsat 8 is an optimal tool for coordinating agricultural applications, soil practices and genetic coding of the crop to produce higher yield as well as have early crop maturity, desirable in northern climates.

  9. Space mutagenesis of genetically engineered bacteria expressing recombinant human interferon α1b and screening of higher yielding strains.

    PubMed

    Wang, Junfeng; Liu, Changting; Liu, Jinyi; Fang, Xiangqun; Xu, Chen; Guo, Yinghua; Chang, De; Su, Longxiang

    2014-03-01

    The aim of this study was to investigate the space mutagenesis of genetically engineered bacteria expressing recombinant human interferon α1b. The genetically engineered bacteria expressing the recombinant interferon α1b were sent into outer space on the Chinese Shenzhou VIII spacecraft. After the 17 day space flight, mutant strains that highly expressed the target gene were identified. After a series of screening of spaceflight-treated bacteria and the quantitative comparison of the mutant strains and original strain, we found five strains that showed a significantly higher production of target proteins, compared with the original strain. Our results support the notion that the outer space environment has unique effects on the mutation breeding of microorganisms, including genetically engineered strains. Mutant strains that highly express the target protein could be obtained through spaceflight-induced mutagenesis.

  10. Over-expression of AtPAP2 in Camelina sativa leads to faster plant growth and higher seed yield

    PubMed Central

    2012-01-01

    Background Lipids extracted from seeds of Camelina sativa have been successfully used as a reliable source of aviation biofuels. This biofuel is environmentally friendly because the drought resistance, frost tolerance and low fertilizer requirement of Camelina sativa allow it to grow on marginal lands. Improving the species growth and seed yield by genetic engineering is therefore a target for the biofuels industry. In Arabidopsis, overexpression of purple acid phosphatase 2 encoded by Arabidopsis (AtPAP2) promotes plant growth by modulating carbon metabolism. Overexpression lines bolt earlier and produce 50% more seeds per plant than wild type. In this study, we explored the effects of overexpressing AtPAP2 in Camelina sativa. Results Under controlled environmental conditions, overexpression of AtPAP2 in Camelina sativa resulted in longer hypocotyls, earlier flowering, faster growth rate, higher photosynthetic rate and stomatal conductance, increased seed yield and seed size in comparison with the wild-type line and null-lines. Similar to transgenic Arabidopsis, activity of sucrose phosphate synthase in leaves of transgenic Camelina was also significantly up-regulated. Sucrose produced in photosynthetic tissues supplies the building blocks for cellulose, starch and lipids for growth and fuel for anabolic metabolism. Changes in carbon flow and sink/source activities in transgenic lines may affect floral, architectural, and reproductive traits of plants. Conclusions Lipids extracted from the seeds of Camelina sativa have been used as a major constituent of aviation biofuels. The improved growth rate and seed yield of transgenic Camelina under controlled environmental conditions have the potential to boost oil yield on an area basis in field conditions and thus make Camelina-based biofuels more environmentally friendly and economically attractive. PMID:22472516

  11. Differential expression of CURS gene during various growth stages, climatic condition and soil nutrients in turmeric (Curcuma longa): Towards site specific cultivation for high curcumin yield.

    PubMed

    Sandeep, I Sriram; Das, Suryasnata; Nasim, Noohi; Mishra, Antaryami; Acharya, Laxmikanta; Joshi, Raj Kumar; Nayak, Sanghamitra; Mohanty, Sujata

    2017-09-01

    Curcuma longa L., accumulates substantial amount of curcumin and essential oil. Little is known about the differential expression of curcumin synthase (CURS) gene and consequent curcumin content variations at different agroclimatic zones. The present study aimed to evaluate the effect of climate, soil and harvesting phase on expression of CURS gene for curcumin yield in two high yielding turmeric cultivars. Expression of CURS gene at different experimental zones as well as at different harvesting phase was studied through transcriptional analysis by qRT-PCR. Curcumin varied from 1.5 to 5% and 1.4-5% in Surama and Roma respectively. The expression of CURS also varied from 0.402 to 5.584 fold in Surama and 0.856-5.217 fold in Roma. Difference in curcumin content at a particular zone varied among different harvesting period from 3.95 to 4.31% in Surama and 3.57-3.83% in Roma. Expression of CURS gene was also effected by harvesting time of the rhizome which varied from 7.389 to 16.882 fold in Surama and 4.41-8.342 fold in Roma. The CURS gene expression was found regardless of variations in curcumin content at different experimental zones. This may be due to the effects of soil and environmental variables. Expression was positively correlated with curcumin content with different harvesting time at a particular zone. This find indicates effect of soil and environment on molecular and biochemical dynamics of curcumin biosynthesis and could be useful in genetic improvement of turmeric. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  12. Negative energy balance and hepatic gene expression patterns in high-yielding dairy cows during the early postpartum period: a global approach

    PubMed Central

    McCarthy, S. D.; Waters, S. M.; Kenny, D. A.; Diskin, M. G.; Fitzpatrick, R.; Patton, J.; Wathes, D. C.

    2010-01-01

    In high-yielding dairy cows the liver undergoes extensive physiological and biochemical changes during the early postpartum period in an effort to re-establish metabolic homeostasis and to counteract the adverse effects of negative energy balance (NEB). These adaptations are likely to be mediated by significant alterations in hepatic gene expression. To gain new insights into these events an energy balance model was created using differential feeding and milking regimes to produce two groups of cows with either a mild (MNEB) or severe NEB (SNEB) status. Cows were slaughtered and liver tissues collected on days 6–7 of the first follicular wave postpartum. Using an Affymetrix 23k oligonucleotide bovine array to determine global gene expression in hepatic tissue of these cows, we found a total of 416 genes (189 up- and 227 downregulated) to be altered by SNEB. Network analysis using Ingenuity Pathway Analysis revealed that SNEB was associated with widespread changes in gene expression classified into 36 gene networks including those associated with lipid metabolism, connective tissue development and function, cell signaling, cell cycle, and metabolic diseases, the three most significant of which are discussed in detail. SNEB cows displayed reduced expression of transcription activators and signal transducers that regulate the expression of genes and gene networks associated with cell signaling and tissue repair. These alterations are linked with increased expression of abnormal cell cycle and cellular proliferation associated pathways. This study provides new information and insights on the effect of SNEB on gene expression in high-yielding Holstein Friesian dairy cows in the early postpartum period. PMID:20716645

  13. Negative energy balance and hepatic gene expression patterns in high-yielding dairy cows during the early postpartum period: a global approach.

    PubMed

    McCarthy, S D; Waters, S M; Kenny, D A; Diskin, M G; Fitzpatrick, R; Patton, J; Wathes, D C; Morris, D G

    2010-11-15

    In high-yielding dairy cows the liver undergoes extensive physiological and biochemical changes during the early postpartum period in an effort to re-establish metabolic homeostasis and to counteract the adverse effects of negative energy balance (NEB). These adaptations are likely to be mediated by significant alterations in hepatic gene expression. To gain new insights into these events an energy balance model was created using differential feeding and milking regimes to produce two groups of cows with either a mild (MNEB) or severe NEB (SNEB) status. Cows were slaughtered and liver tissues collected on days 6-7 of the first follicular wave postpartum. Using an Affymetrix 23k oligonucleotide bovine array to determine global gene expression in hepatic tissue of these cows, we found a total of 416 genes (189 up- and 227 downregulated) to be altered by SNEB. Network analysis using Ingenuity Pathway Analysis revealed that SNEB was associated with widespread changes in gene expression classified into 36 gene networks including those associated with lipid metabolism, connective tissue development and function, cell signaling, cell cycle, and metabolic diseases, the three most significant of which are discussed in detail. SNEB cows displayed reduced expression of transcription activators and signal transducers that regulate the expression of genes and gene networks associated with cell signaling and tissue repair. These alterations are linked with increased expression of abnormal cell cycle and cellular proliferation associated pathways. This study provides new information and insights on the effect of SNEB on gene expression in high-yielding Holstein Friesian dairy cows in the early postpartum period.

  14. Escherichia coli Protein Expression System for Acetylcholine Binding Proteins (AChBPs)

    PubMed Central

    Abraham, Nikita; Paul, Blessy; Ragnarsson, Lotten; Lewis, Richard J.

    2016-01-01

    Nicotinic acetylcholine receptors (nAChR) are ligand gated ion channels, identified as therapeutic targets for a range of human diseases. Drug design for nAChR related disorders is increasingly using structure-based approaches. Many of these structural insights for therapeutic lead development have been obtained from co-crystal structures of nAChR agonists and antagonists with the acetylcholine binding protein (AChBP). AChBP is a water soluble, structural and functional homolog of the extracellular, ligand-binding domain of nAChRs. Currently, AChBPs are recombinantly expressed in eukaryotic expression systems for structural and biophysical studies. Here, we report the establishment of an Escherichia coli (E. coli) expression system that significantly reduces the cost and time of production compared to the existing expression systems. E. coli can efficiently express unglycosylated AChBP for crystallography and makes the expression of isotopically labelled forms feasible for NMR. We used a pHUE vector containing an N-terminal His-tagged ubiquitin fusion protein to facilitate AChBP expression in the soluble fractions, and thus avoid the need to recover protein from inclusion bodies. The purified protein yield obtained from the E. coli expression system is comparable to that obtained from existing AChBP expression systems. E. coli expressed AChBP bound nAChR agonists and antagonists with affinities matching those previously reported. Thus, the E. coli expression system significantly simplifies the expression and purification of functional AChBP for structural and biophysical studies. PMID:27304486

  15. HCHL expression in hairy roots of Beta vulgaris yields a high accumulation of p-hydroxybenzoic acid (pHBA) glucose ester, and linkage of pHBA into cell walls.

    PubMed

    Rahman, Laiq ur; Kouno, Hitomi; Hashiguchi, Yuya; Yamamoto, Hirobumi; Narbad, Arjan; Parr, Adrian; Walton, Nicholas; Ikenaga, Toshihiko; Kitamura, Yoshie

    2009-10-01

    As part of a study to explore the potential for new or modified bio-product formation, Beta vulgaris (sugar beet) has been genetically modified to express in root-organ culture a bacterial gene of phenylpropanoid catabolism. The HCHL gene, encoding p-hydroxycinnamoyl-CoA hydratase/lyase, was introduced into B. vulgaris under the control of a CaMV 35S promoter, using Agrobacterium rhizogenes LBA 9402. Hairy root clones expressing the HCHL gene, together with non-expressing clones, were analysed and revealed that one expression-positive clone accumulated the glucose ester of p-hydroxybenzoic acid (pHBA) at about 14% on a dry weight basis. This is the best yield achieved in plant systems so far. Determination of cell-wall components liberated by alkaline hydrolysis confirmed that the ratio of pHBA to ferulic acid was considerably higher in the HCHL-expressing clones, whereas only ferulic acid was detected in a non-expressing clone. The change in cell-wall components also resulted in a decrease in tensile strength in the HCHL-expressing clones.

  16. The Expression of TaRca2-α Gene Associated with Net Photosynthesis Rate, Biomass and Grain Yield in Bread Wheat (Triticum aestivum L.) under Field Conditions

    PubMed Central

    Saeed, Iqbal; Bachir, Daoura Goudia; Chen, Liang; Hu, Yin-Gang

    2016-01-01

    Improvement in activation of Rubisco by Rubisco activase can potentially enhance CO2 assimilation and photosynthetic efficiency in plants. The three homoeologous copies of TaRca2-α were identified on chromosomes 4AL, 4BS and 4DS (TaRca2-α-4AL, TaRca2-α-4BS, and TaRca2-α-4DS) in bread wheat. Expression patterns of the three copies at heading (Z55), anthesis (Z67) and grain-filling (Z73) stages were investigated through qRT-PCR analyses in a panel of 59 bread wheat genotypes and their effects on net photosynthesis rate (Pn), biomass plant-1 (BMPP) and grain yield plant-1 (GYPP) were further explored. Different but similar expression patterns were observed for the three copies of TaRca2-α at the three growth stages with highest expression at grain-filling stage. TaRca2-α-4BS expressed higher at the three stages than TaRca2-α-4AL and TaRca2-α-4DS. The 59 genotypes could be clustered into three groups as high (7 genotypes), intermediate (41 genotypes) and low (11 genotypes) expression based on the expression of the three copies of TaRca2-α at three growth stages. Significant variations (P<0.01) were observed among the three groups of bread wheat genotypes for Pn, BMPP and GYPP. Generally, the genotypes with higher TaRca2-α expression also showed higher values for Pn, BMPP and GYPP. The expressions of the three copies of TaRca2-α at heading, anthesis and grain-filling stages were positively correlated with Pn, BMPP and GYPP (P<0.01) with stronger association for TaRca2-α-4BS at grain-filling stage. These results revealed that the expression of TaRca2-α contribute substantially to Pn, BMPP and GYPP, and suggested that manipulating TaRca-α expression may efficiently improve Pn, BMPP and GYPP in bread wheat and detecting TaRca-α expression levels with emphasis on TaRca2-α-4BS may be a positive strategy for selection in improving photosynthetic efficiency and grain yield of bread wheat. PMID:27548477

  17. The Expression of TaRca2-α Gene Associated with Net Photosynthesis Rate, Biomass and Grain Yield in Bread Wheat (Triticum aestivum L.) under Field Conditions.

    PubMed

    Saeed, Iqbal; Bachir, Daoura Goudia; Chen, Liang; Hu, Yin-Gang

    2016-01-01

    Improvement in activation of Rubisco by Rubisco activase can potentially enhance CO2 assimilation and photosynthetic efficiency in plants. The three homoeologous copies of TaRca2-α were identified on chromosomes 4AL, 4BS and 4DS (TaRca2-α-4AL, TaRca2-α-4BS, and TaRca2-α-4DS) in bread wheat. Expression patterns of the three copies at heading (Z55), anthesis (Z67) and grain-filling (Z73) stages were investigated through qRT-PCR analyses in a panel of 59 bread wheat genotypes and their effects on net photosynthesis rate (Pn), biomass plant-1 (BMPP) and grain yield plant-1 (GYPP) were further explored. Different but similar expression patterns were observed for the three copies of TaRca2-α at the three growth stages with highest expression at grain-filling stage. TaRca2-α-4BS expressed higher at the three stages than TaRca2-α-4AL and TaRca2-α-4DS. The 59 genotypes could be clustered into three groups as high (7 genotypes), intermediate (41 genotypes) and low (11 genotypes) expression based on the expression of the three copies of TaRca2-α at three growth stages. Significant variations (P<0.01) were observed among the three groups of bread wheat genotypes for Pn, BMPP and GYPP. Generally, the genotypes with higher TaRca2-α expression also showed higher values for Pn, BMPP and GYPP. The expressions of the three copies of TaRca2-α at heading, anthesis and grain-filling stages were positively correlated with Pn, BMPP and GYPP (P<0.01) with stronger association for TaRca2-α-4BS at grain-filling stage. These results revealed that the expression of TaRca2-α contribute substantially to Pn, BMPP and GYPP, and suggested that manipulating TaRca-α expression may efficiently improve Pn, BMPP and GYPP in bread wheat and detecting TaRca-α expression levels with emphasis on TaRca2-α-4BS may be a positive strategy for selection in improving photosynthetic efficiency and grain yield of bread wheat.

  18. Primary breast cancer cell culture yields intra-tumor heterogeneous subpopulations expressing exclusive patterns of receptor tyrosine kinases.

    PubMed

    Esparza-López, José; Ramos-Elías, Pier A; Castro-Sánchez, Andrea; Rocha-Zavaleta, Leticia; Escobar-Arriaga, Elizabeth; Zentella-Dehesa, Alejandro; León-Rodríguez, Eucario; Medina-Franco, Heriberto; Ibarra-Sánchez, María de Jesus

    2016-09-20

    It has become evident that intra-tumor heterogeneity of breast cancer impact on several biological processes such as proliferation, migration, cell death and also might contribute to chemotherapy resistance. The expression of Receptor Tyrosine Kinases (RTKs) has not been analyzed in the context of intra-tumor heterogeneity in a primary breast cancer cell culture. Several subpopulations were isolated from the MBCDF (M serial-breast cancer ductal F line) primary breast cancer cells and were successfully maintained in culture and divided in two groups according to their morphology and RTKs expression pattern, and correlated with biological processes like proliferation, migration, anchorage-independent cell growth, and resistance to cytotoxic chemotherapy drugs and tyrosine kinase inhibitors (TKIs). Subpopulations were isolated from MBCDF primary breast cancer cell culture by limiting dilution. RTKs and hormone receptors were examined by Western blot. Proliferation was measure by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT assay). Cell viability was evaluated by Crystal Violet. Migration was assessed using Boyden chambers. Anchorage-independent cell growth was evaluated by colony formation in soft agar. Several subpopulations were isolated from the MBCDF breast cancer cells that were divided into two groups according to their morphology. Analysis of RTKs expression pattern showed that HER1, HER3, c-Met and VEGFR2 were expressed exclusively in cells from group 1, but not in cells from group 2. PDGFR was expressed only in cells from group 2, but not in cells from group 1. HER2, HER4, c-Kit, IGF1-R were expressed in all subpopulations. Biological processes correlated with the RTKs expression pattern. Group 2 subpopulations present the highest rate of cell proliferation, migration and anchorage-independent cell growth. Analysis of susceptibility to chemotherapy drugs and TKIs showed that only Paclitaxel and Imatinib behaved differently between groups

  19. The separation distance distribution in electron-donor-acceptor systems and the wavelength dependence of free ion yields

    NASA Astrophysics Data System (ADS)

    Zhou, Jinwei; Findley, Bret R.; Braun, Charles L.; Sutin, Norman

    2001-06-01

    We recently reported that free radical ion quantum yields for electron-donor-acceptor (EDA) systems of alkylbenzenes-tetracyanoethylene (TCNE) exhibit a remarkable wavelength dependence in dichloromethane, a medium polarity solvent. We proposed that weak absorption by long-distance, unassociated or "random" D⋯A pairs is mainly responsible for the free radical ion yield. Here a model for the wavelength dependence of the free ion yield is developed for four systems in which differing degrees of EDA complex formation are present: 1,3,5-tri-tert-butylbenzene-TCNE in which only random pairs exist due to the bulky groups on the electron donor, and toluene—TCNE, 1,3,5-triethylbenzene-TCNE and 1,3,5-trimethylbenzene-TCNE. Mulliken-Hush theory is used to determine the excitation distance distribution of unassociated, random pairs at different wavelengths. For each absorption distribution, free radical ion yields at different wavelengths are then calculated using Onsager's result for the ion separation probability. Encouraging agreement between the calculated yields and our experimental results is obtained. As far as we are aware, this is the first time that photoexcitation of unassociated donor/acceptor pairs has been invoked as the source of separated radical ion pairs.

  20. Appropriateness and diagnostic yield of upper gastrointestinal endoscopy in an open-access endoscopy system.

    PubMed

    Aljebreen, Abdulrahman M; Alswat, Khalid; Almadi, Majid A

    2013-01-01

    Open access endoscopy (OAE) decreases the waiting time for patients and clinical burden to gastroenterologist; however, the appropriateness of referrals for endoscopy and thus the diagnostic yield of these endoscopies has become an important issue. The aim of this study was to determine the appropriateness of upper gastrointestinal (GI) endoscopy requests in an OAE system. A retrospective chart review of all consecutive patients who underwent an upper gastroscopy in the year 2008 was performed and was defined as appropriate or inappropriate according to the American Society for Gastrointestinal Endoscopy (ASGE) guidelines. Endoscopic findings were recorded and classified as positive or negative. Referrals were categorized as being from a gastroenterologist, internist, surgeon, primary care physicians or others, and on an inpatient or out-patient basis. A total of 505 consecutive patients were included. The mean age was 45.3 (standard deviation 18.1), 259 (51%) of them were males. 31% of the referrals were thought to be inappropriate. Referrals from primary care physicians were inappropriate in 47% of patients while only 19.5% of gastroenterologists referrals were considered inappropriate. Nearly, 37.8% of the out-patient referrals were inappropriate compared to only 7.8% for inpatients. Abnormal findings were found in 78.5% and 78% of patients referred by gastroenterologists and surgeons respectively while in those referred by primary care physicians it was (49.7%). Inpatients referred for endoscopy had abnormal findings in (81.7%) while in out-patients it was (66.6%). The most common appropriate indications in order of frequency were "upper abdominal distress that persisted despite an appropriate trial of therapy "(78.9%),''persistent vomiting of unknown cause "(19.2%), upper GI bleeding or unexplained iron deficiency anemia (7.6%). The sensitivity and specificity of the ASGE guidelines in our study population was 70.3% and 35% respectively. A large proportion of

  1. Ectopic Expression of CDF3 Genes in Tomato Enhances Biomass Production and Yield under Salinity Stress Conditions

    PubMed Central

    Renau-Morata, Begoña; Molina, Rosa V.; Carrillo, Laura; Cebolla-Cornejo, Jaime; Sánchez-Perales, Manuel; Pollmann, Stephan; Domínguez-Figueroa, José; Corrales, Alba R.; Flexas, Jaume; Vicente-Carbajosa, Jesús; Medina, Joaquín; Nebauer, Sergio G.

    2017-01-01

    Cycling Dof Factor (CDF) transcription factors (TFs) are involved in multiple processes related to plant growth and development. A member of this family, CDF3, has recently been linked in Arabidopsis to the regulation of primary metabolism and abiotic stress responses, but its role in crop production under stress is still unknown. In this study, we characterized tomato plants overexpressing the CDF3 genes from Arabidopsis and tomato and analyzed their effects on growth and yield under salinity, additionally gaining deeper insights into the molecular function of these TFs. Our results provide evidence for higher biomass production and yield in the 35S::AtCDF3 and 35S::SlCDF3 plants, likely due to a higher photosynthetic capacity resulting in increased sucrose availability. Transcriptome analysis revealed that CDF3 genes regulate a set of genes involved in redox homeostasis, photosynthesis performance and primary metabolism that lead to enhanced biomass production. Consistently, metabolomic profiling revealed that CDF3 evokes changes in the primary metabolism triggering enhanced nitrogen assimilation, and disclosed that the amount of some protective metabolites including sucrose, GABA and asparagine were higher in vegetative tissues of CDF3 overexpressing plants. Altogether these changes improved performance of 35S::AtCDF3 and 35S::SlCDF3 plants under salinity conditions. Moreover, the overexpression of CDF3 genes modified organic acid and sugar content in fruits, improving variables related to flavor perception and fruit quality. Overall, our results associate the CDF3 TF with a role in the control of growth and C/N metabolism, and highlight that overexpression of CDF3 genes can substantially improve plant yield. PMID:28515731

  2. Assimilating remote sensing observations of leaf area index and soil moisture for wheat yield estimates: An observing system simulation experiment

    USDA-ARS?s Scientific Manuscript database

    We develop a robust understanding of the effects of assimilating remote sensing observations of leaf area index and soil moisture (in the top 5 cm) on DSSAT-CSM CropSim-Ceres wheat yield estimates. Synthetic observing system simulation experiments compare the abilities of the Ensemble Kalman Filter...

  3. Impact of liquid fertilizers on plant growth, yield, fruit quality and fertigation management in an organic processing blackberry production system

    USDA-ARS?s Scientific Manuscript database

    The impact of organic fertilizer source on the growth, fruit quality, and yield of blackberry cultivars (‘Marion’ and ‘Black Diamond’) grown in machine-harvested, organic production systems for the processed market was evaluated from 2011-13. The planting was established in spring 2010 using approve...

  4. Genetic, Physiological, and Gene Expression Analyses Reveal That Multiple QTL Enhance Yield of Rice Mega-Variety IR64 under Drought

    PubMed Central

    Swamy B. P., Mallikarjuna; Ahmed, Helal Uddin; Henry, Amelia; Mauleon, Ramil; Dixit, Shalabh; Vikram, Prashant; Tilatto, Ram; Verulkar, Satish B.; Perraju, Puvvada; Mandal, Nimai P.; Variar, Mukund; S., Robin; Chandrababu, Ranganath; Singh, Onkar N.; Dwivedi, Jawaharlal L.; Das, Sankar Prasad; Mishra, Krishna K.; Yadaw, Ram B.; Aditya, Tamal Lata; Karmakar, Biswajit; Satoh, Kouji; Moumeni, Ali; Kikuchi, Shoshi; Leung, Hei; Kumar, Arvind

    2013-01-01

    Background Rice (Oryza sativa L.) is a highly drought sensitive crop, and most semi dwarf rice varieties suffer severe yield losses from reproductive stage drought stress. The genetic complexity of drought tolerance has deterred the identification of agronomically relevant quantitative trait loci (QTL) that can be deployed to improve rice yield under drought in rice. Convergent evidence from physiological characterization, genetic mapping, and multi-location field evaluation was used to address this challenge. Methodology/Principal Findings Two pairs of backcross inbred lines (BILs) from a cross between drought-tolerant donor Aday Sel and high-yielding but drought-susceptible rice variety IR64 were produced. From six BC4F3 mapping populations produced by crossing the +QTL BILs with the −QTL BILs and IR64, four major-effect QTL - one each on chromosomes 2, 4, 9, and 10 - were identified. Meta-analysis of transcriptome data from the +QTL/−QTL BILs identified differentially expressed genes (DEGs) significantly associated with QTL on chromosomes 2, 4, 9, and 10. Physiological characterization of BILs showed increased water uptake ability under drought. The enrichment of DEGs associated with root traits points to differential regulation of root development and function as contributing to drought tolerance in these BILs. BC4F3-derived lines with the QTL conferred yield advantages of 528 to 1875 kg ha−1 over IR64 under reproductive-stage drought stress in the targeted ecosystems of South Asia. Conclusions/Significance Given the importance of rice in daily food consumption and the popularity of IR64, the BC4F3 lines with multiple QTL could provide higher livelihood security to farmers in drought-prone environments. Candidate genes were shortlisted for further characterization to confirm their role in drought tolerance. Differential yield advantages of different combinations of the four QTL reported here indicate that future research should include optimizing QTL

  5. Comparison of disease severity and yield potential of rice varieties in two organic cropping systems

    USDA-ARS?s Scientific Manuscript database

    The market demand for organically produced rice has driven the steady increase in acreage of organic rice in the U. S., with Texas and California having the most acreage. Yield potential and disease management are among the principal challenges associated with organic rice production. Organic rice c...

  6. Increasing plant density in eastern United States broccoli production systems to maximize marketable head yields

    USDA-ARS?s Scientific Manuscript database

    Increased demand for fresh market broccoli (Brassica oleracea L. var. italica) has led to increased production along the eastern seaboard of the United States. Maximizing broccoli yields is a primary concern for quickly expanding eastern commercial markets. Thus, a plant density study was carried ...

  7. Disease severity and yield potential of rice cultivars in organic production systems

    USDA-ARS?s Scientific Manuscript database

    The market demand for organically produced rice has driven the steady increase in the acreage of organic rice in the U. S., with Texas and California being the largest states. Yield potential and disease management are among the principal challenges associated with organic rice production. We evalua...

  8. High yields and soluble expression of superoxide dismutases in Escherichia coli due to the HIV-1 Tat peptide via increases in mRNA transcription

    PubMed Central

    Sun, Yangdong; Ye, Qiao; Wu, Min; Wu, Yonghong; Zhang, Chenggang; Yan, Weiqun

    2016-01-01

    This study aimed to validate the high yield and soluble expression of proteins carrying the transactivator of transcription (Tat) peptide tag, and further explored the potential mechanism by which the Tat tag increases expression. Escherichia coli superoxide dismutase (SOD) proteins, including SodA, SodB and SodC, were selected for analysis. As expected, the yields and the solubility of Tat-tagged proteins were higher than those of Tat-free proteins, and similar results were observed for the total SOD enzyme activity. Bacterial cells that overexpressed Tat-tagged proteins exhibited increased anti-paraquat activity compared with those expressing Tat-free proteins that manifested as SodA>SodC>SodB. When compared with an MG1655 wild-type strain, the growth of a ΔSodA mutant strain was found to be inhibited after paraquat treatment; the growth of ΔSodB and ΔSodC mutant strains was also slightly inhibited. The mRNA transcript level of genes encoding Tat-tagged proteins was higher than that of genes encoding Tat-free proteins. Furthermore, the α-helix and turn of Tat-tagged proteins were higher than those of Tat-free proteins, but the β-sheet and random coil content was lower. These results indicated that the incorporation of the Tat core peptide as a significant basic membrane transduction peptide in fusion proteins could increase mRNA transcripts and promote the high yield and soluble expression of heterologous proteins in E. coli. PMID:27741225

  9. High yield of recombinant human Apolipoprotein A-I expressed in Pichia pastoris by using mixed-mode chromatography.

    PubMed

    Narasimhan Janakiraman, Vignesh; Noubhani, Abdelmajid; Venkataraman, Krishnan; Vijayalakshmi, Mookambeswaran; Santarelli, Xavier

    2016-01-01

    A vast majority of the cardioprotective properties exhibited by High-Density Lipoprotein (HDL) is mediated by its major protein component Apolipoprotein A-I (ApoA1). In order to develop a simplified bioprocess for producing recombinant human Apolipoprotein A-I (rhApoA1) in its near-native form, rhApoA1was expressed without the use of an affinity tag in view of its potential therapeutic applications. Expressed in Pichia pastoris at expression levels of 58.2 mg ApoA1 per litre of culture in a reproducible manner, the target protein was purified by mixed-mode chromatography using Capto™ MMC ligand with a purity and recovery of 84% and 68%, respectively. ApoA1 purification was scaled up to Mixed-mode Expanded Bed Adsorption chromatography to establish an 'on-line' process for the efficient capture of rhApoA1 directly from the P. pastoris expression broth. A polishing step using anion exchange chromatography enabled the recovery of ApoA1 up to 96% purity. Purified ApoA1 was identified and verified by RPLC-ESI-Q-TOF mass spectrometry. This two-step process would reduce processing times and therefore costs in comparison to the twelve-step procedure currently used for recovering rhApoA1 from P. pastoris. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Heterologous expression of laccase cDNA from Ceriporiopsis subvermispora yields copper-activated apoprotein and complex isoform patterns

    Treesearch

    Luis F. Larrondo; Marcela Avila; Loreto Salas; Dan Cullen; Rafael Vicuna

    2003-01-01

    Analysis of genomic clones encoding a putative laccase in homokaryon strains of Ceriporiopsis subvermispora led to the identification of an allelic variant of the previously described lcs-1 gene. A cDNA clone corresponding to this gene was expressed in Aspergillus nidulans and in Aspergillus niger. Enzyme assays and Western blots showed that both hosts secreted active...

  11. Effect of chemical and mechanical weed control on cassava yield, soil quality and erosion under cassava cropping system

    NASA Astrophysics Data System (ADS)

    Islami, Titiek; Wisnubroto, Erwin; Utomo, Wani

    2016-04-01

    Three years field experiments were conducted to study the effect of chemical and mechanical weed control on soil quality and erosion under cassava cropping system. The experiment were conducted at University Brawijaya field experimental station, Jatikerto, Malang, Indonesia. The experiments were carried out from 2011 - 2014. The treatments consist of three cropping system (cassava mono culture; cassava + maize intercropping and cassava + peanut intercropping), and two weed control method (chemical and mechanical methods). The experimental result showed that the yield of cassava first year and second year did not influenced by weed control method and cropping system. However, the third year yield of cassava was influence by weed control method and cropping system. The cassava yield planted in cassava + maize intercropping system with chemical weed control methods was only 24 t/ha, which lower compared to other treatments, even with that of the same cropping system used mechanical weed control. The highest cassava yield in third year was obtained by cassava + peanuts cropping system with mechanical weed control method. After three years experiment, the soil of cassava monoculture system with chemical weed control method possessed the lowest soil organic matter, and soil aggregate stability. During three years of cropping soil erosion in chemical weed control method, especially on cassava monoculture, was higher compared to mechanical weed control method. The soil loss from chemical control method were 40 t/ha, 44 t/ha and 54 t/ha for the first, second and third year crop. The soil loss from mechanical weed control method for the same years was: 36 t/ha, 36 t/ha and 38 t/ha. Key words: herbicide, intercropping, soil organic matter, aggregate stability.

  12. Characterisation of endometrial gene expression and metabolic parameters in beef heifers yielding viable or non-viable embryos on Day 7 after insemination.

    PubMed

    Beltman, M E; Forde, N; Furney, P; Carter, F; Roche, J F; Lonergan, P; Crowe, M A

    2010-01-01

    The aim of the present study was to compare the hormonal and metabolic characteristics and endometrial gene expression profiles in beef heifers yielding either a viable or degenerate embryo on Day 7 after insemination as a means to explain differences in embryo survival. Oestrus was synchronised in cross-bred beef heifers (n = 145) using a controlled internal drug release (CIDR)-prostaglandin protocol. Heifers (n = 102) detected in standing oestrus (within 24-48 h after CIDR removal) were inseminated 12-18 h after detection of oestrus (Day 0) with frozen-thawed semen from a single ejaculate of a bull with proven fertility. Blood samples were collected from Day 4 to Day 7 after oestrus to measure progesterone (on Days 4, 5 and 7), insulin and insulin-like growth factor (IGF)-I (on Days 4 and 6) and urea (on Day 7) concentrations. All animals were killed on Day 7. Uterine pH was determined at the time of death. Animals from which an embryo was recovered were classified as either having a viable embryo (morula/blastocyst stage; n = 32) or a retarded embryo (arrested at the two- to 16-cell stage; n = 19). In addition, 14 single-celled unfertilised oocytes were recovered, giving an overall recovery rate of 64%. There was no significant difference in the blood parameters determined or uterine pH at the time of death between heifers with either a viable or retarded embryo. The relative abundance of nine transcripts (i.e. MOGAT1, PFKB2, LYZ2, SVS8, UHRF1, PTGES, AGPAT4, DGKA and HGPD) of 53 tested in the endometrial tissue differed between heifers with a viable or retarded embryo. Both LYZ2 and UHRF1 are associated with regulation of the immune system; PFKFB2 is a mediator in glycolysis; MOGAT, AGPAT4 and DGKA belong to the triglyceride synthesis pathway; and PTGES and HGPD belong to the prostaglandin pathway. Both these metabolic pathways are important for early embryonic development. In conclusion, retarded embryo development in the present study was not related to serum

  13. Effects of polyunsaturated fatty acids from plant oils and algae on milk fat yield and composition are associated with mammary lipogenic and SREBF1 gene expression.

    PubMed

    Angulo, J; Mahecha, L; Nuernberg, K; Nuernberg, G; Dannenberger, D; Olivera, M; Boutinaud, M; Leroux, C; Albrecht, E; Bernard, L

    2012-12-01

    The main aim of the present study was to examine the effects of long-term supplementing diets with saturated or unprotected polyunsaturated fatty acids from two different plant oils rich in either n-3 or n-6 fatty acids (FAs) plus docosahexaenoic acid (DHA)-rich algae on mammary gene expression and milk fat composition in lactating dairy cows. Gene expression was determined from mammary tissue and milk epithelial cells. Eighteen primiparous German Holstein dairy cows in mid-lactation were randomly assigned into three dietary treatments that consist of silage-based diets supplemented with rumen-stable fractionated palm fat (SAT; 3.1% of the basal diet dry matter, DM), or a mixture of linseed oil (2.7% of the basal diet DM) plus DHA-rich algae (LINA; 0.4% of the basal diet DM) or a mixture of sunflower oil (2.7% of the basal diet DM) plus DHA-rich algae (SUNA; 0.4% of the basal diet DM), for a period of 10 weeks. At the end of the experimental period, the cows were slaughtered and mammary tissues were collected to study the gene expression of lipogenic enzymes. During the last week, the milk yield and composition were determined, and milk was collected for FA measurements and the isolation of milk purified mammary epithelial cells (MECs). Supplementation with plant oils and DHA-rich algae resulted in milk fat depression (MFD; yield and percentage). The secretion of de novo FAs in the milk was reduced, whereas the secretion of trans-10,cis-12-CLA and DHA were increased. These changes in FA secretions were associated in mammary tissue with a joint down-regulation of mammary lipogenic enzyme gene expression (stearoyl-CoA desaturase, SCD1; FA synthase, FASN) and expression of the regulatory element binding transcription factor (SREBF1), whereas no effect was observed on lipoprotein lipase (LPL) and glycerol-3-phosphate acyltransferase 1, mitochondrial (GPAM). A positive relationship between mammary SCD1 and SREBF1 mRNA abundances was observed, suggesting a similar

  14. A simple and robust protocol for high-yield expression of perdeuterated proteins in Escherichia coli grown in shaker flasks.

    PubMed

    Cai, Mengli; Huang, Ying; Yang, Renbin; Craigie, Robert; Clore, G M

    2016-10-01

    We present a simple, convenient and robust protocol for expressing perdeuterated proteins in E. coli BL21(DE3) cells in shaker flasks that reduces D2O usage tenfold and d7-glucose usage by 30 %. Using a modified M9 medium and optimized growth conditions, we were able to grow cells in linear log phase to an OD600 of up to 10. Inducing the cells with isopropyl β-D-1-thiogalactopyranoside at an OD600 of 10, instead of less than 1, enabled us to increase the cell mass tenfold per unit volume of cell culture. We show that protein expression levels per cell are the same when induced at an OD600 between 1 and 10 under these growth conditions. Thus, our new protocol can increase protein yield per unit volume of cell culture tenfold. Adaptation of E. coli from H2O-based to D2O-based medium is also key for ensuring high levels of protein expression in D2O. We find that a simple three-step adaptation approach-Luria-Bertani (LB) medium in H2O to LB in D2O to modified-M9 medium in D2O is both simple and reliable. The method increases the yield of perdeuterated proteins by up to tenfold using commonly available air shakers without any requirement for specialized fermentation equipment.

  15. Interspecific Differential Expression Analysis of RNA-Seq Data Yields Insight into Life Cycle Variation in Hydractiniid Hydrozoans.

    PubMed

    Sanders, Steven M; Cartwright, Paulyn

    2015-08-06

    Hydrozoans are known for their complex life cycles, which can alternate between an asexually reproducing polyp stage and a sexually reproducing medusa stage. Most hydrozoan species, however, lack a free-living medusa stage and instead display a developmentally truncated form, called a medusoid or sporosac, which generally remains attached to the polyp. Although evolutionary transitions in medusa truncation and loss have been investigated phylogenetically, little is known about the genes involved in the development and loss of this life cycle stage. Here, we present a new workflow for evaluating differential expression (DE) between two species using short read Illumina RNA-seq data. Through interspecific DE analyses between two hydractiniid hydrozoans, Hydractinia symbiolongicarpus and Podocoryna carnea, we identified genes potentially involved in the developmental, functional, and morphological differences between the fully developed medusa of P. carnea and reduced sporosac of H. symbiolongicarpus. A total of 10,909 putative orthologs of H. symbiolongicarpus and P. carnea were identified from de novo assemblies of short read Illumina data. DE analysis revealed 938 of these are differentially expressed between P. carnea developing and adult medusa, when compared with H. symbiolongicarpus sporosacs, the majority of which have not been previously characterized in cnidarians. In addition, several genes with no corresponding ortholog in H. symbiolongicarpus were expressed in developing medusa of P. carnea. Results presented here show interspecific DE analyses of RNA-seq data to be a sensitive and reliable method for identifying genes and gene pathways potentially involved in morphological and life cycle differences between species.

  16. Use of a stress inducible promoter to drive ectopic AtCBF expression improves potato freezing tolerance while minimizing negative effects on tuber yield.

    PubMed

    Pino, María-Teresa; Skinner, Jeffrey S; Park, Eung-Jun; Jeknić, Zoran; Hayes, Patrick M; Thomashow, Michael F; Chen, Tony H H

    2007-09-01

    Solanum tuberosum is a frost-sensitive species incapable of cold acclimation. A brief exposure to frost can significantly reduce its yields, while hard frosts can completely destroy entire crops. Thus, gains in freezing tolerance of even a few degrees would be of considerable benefit relative to frost damage. The S. tuberosum cv. Umatilla was transformed with three Arabidopsis CBF genes (AtCBF1-3) driven by either a constitutive CaMV35S or a stress-inducible Arabidopsis rd29A promoter. AtCBF1 and AtCBF3 over-expression via the 35S promoter increased freezing tolerance about 2 degrees C, whereas AtCBF2 over-expression failed to increase freezing tolerance. Transgenic plants of AtCBF1 and AtCBF3 driven by the rd29A promoter reached the same level of freezing tolerance as the 35S versions within a few hours of exposure to low but non-freezing temperatures. Constitutive expression of AtCBF genes was associated with negative phenotypes, including smaller leaves, stunted plants, delayed flowering, and reduction or lack of tuber production. While imparting the same degree of freezing tolerance, control of AtCBF expression via the stress-inducible promoter ameliorated these negative phenotypic effects and restored tuber production to levels similar to wild-type plants. These results suggest that use of a stress-inducible promoter to direct CBF transgene expression can yield significant gains in freezing tolerance without negatively impacting agronomically important traits in potato.

  17. Acetate yield increased by gas circulation and fed-batch fermentation in a novel syntrophic acetogenesis and homoacetogenesis coupling system.

    PubMed

    Nie, YanQiu; Liu, He; Du, GuoCheng; Chen, Jian

    2008-05-01

    Gas circulation and fed-batch fermentation were applied for enhancing acetate production by mixed culture in a novel syntrophic acetogenesis and homoacetogenesis coupling system. The results show that the acetate yield in the fed-batch test with gas circulation is about 47% higher than that in the batch test without gas circulation. The fed-batch method helps to increase acetate yield by balancing hydrogen production in the acetogenesis phase (the 1st phase) and hydrogen consumption in the homoacetogenesis phase (the 2nd phase) of the coupling system. Gas circulation enhances mass transfer between different phases of the coupling system, hence resulting in increased homoacetogenesis in the 2nd phase and relief of the products (H2) inhibition to syntrophic acetogenesis in the 1st phase. The effects of gas circulation and fed-batch fermentation on direct glucose conversion to acetate were also investigated.

  18. Alfalfa yield response to inoculation with recombinant strains of Rhizobium meliloti with an extra copy of dctABD and/or modified nifA expression.

    PubMed Central

    Bosworth, A H; Williams, M K; Albrecht, K A; Kwiatkowski, R; Beynon, J; Hankinson, T R; Ronson, C W; Cannon, F; Wacek, T J; Triplett, E W

    1994-01-01

    The construction of rhizobial strains which increase plant biomass under controlled conditions has been previously reported. However, there is no evidence that these newly constructed strains increase legume yield under agricultural conditions. This work tested the hypothesis that carefully manipulating expression of additional copies of nifA and dctABD in strains of Rhizobium meliloti would increase alfalfa yield in the field. The rationale for this hypothesis is based on the positive regulatory role that nifA plays in the expression of the nif regulon and the fact that a supply of dicarboxylic acids from the plant is required as a carbon and energy source for nitrogen fixation by the Rhizobium bacteroids in the nodule. These recombinant strains, as well as the wild-type strains from which they were derived, are ideal tools to examine the effects of modifying or increasing the expression of these genes on alfalfa biomass. The experimental design comprised seven recombinant strains, two wild-type strains, and an uninoculated control. Each treatment was replicated eight times and was conducted at four field sites in Wisconsin. Recombinant strain RMBPC-2, which has an additional copy of both nifA and dctABD, increased alfalfa biomass by 12.9% compared with the yield with the wild-type strain RMBPC and 17.9% over that in the uninoculated control plot at the site where soil nitrogen and organic matter content was lowest. These increases were statistically significant at the 5% confidence interval for each of the three harvests made during the growing season. Strain RMBPC-2 did increase alfalfa biomass at the Hancock site; however, no other significant increases or decreases in alfalfa biomass were observed with the seven other recombinant strains at that site. At three sites where this experiment was conducted, either native rhizobial populations or soil nitrogen concentrations were high. At these sites, none of the recombinant strains affected yield. We conclude that

  19. Comparative Proteomic Analysis of saccharopolyspora spinosa SP06081 and PR2 strains reveals the differentially expressed proteins correlated with the increase of spinosad yield

    PubMed Central

    2011-01-01

    Background Saccharopolyspora spinosa produces the environment-friendly biopesticide spinosad, a mixture of two polyketide-derived macrolide active ingredients called spinosyns A and D. Therefore considerable interest is in the improvement of spinosad production because of its low yield in wild-type S. spinosa. Recently, a spinosad-hyperproducing PR2 strain with stable heredity was obtained from protoplast regeneration of the wild-type S. spinosa SP06081 strain. A comparative proteomic analysis was performed on the two strains during the first rapid growth phase (RG1) in seed medium (SM) by using label-free quantitative proteomics to investigate the underlying mechanism leading to the enhancement of spinosad yield. Results In total, 224 proteins from the SP06081 strain and 204 proteins from the PR2 strain were unambiguously identified by liquid chromatography-tandem mass spectrometry analysis, sharing 140 proteins. A total of 12 proteins directly related to spinosad biosynthesis were identified from the two strains in RG1. Comparative analysis of the shared proteins revealed that approximately 31% of them changed their abundance significantly and fell in all of the functional groups, such as tricarboxylic acid cycles, glycolysis, biosynthetic processes, catabolic processes, transcription, translation, oxidation and reduction. Several key enzymes involved in the synthesis of primary metabolic intermediates used as precursors for spinosad production, energy supply, polyketide chain assembly, deoxysugar methylation, and antioxidative stress were differentially expressed in the same pattern of facilitating spinosad production by the PR2 strain. Real-time reverse transcriptase polymerase chain reaction analysis revealed that four of five selected genes showed a positive correlation between changes at the translational and transcriptional expression level, which further confirmed the proteomic analysis. Conclusions The present study is the first comprehensive and

  20. Crop yield summary for three wetland reservoir subirrigation systems in northwest Ohio

    USDA-ARS?s Scientific Manuscript database

    Wetland Reservoir Subirrigation Systems (WRSIS) are innovative agricultural water management and recycling systems comprised of three main components; a constructed wetland, a water storage reservoir, and cropland containing subsurface drainage pipe systems. Surface runoff and subsurface drainage f...

  1. Interspecific Differential Expression Analysis of RNA-Seq Data Yields Insight into Life Cycle Variation in Hydractiniid Hydrozoans

    PubMed Central

    Sanders, Steven M.; Cartwright, Paulyn

    2015-01-01

    Hydrozoans are known for their complex life cycles, which can alternate between an asexually reproducing polyp stage and a sexually reproducing medusa stage. Most hydrozoan species, however, lack a free-living medusa stage and instead display a developmentally truncated form, called a medusoid or sporosac, which generally remains attached to the polyp. Although evolutionary transitions in medusa truncation and loss have been investigated phylogenetically, little is known about the genes involved in the development and loss of this life cycle stage. Here, we present a new workflow for evaluating differential expression (DE) between two species using short read Illumina RNA-seq data. Through interspecific DE analyses between two hydractiniid hydrozoans, Hydractinia symbiolongicarpus and Podocoryna carnea, we identified genes potentially involved in the developmental, functional, and morphological differences between the fully developed medusa of P. carnea and reduced sporosac of H. symbiolongicarpus. A total of 10,909 putative orthologs of H. symbiolongicarpus and P. carnea were identified from de novo assemblies of short read Illumina data. DE analysis revealed 938 of these are differentially expressed between P. carnea developing and adult medusa, when compared with H. symbiolongicarpus sporosacs, the majority of which have not been previously characterized in cnidarians. In addition, several genes with no corresponding ortholog in H. symbiolongicarpus were expressed in developing medusa of P. carnea. Results presented here show interspecific DE analyses of RNA-seq data to be a sensitive and reliable method for identifying genes and gene pathways potentially involved in morphological and life cycle differences between species. PMID:26251524

  2. Effects of potato-cotton cropping systems and nematicides on plant-parasitic nematodes and crop yields.

    PubMed

    Crow, W T; Weingartner, D P; Dickson, D W

    2000-09-01

    Belonolaimus longicaudatus has been reported as damaging both potato (Solanum tuberosum) and cotton (Gossypium hirsutum). These crops are not normally grown in cropping systems together in areas where the soil is infested with B. longicaudatus. During the 1990s cotton was grown in a potato production region that was a suitable habitat for B. longicaudatus. It was not known how integrating the production of these two crops by rotation or double-cropping would affect the population densities of B. longicaudatus, other plant-parasitic nematodes common in the region, or crop yields. A 3-year field study evaluated the viability of both crops in monocropping, rotation, and double-cropping systems. Viability was evaluated using effects on population densities of plant-parasitic nematodes and yields. Rotation of cotton with potato was found to decrease population densities of B. longicaudatus and Meloidogyne incognita in comparison with continuous potato. Population densities of B. longicaudatus following double-cropping were greater than following continuous cotton. Yields of both potato and cotton in rotation were equivalent to either crop in monocropping. Yields of both crops were lower following double-cropping when nematicides were not used.

  3. Techno-economic analysis of horseradish peroxidase production using a transient expression system in Nicotiana benthamiana.

    PubMed

    Walwyn, David Richard; Huddy, Suzanne M; Rybicki, Edward P

    2015-01-01

    Despite the advantages of plant-based transient expression systems relative to microbial or mammalian cell systems, the commercial production of recombinant proteins using plants has not yet been achieved to any significant extent. One of the challenges has been the lack of published data on the costs of manufacture for products other than biopharmaceuticals. In this study, we report on the techno-economic analysis of the production of a standard commercial enzyme, namely, horseradish peroxidase (HRP), using a transient expression system in Nicotiana benthamiana. Based on the proven plant yield of 240 mg HRP/kg biomass, a biomass productivity of 15-kg biomass/m(2)/year and a process yield of 54 % (mg HRP product/mg HRP in biomass), it is apparent that HRP can be manufactured economically via transient expression in plants in a large-scale facility (>5 kg HRP/year). At this level, the process is competitive versus the existing technology (extraction of the enzyme from horseradish), and the product is of comparable or improved activity, containing only the preferred isoenzyme C. Production scale, protein yield and biomass productivity are found to be the most important determinants of overall viability.

  4. Development and characterization of a high yield transportable pulsed neutron source with efficient and compact pulsed power system

    NASA Astrophysics Data System (ADS)

    Verma, Rishi; Mishra, Ekansh; Dhang, Prosenjit; Sagar, Karuna; Meena, Manraj; Shyam, Anurag

    2016-09-01

    The results of characterization experiments carried out on a newly developed dense plasma focus device based intense pulsed neutron source with efficient and compact pulsed power system are reported. Its high current sealed pseudospark switch based low inductance capacitor bank with maximum stored energy of ˜10 kJ is segregated into four modules of ˜2.5 kJ each and it cumulatively delivers peak current in the range of 400 kA-600 kA (corresponding to charging voltage range of 14 kV-18 kV) in a quarter time period of ˜2 μs. The neutron yield performance of this device has been optimized by discretely varying deuterium filling gas pressure in the range of 6 mbar-11 mbar at ˜17 kV/550 kA discharge. At ˜7 kJ/8.5 mbar operation, the average neutron yield has been measured to be in the order of ˜4 × 109 neutrons/pulse which is the highest ever reported neutron yield from a plasma focus device with the same stored energy. The average forward to radial anisotropy in neutron yield is found to be ˜2. The entire system is contained on a moveable trolley having dimensions 1.5 m × 1 m × 0.7 m and its operation and control (up to the distance of 25 m) are facilitated through optically isolated handheld remote console. The overall compactness of this system provides minimum proximity to small as well as large samples for irradiation. The major intended application objective of this high neutron yield dense plasma focus device development is to explore the feasibility of active neutron interrogation experiments by utilization of intense pulsed neutron sources.

  5. Development and characterization of a high yield transportable pulsed neutron source with efficient and compact pulsed power system.

    PubMed

    Verma, Rishi; Mishra, Ekansh; Dhang, Prosenjit; Sagar, Karuna; Meena, Manraj; Shyam, Anurag

    2016-09-01

    The results of characterization experiments carried out on a newly developed dense plasma focus device based intense pulsed neutron source with efficient and compact pulsed power system are reported. Its high current sealed pseudospark switch based low inductance capacitor bank with maximum stored energy of ∼10 kJ is segregated into four modules of ∼2.5 kJ each and it cumulatively delivers peak current in the range of 400 kA-600 kA (corresponding to charging voltage range of 14 kV-18 kV) in a quarter time period of ∼2 μs. The neutron yield performance of this device has been optimized by discretely varying deuterium filling gas pressure in the range of 6 mbar-11 mbar at ∼17 kV/550 kA discharge. At ∼7 kJ/8.5 mbar operation, the average neutron yield has been measured to be in the order of ∼4 × 10(9) neutrons/pulse which is the highest ever reported neutron yield from a plasma focus device with the same stored energy. The average forward to radial anisotropy in neutron yield is found to be ∼2. The entire system is contained on a moveable trolley having dimensions 1.5 m × 1 m × 0.7 m and its operation and control (up to the distance of 25 m) are facilitated through optically isolated handheld remote console. The overall compactness of this system provides minimum proximity to small as well as large samples for irradiation. The major intended application objective of this high neutron yield dense plasma focus device development is to explore the feasibility of active neutron interrogation experiments by utilization of intense pulsed neutron sources.

  6. Development and characterization of a high yield transportable pulsed neutron source with efficient and compact pulsed power system

    SciTech Connect

    Verma, Rishi E-mail: rishiv@barc.gov.in; Mishra, Ekansh; Dhang, Prosenjit; Sagar, Karuna; Meena, Manraj; Shyam, Anurag

    2016-09-15

    The results of characterization experiments carried out on a newly developed dense plasma focus device based intense pulsed neutron source with efficient and compact pulsed power system are reported. Its high current sealed pseudospark switch based low inductance capacitor bank with maximum stored energy of ∼10 kJ is segregated into four modules of ∼2.5 kJ each and it cumulatively delivers peak current in the range of 400 kA–600 kA (corresponding to charging voltage range of 14 kV–18 kV) in a quarter time period of ∼2 μs. The neutron yield performance of this device has been optimized by discretely varying deuterium filling gas pressure in the range of 6 mbar–11 mbar at ∼17 kV/550 kA discharge. At ∼7 kJ/8.5 mbar operation, the average neutron yield has been measured to be in the order of ∼4 × 10{sup 9} neutrons/pulse which is the highest ever reported neutron yield from a plasma focus device with the same stored energy. The average forward to radial anisotropy in neutron yield is found to be ∼2. The entire system is contained on a moveable trolley having dimensions 1.5 m × 1 m × 0.7 m and its operation and control (up to the distance of 25 m) are facilitated through optically isolated handheld remote console. The overall compactness of this system provides minimum proximity to small as well as large samples for irradiation. The major intended application objective of this high neutron yield dense plasma focus device development is to explore the feasibility of active neutron interrogation experiments by utilization of intense pulsed neutron sources.

  7. Characterization of NoV P particle-based chimeric protein vaccines developed from two different expression systems.

    PubMed

    Fu, Lu; Jin, Hao; Yu, Yongjiao; Yu, Bin; Zhang, Haihong; Wu, Jiaxin; Yin, Yuhe; Yu, Xianghui; Wu, Hui; Kong, Wei

    2017-02-01

    The Norovirus (NoV) P domain, with three surface loops for foreign antigen insertion, has been demonstrated as an excellent platform for antigen presentation and novel vaccine development. The P domain alone can self-assemble into a P dimer, 12-mer small particle or 24-mer P particle, and vaccines based on those particles may elicit different levels of immunogenicity. Currently, P particles are generally produced in soluble expression systems in Escherichia coli, mainly in the 24-mer form. However, the low yield of the soluble protein has hindered further clinical applications of P particle-based protein vaccines. In this study, we inserted the Alzheimer's disease (AD) immunogen Aβ1-6 into the three loops of the P particle to generate an AD protein vaccine. To increase the yield of this chimeric protein, we tested the generation of proteins in a soluble expression system and an inclusion body expression system separately in E. coli. The result showed that the inclusion body expression system could greatly enhance the product yield of the chimeric protein compared with the soluble expression system. The refolded protein from the inclusion bodies was mainly in the 12-mer form, while the protein generated from the soluble supernatant was mainly in the 24-mer form. Moreover, the immunogenicity of soluble proteins was significantly stronger than that of the refolded proteins. Thus, comparisons between the two expression methods suggested that the soluble expression system generated chimeric P particles with better immunogenicity, while inclusion body expression system yielded more P particle proteins.

  8. Optimization of eGFP expression using a modified baculovirus expression system.

    PubMed

    Ge, Jingping; Jin, Liying; Tang, Xiaoyan; Gao, Dongni; An, Qi; Ping, Wenxiang

    2014-03-10

    The baculovirus gene expression system is an efficient and safe protein expression system, since baculoviruses cannot replicate in mammalian cells. In order to improve the transduction efficiency and increase the reporter gene expression levels delivered by baculoviruses, we tested in the baculovirus expression cassette the Woodchuck hepatitis virus response element (WPRE), and AAV-derived inverted terminal repeats (ITRs) and the truncated vesicular stomatitis virus G protein (VSV-GED). The results showed that WPRE and VSV-GED have synergistic effects and could enhance the expression efficiency of enhanced green fluorescence protein (eGFP), and that ITRs effectively extended the duration of eGFP expression. We also demonstrated that the efficiency of eGFP expression varied under the control of the CMV, CBA, EF1-α or WSSV ie1 promoters in different cell lines. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Evaluation of current methods used to analyze the expression profiles of ABC transporters yields an improved drug-discovery database

    PubMed Central

    Orina, Josiah N.; Calcagno, Anna Maria; Wu, Chung-Pu; Varma, Sudhir; Shih, Joanna; Lin, Min; Eichler, Gabriel; Weinstein, John N.; Pommier, Yves; Ambudkar, Suresh V.; Gottesman, Michael M.; Gillet, Jean-Pierre

    2009-01-01

    The development of multidrug resistance (MDR) to chemotherapy remains a major challenge in the treatment of cancer. Resistance exists against every effective anti-cancer drug and can develop by multiple mechanisms. These mechanisms can act individually or synergistically, leading to multidrug resistance (MDR), in which the cell becomes resistant to a variety of structurally and mechanistically unrelated drugs in addition to the drug initially administered. Although extensive work has been done to characterize MDR mechanisms in vitro, the translation of this knowledge to the clinic has not been successful. Therefore, identifying genes and mechanisms critical to the development of MDR in vivo and establishing a reliable method for analyzing highly homologous genes from small amounts of tissue is fundamental to achieving any significant enhancement in our understanding of multidrug resistance mechanisms and could lead to treatments designed to circumvent it. In this study, we use a previously established database that allows the identification of lead compounds in the early stages of drug discovery that are not ABC transporter substrates. We believe this can serve as a model for appraising the accuracy and sensitivity of current methods used to analyze the expression profiles of ABC transporters. We found two platforms to be superior methods for the analysis of expression profiles of highly homologous gene superfamilies. This study also led to an improved database by revealing previously unidentified substrates for ABCB1, ABCC1 and ABCG2, transporters that contribute to multidrug resistance. PMID:19584229

  10. Influence of solid-liquid separation strategy on biogas yield from a stratified swine production system.

    PubMed

    Cestonaro do Amaral, André; Kunz, Airton; Radis Steinmetz, Ricardo Luis; Scussiato, Lucas Antunes; Tápparo, Deisi Cristina; Gaspareto, Taís Carla

    2016-03-01

    As the fourth largest swine producer and exporter, Brazil has increased its participation in the global swine production market. Generally, these units concentrate a large number of animals and generate effluents that must be correctly managed to prevent environmental impacts, being anaerobic digestion is an interesting alternative for treating these effluents. The low-volatile solid concentration in the manure suggests the need for solid-liquid separation as a tool to improve the biogas generation capacity. This study aimed to determine the influence of simplified and inexpensive solid-liquid separation strategies (screening and settling) and the different manures produced during each swine production phase (gestating and farrowing sow houses, nursery houses and finishing houses) on biogas and methane yield. We collected samples in two gestating sow houses (GSH-a and GSH-b), two farrowing sow houses (FSH-a and FSH-b), a nursery house (NH) and a finishing house (FH). Biochemical methane potential (BMP) tests were performed according to international standard procedures. The settled sludge fraction comprised 20-30% of the raw manure volume, which comprises 40-60% of the total methane yield. The methane potential of the settled sludge fraction was approximately two times higher than the methane potential of the supernatant fraction. The biogas yield differed among the raw manures from different swine production phases (GSH-a 326.4 and GSH-b 577.1; FSH-a 860.1 and FSH-b 479.2; NH -970.2; FH 474.5 NmLbiogas.gVS(-1)). The differences were relative to the production phase (feed type and feeding techniques) and the management of the effluent inside the facilities (water management). Brazilian swine production has increased his participation in the global market, been the fourth producer and the fourth exporter. The segregation of swine production in multiple sites has increased its importance, due to the possibilities to have more specialized units. Generally, these units

  11. Towards the development of systems for high-yield production of microbial lipases.

    PubMed

    Turki, Saoussen

    2013-10-01

    Microbial lipases are a versatile and attractive class of biocatalysts for a wide variety of applications. Lipases can be produced by bacteria, yeasts or filamentous fungi. Nevertheless, they are often not optimal for direct use in industrial conditions due to low yields, low specific activities and a limited spectrum of activities. Improvements in the productivity of lipases have been made by genetic manipulation of the cell factory production hosts and by optimizing production media and conditions. Advances in protein engineering technology, ranging from directed evolution to rational design, have also been able to tailor lipases to particular applications. This review describes various approaches used to improve lipase production and applications.

  12. Neutron yield study of direct-drive, low-adiabat cryogenic D{sub 2} implosions on OMEGA laser system

    SciTech Connect

    Hu, S. X.; Radha, P. B.; Marozas, J. A.; Betti, R.; Collins, T. J. B.; Craxton, R. S.; Delettrez, J. A.; Edgell, D. H.; Epstein, R.; Goncharov, V. N.; Igumenshchev, I. V.; Marshall, F. J.; McCrory, R. L.; Meyerhofer, D. D.; Regan, S. P.; Sangster, T. C.; Skupsky, S.; Smalyuk, V. A.; Elbaz, Y.; Shvarts, D.

    2009-11-15

    Neutron yields of direct-drive, low-adiabat ({alpha}{approx_equal}2 to 3) cryogenic D{sub 2} target implosions on the OMEGA laser system [T. R. Boehly et al., Opt. Commun. 133, 495 (1997)] have been systematically investigated using the two-dimensional (2D) radiation hydrodynamics code DRACO[P. B. Radha et al., Phys. Plasmas 12, 056307 (2005)]. Low-mode (l{<=}12) perturbations, including initial target offset, ice-layer roughness, and laser-beam power imbalance, were found to be the primary source of yield reduction for thin-shell (5 {mu}m), low-{alpha}, cryogenic targets. The 2D simulations of thin-shell implosions track experimental measurements for different target conditions and peak laser intensities ranging from 2.5x10{sup 14}-6x10{sup 14} W/cm{sup 2}. Simulations indicate that the fusion yield is sensitive to the relative phases between the target offset and the ice-layer perturbations. The results provide a reasonable good guide to understanding the yield degradation in direct-drive, low-adiabat, cryogenic, thin-shell-target implosions. Thick-shell (10 {mu}m) implosions generally give lower yield over clean than low-l-mode DRACO simulation predictions. Simulations including the effect of laser-beam nonuniformities indicate that high-l-mode perturbations caused by laser imprinting further degrade the neutron yield of thick-shell implosions. To study ICF compression physics, these results suggest a target specification with a {<=}30 {mu}m offset and ice-roughness of {sigma}{sub rms}<3 {mu}m are required.

  13. Neutron Yield Study of Direct-Drive, Low-Adiabat Cryogenic D2 Implosions on OMEGA Laser System

    SciTech Connect

    Hu, S.X.; Radha, P.B.; Marozas, J.A.; Betti, R.; Collins, T.J.B.; Craxton, R.S.; Delettrez, J.A.; Edgell, D.H.; Epstein, R.; Goncharov, V.N.; Igumenshchev, I.V.; Marshall, F.J.; McCrory, R.L.; Meyerhofer, D.D.; Regan, S.P.; Sangster, T.C.; Skupsky, S.; Smalyuk, V.A.; Elbaz, Y.; Shvarts, D.

    2009-11-17

    Neutron yields of direct-drive, low-adiabat (alpha ~~ 2 to 3) cryogenic D2 target implosions on the OMEGA laser system [T. R. Boehly et al., Opt. Commun. 133, 495 (1997)] have been systematically investigated using the two-dimensional (2D) radiation hydrodynamics code DRACO [P. B. Radha et al., Phys. Plasmas 12, 056307 (2005)]. Low-mode (ell <- 12) perturbations, including initial target offset, ice-layer roughness, and laser-beam power imbalance, were found to be the primary source of yield reduction for thin-shell (5 um), low-alpha, cryogenic targets. The 2D simulations of thin-shell implosions track experimental measurements for different target conditions and peak laser intensities ranging from 2.5 x 10^14–6 x 10^14 W/cm^2. Simulations indicate that the fusion yield is sensitive to the relative phases between the target offset and the ice-layer perturbations. The results provide a reasonable good guide to understanding the yield degradation in direct-drive, low-adiabat, cryogenic, thin-shell-target implosions. Thick-shell (10 um) implosions generally give lower yield over clean than low-ell-mode DRACO simulation predictions. Simulations including the effect of laser-beam nonuniformities indicate that high-ell-mode perturbations caused by laser imprinting further degrade the neutron yield of thick-shell implosions. To study ICF compression physics, these results suggest a target specification with a <-30 um offset and ice-roughness of sigma_rms < 3 um are required.

  14. Positive feedback system provides efficient and persistent transgene expression.

    PubMed

    Ochiai, Hiroshi; Harashima, Hideyoshi; Kamiya, Hiroyuki

    2010-08-02

    The two-step transcriptional amplification (TSTA) system, using artificial transcription factors, effectively enhances transgene expression. In this study, a TSTA system-based positive feedback system was developed to achieve efficient and persistent transgene expression. A fusion protein of the sequence-specific DNA binding domain of yeast GAL4 and the transcriptional activation domain of herpes simplex virus VP16 (GAL4-VP16) was used as an "activator" to amplify the expression of the luciferase "reporter" gene. It was found that the introduction of five tandem copies of the GAL4 recognition sequence (G5) into both the upstream and downstream regions of the expression cassette synergistically enhanced the transgene expression. The upstream and downstream G5 sequences were introduced into the expression cassette of the activator itself, and into that of the reporter, to form the positive feedback loop that enabled continuous activator expression. This positive feedback system maintained the expression levels of the reporter for 4 days in HeLa cells and for a week in mouse liver, while those from the usual plasmids decreased by 30- and 50-fold, respectively. These results constitute the first evidence that the positive feedback system is a useful method for long-term transgene expression in cultured cells and in vivo. This system would be applicable to gene therapy, in vivo imaging, and biotechnology.

  15. The silver lining of a viral agent: increasing seed yield and harvest index in Arabidopsis by ectopic expression of the potato leaf roll virus movement protein.

    PubMed

    Kronberg, Kristin; Vogel, Florian; Rutten, Twan; Hajirezaei, Mohammed-Reza; Sonnewald, Uwe; Hofius, Daniel

    2007-11-01

    Ectopic expression of viral movement proteins (MPs) has previously been shown to alter plasmodesmata (PD) function and carbon partitioning in transgenic plants, giving rise to the view of PD being dynamic and highly regulated structures that allow resource allocation to be adapted to environmental and developmental needs. However, most work has been restricted to solanaceous species and the potential use of MP expression to improve biomass and yield parameters has not been addressed in detail. Here we demonstrate that MP-mediated modification of PD function can substantially alter assimilate allocation, biomass production, and reproductive growth in Arabidopsis (Arabidopsis thaliana). These effects were achieved by constitutive expression of the potato leaf roll virus 17-kD MP (MP17) fused to green fluorescent protein (GFP) in different Arabidopsis ecotypes. The resulting transgenic plants were analyzed for PD localization of the MP17:GFP fusion protein and different lines with low to high expression levels were selected for further analysis. Low-level accumulation of MP17 resulted in enhanced sucrose efflux from source leaves and a considerably increased vegetative biomass production. In contrast, high MP17 levels impaired sucrose export, resulting in source leaf-specific carbohydrate accumulation and a strongly reduced vegetative growth. Surprisingly, later during development the MP17-mediated inhibition of resource allocation was reversed, and final seed yield increased in average up to 30% in different transgenic lines as compared to wild-type plants. This resulted in a strongly improved harvest index. The release of the assimilate export block was paralleled by a reduced PD binding of MP17 in senescing leaves, indicating major structural changes of PD during leaf senescence.

  16. Constitutive expression of selected genes from the pentose phosphate and aromatic pathways increases the shikimic acid yield in high-glucose batch cultures of an Escherichia coli strain lacking PTS and pykF

    PubMed Central

    2013-01-01

    Background During the last two decades many efforts have been directed towards obtaining efficient microbial processes for the production of shikimic acid (SA); however, feeding high amounts of substrate to increase the titer of this compound has invariably rendered low conversion yields, leaving room for improvement of the producing strains. In this work we report an alternative platform to overproduce SA in a laboratory-evolved Escherichia coli strain, based on plasmid-driven constitutive expression of six genes selected from the pentose phosphate and aromatic amino acid pathways, artificially arranged as an operon. Production strains also carried inactivated genes coding for phosphotransferase system components (ptsHIcrr), shikimate kinases I and II (aroK and aroL), pyruvate kinase I (pykF) and the lactose operon repressor (lacI). Results The strong and constitutive expression of the constructed operon permitted SA production from the beginning of the cultures, as evidenced in 1 L batch-mode fermentors starting with high concentrations of glucose and yeast extract. Inactivation of the pykF gene improved SA production under the evaluated conditions by increasing the titer, yield and productivity of this metabolite compared to the isogenic pykF+ strain. The best producing strain accumulated up to 43 g/L of SA in 30 h and relatively low concentrations of acetate and aromatic byproducts were detected, with SA accounting for 80% of the produced aromatic compounds. These results were consistent with high expression levels of the glycolytic pathway and synthetic operon genes from the beginning of fermentations, as revealed by transcriptomic analysis. Despite the consumption of 100 g/L of glucose, the yields on glucose of SA and of total aromatic compounds were about 50% and 60% of the theoretical maximum, respectively. The obtained yields and specific production and consumption rates proved to be constant with three different substrate concentrations. Conclusions

  17. Comparative Single-Cell Analysis of Different E. coli Expression Systems during Microfluidic Cultivation

    PubMed Central

    Hilgers, Fabienne; Loeschcke, Anita; Jaeger, Karl-Erich; Kohlheyer, Dietrich; Drepper, Thomas

    2016-01-01

    Recombinant protein production is mostly realized with large-scale cultivations and monitored at the level of the entire population. Detailed knowledge of cell-to-cell variations with respect to cellular growth and product formation is limited, even though phenotypic heterogeneity may distinctly hamper overall production yields, especially for toxic or difficult-to-express proteins. Unraveling phenotypic heterogeneity is thus a key aspect in understanding and optimizing recombinant protein production in biotechnology and synthetic biology. Here, microfluidic single-cell analysis serves as the method of choice to investigate and unmask population heterogeneities in a dynamic and spatiotemporal fashion. In this study, we report on comparative microfluidic single-cell analyses of commonly used E. coli expression systems to uncover system-inherent specifications in the synthetic M9CA growth medium. To this end, the PT7lac/LacI, the PBAD/AraC and the Pm/XylS system were systematically analyzed in order to gain detailed insights into variations of growth behavior and expression phenotypes and thus to uncover individual strengths and deficiencies at the single-cell level. Specifically, we evaluated the impact of different system-specific inducers, inducer concentrations as well as genetic modifications that affect inducer-uptake and regulation of target gene expression on responsiveness and phenotypic heterogeneity. Interestingly, the most frequently applied expression system based on E. coli strain BL21(DE3) clearly fell behind with respect to expression homogeneity and robustness of growth. Moreover, both the choice of inducer and the presence of inducer uptake systems proved crucial for phenotypic heterogeneity. Conclusively, microfluidic evaluation of different inducible E. coli expression systems and setups identified the modified lacY-deficient PT7lac/LacI as well as the Pm/XylS system with conventional m-toluic acid induction as key players for precise and robust

  18. Expression Profiling of Liposarcoma Yields a Multigene Predictor of Patient Outcome and Identifies Genes that Contribute to Liposarcomagenesis

    PubMed Central

    Gobble, Ryan M.; Qin, Li-Xuan; Brill, Elliott R.; Angeles, Christina V.; Ugras, Stacy; O’Connor, Rachael B.; Moraco, Nicole H.; DeCarolis, Penelope L.; Antonescu, Christina; Singer, Samuel

    2011-01-01

    Liposarcomas are the second most common type of soft tissue sarcoma but its genetics are poorly defined. To identify genes that contribute to liposarcomagenesis and serve as prognostic candidates, we undertook expression profiling of 140 primary liposarcoma samples, which were randomly split into training set (n=95) and test set (n=45). A multi-gene predictor for DRFS was developed using the supervised principal component method. Expression levels of the 588 genes in the predictor were used to calculate a risk score for each patient. In validation of the predictor in the test set, patients with low risk score had a 3-year DRFS of 83% vs. 45% for high risk score patients (P=0.001). The hazard ratio for high vs. low score, adjusted for histologic subtype, was 4.42 (95% confidence interval 1.26–15.55; P=0.021). The concordance probability for risk score was 0.732. In contrast, the concordance probability for histologic subtype, which had been considered the best predictor of outcome in liposarcoma, was 0.669. Genes related to adipogenesis, DNA replication, mitosis, and spindle assembly checkpoint control were all highly represented in the multi-gene predictor. Three genes from the predictor, TOP2A, PTK7, and CHEK1, were found to be overexpressed in liposarcoma samples of all five subtypes and in liposarcoma cell lines. RNAi-mediated knockdown of these genes in liposarcoma cell lines reduced proliferation and invasiveness and increased apoptosis. Taken together, our findings identify genes that appear to be involved liposarcomagenesis and have promise as therapeutic targets, and support the use of this multi-gene predictor to improve risk stratification for individual patients with liposarcoma. PMID:21335544

  19. [Effects of maize plant types on dry matter accumulation characteristics and yield of soybean in maize-soybean intercropping systems].

    PubMed

    Cui, Liang; Yang, Wen-yu; Huang, Ni; Liu, Jiang; Wang, Yan-ling; Wang, Xiao-hui; Liu, Yang; Yan, Shou

    2015-08-01

    In order to explore the effects of maize plant types on dry matter accumulation and yield of soybean, a field experiment was conducted in 2013, including three maize-soybean relay strip intercropping systems. The relay strip intercropping systems were designed as soybean (Gongxuan 1) intercropped with Denghai 605 (RI1), Chuandan 418 (RI2) or Yayu 13 ( RI3), and the monocultured soybean was used as control. The results demonstrated that the dry matter accumulation rates of intercropped soybean in RI2 and RI3 treatments were lower than in RI1 treatment, and the leaf, stem and pod dry matter accumulation of intercropped soybean in RI1 treatment was 17.6%, 16.5% and 13.7% higher than that in RI2 treatment, and 34.6%, 33.1% and 28.4% higher than that in RI3 treatment, respectively. The distribution proportion of leaf and stem of intercropped soybean was in the order of RI1 > RI2 > RI3. However, the trend of the distribution proportion of pod was opposite. Compared with RI2 and RI3, the dry matter translocation amount, translocation proportion, contribution proportion of soybean vegetative organs to pod of soybean were improved in RI, treatment, and the pod per plant, seeds per plant, seeds per pod, yield per plant and yield of soybean in RI, were higher than RI2 and RI3 by 6.8%, 11.5%, 4.4%, 15.9%, 15.6% and 14.3%, 22.2%, 6.7%, 33.4%, 36.8%, respectively. The results showed that the yield was positively related with the accumulation rate of dry matter, dry matter translocation, dry matter translocation ratio and the contribution of dry matter accumulation, and these indices were highest in RI treatment. The results indicated that the compact maize relay intercropped with soybean could effectively regulate the dry matter accumulation, translocation and distribution, and improve the yield of soybean.

  20. Regulated expression of a cytokinin biosynthesis gene IPT delays leaf senescence and improves yield under rainfed and irrigated conditions in canola (Brassica napus L.).

    PubMed

    Kant, Surya; Burch, David; Badenhorst, Pieter; Palanisamy, Rajasekaran; Mason, John; Spangenberg, German

    2015-01-01

    Delay of leaf senescence through genetic modification can potentially improve crop yield, through maintenance of photosynthetically active leaves for a longer period. Plant growth hormones such as cytokinin regulate and delay leaf senescence. Here, the structural gene (IPT) encoding the cytokinin biosynthetic enzyme isopentenyltransferase was fused to a functionally active fragment of the AtMYB32 promoter and was transformed into canola plants. Expression of the AtMYB32xs::IPT gene cassette delayed the leaf senescence in transgenic plants grown under controlled environment conditions and field experiments conducted for a single season at two geographic locations. The transgenic canola plants retained higher chlorophyll levels for an extended period and produced significantly higher seed yield with similar growth and phenology compared to wild type and null control plants under rainfed and irrigated treatments. The yield increase in transgenic plants was in the range of 16% to 23% and 7% to 16% under rainfed and irrigated conditions, respectively, compared to control plants. Most of the seed quality parameters in transgenic plants were similar, and with elevated oleic acid content in all transgenic lines and higher oil content and lower glucosinolate content in one specific transgenic line as compared to control plants. The results suggest that by delaying leaf senescence using the AtMYB32xs::IPT technology, productivity in crop plants can be improved under water stress and well-watered conditions.

  1. Develop a field grid system for yield mapping and machine control. Quarterly report, July 1, 1995--September 30, 1995

    SciTech Connect

    Hart, F.; Windish, J.

    1995-10-01

    Build and test the Field Grid Sense system for yield mapping and machine control during harvesting. Secondly, use Field Grid Sense with chemical application equipment to demonstrate a workable in-field system. More specifically, the operation of the patented hardware/software Field Grid Sense (FGS) system will be tested in crop harvesting to demonstrate the system`s utility and to analyze the flexibility of operation under true field conditions. Additionally, FGS will again be used with chemical application equipment - equipment that needs modification to correct one or two slight shortcomings. This action will create improved systems and establish the worthiness, efficiency and necessity of chemical application equipment that is controlled and directed via the FGS package.

  2. Improving potato cropping systems: longer-term effects on diseases and yield

    USDA-ARS?s Scientific Manuscript database

    The development of effective cropping systems can provide the structural basis for enhanced crop production and sustainability through the conservation, maintenance, and replenishment of various soil resources. In 2004, field trials evaluating potato cropping systems focused on different specific cr...

  3. Rectal site and suboptimal nodal yield predict systemic recurrence in resected colorectal carcinoma: a case-control study.

    PubMed

    Hayes, Brian D; O'Riordan, James M; Stuart, Charlotte; Muldoon, Cian

    2014-09-01

    We assessed the contribution of histopathological features to systemic recurrence (SR) in patients with colorectal cancer, using a case-control design: 71 cases and 184 controls were included, with a mean time until SR of 1.4 ± 0.1 years and a mean follow-up of controls of 1.6 ± 0.06 years. Cases had significantly greater odds of rectal site (odds ratio [OR] = 1.82), stage ≥ pT3 (OR = 2.11), suboptimal (<12) lymph node yield (OR = 4.6), stage ≥ pN1 (OR = 2.46), KRAS mutation (OR = 2.76), and extramural venous invasion (OR = 1.97). By multiple regression analysis, rectal site, stage ≥ pT3, suboptimal lymph node yield, and lymph node positivity independently predicted SR. Rectal cancers were more likely to have a suboptimal node yield than nonrectal cancers (relative risk = 1.6) among the entire cohort. We conclude that rectal cancers have greater risk of SR than colon cancers. A lower yield of lymph nodes in rectal cancer specimens may contribute to this.

  4. Homologous expression of cytosolic dehydroascorbate reductase increases grain yield and biomass under paddy field conditions in transgenic rice (Oryza sativa L. japonica).

    PubMed

    Kim, Young-Saeng; Kim, Il-Sup; Bae, Mi-Jung; Choe, Yong-Hoe; Kim, Yul-Ho; Park, Hyang-Mi; Kang, Hong-Gyu; Yoon, Ho-Sung

    2013-06-01

    Dehydroascorbate reductase (DHAR, EC 1.8.5.1) maintains redox pools of ascorbate (AsA) by recycling oxidized AsA to reduced AsA. To investigate whether DHAR affects rice yield under normal environmental conditions, cDNA-encoding DHAR (OsDHAR1) was isolated from rice and used to develop OsDHAR1-overexpressing transgenic rice plants, under the regulation of a maize ubiquitin promoter. Incorporation and expression of the transgene in transgenic rice plants was confirmed by genomic polymerase chain reaction (PCR), semi-quantitative reverse transcription PCR (RT-PCR), western blot, and enzyme activity. The expression levels were at least twofold higher in transgenic (TG) rice plants than in control wild-type (WT) rice plants. In addition, OsDHAR1-overexpression in seven-independent homologous transgenic plants, as compared to WT plants, increased photosynthetic capacity and antioxidant enzyme activities under paddy field conditions, which led to an improved AsA pool and redox homeostasis. Furthermore, OsDHAR1 overexpression significantly improved grain yield and biomass due to the increase of culm and root weights and to enhance panicle and spikelet numbers in the same seven independent TG rice plants during the farming season (2010 and 2011) in South Korea. The OsDHAR protein contained the redox-active site (Cys20), as well as the conserved GSH-binding region, GSH-binding motif, glutathione-S-transferase (GST) N-terminal domain, C-terminal domain interface, and GST C-terminal domain. Therefore, our results indicate that OsDHAR1 overexpression, capable of functioning in AsA recycling, and protein folding increases environmental adaptation to paddy field conditions by the improving AsA pool and redox homeostasis, which enhances rice grain yield and biomass.

  5. Expression of functional Plasmodium falciparum enzymes using a wheat germ cell-free system.

    PubMed

    Mudeppa, Devaraja G; Rathod, Pradipsinh K

    2013-12-01

    One decade after the sequencing of the Plasmodium falciparum genome, 95% of malaria proteins in the genome cannot be expressed in traditional cell-based expression systems, and the targets of the best new leads for antimalarial drug discovery are either not known or not available in functional form. For a disease that kills up to 1 million people per year, routine expression of recombinant malaria proteins in functional form is needed both for the discovery of new therapeutics and for identification of targets of new drugs. We tested the general utility of cell-free systems for expressing malaria enzymes. Thirteen test enzyme sequences were reverse amplified from total RNA, cloned into a plant-like expression vector, and subjected to cell-free expression in a wheat germ system. Protein electrophoresis and autoradiography confirmed the synthesis of products of expected molecular masses. In rare problematic cases, truncated products were avoided by using synthetic genes carrying wheat codons. Scaled-up production generated 39 to 354 μg of soluble protein per 10 mg of translation lysate. Compared to rare proteins where cell-based systems do produce functional proteins, the cell-free yields are comparable or better. All 13 test products were enzymatically active, without failure. This general path to produce functional malaria proteins should now allow the community to access new tools, such as biologically active protein arrays, and lead to the discovery of new chemical functions, structures, and inhibitors of previously inaccessible malaria gene products.

  6. A Comparative Analysis of Recombinant Protein Expression in Different Biofactories: Bacteria, Insect Cells and Plant Systems

    PubMed Central

    Brozzetti, Annalisa; Falorni, Alberto; Pezzotti, Mario; Avesani, Linda

    2015-01-01

    Plant-based systems are considered a valuable platform for the production of recombinant proteins as a result of their well-documented potential for the flexible, low-cost production of high-quality, bioactive products. In this study, we compared the expression of a target human recombinant protein in traditional fermenter-based cell cultures (bacterial and insect) with plant-based expression systems, both transient and stable. For each platform, we described the set-up, optimization and length of the production process, the final product quality and the yields and we evaluated provisional production costs, specific for the selected target recombinant protein. Overall, our results indicate that bacteria are unsuitable for the production of the target protein due to its accumulation within insoluble inclusion bodies. On the other hand, plant-based systems are versatile platforms that allow the production of the selected protein at lower-costs than Baculovirus/insect cell system. In particular, stable transgenic lines displayed the highest-yield of the final product and transient expressing plants the fastest process development. However, not all recombinant proteins may benefit from plant-based systems but the best production platform should be determined empirically with a case-by-case approach, as described here. PMID:25867956

  7. An IPTG Inducible Conditional Expression System for Mycobacteria

    PubMed Central

    Ravishankar, Sudha; Ambady, Anisha; Ramu, Haripriya; Mudugal, Naina Vinay; Tunduguru, Ragadeepthi; Anbarasu, Anand; Sharma, Umender K.; Sambandamurthy, Vasan K.; Ramaiah, Sudha

    2015-01-01

    Conditional expression strains serve as a valuable tool to study the essentiality and to establish the vulnerability of a target under investigation in a drug discovery program. While essentiality implies an absolute requirement of a target function, vulnerability provides valuable information on the extent to which a target function needs to be depleted to achieve bacterial growth inhibition followed by cell death. The critical feature of an ideal conditional expression system is its ability to tightly regulate gene expression to achieve the full spectrum spanning from a high level of expression in order to support growth and near zero level of expression to mimic conditions of gene knockout. A number of bacterial conditional expression systems have been reported for use in mycobacteria. The utility of an isopropylthiogalactoside (IPTG) inducible system in mycobacteria has been reported for protein overexpression and anti-sense gene expression from a replicating multi-copy plasmid. Herein, we report the development of a versatile set of non-replicating IPTG inducible vectors for mycobacteria which can be used for generation of conditional expression strains through homologous recombination. The role of a single lac operator versus a double lac operator to regulate gene expression was evaluated by monitoring the expression levels of β-galactosidase in Mycobacterium smegmatis. These studies indicated a significant level of leaky expression from the vector with a single lac operator but none from the vector with double lac operator. The significance of the double lac operator vector for target validation was established by monitoring the growth kinetics of an inhA, a rpoB and a ftsZ conditional expression strain grown in the presence of different concentrations of IPTG. The utility of this inducible system in identifying target specific inhibitors was established by screening a focussed library of small molecules using an inhA and a rpoB conditional expression

  8. Solid-phase fermentation and juice expression systems for sweet sorghum

    SciTech Connect

    Bryan, W.L.; Monroe, G.E.; Caussariel, P.M.

    1985-01-01

    Two systems to recover fermented juice from variety M 81E sweet sorghum stalks that contained about 11% fermentable sugar were compared. (a) Stalks with leaves and tops removed were chopped and inoculated with 0.2% yeast in a forage harvester, stored under anaerobic conditions for 75 hours in insulated fermentors and pressed in a screw press to recover fermented juice (5-6% ethanol). (b) Mechanically harvested sweet sorghum billets (30 cm length) without leaves or seed heads were shredded and milled in a 3-roll mill; and bagasse was inoculated with 0.2% yeast, fermented for 100 h and pressed to recover fermented juice (4 to 5% ethanol). Potential ethanol yields were 75% of theoretical for the forage harvest system and 78% for the shredder mill system, based on 95% of theoretical ethanol yield from juice expressed during milling and no loss of ethanol during fermentation, handling and pressing in the screw press. 20 references.

  9. Decomposing global crop yield variability

    NASA Astrophysics Data System (ADS)

    Ben-Ari, Tamara; Makowski, David

    2014-11-01

    Recent food crises have highlighted the need to better understand the between-year variability of agricultural production. Although increasing future production seems necessary, the globalization of commodity markets suggests that the food system would also benefit from enhanced supplies stability through a reduction in the year-to-year variability. Here, we develop an analytical expression decomposing global crop yield interannual variability into three informative components that quantify how evenly are croplands distributed in the world, the proportion of cultivated areas allocated to regions of above or below average variability and the covariation between yields in distinct world regions. This decomposition is used to identify drivers of interannual yield variations for four major crops (i.e., maize, rice, soybean and wheat) over the period 1961-2012. We show that maize production is fairly spread but marked by one prominent region with high levels of crop yield interannual variability (which encompasses the North American corn belt in the USA, and Canada). In contrast, global rice yields have a small variability because, although spatially concentrated, much of the production is located in regions of below-average variability (i.e., South, Eastern and South Eastern Asia). Because of these contrasted land use allocations, an even cultivated land distribution across regions would reduce global maize yield variance, but increase the variance of global yield rice. Intermediate results are obtained for soybean and wheat for which croplands are mainly located in regions with close-to-average variability. At the scale of large world regions, we find that covariances of regional yields have a negligible contribution to global yield variance. The proposed decomposition could be applied at any spatial and time scales, including the yearly time step. By addressing global crop production stability (or lack thereof) our results contribute to the understanding of a key

  10. Stormwater treatment: do constructed wetlands yield improved pollutant management performance over a detention pond system?

    PubMed

    Bavor, H J; Davies, C M; Sakadevan, K

    2001-01-01

    Constructed wetland systems have been proposed as representing an improved ecotechnological option over detention basins, in terms of their abilities to reduce stormwater bacterial and nutrient loads to receiving waters. Concentrations of microbial and pollutants were determined in inflow and outflow samples collected from each type of system. Removal efficiencies for the wetland although higher than for the pond, were lower than some previously reported values for the treatment of municipal wastewater by constructed wetlands. Performance of a number of constructed wetland systems for stormwater treatment is evaluated considering the functional components of the systems.

  11. Optimizing Hill Seeding Density for High-Yielding Hybrid Rice in a Single Rice Cropping System in South China

    PubMed Central

    Wang, Danying; Chen, Song; Wang, Zaiman; Ji, Chenglin; Xu, Chunmei; Zhang, Xiufu; Chauhan, Bhagirath Singh

    2014-01-01

    Mechanical hill direct seeding of hybrid rice could be the way to solve the problems of high seeding rates and uneven plant establishment now faced in direct seeded rice; however, it is not clear what the optimum hill seeding density should be for high-yielding hybrid rice in the single-season rice production system. Experiments were conducted in 2010 and 2011 to determine the effects of hill seeding density (25 cm×15 cm, 25 cm×17 cm, 25 cm×19 cm, 25 cm×21 cm, and 25 cm×23 cm; three to five seeds per hill) on plant growth and grain yield of a hybrid variety, Nei2you6, in two fields with different fertility (soil fertility 1 and 2). In addition, in 2012 and 2013, comparisons among mechanical hill seeding, broadcasting, and transplanting were conducted with three hybrid varieties to evaluate the optimum seeding density. With increases in seeding spacing from 25 cm×15 cm to 25 cm×23 cm, productive tillers per hill increased by 34.2% and 50.0% in soil fertility 1 and 2. Panicles per m2 declined with increases in seeding spacing in soil fertility 1. In soil fertility 2, no difference in panicles per m2 was found at spacing ranging from 25 cm×17 cm to 25 cm×23 cm, while decreases in the area of the top three leaves and aboveground dry weight per shoot at flowering were observed. Grain yield was the maximum at 25 cm×17 cm spacing in both soil fertility fields. Our results suggest that a seeding density of 25 cm×17 cm was suitable for high-yielding hybrid rice. These results were verified through on-farm demonstration experiments, in which mechanical hill-seeded rice at this density had equal or higher grain yield than transplanted rice. PMID:25290342

  12. Optimizing hill seeding density for high-yielding hybrid rice in a single rice cropping system in South China.

    PubMed

    Wang, Danying; Chen, Song; Wang, Zaiman; Ji, Chenglin; Xu, Chunmei; Zhang, Xiufu; Chauhan, Bhagirath Singh

    2014-01-01

    Mechanical hill direct seeding of hybrid rice could be the way to solve the problems of high seeding rates and uneven plant establishment now faced in direct seeded rice; however, it is not clear what the optimum hill seeding density should be for high-yielding hybrid rice in the single-season rice production system. Experiments were conducted in 2010 and 2011 to determine the effects of hill seeding density (25 cm×15 cm, 25 cm×17 cm, 25 cm×19 cm, 25 cm×21 cm, and 25 cm×23 cm; three to five seeds per hill) on plant growth and grain yield of a hybrid variety, Nei2you6, in two fields with different fertility (soil fertility 1 and 2). In addition, in 2012 and 2013, comparisons among mechanical hill seeding, broadcasting, and transplanting were conducted with three hybrid varieties to evaluate the optimum seeding density. With increases in seeding spacing from 25 cm×15 cm to 25 cm×23 cm, productive tillers per hill increased by 34.2% and 50.0% in soil fertility 1 and 2. Panicles per m2 declined with increases in seeding spacing in soil fertility 1. In soil fertility 2, no difference in panicles per m2 was found at spacing ranging from 25 cm×17 cm to 25 cm×23 cm, while decreases in the area of the top three leaves and aboveground dry weight per shoot at flowering were observed. Grain yield was the maximum at 25 cm×17 cm spacing in both soil fertility fields. Our results suggest that a seeding density of 25 cm×17 cm was suitable for high-yielding hybrid rice. These results were verified through on-farm demonstration experiments, in which mechanical hill-seeded rice at this density had equal or higher grain yield than transplanted rice.

  13. Yeast-expressed human membrane protein aquaporin-1 yields excellent resolution of solid-state MAS NMR spectra.

    PubMed

    Emami, Sanaz; Fan, Ying; Munro, Rachel; Ladizhansky, Vladimir; Brown, Leonid S

    2013-02-01

    One of the biggest challenges in solid-state NMR studies of membrane proteins is to obtain a homogeneous natively folded sample giving high spectral resolution sufficient for structural studies. Eukaryotic membrane proteins are especially difficult and expensive targets in this respect. Methylotrophic yeast Pichia pastoris is a reliable producer of eukaryotic membrane proteins for crystallography and a promising economical source of isotopically labeled proteins for NMR. We show that eukaryotic membrane protein human aquaporin 1 can be doubly ((13)C/(15)N) isotopically labeled in this system and functionally reconstituted into phospholipids, giving excellent resolution of solid-state magic angle spinning NMR spectra.

  14. Current status of viral expression systems in plants and perspectives for oral vaccines development.

    PubMed

    Salazar-González, Jorge A; Bañuelos-Hernández, Bernardo; Rosales-Mendoza, Sergio

    2015-02-01

    During the last 25 years, the technology to produce recombinant vaccines in plant cells has evolved from modest proofs of the concept to viable technologies adopted by some companies due to significant improvements in the field. Viral-based expression strategies have importantly contributed to this success owing to high yields, short production time (which is in most cases free of tissue culture steps), and the implementation of confined processes for production under GMPs. Herein the distinct expression systems based on viral elements are analyzed. This review also presents the outlook on how these technologies have been successfully applied to the development of plant-based vaccines, some of them being in advanced stages of development. Perspectives on how viral expression systems could allow for the development of innovative oral vaccines constituted by minimally-processed plant biomass are discussed.

  15. An Individual-Tree Growth and Yield Prediction System for Uneven-Aged Shortleaf Pine Stands

    Treesearch

    Michael M. Huebschmann; Lawrence R. Gering; Thomas B. Lynch; Onesphore Bitoki; Paul A. Murphy

    2000-01-01

    A system of equations modeling the growth and development of uneven-aged shortleaf pine (Pinus echinata Mill.) stands is described. The prediction system consists of two main components: (1) a distance-independent, individual-tree simulator containing equations that forecast ingrowth, basal-area growth, probability of survival, total and...

  16. Nitrous oxide, methane emission, and yield-scaled emission from organically and conventionally managed systems

    USDA-ARS?s Scientific Manuscript database

    There is a gap in empirical greenhouse gas (GHG) data from many regions of the USA including the northern Corn Belt. This study compared nitrous oxide (N2O) emission in diverse, low-tillage conventionally- and organically-managed systems. Both systems had a four-year rotation, mole-knife, strip till...

  17. Dependency of global primary bioenergy crop potentials in 2050 on food systems, yields, biodiversity conservation and political stability.

    PubMed

    Erb, Karl-Heinz; Haberl, Helmut; Plutzar, Christoph

    2012-08-01

    The future bioenergy crop potential depends on (1) changes in the food system (food demand, agricultural technology), (2) political stability and investment security, (3) biodiversity conservation, (4) avoidance of long carbon payback times from deforestation, and (5) energy crop yields. Using a biophysical biomass-balance model, we analyze how these factors affect global primary bioenergy potentials in 2050. The model calculates biomass supply and demand balances for eleven world regions, eleven food categories, seven food crop types and two livestock categories, integrating agricultural forecasts and scenarios with a consistent global land use and NPP database. The TREND scenario results in a global primary bioenergy potential of 77 EJ/yr, alternative assumptions on food-system changes result in a range of 26-141 EJ/yr. Exclusion of areas for biodiversity conservation and inaccessible land in failed states reduces the bioenergy potential by up to 45%. Optimistic assumptions on future energy crop yields increase the potential by up to 48%, while pessimistic assumptions lower the potential by 26%. We conclude that the design of sustainable bioenergy crop production policies needs to resolve difficult trade-offs such as food vs. energy supply, renewable energy vs. biodiversity conservation or yield growth vs. reduction of environmental problems of intensive agriculture.

  18. Assimilating Remote Sensing Observations of Leaf Area Index and Soil Moisture for Wheat Yield Estimates: An Observing System Simulation Experiment

    NASA Technical Reports Server (NTRS)

    Nearing, Grey S.; Crow, Wade T.; Thorp, Kelly R.; Moran, Mary S.; Reichle, Rolf H.; Gupta, Hoshin V.

    2012-01-01

    Observing system simulation experiments were used to investigate ensemble Bayesian state updating data assimilation of observations of leaf area index (LAI) and soil moisture (theta) for the purpose of improving single-season wheat yield estimates with the Decision Support System for Agrotechnology Transfer (DSSAT) CropSim-Ceres model. Assimilation was conducted in an energy-limited environment and a water-limited environment. Modeling uncertainty was prescribed to weather inputs, soil parameters and initial conditions, and cultivar parameters and through perturbations to model state transition equations. The ensemble Kalman filter and the sequential importance resampling filter were tested for the ability to attenuate effects of these types of uncertainty on yield estimates. LAI and theta observations were synthesized according to characteristics of existing remote sensing data, and effects of observation error were tested. Results indicate that the potential for assimilation to improve end-of-season yield estimates is low. Limitations are due to a lack of root zone soil moisture information, error in LAI observations, and a lack of correlation between leaf and grain growth.

  19. Dependency of global primary bioenergy crop potentials in 2050 on food systems, yields, biodiversity conservation and political stability

    PubMed Central

    Erb, Karl-Heinz; Haberl, Helmut; Plutzar, Christoph

    2012-01-01

    The future bioenergy crop potential depends on (1) changes in the food system (food demand, agricultural technology), (2) political stability and investment security, (3) biodiversity conservation, (4) avoidance of long carbon payback times from deforestation, and (5) energy crop yields. Using a biophysical biomass-balance model, we analyze how these factors affect global primary bioenergy potentials in 2050. The model calculates biomass supply and demand balances for eleven world regions, eleven food categories, seven food crop types and two livestock categories, integrating agricultural forecasts and scenarios with a consistent global land use and NPP database. The TREND scenario results in a global primary bioenergy potential of 77 EJ/yr, alternative assumptions on food-system changes result in a range of 26–141 EJ/yr. Exclusion of areas for biodiversity conservation and inaccessible land in failed states reduces the bioenergy potential by up to 45%. Optimistic assumptions on future energy crop yields increase the potential by up to 48%, while pessimistic assumptions lower the potential by 26%. We conclude that the design of sustainable bioenergy crop production policies needs to resolve difficult trade-offs such as food vs. energy supply, renewable energy vs. biodiversity conservation or yield growth vs. reduction of environmental problems of intensive agriculture. PMID:23576836

  20. Maintaining yields and reducing nitrogen loss in rice-wheat rotation system in Taihu Lake region with proper fertilizer management

    NASA Astrophysics Data System (ADS)

    Xue, Lihong; Yu, Yingliang; Yang, Linzhang

    2014-11-01

    In the Tailake region of China, heavy nitrogen (N) loss of rice-wheat rotation systems, due to high fertilizer-N input with low N use efficiency (NUE), was widely reported. To alleviate the detrimental impacts caused by N loss, it is necessary to improve the fertilizer management practices. Therefore, a 3 yr field experiments with different N managements including organic combined chemical N treatment (OCN, 390 kg N ha-1 yr-1, 20% organic fertilizer), control-released urea treatment (CRU, 390 kg N ha-1 yr-1, 70% resin-coated urea), reduced chemical N treatment (RCN, 390 kg N ha-1 yr-1, all common chemical fertilizer), and site-specific N management (SSNM, 333 kg N ha-1 yr-1, all common chemical fertilizer) were conducted in the Taihu Lake region with the ‘farmer’s N’ treatment (FN, 510 kg N ha-1 yr-1, all common chemical fertilizer) as a control. Grain yield, plant N uptake (PNU), NUE, and N losses via runoff, leaching, and ammonia volatilization were assessed. In the rice season, the FN treatment had the highest N loss and lowest NUE, which can be attributed to an excessive rate of N application. Treatments of OCN and RCN with a 22% reduced N rate from FN had no significant effect on PNU nor the yield of rice in the 3 yr; however, the NUE was improved and N loss was reduced 20-32%. OCN treatment achieved the highest yield, while SSNM has the lowest N loss and highest NUE due to the lowest N rate. In wheat season, N loss decreased about 28-48% with the continuous reduction of N input, but the yield also declined, with the exception of OCN treatment. N loss through runoff, leaching and ammonia volatilization was positively correlated with the N input rate. When compared with the pure chemical fertilizer treatment of RCN under the same N input, OCN treatment has better NUE, better yield, and lower N loss. 70% of the urea replaced with resin-coated urea had no significant effect on yield and NUE improvement, but decreased the ammonia volatilization loss. Soil

  1. Current warming will reduce yields unless maize breeding and seed systems adapt immediately

    NASA Astrophysics Data System (ADS)

    Challinor, A. J.; Koehler, A.-K.; Ramirez-Villegas, J.; Whitfield, S.; Das, B.

    2016-10-01

    The development of crop varieties that are better suited to new climatic conditions is vital for future food production. Increases in mean temperature accelerate crop development, resulting in shorter crop durations and reduced time to accumulate biomass and yield. The process of breeding, delivery and adoption (BDA) of new maize varieties can take up to 30 years. Here, we assess for the first time the implications of warming during the BDA process by using five bias-corrected global climate models and four representative concentration pathways with realistic scenarios of maize BDA times in Africa. The results show that the projected difference in temperature between the start and end of the maize BDA cycle results in shorter crop durations that are outside current variability. Both adaptation and mitigation can reduce duration loss. In particular, climate projections have the potential to provide target elevated temperatures for breeding. Whilst options for reducing BDA time are highly context dependent, common threads include improved recording and sharing of data across regions for the whole BDA cycle, streamlining of regulation, and capacity building. Finally, we show that the results have implications for maize across the tropics, where similar shortening of duration is projected.

  2. Whole intact rapeseeds or sunflower oil in high-forage or high-concentrate diets affects milk yield, milk composition, and mammary gene expression profile in goats.

    PubMed

    Ollier, S; Leroux, C; de la Foye, A; Bernard, L; Rouel, J; Chilliard, Y

    2009-11-01

    This study aimed to ascertain the response of goat mammary metabolic pathways to concentrate and lipid feeding in relation to milk fatty acid (FA) composition and secretion. Sixteen midlactation multiparous goats received diets differing in forage-to-concentrate ratio [high forage (HF) 64:36, and low forage (LF) 43:57] supplemented or not with lipids [HF with 130 g/d of oil from whole intact rapeseeds (RS) and LF with 130 g/d of sunflower oil (SO)] in a 4 x 4 Latin square design. Milk yield, milk composition, FA profile, and FA secretion were measured, as well as the expression profiles of key genes in mammary metabolism and of 8,382 genes, using a bovine oligonucleotide microarray. After 3 wk of treatment, milk, lactose, and protein yields were lower with HF-RS than with the other diets, whereas treatment had no effect on milk protein content. Milk fat content was higher with the HF-RS and LF-SO diets than with the HF and LF diets, and SO supplementation increased milk fat yield compared with the LF diet. Decreasing the forage-to-concentrate ratio from 64:36 to 43:57 had a limited effect on goat milk FA concentrations and secretions. Supplementing the LF diet with SO changed almost all the FA concentrations, including decreases in medium-chain saturated FA and large increases in trans C18:1 and C18:2 isomers (particularly trans-11 C18:1 and cis-9, trans-11 conjugated linoleic acid), without significant changes in C18:0 and cis-9 C18:1, whereas supplementing the HF diet with RS led to a strong decrease in short- and medium-chain saturated FA and a very strong increase in C18:0 and cis-9 C18:1, without significant changes in trans C18:1 and conjugated linoleic acid. Despite the decreases in milk lactose and protein yields observed with HF-RS, and despite the decrease in milk medium-chain FA and the increase in C18 FA secretion with RS or SO supplementation, none of the dietary treatments had any effect on mammary mRNA expression of the key genes involved in lactose

  3. Ethylene stimulation of latex yield depends on the expression of a sucrose transporter (HbSUT1B) in rubber tree (Hevea brasiliensis).

    PubMed

    Dusotoit-Coucaud, Anaïs; Kongsawadworakul, Panida; Maurousset, Laurence; Viboonjun, Unshira; Brunel, Nicole; Pujade-Renaud, Valérie; Chrestin, Hervé; Sakr, Soulaïman

    2010-12-01

    Hevea brasiliensis is an important industrial crop for natural rubber production. Latex biosynthesis occurs in the cytoplasm of highly specialized latex cells and requires sucrose as the unique precursor. Ethylene stimulation of latex production results in high sugar flow from the surrounding cells of inner bark towards the latex cells. The aim of this work was to understand the role of seven sucrose transporters (HbSUTs) and one hexose transporter (HbHXT1) in this process. Two Hevea clones were used: PB217 and PB260, respectively described as high and low yielding clones. The expression pattern of these sugar transporters (HbSUTs and HbHXT1) was monitored under different physiological conditions and found to be maximal in latex cells. HbSUT1, one of the most abundant isoforms, displayed the greatest response to ethylene treatment. In clone PB217, ethylene treatment led to a higher accumulation of HbSUT1B in latex cells than in the inner bark tissues. Conversely, stronger expression of HbSUT1B was observed in inner bark tissues than in latex cells of PB260. A positive correlation with HbSUT1B transcript accumulation and increased latex production was further supported by its lower expression in latex cells of the virgin clone PB217.

  4. Row Spacing, Tillage System, and Herbicide Technology Affects Cotton Plant Growth and Yield

    USDA-ARS?s Scientific Manuscript database

    Cotton (Gossypium hirsutum L.) producers are faced with numerous production choices including cotton varieties, herbicide technology, tillage systems, and row spacing. A study was conducted to compare cotton production across conventional, glyphosate tolerant, and glufosinate tolerant varieties in ...

  5. Crop diversification, tillage, and management system influences on spring wheat yield and soil water use

    USDA-ARS?s Scientific Manuscript database

    Depleted soil quality, decreased water availability, and increased weed competition constrain spring wheat production in the northern Great Plains. Integrated crop management systems are necessary for improved crop productivity. We conducted a field experiment from 2004-2010 comparing productivity...

  6. Wheat germ systems for cell-free protein expression.

    PubMed

    Harbers, Matthias

    2014-08-25

    Cell-free protein expression plays an important role in biochemical research. However, only recent developments led to new methods to rapidly synthesize preparative amounts of protein that make cell-free protein expression an attractive alternative to cell-based methods. In particular the wheat germ system provides the highest translation efficiency among eukaryotic cell-free protein expression approaches and has a very high success rate for the expression of soluble proteins of good quality. As an open in vitro method, the wheat germ system is a preferable choice for many applications in protein research including options for protein labeling and the expression of difficult-to-express proteins like membrane proteins and multiple protein complexes. Here I describe wheat germ cell-free protein expression systems and give examples how they have been used in genome-wide expression studies, preparation of labeled proteins for structural genomics and protein mass spectroscopy, automated protein synthesis, and screening of enzymatic activities. Future directions for the use of cell-free expression methods are discussed.

  7. Relationships between Written Expression Achievement and the Cognitive Assessment System.

    ERIC Educational Resources Information Center

    Johnson, Judy A.; Bardos, Achilles N.; Tayebi, Kandi A.

    The purpose of this study was to explore the relationships between the Cognitive Assessment System (CAS) and writing achievement in junior high students (aged 11-15 years) with and without written expression disabilities. Ninety-six students with (n=48) and without (n=48) written expression disabilities were administered the Das-Naglieri:…

  8. The Facial Expression Coding System (FACES): Development, Validation, and Utility

    ERIC Educational Resources Information Center

    Kring, Ann M.; Sloan, Denise M.

    2007-01-01

    This article presents information on the development and validation of the Facial Expression Coding System (FACES; A. M. Kring & D. Sloan, 1991). Grounded in a dimensional model of emotion, FACES provides information on the valence (positive, negative) of facial expressive behavior. In 5 studies, reliability and validity data from 13 diverse…

  9. Local and Systemic Cytokine Expression in Patients with Postherpetic Neuralgia

    PubMed Central

    Üçeyler, Nurcan; Valet, Michael; Kafke, Waldemar; Tölle, Thomas R.; Sommer, Claudia

    2014-01-01

    Background Postherpetic neuralgia (PHN) is the painful complication of a varicella zoster virus reactivation. We investigated the systemic and local gene expression of pro- and anti-inflammatory cytokine expression in patients with PHN. Methods Thirteen patients with PHN at the torso (Th4-S1) were recruited. Skin punch biopsies were obtained from the painful and the contralateral painless body area for intraepidermal nerve fiber density (IENFD) and cytokine profiling. Additionally, blood was withdrawn for systemic cytokine expression and compared to blood values of healthy controls. We analyzed the gene expression of selected pro- and anti-inflammatory cytokines (tumor necrosis factor-alpha [TNF] and interleukins [IL]-1β, IL-2, and IL-8). Results IENFD was lower in affected skin compared to unaffected skin (p<0.05), while local gene expression of pro- and anti-inflammatory cytokines did not differ except for two patients who had 7fold higher IL-6 and 10fold higher IL-10 gene expression in the affected skin compared to the contralateral unaffected skin sample. Also, the systemic expression of cytokines in patients with PHN and in healthy controls was similar. Conclusion While the systemic and local expression of the investigated pro- and anti-inflammatory cytokines was not different from controls, this may have been influenced by study limitations like the low number of patients and different disease durations. Furthermore, other cytokines or pain mediators need to be considered. PMID:25127283

  10. The Facial Expression Coding System (FACES): Development, Validation, and Utility

    ERIC Educational Resources Information Center

    Kring, Ann M.; Sloan, Denise M.

    2007-01-01

    This article presents information on the development and validation of the Facial Expression Coding System (FACES; A. M. Kring & D. Sloan, 1991). Grounded in a dimensional model of emotion, FACES provides information on the valence (positive, negative) of facial expressive behavior. In 5 studies, reliability and validity data from 13 diverse…

  11. Expression of monellin in a food-grade delivery system in Saccharomyces cerevisiae.

    PubMed

    Liu, Jun; Yan, Da-zhong; Zhao, Sheng-jun

    2015-10-01

    Genetically modified (GM) foods have caused much controversy. Construction of a food-grade delivery system is a desirable technique with presumptive impact on industrial applications from the perspective of bio-safety. The aim of this study was to construct a food-grade delivery system for Saccharomyces cerevisiae and to study the expression of monellin from the berries of the West African forest plant Dioscoreophyllum cumminsii in this system. A food-grade system for S. cerevisiae was constructed based on ribosomal DNA (rDNA)-mediated homologous recombination to enable high-copy-number integration of the expression cassette inserted into the rDNA locus. A copper resistance gene (CUP1) was used as the selection marker for yeast transformation. Because variants of transformants containing different copy numbers at the CUP1 locus can be readily selected after growth in the presence of elevated copper levels, we suggest that this system would prove useful in the generation of tandemly iterated gene clusters. Using this food-grade system, a single-chain monellin gene was heterologously expressed. The yield of monellin reached a maximum of 675 mg L(-1) . This system harbors exclusively S. cerevisiae DNA with no antibiotic resistance genes, and it should therefore be appropriate for safe use in the food industry. Monellin was shown to be expressed in this food-grade delivery system. To our knowledge, this is the first report so far on expression of monellin in a food-grade expression system in S. cerevisiae. © 2014 Society of Chemical Industry.

  12. Expression of the lysostaphin gene of Staphylococcus simulans in a eukaryotic system.

    PubMed Central

    Williamson, C M; Bramley, A J; Lax, A J

    1994-01-01

    The lysostaphin gene of Staphylococcus simulans was cloned into Escherichia coli. The 5' end of the gene was modified to include a eukaryotic start codon, the Kozak expression start site consensus sequence, and an enzyme site to facilitate manipulation of the gene. Transcription of the modified gene in vitro yielded an RNA transcript which, when added to a rabbit reticulocyte cell-free translation system, directed the synthesis of several products. The largest product, migrating at approximately 93 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was probably preprolysostaphin, since it was cleaved in the presence of an S. simulans culture supernatant to yield a polypeptide of a size similar to that of mature lysostaphin. When canine pancreatic microsomal vesicles were added to the translation system, translocation of the newly synthesized polypeptides occurred, as judged by protection from proteolysis. The gene was also expressed transiently from the human cytomegalovirus promoter in COS-7 cells. Active enzyme could be detected in the cell lysate, and the prokaryotic signal appeared to target secretion of active enzyme to the culture medium. The successful expression of the lysostaphin gene and processing of the precursor to produce active secreted enzyme open up the possibility of controlling staphylococcal mastitis by targeting expression of this gene to the mammary glands of transgenic animals. Images PMID:8161174

  13. Development of an efficient autoinducible expression system by promoter engineering in Bacillus subtilis.

    PubMed

    Guan, Chengran; Cui, Wenjing; Cheng, Jintao; Zhou, Li; Liu, Zhongmei; Zhou, Zhemin

    2016-04-25

    Bacillus subtilis, a Gram-positive organism, has been developed to be an attractive expression platform to produce both secreted and cytoplasmic proteins owing to its prominent biological characteristics. We previously developed an auto-inducible expression system containing the srfA promoter (PsrfA) which was activated by the signal molecules acting in the quorum-sensing pathway for competence. The P srfA promoter exhibited the unique property of inducer-free activity that is closely correlated with cell density. To improve the PsrfA-mediated expression system to the high-cell-density fermentation for industrial production in the B. subtilis mutant strain that is unable to sporulate, a spore mutant strain BSG1682 was developed, and the PsrfA promoter was enhanced by promoter engineering. Using green fluorescent protein (GFP) as the reporter, higher fluorescent intensity was observed in BSG1682 with expression from either plasmid or chromosome than that of the wild type B. subtilis 168. Thereafter, the PsrfA was engineered, yielding a library of PsrfA derivatives varied in the strength of GFP expression. The P23 promoter exhibited the best performance, almost twofold stronger than that of P srfA. Two heterologous proteins, aminopeptidase (AP) and nattokinase (NK), were successfully overproduced under the control of P23 in BSG1682. Finally, the capacity of the expression system was demonstrated in batch fermentation in a 5-L fermenter. The expression system demonstrates prominence in the activity of the auto-inducible promoter. Desired proteins could be highly and stably produced by integrating the corresponding genes downstream of the promoter on the plasmid or the chromosome in strain BSG1682. The expression system is conducive to the industrial production of pharmaceuticals and heterologous proteins in high-cell-density fermentation in BSG1682.

  14. High-throughput, genome-scale protein production method based on the wheat germ cell-free expression system.

    PubMed

    Endo, Yaeta; Sawasaki, Tatsuya

    2004-01-01

    Current cell-free protein expression systems are capable of synthesizing proteins with high speed and accuracy; however, the yields are low due to their instability over time. Escherichia coli based systems are not always sufficient for expression of eukaryotic proteins. This report reviews a high-throughput protein production method based on the cell-free system prepared from eukaryote, wheat embryos. We first demonstrate a method for preparation of this extract that exhibited a high degree of stability and activity. To maximize translation yield and throughput, we address and resolve the following issues: (1) optimization of the ORF flanking regions; (2) PCR-based generation of DNA for mRNA production; (3) expression vectors for large-scale protein production; and (4) a translation reaction that does not require a membrane. The combination of these elemental processes with robotic automation resulted in high-throughput protein synthesis.

  15. Inter-comparison of laboratory smog chamber and flow reactor systems on organic aerosol yield and composition

    NASA Astrophysics Data System (ADS)

    Bruns, E. A.; El Haddad, I.; Keller, A.; Klein, F.; Kumar, N. K.; Pieber, S. M.; Corbin, J. C.; Slowik, J. G.; Brune, W. H.; Baltensperger, U.; Prévôt, A. S. H.

    2015-06-01

    A variety of tools are used to simulate atmospheric aging, including smog chambers and flow reactors. Traditional, large-scale smog chambers age emissions over the course of hours to days, whereas flow reactors rapidly age emissions using high oxidant concentrations to reach higher degrees of oxygenation than typically attained in smog chamber experiments. The atmospheric relevance of the products generated under such rapid oxidation warrants further study. However, no previously published studies have compared the yields and chemical composition of products generated in flow reactors and smog chambers from the same starting mixture. The yields and composition of the organic aerosol formed from the photo-oxidation of α-pinene and of wood-combustion emissions in a smog chamber (SC) and two flow reactors: a potential aerosol mass reactor (PAM) and a micro-smog chamber (MSC), were determined using aerosol mass spectrometry. Reactants were sampled from the SC and aged in the MSC and the PAM using a range of hydroxyl radical (OH) concentrations and then photo-chemically aged in the SC. The chemical composition, as well as the maximum yields and emission factors, of the products in both the α-pinene and wood-combustion systems determined with the PAM and the SC agreed reasonably well. High OH exposures have been shown previously to lower yields by breaking carbon-carbon bonds and forming higher volatility species, which reside largely in the gas phase; however, fragmentation in the PAM was not observed. The yields determined using the PAM for the α-pinene system were slightly lower than in the SC, possibly from increased wall losses of gas phase species due to the higher surface area to volume ratios in the PAM, even when offset with better isolation of the sampled flow from the walls. The α-pinene SOA results for the MSC were not directly comparable, as particles were smaller than the optimal AMS transmission range. The higher supersaturation in the flow reactors

  16. Plant development and yield of four sugarcane varieties irrigated by a subsurface drip irrigation system in Campinas, Brazil

    NASA Astrophysics Data System (ADS)

    Silva, André Luiz Barros de O.; Célia de Matos Pires, Regina; Yukitaka Pessinati Ohashi, Augusto; Vasconcelos Ribeiro, Rafael; Landell, Marcos Guimarães de Andrade; Aparecida Creste Dias de Souza, Silvana

    2013-04-01

    The biofuel production is a growing concern on modern society due to the agricultural sustainability, in which both food and energy supply should be taken into account. The agroclimatic zoning indicates that sugarcane expansion in Brazil can only take place in marginal lands, where water deficit occurs and irrigation is necessary. The use of subsurface drip irrigation (SDI) in sugarcane cultivation is an interesting cultural practice to improve production and allow cultivation in marginal lands due to water deficit conditions or to attain high yield and to increase longevity of plants. In this context it is necessary to investigate responses of different varieties to water supply. The aim of this work was to evaluate the plant development and yield of four sugarcane varieties irrigated by a subsurface drip irrigation system in Campinas, Brazil in the 1st cane ratoon cycle. The field experiment was carried out in Campinas SP Brazil, with IACSP95-5000, IACSP94-2094, IACSP94-2101 and SP79-1011 cultivars in the 1st cane ratoon cycle, from January (after the harvest of cane plant cycle) to October (harvest the 1st cane ratoon cycle). The plant spacing was 1.5 m between rows. Each cultivar was planted in an area of 0.4 hectares. The irrigation was done by a subsuperficial drip system with one drip line in each plant row installed at 0.25 m deep. During the 1st cane ratoon cycle the parameters were analysed on the 33rd, 123rd, 185th and 277th day. The analysed parameters were: plant yield (m), leaf area index (LAI) and yield (tons per hectare). According to the results from the second sampling (123rd day) the varieties IACSP95-5000 and IACSP94-2101 showed higher plant height when compared to the other varieties. However, from the third sampling (185th day) on the IACSP95-5000 variety grew considerably taller than the other varieties. The varieties SP79-1011and IACSP94-2101 presented lower values of LAI throughout the crop cycle when compared to other varieties. But on the

  17. Enhancing digestibility and ethanol yield of Populus wood via expression of an engineered monolignol 4-O-methyltransferase

    SciTech Connect

    Cai, Yuanheng; Zhang, Kewei; Kim, Hoon; Hou, Guichuan; Zhang, Xuebin; Yang, Huijun; Feng, Huan; Miller, Lisa; Ralph, John; Liu, Chang -Jun

    2016-06-28

    Producing cellulosic biofuels and bio-based chemicals from woody biomass is impeded by the presence of lignin polymer in the plant cell wall. Manipulating the monolignol biosynthetic pathway offers a promising approach to improved processability, but often impairs plant growth and development. Here, we show that expressing an engineered 4-O-methyltransferase that chemically modifies the phenolic moiety of lignin monomeric precursors, thus preventing their incorporation into the lignin polymer, substantially alters hybrid aspens’ lignin content and structure. Woody biomass derived from the transgenic aspens shows a 62% increase in the release of simple sugars and up to a 49% increase in the yield of ethanol when the woody biomass is subjected to enzymatic digestion and yeast-mediated fermentation. Furthermore, the cell wall structural changes do not affect growth and biomass production of the trees. Our study provides a useful strategy for tailoring woody biomass for bio-based applications.

  18. Comparison of cellulosic ethanol yields from midwestern maize and reconstructed tallgrass prairie systems managed for bioenergy

    USDA-ARS?s Scientific Manuscript database

    Maize- and prairie-based systems were investigated as cellulosic feedstocks by conducting a 9 ha side-by-side comparison on fertile soils in the Midwestern United States. Maize was grown continuously with adequate fertilization over years both with and without a winter rye cover crop, and the 31-spe...

  19. Object-Oriented Agricultural System Modeling: Component-Driven Nutrient Dynamics and Crop Yield Simulations

    USDA-ARS?s Scientific Manuscript database

    Challenges in agro-ecosystem conservation management have created demand for state-of-the-art, integrated, and flexible modeling tools. For example, agricultural system modeling tools are needed which are robust and fast enough to be applied on large watershed scales, but which are also able to sim...

  20. Grazing winter rye cover crop in a cotton no-till system: yield and economics

    USDA-ARS?s Scientific Manuscript database

    Winter cover crop adoption in conservation management systems continues to be limited in the US but could be encouraged if establishment costs could be offset. A 4-yr field experiment was conducted near Watkinsville, Georgia in which a rye (Secale cereale L.) cover crop was either grazed by catt...

  1. Yields in stripper header vs conventional header in dryland cropping systems

    USDA-ARS?s Scientific Manuscript database

    Differences in crop residue quality can impact the amount of soil water storage in semi-arid no-till systems of the West Central Great Plains. Using a stripper header as opposed to a conventional-reel type header to harvest small grains impacts the quality of the crop residue left in the field. Pr...

  2. Winter Cereal Termination and Conservation Agriculture Cotton Yield Following Mechanical and Chemical Management Systems

    USDA-ARS?s Scientific Manuscript database

    An integral component of conservation-tillage systems in cotton is the use of a high-residue winter cover crop; however, managing such cover crops is a challenge. Black oat (Avena strigosa Schreb.), rye (Secale cereale L.), and wheat (Triticum aestivum L.) winter cover crops were established in ear...

  3. Linking stomatal traits and expression of slow anion channel genes HvSLAH1 and HvSLAC1 with grain yield for increasing salinity tolerance in barley.

    PubMed

    Liu, Xiaohui; Mak, Michelle; Babla, Mohammad; Wang, Feifei; Chen, Guang; Veljanoski, Filip; Wang, Gang; Shabala, Sergey; Zhou, Meixue; Chen, Zhong-Hua

    2014-01-01

    Soil salinity is an environmental and agricultural problem in many parts of the world. One of the keys to breeding barley for adaptation to salinity lies in a better understanding of the genetic control of stomatal regulation. We have employed a range of physiological (stomata assay, gas exchange, phylogenetic analysis, QTL analysis), and molecular techniques (RT-PCR and qPCR) to investigate stomatal behavior and genotypic variation in barley cultivars and a genetic population in four experimental trials. A set of relatively efficient and reliable methods were developed for the characterization of stomatal behavior of a large number of varieties and genetic lines. Furthermore, we found a large genetic variation of gas exchange and stomatal traits in barley in response to salinity stress. Salt-tolerant cultivar CM72 showed significantly larger stomatal aperture under 200 mM NaCl treatment than that of salt-sensitive cultivar Gairdner. Stomatal traits such as aperture width/length were found to significantly correlate with grain yield under salt treatment. Phenotypic characterization and QTL analysis of a segregating double haploid population of the CM72/Gairdner resulted in the identification of significant stomatal traits-related QTLs for salt tolerance. Moreover, expression analysis of the slow anion channel genes HvSLAH1 and HvSLAC1 demonstrated that their up-regulation is linked to higher barley grain yield in the field.

  4. Linking stomatal traits and expression of slow anion channel genes HvSLAH1 and HvSLAC1 with grain yield for increasing salinity tolerance in barley

    PubMed Central

    Liu, Xiaohui; Mak, Michelle; Babla, Mohammad; Wang, Feifei; Chen, Guang; Veljanoski, Filip; Wang, Gang; Shabala, Sergey; Zhou, Meixue; Chen, Zhong-Hua

    2014-01-01

    Soil salinity is an environmental and agricultural problem in many parts of the world. One of the keys to breeding barley for adaptation to salinity lies in a better understanding of the genetic control of stomatal regulation. We have employed a range of physiological (stomata assay, gas exchange, phylogenetic analysis, QTL analysis), and molecular techniques (RT-PCR and qPCR) to investigate stomatal behavior and genotypic variation in barley cultivars and a genetic population in four experimental trials. A set of relatively efficient and reliable methods were developed for the characterization of stomatal behavior of a large number of varieties and genetic lines. Furthermore, we found a large genetic variation of gas exchange and stomatal traits in barley in response to salinity stress. Salt-tolerant cultivar CM72 showed significantly larger stomatal aperture under 200 mM NaCl treatment than that of salt-sensitive cultivar Gairdner. Stomatal traits such as aperture width/length were found to significantly correlate with grain yield under salt treatment. Phenotypic characterization and QTL analysis of a segregating double haploid population of the CM72/Gairdner resulted in the identification of significant stomatal traits-related QTLs for salt tolerance. Moreover, expression analysis of the slow anion channel genes HvSLAH1 and HvSLAC1 demonstrated that their up-regulation is linked to higher barley grain yield in the field. PMID:25505473

  5. Engineering of recombinant Escherichia coli cells co-expressing poly-γ-glutamic acid (γ-PGA) synthetase and glutamate racemase for differential yielding of γ-PGA.

    PubMed

    Cao, Mingfeng; Geng, Weitao; Zhang, Wei; Sun, Jibin; Wang, Shufang; Feng, Jun; Zheng, Ping; Jiang, Anna; Song, Cunjiang

    2013-11-01

    Poly-γ-glutamic acid (γ-PGA) is a promising environmental-friendly material with outstanding water solubility, biocompatibility and degradability. However, it is tough to determine the relationship between functional synthetic enzyme and the strains' yield or substrate dependency. We cloned γ-PGA synthetase genes pgsBCA and glutamate racemase gene racE from both L-glutamate-dependent γ-PGA-producing Bacillus licheniformis NK-03 and L-glutamate-independent B. amyloliquefaciens LL3 strains. The deduced RacE and PgsA from the two strains shared the identity of 84.5% and 78.53%, while PgsB and PgsC possessed greater similarity with 93.13% and 93.96%. The induced co-expression of pgsBCA and racE showed that the engineered Escherichia coli strains had the capacity of synthesizing γ-PGA, and LL3 derived PgsBCA had higher catalytic activity and enhanced productivity than NK-03 in Luria-Bertani medium containing glucose or L-glutamate. However, the differential effect was weakened when providing sufficient immediateness L-glutamate substrate, that is, the supply of substrate could be served as the ascendance upon γ-PGA production. Furthermore, RacE integration could enhance γ-PGA yield through improving the preferred d-glutamate content. This is the first report about co-expression of pgsBCA and racE from the two Bacillus strains, which will be of great value for the determination of the biosynthetic mechanism of γ-PGA.

  6. High-yield soluble expression, purification and characterization of human steroidogenic acute regulatory protein (StAR) fused to a cleavable Maltose-Binding Protein (MBP).

    PubMed

    Sluchanko, Nikolai N; Tugaeva, Kristina V; Faletrov, Yaroslav V; Levitsky, Dmitrii I

    2016-03-01

    Steroidogenic acute regulatory protein (StAR) is responsible for the rapid delivery of cholesterol to mitochondria where the lipid serves as a source for steroid hormones biosynthesis in adrenals and gonads. Despite many successful investigations, current understanding of the mechanism of StAR action is far from being completely clear. StAR was mostly obtained using denaturation/renaturation or in minor quantities in a soluble form at decreased temperatures that, presumably, limited the possibilities for its consequent detailed exploration. In our hands, existing StAR expression constructs could be bacterially expressed almost exclusively as insoluble forms, even upon decreased expression temperatures and in specific strains of Escherichia coli, and isolated protein tended to aggregate and was difficult to handle. To maximize the yield of soluble protein, optimized StAR sequence encompassing functional domain STARD1 (residues 66-285) was fused to the C-terminus of His-tagged Maltose-Binding Protein (MBP) with the possibility to cleave off the whole tag by 3C protease. The developed protocol of expression and purification comprising of a combination of subtractive immobilized metal affinity chromatography (IMAC) and size-exclusion chromatography allowed us to obtain up to 25 mg/1 L culture of completely soluble StAR protein, which was (i) homogenous according to SDS-PAGE, (ii) gave a single symmetrical peak on a gel-filtration, (iii) showed the characteristic CD spectrum and (iv) pH-dependent ability to bind a fluorescently-labeled cholesterol analogue. We conclude that our strategy provides fully soluble and native StAR protein which in future could be efficiently used for biotechnology and drug discovery aimed at modulation of steroids production. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system*

    PubMed Central

    Gong, Xing-guo; Ji, Jing; Xie, Jie; Zhou, Yuan; Zhang, Jun-yan; Zhong, Wen-tao

    2006-01-01

    v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present here the expression, purification, and bioactivity of a GST (glutathione S-transferase)-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl β-D-thiogalactopyranoside (IPTG)-regulated expression, and the fused protein was purified using GSH (glutathione) affinity chromatography. ELISA (enzyme-linked immunosorbent assay) was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems employed in earlier Src expression. PMID:16365920

  8. Applied studies of plant meridian system: I. The effect of agri-wave technology on yield and quality of tomato.

    PubMed

    Hou, T Z; Mooneyham, R E

    1999-01-01

    Agri-wave technology is a new agricultural technology based on the plant meridian system, that focuses on measurement of plant sound characteristics. The basic principle of agri-wave technology is to improve the yield and quality of plants such as vegetables, flowers, and fruit trees by broadcasting sound waves of certain frequencies and spraying a compound microelement fertilizer on the leaves. The application of agri-wave technology on tomatoes remarkably stimulates growth of their seedlings. Fresh weight of the branch, stems, and leaves of the treated tomatoes is significantly (59.53%, P < 0.0001) higher than that of the control group. Sampling survey results indicate that agri-wave technology accelerates the ripeness of tomatoes. The fresh weight of ripe tomatoes treated with this technique is 30.73% higher than that of the untreated (P = 0.0018), while the fresh weight of the treated unripe tomatoes is 27.29% lower than that of the untreated unripe group (P = 0.0020). Yield surveys show that the yield of treated plants is 13.89% (p < 0.0001) higher than that of the control group. Moreover, with agri-wave technology treatment the storage period of tomatoes is almost doubled. Analysis of tomato nutrition shows that agri-wave technology has increased their sugar content by 26.19%, vitamin A and niacin (an antifavours vitamin) by 55.39% and 92.31% respectively. There is no difference concerning vitamin B1, B2, and D content between the two groups, and vitamin C and E contents decreased by 2.10% and 12.69%, respectively. Among the analyzed 33 minerals of tomatoes, 26 increased in content, while 7 decreased. In conclusion, agri-wave technology has promoted the growth of the tomato, increased its yield, and improved its quality.

  9. Avalanches in 2D dislocation systems: plastic yielding is not depinning.

    PubMed

    Ispánovity, Péter Dusán; Laurson, Lasse; Zaiser, Michael; Groma, István; Zapperi, Stefano; Alava, Mikko J

    2014-06-13

    We study the properties of strain bursts (dislocation avalanches) occurring in two-dimensional discrete dislocation dynamics models under quasistatic stress-controlled loading. Contrary to previous suggestions, the avalanche statistics differ fundamentally from predictions obtained for the depinning of elastic manifolds in quenched random media. Instead, we find an exponent τ=1 of the power-law distribution of slip or released energy, with a cutoff that increases exponentially with the applied stress and diverges with system size at all stresses. These observations demonstrate that the avalanche dynamics of 2D dislocation systems is scale-free at every applied stress and, therefore, cannot be envisaged in terms of critical behavior associated with a depinning transition.

  10. Biomass yield and composition of sweetpotato grown in a nutrient film technique system.

    PubMed

    Almazan, A M; Zhou, X

    1997-01-01

    Sweetpotato cultivar TU-82-155 grown in a nutrient film technique system and separated into foliage, tips, fibrous, string and storage roots at harvest had a total dry biomass of 89.9 g per plant with 38.4% inedible portion. Tips and storage roots, the traditional edible parts, were analyzed for dry matter, protein, fat, ash, minerals (Ca, Fe, K, Mg, Na, Zn), vitamins (carotene, ascorbic acid, thiamin), oxalic and tannic acids, and trypsin and chymotrypsin inhibitors to determine their nutritional quality. Water soluble matter, minerals (Ca, Fe, K, Mg, Na, Zn), cellulose, hemicellulose and lignin concentrations in the edible and inedible parts were obtained to provide information needed for the selection of appropriate bioconversion processes of plant wastes into food or forms suitable for crop production in a controlled biological life support system.

  11. Vestigial expression in the Drosophila embryonic central nervous system.

    PubMed

    Guss, Kirsten A; Mistry, Hemlata; Skeath, James B

    2008-09-01

    The Drosophila central nervous system is an excellent model system in which to resolve the genetic and molecular control of neuronal differentiation. Here we show that the wing selector vestigial is expressed in discrete sets of neurons. We track the axonal trajectories of VESTIGIAL-expressing cells in the ventral nerve cord and show that these cells descend from neuroblasts 1-2, 5-1, and 5-6. In addition, along the midline, VESTIGIAL is expressed in ventral unpaired median motorneurons and cells that may descend from the median neuroblast. These studies form the requisite descriptive foundation for functional studies addressing the role of vestigial during interneuron differentiation.

  12. Combined electron and focused ion beam system for improvement of secondary ion yield in secondary ion mass spectrometry instrument

    SciTech Connect

    Ji, L.; Ji, Q.; Leung, K.-N.; Gough, R. A.

    2006-10-16

    Using a combined electron and focused ion beam system to improve performance of secondary ion mass spectrometry instruments has been investigated experimentally. The secondary ion yield for an Al target has been enhanced to about one order of magnitude higher with the postionization induced by the low energy electrons in the combined beam. It can be further improved with the increase of electron beam current. When the combined beam is applied to insulating targets, sample charging is also eliminated. For Teflon targets, the secondary ion signal is increased by more than a factor of 20.

  13. Conjunctive-management models for sustained yield of stream-aquifer systems

    USGS Publications Warehouse

    Barlow, P.M.; Ahlfeld, D.P.; Dickerman, D.C.

    2003-01-01

    Conjunctive-management models that couple numerical simulation with linear optimization were developed to evaluate trade-offs between groundwater withdrawals and streamflow depletions for alluvial-valley stream-aquifer systems representative of those of the northeastern United States. A conjunctive-management model developed for a hypothetical stream-aquifer system was used to assess the effect of interannual hydrologic variability on minimum monthly streamflow requirements. The conjunctive-management model was applied to the Hunt-Annaquatucket-Pettaquamscutt stream-aquifer system of central Rhode Island. Results show that it is possible to increase the amount of current withdrawal from the aquifer by as much as 50% by modifying current withdrawal schedules, modifying the number and configuration of wells in the supply-well network, or allowing increased streamflow depletion in the Annaquatucket and Pettaquamscutt rivers. Alternatively, it is possible to reduce current rates of streamflow depletion in the Hunt River by as much as 35% during the summer, but such reductions would result increases in groundwater withdrawals.

  14. High yield expression in a recombinant E. coli of a codon optimized chicken anemia virus capsid protein VP1 useful for vaccine development.

    PubMed

    Lee, Meng-Shiou; Hseu, You-Cheng; Lai, Guan-Hua; Chang, Wen-Te; Chen, Hsi-Jien; Huang, Chi-Hung; Lee, Meng-Shiunn; Wang, Min-Ying; Kao, Jung-Yie; You, Bang-Jau; Lin, Wen- Hsin; Lien, Yi-Yang; Lin, Ming-Kuem

    2011-07-23

    Chicken anemia virus (CAV), the causative agent chicken anemia, is the only member of the genus Gyrovirus of the Circoviridae family. CAV is an immune suppressive virus and causes anemia, lymph organ atrophy and immunodeficiency. The production and biochemical characterization of VP1 protein and its use in a subunit vaccine or as part of a diagnostic kit would be useful to CAV infection prevention. Significantly increased expression of the recombinant full-length VP1 capsid protein from chicken anemia virus was demonstrated using an E. coli expression system. The VP1 gene was cloned into various different expression vectors and then these were expressed in a number of different E. coli strains. The expression of CAV VP1 in E. coli was significantly increased when VP1 was fused with GST protein rather than a His-tag. By optimizing the various rare amino acid codons within the N-terminus of the VP1 protein, the expression level of the VP1 protein in E. coli BL21(DE3)-pLysS was further increased significantly. The highest protein expression level obtained was 17.5 g/L per liter of bacterial culture after induction with 0.1 mM IPTG for 2 h. After purification by GST affinity chromatography, the purified full-length VP1 protein produced in this way was demonstrated to have good antigenicity and was able to be recognized by CAV-positive chicken serum in an ELISA assay. Purified recombinant VP1 protein with the gene's codons optimized in the N-terminal region has potential as chimeric protein that, when expressed in E. coli, may be useful in the future for the development of subunit vaccines and diagnostic tests.

  15. High-yield isolation of extracellular vesicles using aqueous two-phase system

    PubMed Central

    Shin, Hyunwoo; Han, Chungmin; Labuz, Joseph M.; Kim, Jiyoon; Kim, Jongmin; Cho, Siwoo; Gho, Yong Song; Takayama, Shuichi; Park, Jaesung

    2015-01-01

    Extracellular vesicles (EVs) such as exosomes and microvesicles released from cells are potential biomarkers for blood-based diagnostic applications. To exploit EVs as diagnostic biomarkers, an effective pre-analytical process is necessary. However, recent studies performed with blood-borne EVs have been hindered by the lack of effective purification strategies. In this study, an efficient EV isolation method was developed by using polyethylene glycol/dextran aqueous two phase system (ATPS). This method provides high EV recovery efficiency (~70%) in a short time (~15 min). Consequently, it can significantly increase the diagnostic applicability of EVs. PMID:26271727

  16. Efficient silkworm expression of human GPCR (nociceptin receptor) by a Bombyx mori bacmid DNA system

    SciTech Connect

    Kajikawa, Mizuho; Sasaki, Kaori; Wakimoto, Yoshitaro; Toyooka, Masaru; Motohashi, Tomoko; Shimojima, Tsukasa; Takeda, Shigeki; Park, Enoch Y.; Maenaka, Katsumi

    2009-07-31

    Guanine nucleotide-binding protein (G protein) coupled receptors (GPCRs) are frequently expressed by a baculovirus expression vector system (BEVS). We recently established a novel BEVS using the bacmid system of Bombyx mori nucleopolyhedrovirus (BmNPV), which is directly applicable for protein expression in silkworms. Here, we report the first example of GPCR expression in silkworms by the simple injection of BmNPV bacmid DNA. Human nociceptin receptor, an inhibitory GPCR, and its fusion protein with inhibitory G protein alpha subunit (G{sub i}{alpha}) were both successfully expressed in the fat bodies of silkworm larvae as well as in the BmNPV viral fraction. Its yield was much higher than that from Sf9 cells. The microsomal fractions including the nociceptin receptor fusion, which are easily prepared by only centrifugation steps, exhibited [{sup 35}S]GTP{gamma}S-binding activity upon specific stimulation by nociceptin. Therefore, this rapid method is easy-to-use and has a high expression level, and thus will be an important tool for human GPCR production.

  17. Hubble Discovery of Runaway Star Yields Clues to Breakup of Multiple-Star System

    NASA Image and Video Library

    2017-09-28

    NASA's Hubble Space Telescope has helped astronomers find the final piece of a celestial puzzle by nabbing a third runaway star. As British royal families fought the War of the Roses in the 1400s for control of England's throne, a grouping of stars was waging its own contentious skirmish — a star war far away in the Orion Nebula. The stars were battling each other in a gravitational tussle, which ended with the system breaking apart and at least three stars being ejected in different directions. The speedy, wayward stars went unnoticed for hundreds of years until, over the past few decades, two of them were spotted in infrared and radio observations, which could penetrate the thick dust in the Orion Nebula. Read more: go.nasa.gov/2ni3EZX NASA image use policy. NASA Goddard Space Flight Center enables NASA’s mission through four scientific endeavors: Earth Science, Heliophysics, Solar System Exploration, and Astrophysics. Goddard plays a leading role in NASA’s accomplishments by contributing compelling scientific knowledge to advance the Agency’s mission. Follow us on Twitter Like us on Facebook Find us on Instagram

  18. Bacterial expression system with tightly regulated gene expression and plasmid copy number.

    PubMed

    Bowers, Lisa M; Lapoint, Kathleen; Anthony, Larry; Pluciennik, Anna; Filutowicz, Marcin

    2004-09-29

    A new Escherichia coli host/vector system has been engineered to allow tight and uniform modulation of gene expression and gamma origin (ori) plasmid copy number. Regulation of gamma ori plasmid copy number is achieved through arabinose-inducible expression of the necessary Rep protein, pi, whose gene was integrated into the chromosome of the host strain under control of the P(BAD) promoter. gamma ori replication can be uniformly modulated over 100-fold by changing the concentration of l-arabinose in the growth medium. This strain avoids the problem of all-or-nothing induction of P(BAD) because it is deficient in both arabinose uptake and degradation genes. Arabinose enters the cell by a mutant LacY transporter, LacYA177C, which is expressed from the host chromosome. Although this strain could be compatible with any gamma ori plasmid, we describe the utility of a gamma ori expression vector that allows especially tight regulation of gene expression. With this host/vector system, it is possible to independently modulate gene expression and gene dosage, facilitating the cloning and overproduction of toxic gene products. We describe the successful use of this system for cloning a highly potent toxin, Colicin E3, in the absence of its cognate immunity protein. This system could be useful for cloning genes encoding other potent toxins, screening libraries for potential toxins, and maintaining any gamma ori vector at precise copy levels in a cell.

  19. Improved Production Efficiency of Virus-Like Particles by the Baculovirus Expression Vector System.

    PubMed

    López-Vidal, Javier; Gómez-Sebastián, Silvia; Bárcena, Juan; Nuñez, Maria del Carmen; Martínez-Alonso, Diego; Dudognon, Benoit; Guijarro, Eva; Escribano, José M

    2015-01-01

    Vaccines based on virus-like particles (VLPs) have proven effective in humans and animals. In this regard, the baculovirus expression vector system (BEVS) is one of the technologies of choice to generate such highly immunogenic vaccines. The extended use of these vaccines for human and animal populations is constrained because of high production costs, therefore a significant improvement in productivity is crucial to ensure their commercial viability. Here we describe the use of the previously described baculovirus expression cassette, called TB, to model the production of two VLP-forming vaccine antigens in insect cells. Capsid proteins from porcine circovirus type 2 (PCV2 Cap) and from the calicivirus that causes rabbit hemorrhagic disease (RHDV VP60) were expressed in insect cells using baculoviruses genetically engineered with the TB expression cassette. Productivity was compared to that obtained using standard counterpart vectors expressing the same proteins under the control of the polyhedrin promoter. Our results demonstrate that the use of the TB expression cassette increased the production yields of these vaccine antigens by around 300% with respect to the standard vectors. The recombinant proteins produced by TB-modified vectors were fully functional, forming VLPs identical in size and shape to those generated by the standard baculoviruses, as determined by electron microscopy analysis. The use of the TB expression cassette implies a simple modification of the baculovirus vectors that significantly improves the cost efficiency of VLP-based vaccine production, thereby facilitating the commercial viability and broad application of these vaccines for human and animal health.

  20. Free range and deep litter poultry production systems: effect on performance, carcass yield and meat composition of cockerel chickens.

    PubMed

    Sogunle, Olajide Mark; Olaniyi, Olagoke Ayobami; Egbeyale, Lawrence Tokunbo; Akinola, Olufemi Sunday; Shittu, Taofeek A; Abiola, Samuel Soladoye; Ladokun, Abimbola O; Sobayo, Richard Abayomi

    2013-01-01

    This study was carried out on 150 cockerel chickens each of Harco Black and Novogen strains to determine their performance, carcass yield and meat composition on free range and deep litter production systems. The birds were brooded for 4 weeks and thereafter allotted to the different production systems for a period of 12 weeks. Each production system was allotted 150 chicks (75 chicks per strain) with three replicates of 25 chicks. The birds on deep litter production system were fed ad libitum while each bird on free range was fed 50 % of its daily feed requirement. On the 84 th day, a total of 36 birds were randomly selected for analysis of the carcass yield and meat composition. The data generated were subjected to a two-way analysis of variance in a 2 × 2 factorial experimental arrangement. Novogen strain consumed less feed (P < 0.05) on free range and had the best feed/gain (2.72). A higher (P < 0.05) shear force value (3.74 N) was obtained in the thigh muscle for birds on free range. The tibia proximal length and breadth, and tibia distal length and breadth were significantly (P < 0.05) affected by the production systems and strains. On free range, Harco black had more meat (85.69 g) than bone (18.07 g) in the breast while Novogen had the lowest meat/bone (2.38). Conclusively, Novogen strain should be raised on free range for a better performance in terms of feed/gain, but for higher meat composition, Harco black is a better strain.

  1. Effect of materials for micro-electro-mechanical systems on PCR yield.

    PubMed

    Potrich, Cristina; Lunelli, Lorenzo; Forti, Stefania; Vozzi, Diego; Pasquardini, Laura; Vanzetti, Lia; Panciatichi, Cristina; Anderle, Mariano; Pederzolli, Cecilia

    2010-05-01

    In this study we analyzed the surface properties of different silicon-based materials used for micro-electro-mechanical systems (MEMS) production, such as thermally grown silicon oxide, plasma-enhanced chemical vapor deposition (PECVD)-treated silicon oxide, reactive-ion etch (RIE)-treated silicon oxide, and Pyrex. Substrates were characterized by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) to define the surface chemical and morphological properties, and by fluorescence microscopy to directly assess the absorption of the different polymerase chain reaction (PCR) components. By using microchips fabricated with the same materials we investigated their compatibility with PCR reactions, exploiting the use of different enzymes and reagents or proper surface treatments. We established the best conditions for DNA amplification in silicon/Pyrex microdevices depending on the type of device and fabrication method used and the quality of reagents, rather than on the passivation treatment or increment in standard Taq polymerase concentration.

  2. Development of high yielding photonic light delivery system for photodynamic therapy of esophageal carcinomas

    NASA Astrophysics Data System (ADS)

    Premasiri, Amaranath; Happawana, Gemunu; Rosen, Arye

    2007-02-01

    Photodynamic therapy (PDT) is an approved treatment modality for Barrett's and invasive esophageal carcinoma. Proper Combination of photosentizing agent, oxygen, and a specific wavelength of light to activate the photosentizing agents is necessary for the cytotoxic destruction of cancerous cells by PDT. As a light source expensive solid-state laser sources currently are being used for the treatment. Inexpensive semiconductor lasers have been suggested for the light delivery system, however packaging of semiconductor lasers for optimal optical power output is challenging. In this paper, we present a multidirectional direct water-cooling of semiconductor lasers that provides a better efficiency than the conventional unidirectional cooling. AlGaAsP lasers were tested under de-ionized (DI) water and it is shown that the optical power output of the lasers under the DI water is much higher than that of the uni-directional cooling of lasers. Also, in this paper we discuss how direct DI water-cooling can optimize power output of semiconductor lasers. Thereafter an optimal design of the semiconductor laser package is shown with the DI water-cooling system. Further, a microwave antenna is designed which is to be imprinted on to a balloon catheter in order to provide local heating of esophagus, leading to an increase in local oxygenation of the tumor to generate an effective level of singlet oxygen for cellular death. Finally the optimal level of light energy that is required to achieve the expected level of singlet oxygen is modeled to design an efficient PDT protocol.

  3. Optimization of transient gene expression system in Gerbera jemosonii petals.

    PubMed

    Hussein, Gihan M; Abu El-Heba, Ghada A; Abdou, Sara M; Abdallah, Naglaa A

    2013-01-01

    Low transformation efficiency and long generation time for production of transgenic Gerbera jemosonii plants leads to vulnerable gene function studies. Thus, transient expression of genes would be an efficient alternative. In this investigation, a transient expression system for gerbera petals based on the Agrobacterium infiltration protocol was developed using the reporter genes β-glucuronidase (gus) and green florescence protein (gfp). Results revealed the incapability of using the gfp gene as a reporter gene for transient expression study in gerbera flowers due to the detection of green fluorescent color in the non-infiltrated gerbera flower petals. However, the gus reporter gene was successfully utilized for optimizing and obtaining the suitable agroinfiltration system in gerbera flowers. The expression of GUS was detectable after three days of agroinfiltration in gerbera cultivars "Express" and "White Grizzly" with dark pink and white flower colors, respectively. The vacuum agroinfiltration protocol has been applied on the cultivar "Express" for evaluating the transient expression of the two genes involved in the anthocyanin pathway (iris-dfr and petunia-f3' 5'h), which is responsible for the color in flowers. In comparison to the control, transient expression results showed change in the anthocyanin pigment in all infiltrated flowers with color genes. Additionally, blue color was detected in the stigma and pollen grains in the infiltrated flowers. Moreover, blue colors with variant intensities were observed in produced calli during the routine work of stable transformation with f3' 5'h gene.

  4. Enhanced Gene Expression Rather than Natural Polymorphism in Coding Sequence of the OsbZIP23 Determines Drought Tolerance and Yield Improvement in Rice Genotypes

    PubMed Central

    Dey, Avishek; Samanta, Milan Kumar; Gayen, Srimonta; Sen, Soumitra K.; Maiti, Mrinal K.

    2016-01-01

    Drought is one of the major limiting factors for productivity of crops including rice (Oryza sativa L.). Understanding the role of allelic variations of key regulatory genes involved in stress-tolerance is essential for developing an effective strategy to combat drought. The bZIP transcription factors play a crucial role in abiotic-stress adaptation in plants via abscisic acid (ABA) signaling pathway. The present study aimed to search for allelic polymorphism in the OsbZIP23 gene across selected drought-tolerant and drought-sensitive rice genotypes, and to characterize the new allele through overexpression (OE) and gene-silencing (RNAi). Analyses of the coding DNA sequence (CDS) of the cloned OsbZIP23 gene revealed single nucleotide polymorphism at four places and a 15-nucleotide deletion at one place. The single-copy OsbZIP23 gene is expressed at relatively higher level in leaf tissues of drought-tolerant genotypes, and its abundance is more in reproductive stage. Cloning and sequence analyses of the OsbZIP23-promoter from drought-tolerant O. rufipogon and drought-sensitive IR20 cultivar showed variation in the number of stress-responsive cis-elements and a 35-nucleotide deletion at 5’-UTR in IR20. Analysis of the GFP reporter gene function revealed that the promoter activity of O. rufipogon is comparatively higher than that of IR20. The overexpression of any of the two polymorphic forms (1083 bp and 1068 bp CDS) of OsbZIP23 improved drought tolerance and yield-related traits significantly by retaining higher content of cellular water, soluble sugar and proline; and exhibited decrease in membrane lipid peroxidation in comparison to RNAi lines and non-transgenic plants. The OE lines showed higher expression of target genes-OsRab16B, OsRab21 and OsLEA3-1 and increased ABA sensitivity; indicating that OsbZIP23 is a positive transcriptional-regulator of the ABA-signaling pathway. Taken together, the present study concludes that the enhanced gene expression rather

  5. pOsNAR2.1:OsNAR2.1 expression enhances nitrogen uptake efficiency and grain yield in transgenic rice plants.

    PubMed

    Chen, Jingguang; Fan, Xiaoru; Qian, Kaiyun; Zhang, Yong; Song, Miaoquan; Liu, Yu; Xu, Guohua; Fan, Xiaorong

    2017-02-22

    The nitrate (NO3-) transporter has been selected as an important gene maker in the process of environmental adoption in rice cultivars. In this work, we transferred another native OsNAR2.1 promoter with driving OsNAR2.1 gene into rice plants. The transgenic lines with exogenous pOsNAR2.1:OsNAR2.1 constructs showed enhanced OsNAR2.1 expression level, compared with wild type (WT), and (15) N influx in roots increased 21%-32% in response to 0.2 mm and 2.5 mm 15NO3- and 1.25 mm (15) NH4(15) NO3 . Under these three N conditions, the biomass of the pOsNAR2.1:OsNAR2.1 transgenic lines increased 143%, 129% and 51%, and total N content increased 161%, 242% and 69%, respectively, compared to WT. Furthermore in field experiments we found the grain yield, agricultural nitrogen use efficiency (ANUE), and dry matter transfer of pOsNAR2.1:OsNAR2.1 plants increased by about 21%, 22% and 21%, compared to WT. We also compared the phenotypes of pOsNAR2.1:OsNAR2.1 and pOsNAR2.1:OsNRT2.1 transgenic lines in the field, found that postanthesis N uptake differed significantly between them, and in comparison with the WT. Postanthesis N uptake (PANU) increased approximately 39% and 85%, in the pOsNAR2.1:OsNAR2.1 and pOsNAR2.1:OsNRT2.1 transgenic lines, respectively, possibly because OsNRT2.1 expression was less in the pOsNAR2.1:OsNAR2.1 lines than in the pOsNAR2.1:OsNRT2.1 lines during the late growth stage. These results show that rice NO3(-) uptake, yield and NUE were improved by increased OsNAR2.1 expression via its native promoter.

  6. Enhanced Gene Expression Rather than Natural Polymorphism in Coding Sequence of the OsbZIP23 Determines Drought Tolerance and Yield Improvement in Rice Genotypes.

    PubMed

    Dey, Avishek; Samanta, Milan Kumar; Gayen, Srimonta; Sen, Soumitra K; Maiti, Mrinal K

    2016-01-01

    Drought is one of the major limiting factors for productivity of crops including rice (Oryza sativa L.). Understanding the role of allelic variations of key regulatory genes involved in stress-tolerance is essential for developing an effective strategy to combat drought. The bZIP transcription factors play a crucial role in abiotic-stress adaptation in plants via abscisic acid (ABA) signaling pathway. The present study aimed to search for allelic polymorphism in the OsbZIP23 gene across selected drought-tolerant and drought-sensitive rice genotypes, and to characterize the new allele through overexpression (OE) and gene-silencing (RNAi). Analyses of the coding DNA sequence (CDS) of the cloned OsbZIP23 gene revealed single nucleotide polymorphism at four places and a 15-nucleotide deletion at one place. The single-copy OsbZIP23 gene is expressed at relatively higher level in leaf tissues of drought-tolerant genotypes, and its abundance is more in reproductive stage. Cloning and sequence analyses of the OsbZIP23-promoter from drought-tolerant O. rufipogon and drought-sensitive IR20 cultivar showed variation in the number of stress-responsive cis-elements and a 35-nucleotide deletion at 5'-UTR in IR20. Analysis of the GFP reporter gene function revealed that the promoter activity of O. rufipogon is comparatively higher than that of IR20. The overexpression of any of the two polymorphic forms (1083 bp and 1068 bp CDS) of OsbZIP23 improved drought tolerance and yield-related traits significantly by retaining higher content of cellular water, soluble sugar and proline; and exhibited decrease in membrane lipid peroxidation in comparison to RNAi lines and non-transgenic plants. The OE lines showed higher expression of target genes-OsRab16B, OsRab21 and OsLEA3-1 and increased ABA sensitivity; indicating that OsbZIP23 is a positive transcriptional-regulator of the ABA-signaling pathway. Taken together, the present study concludes that the enhanced gene expression rather than

  7. Environmental controls, sediment sources and spatiotemporal variability of suspended sediment yields in partly glacierized catchment systems in western Norway

    NASA Astrophysics Data System (ADS)

    Beylich, Achim A.; Laute, Katja; Storms, Joep E. A.

    2016-04-01

    This work focuses on environmental controls, sediment sources and the spatiotemporal variability of suspended sediment yields in the neighboring, partly glacierized and steep Erdalen (79.5 km2) and Bødalen (60.1 km2) catchment systems in the fjord landscape of the inner Nordfjord in western Norway. Field work, including extended samplings and measurements, was carried out since 2004 in Erdalen and since 2008 in Bødalen. Fluvial suspended sediment transport in the inner Nordfjord is altogether supply-limited and larger thermally and/or pluvially generated runoff events occurring mostly during the period April-November are needed to mobilize and transport significant amounts of suspended sediments. The distinct intra- and inter-annual temporal variability of suspended sediment transport found is mostly controlled by meteorological events, with most suspended sediment transport occurring during pluvial events in autumn (September-November), followed by mostly thermally determined glacier melt in summer (July-August), and by mostly thermally determined snowmelt in spring (April-June). Extreme rainfall events (>70 mm/d) in autumn can trigger relevant debris-flow activity that can cause significant transfers of suspended sediments from ice-free surface areas with sedimentary covers into main stream channels and is particularly important for fluvial suspended sediment transport. In years with occurring relevant debris-flow activity the total annual drainage-basin wide suspended sediment yields are strongly determined by these single extreme events. The share of glacier coverage, followed by steepness of slopes, and degree of vegetation cover in ice-free surface areas with sedimentary covers are the main controls of the detected spatial variability of suspended sediment yields. The contemporary sediment delivery from glacierized surface areas through different outlet glaciers shows a high spatial variability which is mostly explained by a spatially variable availability

  8. Effect of agitation and terminal subcultures on yield and speed of detection of the Oxoid Signal blood culture system versus the BACTEC radiometric system

    SciTech Connect

    Weinstein, M.P.; Mirrett, S.; Reimer, L.G.; Reller, L.B.

    1989-03-01

    In an initial evaluation, we found the Oxoid Signal blood culture system inferior to the BACTEC radiometric system for detection of some microorganisms causing septicemia. To determine whether modified processing of the Oxoid Signal blood culture system could improve its yield and speed of detecting positive cultures relative to the BACTEC radiometric system, we agitated all Oxoid bottles during the first 24 to 48 h of incubation and performed aerobic and anaerobic subcultures of all Oxoid bottles negative after 7 days of incubation. These modifications improved the overall performance of the Oxoid system, particularly with regard to the yield of streptococci, members of the family Enterobacteriaceae, and Haemophilus, Neisseria, and Acinetobacter spp. The speed of detecting positive cultures also was improved, especially within the first 24 h of incubation. However, the BACTEC system still detected more positive cultures (P less than 0.005), especially of obligate aerobes such as Pseudomonas aeruginosa (P less than 0.05) and yeasts (P less than 0.005). The BACTEC system also detected positive cultures earlier than the Oxoid system (e.g., at 24 h of incubation, 70.5% of BACTEC positive cultures detected versus 62.1% of Oxoid positive cultures detected). Further modifications of the Oxoid system which might include a revised medium, additional processing modifications, altered headspace atmosphere, or a complementary second broth medium should be considered, since the system is attractive in concept and is easy to use in the clinical laboratory.

  9. Smoothed Biasing Forces Yield Unbiased Free Energies with the Extended-System Adaptive Biasing Force Method.

    PubMed

    Lesage, Adrien; Lelièvre, Tony; Stoltz, Gabriel; Hénin, Jérôme

    2016-12-27

    We report a theoretical description and numerical tests of the extended-system adaptive biasing force method (eABF), together with an unbiased estimator of the free energy surface from eABF dynamics. Whereas the original ABF approach uses its running estimate of the free energy gradient as the adaptive biasing force, eABF is built on the idea that the exact free energy gradient is not necessary for efficient exploration, and that it is still possible to recover the exact free energy separately with an appropriate estimator. eABF does not directly bias the collective coordinates of interest, but rather fictitious variables that are harmonically coupled to them; therefore is does not require second derivative estimates, making it easily applicable to a wider range of problems than ABF. Furthermore, the extended variables present a smoother, coarse-grain-like sampling problem on a mollified free energy surface, leading to faster exploration and convergence. We also introduce CZAR, a simple, unbiased free energy estimator from eABF trajectories. eABF/CZAR converges to the physical free energy surface faster than standard ABF for a wide range of parameters.

  10. Early safety assessment using cellular systems biology yields insights into mechanisms of action.

    PubMed

    Giuliano, Kenneth A; Gough, Albert H; Taylor, D Lansing; Vernetti, Lawrence A; Johnston, Patricia A

    2010-08-01

    The integration of high-content screening (HCS) readers with organ-specific cell models, panels of functional biomarkers, and advanced informatics is a powerful approach to identifying the toxic liabilities of compounds early in the development process and forms the basis of "early safety assessment." This cellular systems biology (CSB) approach (CellCiphr profile) has been used to integrate rodent and human cellular hepatic models with panels of functional biomarkers measured at multiple time points to profile both the potency and specificity of the cellular toxicological response. These profiles also provide initial insights on the mechanism of the toxic response. The authors describe here mechanistic assay profiles designed to further dissect the toxic mechanisms of action and elucidate subtle effects apparent in subpopulations of cells. They measured 8 key mechanisms of toxicity with multiple biomarker feature measurements made simultaneously in populations of living primary hepatocytes and HepG2 cells. Mining the cell population response from these mechanistic profiles revealed the concentration dependence and nature of the heterogeneity of the response, as well as relationships between the functional responses. These more detailed mechanistic profiles define differences in compound activities that are not apparent in the average population response. Because cells and tissues encounter wide ranges of drug doses in space and time, these mechanistic profiles build on the CellCiphr profile and better reflect the complexity of the response in vivo.

  11. Linking climate projections to performance: A yield-based decision scaling assessment of a large urban water resources system

    NASA Astrophysics Data System (ADS)

    Turner, Sean W. D.; Marlow, David; Ekström, Marie; Rhodes, Bruce G.; Kularathna, Udaya; Jeffrey, Paul J.

    2014-04-01

    Despite a decade of research into climate change impacts on water resources, the scientific community has delivered relatively few practical methodological developments for integrating uncertainty into water resources system design. This paper presents an application of the "decision scaling" methodology for assessing climate change impacts on water resources system performance and asks how such an approach might inform planning decisions. The decision scaling method reverses the conventional ethos of climate impact assessment by first establishing the climate conditions that would compel planners to intervene. Climate model projections are introduced at the end of the process to characterize climate risk in such a way that avoids the process of propagating those projections through hydrological models. Here we simulated 1000 multisite synthetic monthly streamflow traces in a model of the Melbourne bulk supply system to test the sensitivity of system performance to variations in streamflow statistics. An empirical relation was derived to convert decision-critical flow statistics to climatic units, against which 138 alternative climate projections were plotted and compared. We defined the decision threshold in terms of a system yield metric constrained by multiple performance criteria. Our approach allows for fast and simple incorporation of demand forecast uncertainty and demonstrates the reach of the decision scaling method through successful execution in a large and complex water resources system. Scope for wider application in urban water resources planning is discussed.

  12. Statistical analysis of features associated with protein expression/solubility in an in vivo Escherichia coli expression system and a wheat germ cell-free expression system.

    PubMed

    Hirose, Shuichi; Kawamura, Yoshifumi; Yokota, Kiyonobu; Kuroita, Toshihiro; Natsume, Tohru; Komiya, Kazuo; Tsutsumi, Takeshi; Suwa, Yorimasa; Isogai, Takao; Goshima, Naoki; Noguchi, Tamotsu

    2011-07-01

    Recombinant protein technology is an important tool in many industrial and pharmacological applications. Although the success rate of obtaining soluble proteins is relatively low, knowledge of protein expression/solubility under 'standard' conditions may increase the efficiency and reduce the cost of proteomics studies. In this study, we conducted a genome-scale experiment to assess the overexpression and the solubility of human full-length cDNA in an in vivo Escherichia coli expression system and a wheat germ cell-free expression system. We evaluated the influences of sequence and structural features on protein expression/solubility in each system and estimated a minimal set of features associated with them. A comparison of the feature sets related to protein expression/solubility in the in vivo Escherichia coli expression system revealed that the structural information was strongly associated with protein expression, rather than protein solubility. Moreover, a significant difference was found in the number of features associated with protein solubility in the two expression systems.

  13. Development of an anhydrotetracycline-inducible expression system for expression of a neopullulanase in B. subtilis.

    PubMed

    Heravi, Kambiz Morabbi; Watzlawick, Hildegard; Altenbuchner, Josef

    2015-11-01

    Bacillus subtilis is a widely used bacterium for production of heterologous and homologous proteins. The primary challenge in the production of proteins in B. subtilis is choosing a relevant expression system. In this study, we developed a robust expression system based on optimized PtetR of transposon Tn1721, which is repressible by its specific repressor, TetR. The first step of this work was focused on the optimization of structure and core elements of Tn1721 anhydrotetracycline-inducible promoters, PtetA and PtetR. Both promoters were inserted upstream of eGFP on a pUB110-derivative with high copy number. Reduction of the 18 bp spacer region of both PtetA and PtetR to 17 bp significantly increased their strength in B. subtilis. Nevertheless, only the optimized PtetR with 17 bp spacer region (PtetR2) directed high level of eGFP expression. In the second step, regulation of the system was optimized by testing the expression of tetR using well-known promoters, such as PmtlA, PmtlR, PptsG and PpenP. Expression of tetR by PptsG resulted in a tight regulation of PtetR2-eGFP showing 44-fold induction. By using the final expression plasmid in B. subtilis, neopullulanase was produced up to 15% of the total soluble protein. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Modelling Pasture-based Automatic Milking System Herds: The Impact of Large Herd on Milk Yield and Economics.

    PubMed

    Islam, M R; Clark, C E F; Garcia, S C; Kerrisk, K L

    2015-07-01

    The aim of this modelling study was to investigate the effect of large herd size (and land areas) on walking distances and milking interval (MI), and their impact on milk yield and economic penalties when 50% of the total diets were provided from home grown feed either as pasture or grazeable complementary forage rotation (CFR) in an automatic milking system (AMS). Twelve scenarios consisting of 3 AMS herds (400, 600, 800 cows), 2 levels of pasture utilisation (current AMS utilisation of 15.0 t dry matter [DM]/ha, termed as 'moderate'; optimum pasture utilisation of 19.7 t DM/ha, termed as 'high') and 2 rates of incorporation of grazeable complementary forage system (CFS: 0, 30%; CFS = 65% farm is CFR and 35% of farm is pasture) were investigated. Walking distances, energy loss due to walking, MI, reduction in milk yield and income loss were calculated for each treatment based on information available in the literature. With moderate pasture utilisation and 0% CFR, increasing the herd size from 400 to 800 cows resulted in an increase in total walking distances between the parlour and the paddock from 3.5 to 6.3 km. Consequently, MI increased from 15.2 to 16.4 h with increased herd size from 400 to 800 cows. High pasture utilisation (allowing for an increased stocking density) reduced the total walking distances up to 1 km, thus reduced the MI by up to 0.5 h compared to the moderate pasture, 800 cow herd combination. The high pasture utilisation combined with 30% of the farm in CFR in the farm reduced the total walking distances by up to 1.7 km and MI by up to 0.8 h compared to the moderate pasture and 800 cow herd combination. For moderate pasture utilisation, increasing the herd size from 400 to 800 cows resulted in more dramatic milk yield penalty as yield increasing from c.f. 2.6 and 5.1 kg/cow/d respectively, which incurred a loss of up to $AU 1.9/cow/d. Milk yield losses of 0.61 kg and 0.25 kg for every km increase in total walking distance (voluntary return

  15. Modelling Pasture-based Automatic Milking System Herds: The Impact of Large Herd on Milk Yield and Economics

    PubMed Central

    Islam, M. R.; Clark, C. E. F.; Garcia, S. C.; Kerrisk, K. L.

    2015-01-01

    The aim of this modelling study was to investigate the effect of large herd size (and land areas) on walking distances and milking interval (MI), and their impact on milk yield and economic penalties when 50% of the total diets were provided from home grown feed either as pasture or grazeable complementary forage rotation (CFR) in an automatic milking system (AMS). Twelve scenarios consisting of 3 AMS herds (400, 600, 800 cows), 2 levels of pasture utilisation (current AMS utilisation of 15.0 t dry matter [DM]/ha, termed as ‘moderate’; optimum pasture utilisation of 19.7 t DM/ha, termed as ‘high’) and 2 rates of incorporation of grazeable complementary forage system (CFS: 0, 30%; CFS = 65% farm is CFR and 35% of farm is pasture) were investigated. Walking distances, energy loss due to walking, MI, reduction in milk yield and income loss were calculated for each treatment based on information available in the literature. With moderate pasture utilisation and 0% CFR, increasing the herd size from 400 to 800 cows resulted in an increase in total walking distances between the parlour and the paddock from 3.5 to 6.3 km. Consequently, MI increased from 15.2 to 16.4 h with increased herd size from 400 to 800 cows. High pasture utilisation (allowing for an increased stocking density) reduced the total walking distances up to 1 km, thus reduced the MI by up to 0.5 h compared to the moderate pasture, 800 cow herd combination. The high pasture utilisation combined with 30% of the farm in CFR in the farm reduced the total walking distances by up to 1.7 km and MI by up to 0.8 h compared to the moderate pasture and 800 cow herd combination. For moderate pasture utilisation, increasing the herd size from 400 to 800 cows resulted in more dramatic milk yield penalty as yield increasing from c.f. 2.6 and 5.1 kg/cow/d respectively, which incurred a loss of up to $AU 1.9/cow/d. Milk yield losses of 0.61 kg and 0.25 kg for every km increase in total walking distance (voluntary

  16. Near-infrared spectroscopy of a hydroecological indicator: new tool for determining sustainable yield for Floridan aquifer system

    NASA Astrophysics Data System (ADS)

    Bacchus, Sydney T.; Archibald, Douglas D.; Brook, George A.; Britton, Kerry O.; Haines, Bruce L.; Rathbun, Stephen L.; Madden, Marguerite

    2003-06-01

    Pond-cypress (Taxodium ascendens Brong.) is a dominant canopy species in depressional wetlands of the south-eastern Coastal Plain. Unsustainable withdrawals from the karst Floridan aquifer system have caused premature decline and death of pond-cypress trees, presumably owing to altered hydroperiods (which alter the flow of water and nutrients in trees). There has been no scientifically based means to determine sustainable yield from this regional aquifer system or to detect early stages of physical/ecological damage associated with groundwater mining and aquifer storage and recovery (ASR, which also can alter natural hydroperiods). In this study, the relationship between visual symptoms (indicators) of stress or premature decline, and spectral reflectance was evaluated using dried, milled branch tips collected from natural stands of mature pond-cypress. Depressional systems evaluated represented four of the six aquifer system subregions where subsurface perturbations from groundwater mining: (i) were presumed not to be occurring (reference wetlands); (ii) may be occurring but are not documented; and (iii) have been confirmed. Sampled trees were assigned to one of three stress classes (1, no/minimal; 2, moderate; 3, severe) based on the visual indicators. Partial least squares-linear discriminant analysis of second derivative spectral transformations in the visible/shortwave near-infrared (NIR) region (400-1100 nm) and the NIR region (1100-2500 nm) was used to evaluate the samples in assigned classes.Class 1 samples were discriminated from combined class 2 and 3 samples in the NIR region with 100% and 97% accuracy for consecutive winter sample periods (before bud-break). The percentage of correctly classified samples in this spectral region was lower (85%) for summer samples (full leaf-out). Second-derivative models for the NIR region developed from the winter data sets predicted assigned classes for alternate winter's samples with an accuracy of 97% and 100%. High

  17. Recombinants proteins for industrial uses: utilization of Pichia pastoris expression system.

    PubMed

    Rabert, Claudia; Weinacker, Daniel; Pessoa, Adalberto; Farías, Jorge G

    2013-01-01

    The innovation in industrial process with impact in the efficient production is the major challenge for actual industry. A high numerous of enzymes are utilized in at different level of process; the search for new alternatives with better characteristic has become a field of study of great interest, the recombinant protein achievement in a different host system is an alternative widely assessed for production of this. The microorganism Pichia pastoris has been used like a successful expression system in diverse areas, improved the yield and extraction-recovery of the product expressed. The reported of diverse authors in the production of enzymes with different application in industry is varied, in this review the different industry areas and the characteristic of the enzymes produced are detailed.

  18. Recombinants proteins for industrial uses: utilization of Pichia pastoris expression system

    PubMed Central

    Rabert, Claudia; Weinacker, Daniel; Pessoa, Adalberto; Farías, Jorge G.

    2013-01-01

    The innovation in industrial process with impact in the efficient production is the major challenge for actual industry. A high numerous of enzymes are utilized in at different level of process; the search for new alternatives with better characteristic has become a field of study of great interest, the recombinant protein achievement in a different host system is an alternative widely assessed for production of this. The microorganism Pichia pastoris has been used like a successful expression system in diverse areas, improved the yield and extraction-recovery of the product expressed. The reported of diverse authors in the production of enzymes with different application in industry is varied, in this review the different industry areas and the characteristic of the enzymes produced are detailed. PMID:24294221

  19. Thiamine supplementation facilitates thiamine transporter expression in the rumen epithelium and attenuates high-grain-induced inflammation in low-yielding dairy cows.

    PubMed

    Pan, X H; Yang, L; Beckers, Y; Xue, F G; Tang, Z W; Jiang, L S; Xiong, B H

    2017-07-01

    An experiment was conducted to uncover the effects of increasing dietary grain levels on expression of thiamine transporters in ruminal epithelium, and to assess the protective effects of thiamine against high-grain-induced inflammation in dairy cows. Six rumen-fistulated, lactating Holstein dairy cows (627 ± 16.9 kg of body weight, 180 ± 6 d in milk; mean ± standard deviation) were randomly assigned to a replicated 3 × 3 Latin square design trial. Three treatments were control (20% dietary starch, dry matter basis), high-grain diet (HG, 33.2% dietary starch, DM basis), and HG diet supplemented with 180 mg of thiamine/kg of dry matter intake. On d 19 and 20 of each period, milk performance was measured. On d 21, ruminal pH, endotoxic lipopolysaccharide (LPS), and thiamine contents in rumen and blood, and plasma inflammatory cytokines were detected; a rumen papillae biopsy was taken on d 21 to determine the gene and protein expression of toll-like receptor 4 (TLR4) signaling pathways. The HG diet decreased ruminal pH (5.93 vs. 6.49), increased milk yield from 17.9 to 20.2 kg/d, and lowered milk fat and protein from 4.28 to 3.83%, and from 3.38 to 3.11%, respectively. The HG feeding reduced thiamine content in rumen (2.89 vs. 8.97 μg/L) and blood (11.66 vs. 17.63 μg/L), and the relative expression value of thiamine transporter-2 (0.37-fold) and mitochondrial thiamine pyrophosphate transporter (0.33-fold) was downregulated by HG feeding. The HG-fed cows exhibited higher endotoxin LPS in rumen fluid (134,380 vs. 11,815 endotoxin units/mL), and higher plasma concentrations of lipopolysaccharide binding protein and pro-inflammatory cytokines when compared with the control group. The gene and protein expression of tumor necrosis factor α (TNFα), IL1B, and IL6 in rumen epithelium increased when cows were fed the HG diet, indicating that local inflammation occurred. The depressions in ruminal pH, milk fat, and protein of HG-fed cows were reversed by thiamine

  20. Development and application of a T7 RNA polymerase-dependent expression system for antibiotic production improvement in Streptomyces.

    PubMed

    Wei, Junhong; Tian, Jinjin; Pan, Guoqing; Xie, Jie; Bao, Jialing; Zhou, Zeyang

    2017-06-01

    To develop a reliable and easy to use expression system for antibiotic production improvement of Streptomyces. A two-compound T7 RNA polymerase-dependent gene expression system was developed to fulfill this demand. In this system, the T7 RNA polymerase coding sequence was optimized based on the codon usage of Streptomyces coelicolor. To evaluate the functionality of this system, we constructed an activator gene overexpression strain for enhancement of actinorhodin production. By overexpression of the positive regulator actII-ORF4 with this system, the maximum actinorhodin yield of engineered strain was 15-fold higher and the fermentation time was decreased by 48 h. The modified two-compound T7 expression system improves both antibiotic production and accelerates the fermentation process in Streptomyces. This provides a general and useful strategy for strain improvement of important antibiotic producing Streptomyces strains.

  1. Tree growth and management in Ugandan agroforestry systems: effects of root pruning on tree growth and crop yield.

    PubMed

    Wajja-Musukwe, Tellie-Nelson; Wilson, Julia; Sprent, Janet I; Ong, Chin K; Deans, J Douglas; Okorio, John

    2008-02-01

    Tree root pruning is a potential tool for managing belowground competition when trees and crops are grown together in agroforestry systems. We investigated the effects of tree root pruning on shoot growth and root distribution of Alnus acuminata (H.B. & K.), Casuarina equisetifolia L., Grevillea robusta A. Cunn. ex R. Br., Maesopsis eminii Engl. and Markhamia lutea (Benth.) K. Schum. and on yield of adjacent crops in sub-humid Uganda. The trees were 3 years old at the commencement of the study, and most species were competing strongly with crops. Tree roots were pruned 41 months after planting by cutting and back-filling a trench to a depth of 0.3 m, at a distance of 0.3 m from the trees, on one side of the tree row. The trench was reopened and roots recut at 50 and 62 months after planting. We assessed the effects on tree growth and root distribution over a 3 year period, and crop yield after the third root pruning at 62 months. Overall, root pruning had only a slight effect on aboveground tree growth: height growth was unaffected and diameter growth was reduced by only 4%. A substantial amount of root regrowth was observed by 11 months after pruning. Tree species varied in the number and distribution of roots, and C. equisetifolia and M. lutea had considerably more roots per unit of trunk volume than the other species, especially in the surface soil layers. Casuarina equisetifolia and M. eminii were the tree species most competitive with crops and G. robusta and M. lutea the least competitive. Crop yield data provided strong evidence of the redistribution of root activity following root pruning, with competition increasing on the unpruned side of tree rows. Thus, one-sided root pruning will be useful in only a few circumstances.

  2. Constitutive expression of McCHIT1-PAT enhances resistance to rice blast and herbicide, but does not affect grain yield in transgenic glutinous rice.

    PubMed

    Zeng, Xiao-Fang; Li, Lei; Li, Jian-Rong; Zhao, De-Gang

    2016-01-01

    To produce new rice blast- and herbicide-resistant transgenic rice lines, the McCHIT1 gene encoding the class I chitinase from Momordica charantia and the herbicide resistance gene PAT were introduced into Lailong (Oryza sativa L. ssp. Japonica), a glutinous local rice variety from Guizhou Province, People's Republic of China. Transgenic lines were identified by ß-glucuronidase (GUS) histochemical staining, PCR, and Southern blot analyses. Agronomic traits, resistance to rice blast and herbicide, chitinase activities, and transcript levels of McCHIT1 were assessed in the T2 progeny of three transgenic lines (L1, L8, and L10). The results showed that the introduction of McCHIT1-PAT into Lailong significantly enhanced herbicide and blast resistance. After infection with the blast fungus Magnaporthe oryzae, all of the T2 progeny exhibited less severe lesion symptoms than those of wild type. The disease indices were 100% for wild type, 65.66% for T2 transgenic line L1, 59.69% for T2 transgenic line L8, and 79.80% for T2 transgenic line L10. Transgenic lines expressing McCHIT1-PAT did not show a significant difference from wild type in terms of malondialdehyde (MDA) content, polyphenol oxidase (PPO) activity, and superoxide dismutase (SOD) activity in the leaves. However, after inoculation with M. oryzae, transgenic plants showed significantly higher SOD and PPO activities and lower MDA contents in leaves, compared with those in wild-type leaves. The transgenic and the wild-type plants did not show significant differences in grain yield parameters including plant height, panicles per plant, seeds per panicle, and 1000-grain weight. Therefore, the transgenic plants showed increased herbicide and blast resistance, with no yield penalty.

  3. Brevibacillus expression system: host-vector system for efficient production of secretory proteins.

    PubMed

    Mizukami, Makoto; Hanagata, Hiroshi; Miyauchi, Akira

    2010-04-01

    Brevibacillus expression system is an effective bacterial expression system for secretory proteins. The host bacterium, Brevibacillus choshinensis, a gram-positive bacterium, has strong capacity to secrete a large amount of proteins (approximately 30 g/L), which mostly consist of cell wall protein. A host-vector system that utilizes such high expression capacity has been constructed for the production of secretory proteins and tested for various heterologous proteins, including cytokines, enzymes, antigens, and adjuvants.

  4. Stepwise optimization of a low-temperature Bacillus subtilis expression system for "difficult to express" proteins.

    PubMed

    Welsch, Norma; Homuth, Georg; Schweder, Thomas

    2015-08-01

    In order to improve the overproduction of "difficult to express" proteins, a low-temperature expression system for Bacillus subtilis based on the cold-inducible promoter of the desaturase-encoding des gene was constructed. Selected regulatory DNA sequence elements from B. subtilis genes known to be cold-inducible were fused to different model genes. It could be demonstrated that these regulatory elements are able to mediate increased heterologous gene expression, either by improved translation efficiency or by higher messenger RNA (mRNA) stability. In case of a cold-adapted β-galactosidase from Pseudoalteromonas haloplanktis TAE79A serving as the model, significantly higher expression was achieved by fusing its coding sequence to the so-called "downstream box" sequence of cspB encoding the major B. subtilis cold-shock protein. The combination of this fusion with a cspB 5'-UTR stem-loop structure resulted in further enhancement of the β-galactosidase expression. In addition, integration of the transcription terminator of the B. subtilis cold-inducible bkd operon downstream of the target genes caused a higher mRNA stability and enabled thus a further significant increase in expression. Finally, the fully optimized expression system was validated by overproducing a B. subtilis xylanase as well as an α-glucosidase from Saccharomyces cerevisiae, the latter known for tending to form inclusion bodies. These analyses verified the applicability of the engineered expression system for extracellular and intracellular protein synthesis in B. subtilis, thereby confirming the suitability of this host organism for the overproduction of critical, poorly soluble proteins.

  5. Hormonal treatment of the bark of rubber trees (Hevea brasiliensis) increases latex yield through latex dilution in relation with the differential expression of two aquaporin genes.

    PubMed

    Tungngoen, Kessarin; Viboonjun, Unchera; Kongsawadworakul, Panida; Katsuhara, Maki; Julien, Jean-Louis; Sakr, Soulaiman; Chrestin, Hervé; Narangajavana, Jarunya

    2011-02-15

    Natural rubber is synthesized in laticifers in the inner liber of the rubber tree (Hevea brasiliensis). Upon bark tapping, the latex is expelled due to liber turgor pressure. The mature laticifers are devoid of plasmodesmata; therefore a corresponding decrease in the total latex solid content is likely to occur due to water influx inside the laticifers. Auxins and ethylene used as efficient yield stimulants in mature untapped rubber trees, but, bark treatments with abscisic acid (ABA) and salicylic acid (SA) could also induce a transient increase latex yield. We recently reported that there are three aquaporin genes, HbPIP2;1, HbTIP1;1 and HbPIP1;1, that are regulated differentially after ethylene bark treatment. HbPIP2;1 was up-regulated in both the laticifers and the inner liber tissues, whereas HbTIP1;1 was up-regulated in the latex cells, but very markedly down-regulated in the inner liber tissues. Conversely, HbPIP1;1 was down-regulated in both tissues. In the present study, HbPIP2;1 and HbTIP1;1 showed a similar expression in response to auxin, ABA and SA, as seen in ethylene stimulation, while HbPIP1;1 was slightly regulated by auxin, but neither by ABA nor SA. The analysis of the HbPIP1;1 promoter region indicated the presence of only ethylene and auxin responsive elements. In addition, the poor efficiency of this HbPIP1;1 in increasing plasmalemma water conductance was confirmed in Xenopus oocytes. Thus, an increase in latex yield in response to all of these hormones was proposed to be the major function of aquaporins, HbPIP2;1 and HbTIP1;1. This study emphasized that the circulation of water between the laticifers and their surrounding tissues that result in latex dilution, as well as the probable maintenance of the liber tissues turgor pressure, favor the prolongation of latex flow.

  6. Unmanned aircraft system-derived crop height and normalized difference vegetation index metrics for sorghum yield and aphid stress assessment

    NASA Astrophysics Data System (ADS)

    Stanton, Carly; Starek, Michael J.; Elliott, Norman; Brewer, Michael; Maeda, Murilo M.; Chu, Tianxing

    2017-04-01

    A small, fixed-wing unmanned aircraft system (UAS) was used to survey a replicated small plot field experiment designed to estimate sorghum damage caused by an invasive aphid. Plant stress varied among 40 plots through manipulation of aphid densities. Equipped with a consumer-grade near-infrared camera, the UAS was flown on a recurring basis over the growing season. The raw imagery was processed using structure-from-motion to generate normalized difference vegetation index (NDVI) maps of the fields and three-dimensional point clouds. NDVI and plant height metrics were averaged on a per plot basis and evaluated for their ability to identify aphid-induced plant stress. Experimental soil signal filtering was performed on both metrics, and a method filtering low near-infrared values before NDVI calculation was found to be the most effective. UAS NDVI was compared with NDVI from sensors onboard a manned aircraft and a tractor. The correlation results showed dependence on the growth stage. Plot averages of NDVI and canopy height values were compared with per-plot yield at 14% moisture and aphid density. The UAS measures of plant height and NDVI were correlated to plot averages of yield and insect density. Negative correlations between aphid density and NDVI were seen near the end of the season in the most damaged crops.

  7. Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system.

    PubMed

    Lombana, T Noelle; Dillon, Michael; Bevers, Jack; Spiess, Christoph

    2015-12-03

    Rapid identification of residues that influence antibody expression and thermostability is often needed to move promising therapeutics into the clinic. To establish a method that can assess small expression differences, we developed a Bacterial Antibody Display (BAD) system that overcomes previous limitations, enabling the use of full-length formats for antibody and antigen in a live cell setting. We designed a unique library of individual framework variants using natural diversity introduced by somatic hypermutation, and screened half-antibodies for increased expression using BAD. We successfully identify variants that dramatically improve expression yields and in vitro thermostability of two therapeutically relevant antibodies in E. coli and mammalian cells. While we study antibody expression, bacterial display can now be expanded to examine the processes of protein folding and translocation. Additionally, our natural library design strategy could be applied during antibody humanization and library design for in vitro display methods to maintain expression and formulation stability.

  8. Expression and bioactivity of human α-fetoprotein in a Bac-to-Bac system

    PubMed Central

    Lin, Bo; Liu, Kun; Wang, Wenting; Li, Wei; Dong, Xu; Chen, Yi; Lu, Yan; Guo, Junli; Li, Mengsen

    2016-01-01

    α-fetoprotein (AFP) is an early serum growth factor in foetal embryonic development and hepatic oncogenesis. A growing number of investigations of AFP as a tumour-specific biomarker have concluded that AFP is an important target for cancer treatment. AFP also plays an immunomodulatory role in the treatment of several autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis, myasthenia gravis and thyroiditis. In an effort to support biochemical screening and drug design and discovery, we attempted to express and purify human AFP in a Bac-to-Bac system. Two key factors affecting the expression of recombinant human AFP (R-AFP), namely the infectious baculovirus inoculum volume and the culturing time post-infection, were optimized to maximize the yield. We achieved a high yield of approximately 1.5 mg/l of harvested medium with a 72–96 h incubation period after infection and an inoculum volume ratio of 1:100. We also assessed the role of R-AFP in the proliferation of the human liver cancer cell line Bel 7402, and the results indicated that R-AFP promoted the growth of hepatoma cells. We concluded that this method can produce high yields of R-AFP, which can be used for studies related to AFP. PMID:27913752

  9. Expression and bioactivity of human α-fetoprotein in a Bac-to-Bac system.

    PubMed

    Lin, Bo; Liu, Kun; Wang, Wenting; Li, Wei; Dong, Xu; Chen, Yi; Lu, Yan; Guo, Junli; Zhu, Mingyue; Li, Mengsen

    2017-02-28

    α-fetoprotein (AFP) is an early serum growth factor in foetal embryonic development and hepatic oncogenesis. A growing number of investigations of AFP as a tumour-specific biomarker have concluded that AFP is an important target for cancer treatment. AFP also plays an immunomodulatory role in the treatment of several autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis, myasthenia gravis and thyroiditis. In an effort to support biochemical screening and drug design and discovery, we attempted to express and purify human AFP in a Bac-to-Bac system. Two key factors affecting the expression of recombinant human AFP (R-AFP), namely the infectious baculovirus inoculum volume and the culturing time post-infection, were optimized to maximize the yield. We achieved a high yield of approximately 1.5 mg/l of harvested medium with a 72-96 h incubation period after infection and an inoculum volume ratio of 1:100. We also assessed the role of R-AFP in the proliferation of the human liver cancer cell line Bel 7402, and the results indicated that R-AFP promoted the growth of hepatoma cells. We concluded that this method can produce high yields of R-AFP, which can be used for studies related to AFP.

  10. Genetic selection system for improving recombinant membrane protein expression in E. coli

    PubMed Central

    Massey-Gendel, Elizabeth; Zhao, Anni; Boulting, Gabriella; Kim, Hye-Yeon; Balamotis, Michael A; Seligman, Len M; Nakamoto, Robert K; Bowie, James U

    2009-01-01

    A major barrier to the physical characterization and structure determination of membrane proteins is low yield in recombinant expression. To address this problem, we have designed a selection strategy to isolate mutant strains of Escherichia coli that improve the expression of a targeted membrane protein. In this method, the coding sequence of the membrane protein of interest is fused to a C-terminal selectable marker, so that the production of the selectable marker and survival on selective media is linked to expression of the targeted membrane protein. Thus, mutant strains with improved expression properties can be directly selected. We also introduce a rapid method for curing isolated strains of the plasmids used during the selection process, in which the plasmids are removed by in vivo digestion with the homing endonuclease I-CreI. We tested this selection system on a rhomboid family protein from Mycobacterium tuberculosis (Rv1337) and were able to isolate mutants, which we call EXP strains, with up to 75-fold increased expression. The EXP strains also improve the expression of other membrane proteins that were not the target of selection, in one case roughly 90-fold. PMID:19165721

  11. Development of a Heat-Shock Inducible Gene Expression System in the Red Alga Cyanidioschyzon merolae

    PubMed Central

    Sumiya, Nobuko; Fujiwara, Takayuki; Kobayashi, Yusuke; Misumi, Osami; Miyagishima, Shin-ya

    2014-01-01

    The cell of the unicellular red alga Cyanidioschyzon merolae contains a single chloroplast and mitochondrion, the division of which is tightly synchronized by a light/dark cycle. The genome content is extremely simple, with a low level of genetic redundancy, in photosynthetic eukaryotes. In addition, transient transformation and stable transformation by homologous recombination have been reported. However, for molecular genetic analyses of phenomena that are essential for cellular growth and survival, inducible gene expression/suppression systems are needed. Here, we report the development of a heat-shock inducible gene expression system in C. merolae. CMJ101C, encoding a small heat shock protein, is transcribed only when cells are exposed to an elevated temperature. Using a superfolder GFP as a reporter protein, the 200-bp upstream region of CMJ101C orf was determined to be the optimal promoter for heat-shock induction. The optimal temperature to induce expression is 50°C, at which C. merolae cells are able to proliferate. At least a 30-min heat shock is required for the expression of a protein of interest and a 60-min heat shock yields the maximum level of protein expression. After the heat shock, the mRNA level decreases rapidly. As an example of the system, the expression of a dominant negative form of chloroplast division DRP5B protein, which has a mutation in the GTPase domain, was induced. Expression of the dominant negative DRP5B resulted in the appearance of aberrant-shaped cells in which two daughter chloroplasts and the cells are still connected by a small DRP5B positive tube-like structure. This result suggests that the dominant negative DRP5B inhibited the final scission of the chloroplast division site, but not the earlier stages of division site constriction. It is also suggested that cell cycle progression is not arrested by the impairment of chloroplast division at the final stage. PMID:25337786

  12. Comparative analysis of eukaryotic cell-free expression systems.

    PubMed

    Hartsough, Emily M; Shah, Pankti; Larsen, Andrew C; Chaput, John C

    2015-09-01

    Cell-free protein synthesis (CFPS) allows researchers to rapidly generate functional proteins independent of cell culture. Although advances in eukaryotic lysates have increased the amount of protein that can be produced, the nuances of different translation systems lead to variability in protein production. To help overcome this problem, we have compared the relative yield and template requirements for three commonly used commercial cell-free translation systems: wheat germ extract (WGE), rabbit reticulocyte lysate (RRL), and HeLa cell lysate (HCL). Our results provide a general guide for researchers interested in using cell-free translation to generate recombinant protein for biomedical applications.

  13. Expression of SOD and APX genes positively regulates secondary cell wall biosynthesis and promotes plant growth and yield in Arabidopsis under salt stress.

    PubMed

    Shafi, Amrina; Chauhan, Rohit; Gill, Tejpal; Swarnkar, Mohit K; Sreenivasulu, Yelam; Kumar, Sanjay; Kumar, Neeraj; Shankar, Ravi; Ahuja, Paramvir Singh; Singh, Anil Kumar

    2015-04-01

    Abiotic stresses cause accumulation of reactive oxygen species (ROS), such as hydrogen peroxide (H2O2) in plants. Sophisticated mechanisms are required to maintain optimum level of H2O2 that acts as signalling molecule regulating adaptive response to salt stress. CuZn-superoxide dismutase (CuZn-SOD) and ascorbate peroxidase (APX) constitute first line of defence against oxidative stress. In the present study, PaSOD and RaAPX genes from Potentilla atrosanguinea and Rheum australe, respectively were overexpressed individually as well as in combination in Arabidopsis thaliana. Interestingly, PaSOD and dual transgenic lines exhibit enhanced lignin deposition in their vascular bundles with altered S:G ratio under salt stress. RNA-seq analysis revealed that expression of PaSOD gene in single and dual transgenics positively regulates expression of lignin biosynthesis genes and transcription factors (NACs, MYBs, C3Hs and WRKY), leading to enhanced and ectopic deposition of lignin in vascular tissues with larger xylem fibres and alters S:G ratio, as well. In addition, transgenic plants exhibit growth promotion, higher biomass production and increased yield under salt stress as compared to wild type plants. Our results suggest that in dual transgenics, ROS generated during salt stress gets converted into H2O2 by SOD and its optimum level was maintained by APX. This basal level of H2O2 acts as messenger for transcriptional activation of lignin biosynthesis in vascular tissue, which provides mechanical strength to plants. These findings reveal an important role of PaSOD and RaAPX in enhancing salt tolerance of transgenic Arabidopsis via increased accumulation of compatible solutes and by regulating lignin biosynthesis.

  14. Novel T7-like expression systems used for Halomonas.

    PubMed

    Zhao, Han; Zhang, Haoqian M; Chen, Xiangbin; Li, Teng; Wu, Qiong; Ouyang, Qi; Chen, Guo-Qiang

    2017-01-01

    To engineer non-model organisms, suitable genetic parts must be available. However, biological parts are often host strain sensitive. It is therefore necessary to develop genetic parts that are functional regardless of host strains. Here we report several novel phage-derived expression systems used for transcriptional control in non-model bacteria. Novel T7-like RNA polymerase-promoter pairs were obtained by mining phage genomes, followed by in vivo characterization in non-model strains Halomonas spp TD01 and Pseudomonas entomophila. Three expression systems, namely, MmP1, VP4, and K1F, were developed displaying orthogonality (crosstalk<0.7%), tight regulation (3085-fold induction), and high efficiency (2.5-fold of Ptac) in Halomonas sp. TD01, a chassis strain with a high industrial value. The expression under the corresponding T7-like promoter libraries persisted with striking correlations (R(2) >0.94) between Escherichia coli and Halomonas sp. TD01, implying suitability of broad-host range. Three Halomonas TD strains were then constructed based upon these expression systems that enabled interchangeable and controllable gene expression. One of the strains termed Halomonas TD-MmP1 was used to express the cell-elongation cassette (minCD genes) and polyhydroxybutyrate (PHB) biosynthetic pathway, resulting in a 100-fold increase in cell lengths and high levels of PHB production (up to 92% of cell dry weight), respectively. We envision these T7-like expression systems to benefit metabolic engineering in other non-model organisms. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  15. 50 year trends in nitrogen use efficiency of world cropping systems: the relationship between yield and nitrogen input to cropland

    NASA Astrophysics Data System (ADS)

    Lassaletta, Luis; Billen, Gilles; Grizzetti, Bruna; Anglade, Juliette; Garnier, Josette

    2014-10-01

    Nitrogen (N) is crucial for crop productivity. However, nowadays more than half of the N added to cropland is lost to the environment, wasting the resource, producing threats to air, water, soil and biodiversity, and generating greenhouse gas emissions. Based on FAO data, we have reconstructed the trajectory followed, in the past 50 years, by 124 countries in terms of crop yield and total nitrogen inputs to cropland (manure, synthetic fertilizer, symbiotic fixation and atmospheric deposition). During the last five decades, the response of agricultural systems to increased nitrogen fertilization has evolved differently in the different world countries. While some countries have improved their agro-environmental performances, in others the increased fertilization has produced low agronomical benefits and higher environmental losses. Our data also suggest that, in general, those countries using a higher proportion of N inputs from symbiotic N fixation rather than from synthetic fertilizer have a better N use efficiency.

  16. Testing DAYCENT model simulations of corn yields and nitrous oxide emissions in irrigated tillage systems in Colorado.

    PubMed

    Del Grosso, S J; Halvorson, A D; Parton, W J

    2008-01-01

    Agricultural soils are responsible for the majority of nitrous oxide (N(2)O) emissions in the USA. Irrigated cropping, particularly in the western USA, is an important source of N(2)O emissions. However, the impacts of tillage intensity and N fertilizer amount and type have not been extensively studied for irrigated systems. The DAYCENT biogeochemical model was tested using N(2)O, crop yield, soil N and C, and other data collected from irrigated cropping systems in northeastern Colorado during 2002 to 2006. DAYCENT uses daily weather, soil texture, and land management information to simulate C and N fluxes between the atmosphere, soil, and vegetation. The model properly represented the impacts of tillage intensity and N fertilizer amount on crop yields, soil organic C (SOC), and soil water content. DAYCENT N(2)O emissions matched the measured data in that simulated emissions increased as N fertilization rates increased and emissions from no-till (NT) tended to be lower on average than conventional-till (CT). However, the model overestimated N(2)O emissions. Lowering the amount of N(2)O emitted per unit of N nitrified from 2 to 1% helped improve model fit but the treatments receiving no N fertilizer were still overestimated by more than a factor of 2. Both the model and measurements showed that soil NO(3)(-) levels increase with N fertilizer addition and with tillage intensity, but DAYCENT underestimated NO(3)(-) levels, particularly for the treatments receiving no N fertilizer. We suggest that DAYCENT could be improved by reducing the background nitrification rate and by accounting for the impact of changes in microbial community structure on denitrification rates.

  17. Combined Laboratory and Numerical Study on Thermal Plumes in Yield-Stress Fluids : Implications for Midocean-Ridge Systems

    NASA Astrophysics Data System (ADS)

    Massmeyer, A.; Davaille, A.; Rolf, T.; Tackley, P.; Di Giuseppe, E.

    2012-04-01

    Diapirs form by hot magma migrating through the lithosphere. While hot instabilities in a Newtonian mantle are known to rise as a mushroom-shaped plume, the upwards migration of hot material in "soft" geological materials (presenting solid as well as viscous properties) is still far from understood. We are therefore studying this problem experimentally and numerically, to shed some light into this problem. The results give an idea about which basic properties are needed in mid-ocean ridge systems. The Laboratory setup consists of a localized heat-source placed in a plexiglas-tank containing a mixture of water, glycerine and Carbopol. Due to the existence of a continuous network of interacting micron-sized hydrogel particles, the solution presents a yield stress and is shear-thinning. The rheology is described by a Herschel-Bulkley model. The fluid's rheological properties can be controlled by changing either the proportion of Glycerol and water or the Carbopol concentration. In our experiments we systematically studied the influence of rheological properties and supplied heat. Depending on the Yield parameter Y 0, which compares the thermally-induced stress to the yield stress, three different regimes are observed. For low Y 0, no convection develops. For intermediate values, a small-scale convection cell appears and remains confined around the heater. Only for high Y 0, a thermal plume develops. The morphology differs from the mushroom-shape typically encountered in Newtonian or in purely shear-thinning fluids. The experiments show that the onset time is delayed compared to a Newtonian case and depends on Y 0. Combined temperature and velocity field measurements show that a plug flow develops within the plume thermal anomaly, therefore producing a rising finger-shape. Moreover, although the heat supply is continuous through time, the uplift of such plumes is not: the thermal anomaly marks some "pauses" as it rises through the gel column. In addition we implemented

  18. Effects of prebiotics on immune system and cytokine expression.

    PubMed

    Shokryazdan, Parisa; Faseleh Jahromi, Mohammad; Navidshad, Bahman; Liang, Juan Boo

    2017-02-01

    Nowadays, use of prebiotics as feed and food additives has received increasing interest because of the beneficial effects of prebiotics on the health of animals and humans. One of the beneficial effects of prebiotics is stimulation of immune system, which can be direct or indirect through increasing population of beneficial microbes or probiotics, especially lactic acid bacteria and bifidobacteria, in the gut. An important mechanism of action of probiotics and prebiotics, by which they can affect the immune system, is changing the expression of cytokines. The present review tried to summarize the findings of studies that investigated the effects of prebiotics on immune system with focusing on their effects on cytokine expression. Generally, most of reviewed studies indicated beneficial effects for prebiotics in terms of improving immune system, by increasing the expression of anti-inflammatory cytokines, while reducing the expressions of proinflammatory cytokines. However, most of studies mainly considered the indirect effects of prebiotics on the immune system (through changing the composition and population of gut microbiota), and their direct effects still need to be further studied using prebiotics with different degree of polymerization in different hosts.

  19. Tet-On Systems For Doxycycline-inducible Gene Expression

    PubMed Central

    Das, Atze T.; Tenenbaum, Liliane; Berkhout, Ben

    2016-01-01

    The tetracycline-controlled Tet-Off and Tet-On gene expression systems are used to regulate the activity of genes in eukaryotic cells in diverse settings, varying from basic biological research to biotechnology and gene therapy applications. These systems are based on regulatory elements that control the activity of the tetracycline-resistance operon in bacteria. The Tet-Off system allows silencing of gene expression by administration of tetracycline (Tc) or tetracycline-derivatives like doxycycline (dox), whereas the Tet-On system allows activation of gene expression by dox. Since the initial design and construction of the original Tet-system, these bacterium-derived systems have been significantly improved for their function in eukaryotic cells. We here review how a dox-controlled HIV-1 variant was designed and used to greatly improve the activity and dox-sensitivity of the rtTA transcriptional activator component of the Tet-On system. These optimized rtTA variants require less dox for activation, which will reduce side effects and allow gene control in tissues where a relatively low dox level can be reached, such as the brain. PMID:27216914

  20. Crop monitoring & yield forecasting system based on Synthetic Aperture Radar (SAR) and process-based crop growth model: Development and validation in South and South East Asian Countries

    NASA Astrophysics Data System (ADS)

    Setiyono, T. D.

    2014-12-01

    Accurate and timely information on rice crop growth and yield helps governments and other stakeholders adapting their economic policies and enables relief organizations to better anticipate and coordinate relief efforts in the wake of a natural catastrophe. Such delivery of rice growth and yield information is made possible by regular earth observation using space-born Synthetic Aperture Radar (SAR) technology combined with crop modeling approach to estimate yield. Radar-based remote sensing is capable of observing rice vegetation growth irrespective of cloud coverage, an important feature given that in incidences of flooding the sky is often cloud-covered. The system allows rapid damage assessment over the area of interest. Rice yield monitoring is based on a crop growth simulation and SAR-derived key information, particularly start of season and leaf growth rate. Results from pilot study sites in South and South East Asian countries suggest that incorporation of SAR data into crop model improves yield estimation for actual yields. Remote-sensing data assimilation into crop model effectively capture responses of rice crops to environmental conditions over large spatial coverage, which otherwise is practically impossible to achieve. Such improvement of actual yield estimates offers practical application such as in a crop insurance program. Process-based crop simulation model is used in the system to ensure climate information is adequately captured and to enable mid-season yield forecast.

  1. Human SUMO fusion systems enhance protein expression and solubility.

    PubMed

    Wang, Zhongyuan; Li, Haolong; Guan, Wei; Ling, Haili; Wang, Zhiyong; Mu, Tianyang; Shuler, Franklin D; Fang, Xuexun

    2010-10-01

    A major challenge associated with recombinant protein production in Escherichia coli is generation of large quantities of soluble, functional protein. Yeast SUMO (small ubiquitin-related modifier), has been shown to enhance heterologous protein expression and solubility as fusion tag, however, the effects of human SUMOs on protein expression have not been investigated. Here we describe the use of human SUMO1 and SUMO2 as a useful gene fusion technology. Human SUMO1 and SUMO2 fusion expression vectors were constructed and tested in His-tag and ubiquitin fusion expression systems. Two difficult-to-express model proteins, matrix metalloprotease-13 (MMP13) and enhanced green fluorescence protein (eGFP) were fused to the C-terminus of the human SUMO1 and SUMO2 expression vectors. These constructs were expressed in E. coli and evaluation of MMP13 and eGFP expression and solubility was conducted. We found that both SUMO1 and SUMO2 had the ability to enhance the solubility of MMP13 and eGFP, with the SUMO2 tag having a more significant effect. Since fusion tags produce varying quantities of soluble proteins, we assessed the effect of SUMO2 coupled with ubiquitin (Ub). SUMO2-ubiquitin and ubiquitin-SUMO2 fusion expression plasmids were constructed with eGFP as a passenger protein. Following expression in E. coli, both plasmids could improve eGFP expression and solubility similar to the SUMO2 fusion and better than the ubiquitin fusion. The sequential order of SUMO2 and ubiquitin had little effect on expression and solubility of eGFP. Purification of eGFP from the gene fusion product, SUMO2-ubiquitin-eGFP, involved cleavage by a deubiquitinase (Usp2-cc) and Ni-Sepharose column chromatography. The eGFP protein was purified to high homogeneity. In summary, human SUMO1 and SUMO2 are useful gene fusion technologies enhancing the expression, solubility and purification of model heterologous proteins. Copyright 2010 Elsevier Inc. All rights reserved.

  2. Maximum energy yield approach for CPV tracker design

    NASA Astrophysics Data System (ADS)

    Aldaiturriaga, E.; González, O.; Castro, M.

    2012-10-01

    Foton HC Systems has developed a new CPV tracker model, specially focused on its tracking efficiency and the effect of the tracker control techniques on the final energy yield of the system. This paper presents the theoretical work carried out into determining the energy yield for a CPV system, and illustrates the steps involved in calculating and understanding how energy consumption for tracking is opposed to tracker pointing errors. Additionally, the expressions to compute the optimum parameters are presented and discussed.

  3. Power system comparison for the Pluto Express mission

    SciTech Connect

    Harty, R.B.

    1995-12-31

    This paper presents a comparison of three advanced radioisotope power systems, along with a down sized RTG for the Pluto Express mission. These three advanced radioisotope power systems were the Radioisotope Alkali Metal Thermal--to-Electric Converter (RAMTEC), Radioisotope Stirling, and Radioisotope Thermophotovoltaic (RTPV). For the Pluto Express mission, the power requirement at the end of the 10-y mission is 74 We. It was found that all three advanced power systems could meet the required end of mission power with two General Purpose Heat Source (GPHS) modules. The RTG required six modules to meet the power requirement. Only the RAMTEC and RTPV met the mass goal of 9.5 kg. The AMTEC has a radiator area more than a factor of 10 lower than the Stirling and RTPV power systems, which simplifies spacecraft integration.

  4. Neuromuscular study of early branching Diuronotus aspetos (Paucitubulatina) yields insights into the evolution of organs systems in Gastrotricha.

    PubMed

    Bekkouche, Nicolas; Worsaae, Katrine

    2016-01-01

    Diuronotus is one of the most recently described genera of Paucitubulatina, one of the three major clades in Gastrotricha. Its morphology suggests that Diuronotus is an early branch of Paucitubulatina, making it a key taxon for understanding the evolution of this morphologically understudied group. Here we test its phylogenetic position employing molecular data, and provide detailed descriptions of the muscular, nervous, and ciliary systems of Diuronotus aspetos, using immunohistochemistry and confocal laser scanning microscopy. We confirm the proposed position of D. aspetos within Muselliferidae, and find this family to be the sister group to Xenotrichulidae. The muscular system, revealed by F-actin staining, shows a simple, but unique organization of the trunk musculature with a reduction to three pairs of longitudinal muscles and addition of up to five pairs of dorso-ventral muscles, versus the six longitudinal and two dorso-ventral pairs found in most Paucitubulatina. Using acetylated α-tubulin immunoreactivity, we describe the pharynx in detail, including new nervous structures, two pairs of sensory cilia, and a unique canal system. The central nervous system, as revealed by immunohistochemistry, shows the general pattern of Gastrotricha having a bilobed brain and a pair of ventro-longitudinal nerve cords. However, in addition are found an anterior nerve ring, several anterior longitudinal nerves, and four ventral commissures (pharyngeal, trunk, pre-anal, and terminal). Two pairs of protonephridia are documented, while other Paucitubulatina have one. Moreover, the precise arrangement of multiciliated cells is unraveled, yielding a pattern of possibly systematic importance. Several neural structures of Diuronotus resemble those found in Xenotrichula (Xenotrichulidae) and may constitute new apomorphies of Paucitubulatina, or even Gastrotricha. In order to test these new evolutionary hypotheses, comparable morphological data from other understudied gastrotrich

  5. VITELLOGENIN EXPRESSION IN SHEEPSHEAD MINNOWS FROM THE PENSACOLA BAY SYSTEM

    EPA Science Inventory

    Hemmer, M.J., B.L. Hemmer, S.D. Friedman and P.S. Harris. In press. Vitellogenin Expression in Populations of Sheepshead Minnows from the Pensacola Bay System (Abstract). To be presented at the SETAC Fourth World Congress, 14-18 November 2004, Portland, OR. 1 p. (ERL,GB R1015). <...

  6. VITELLOGENIN EXPRESSION IN SHEEPSHEAD MINNOWS FROM THE PENSACOLA BAY SYSTEM

    EPA Science Inventory

    Hemmer, M.J., B.L. Hemmer, S.D. Friedman and P.S. Harris. In press. Vitellogenin Expression in Populations of Sheepshead Minnows from the Pensacola Bay System (Abstract). To be presented at the SETAC Fourth World Congress, 14-18 November 2004, Portland, OR. 1 p. (ERL,GB R1015). <...

  7. Protoplast Transformation as a Plant-Transferable Transient Expression System.

    PubMed

    Duarte, Patrícia; Ribeiro, Diana; Carqueijeiro, Inês; Bettencourt, Sara; Sottomayor, Mariana

    2016-01-01

    The direct uptake of DNA by naked plant cells (protoplasts) provides an expression system of exception for the quickly growing research in non-model plants, fuelled by the power of next-generation sequencing to identify novel candidate genes. Here, we describe a simple and effective method for isolation and transformation of protoplasts, and illustrate its application to several plant materials.

  8. Large increase in brazzein expression achieved by changing the plasmid /strain combination of the NICE system in Lactococcus lactis.

    PubMed

    Berlec, A; Strukelj, B

    2009-06-01

    To evaluate brazzein production in Lactococcus lactis using the nisin-controlled expression (NICE) system. The approach is through analysis of different plasmid/strain combinations. Two plasmid/strain combinations of the NICE system were used in brazzein expression: L. lactis NZ9000 harbouring plasmid pNZ8148, and L. lactis IL1403 harbouring plasmid pMSP3545. The former combination proved superior, with a >800-fold increase in His-tagged brazzein expression (to 1.65 mg l(-1) of fermentation broth), comparable to expression levels in Escherichia coli. Improved expression resulted in a minor increase in secretion to the medium with the use of the Usp45 signal peptide. The yield of wild-type brazzein corresponded to that of His-tagged brazzein. Wild-type brazzein was partially soluble and low-intensity sweetness was detected. The plasmid/strain combination of the NICE system has a significant impact on the expression of brazzein where a >800-fold increase was achieved. The greatly increased expression of brazzein resulted in minor improvement in secretion and low-intensity sweetness. The choice of the plasmid/strain combination of the NICE system was shown to be of extreme importance in brazzein expression.

  9. Antimicrobial peptide production and plant-based expression systems for medical and agricultural biotechnology.

    PubMed

    Holaskova, Edita; Galuszka, Petr; Frebort, Ivo; Oz, M Tufan

    2015-11-01

    Antimicrobial peptides (AMPs) are vital components of the innate immune system of nearly all living organisms. They generally act in the first line of defense against various pathogenic bacteria, parasites, enveloped viruses and fungi. These low molecular mass peptides are considered prospective therapeutic agents due to their broad-spectrum rapid activity, low cytotoxicity to mammalian cells and unique mode of action which hinders emergence of pathogen resistance. In addition to medical use, AMPs can also be employed for development of innovative approaches for plant protection in agriculture. Conferred disease resistance by AMPs might help us surmount losses in yield, quality and safety of agricultural products due to plant pathogens. Heterologous expression in plant-based systems, also called plant molecular farming, offers cost-effective large-scale production which is regarded as one of the most important factors for clinical or agricultural use of AMPs. This review presents various types of AMPs as well as plant-based platforms ranging from cell suspensions to whole plants employed for peptide production. Although AMP production in plants holds great promises for medicine and agriculture, specific technical limitations regarding product yield, function and stability still remain. Additionally, establishment of particular stable expression systems employing plants or plant tissues generally requires extended time scale for platform development compared to certain other heterologous systems. Therefore, fast and promising tools for evaluation of plant-based expression strategies and assessment of function and stability of the heterologously produced AMPs are critical for molecular farming and plant protection. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Integrative Food-Grade Expression System Based on the Lactose Regulon of Lactobacillus casei

    PubMed Central

    Gosalbes, María José; Esteban, Carlos David; Galán, José Luis; Pérez-Martínez, Gaspar

    2000-01-01

    The lactose operon from Lactobacillus casei is regulated by very tight glucose repression and substrate induction mechanisms, which made it a tempting candidate system for the expression of foreign genes or metabolic engineering. An integrative vector was constructed, allowing stable gene insertion in the chromosomal lactose operon of L. casei. This vector was based on the nonreplicative plasmid pRV300 and contained two DNA fragments corresponding to the 3′ end of lacG and the complete lacF gene. Four unique restriction sites were created, as well as a ribosome binding site that would allow the cloning and expression of new genes between these two fragments. Then, integration of the cloned genes into the lactose operon of L. casei could be achieved via homologous recombination in a process that involved two selection steps, which yielded highly stable food-grade mutants. This procedure has been successfully used for the expression of the E. coli gusA gene and the L. lactis ilvBN genes in L. casei. Following the same expression pattern as that for the lactose genes, β-glucuronidase activity and diacetyl production were repressed by glucose and induced by lactose. This integrative vector represents a useful tool for strain improvement in L. casei that could be applied to engineering fermentation processes or used for expression of genes for clinical and veterinary uses. PMID:11055930

  11. Lipase expression in Pseudomonas alcaligenes is under the control of a two-component regulatory system.

    PubMed

    Krzeslak, Joanna; Gerritse, Gijs; van Merkerk, Ronald; Cool, Robbert H; Quax, Wim J

    2008-03-01

    Preliminary observations in a large-scale fermentation process suggested that the lipase expression of Pseudomonas alcaligenes can be switched on by the addition of certain medium components, such as soybean oil. In an attempt to elucidate the mechanism of induction of lipase expression, we have set up a search method for genes controlling lipase expression by use of a cosmid library containing fragments of P. alcaligenes genomic DNA. A screen for lipase hyperproduction resulted in the selection of multiple transformants, of which the best-producing strains comprised cosmids that shared an overlapping genomic fragment. Within this fragment, two previously unidentified genes were found and named lipQ and lipR. Their encoded proteins belong to the NtrBC family of regulators that regulate gene expression via binding to a specific upstream activator sequence (UAS). Such an NtrC-like UAS was identified in a previous study in the P. alcaligenes lipase promoter, strongly suggesting that LipR acts as a positive regulator of lipase expression. The regulating role could be confirmed by down-regulated lipase expression in a strain with an inactivated lipR gene and a threefold increase in lipase yield in a large-scale fermentation when expressing the lipQR operon from the multicopy plasmid pLAFR3. Finally, cell extracts of a LipR-overexpressing strain caused a retardation of the lipase promoter fragment in a band shift assay. Our results indicate that lipase expression in Pseudomonas alcaligenes is under the control of the LipQR two-component system.

  12. Hierarchical Recognition Scheme for Human Facial Expression Recognition Systems

    PubMed Central

    Siddiqi, Muhammad Hameed; Lee, Sungyoung; Lee, Young-Koo; Khan, Adil Mehmood; Truc, Phan Tran Ho

    2013-01-01

    Over the last decade, human facial expressions recognition (FER) has emerged as an important research area. Several factors make FER a challenging research problem. These include varying light conditions in training and test images; need for automatic and accurate face detection before feature extraction; and high similarity among different expressions that makes it difficult to distinguish these expressions with a high accuracy. This work implements a hierarchical linear discriminant analysis-based facial expressions recognition (HL-FER) system to tackle these problems. Unlike the previous systems, the HL-FER uses a pre-processing step to eliminate light effects, incorporates a new automatic face detection scheme, employs methods to extract both global and local features, and utilizes a HL-FER to overcome the problem of high similarity among different expressions. Unlike most of the previous works that were evaluated using a single dataset, the performance of the HL-FER is assessed using three publicly available datasets under three different experimental settings: n-fold cross validation based on subjects for each dataset separately; n-fold cross validation rule based on datasets; and, finally, a last set of experiments to assess the effectiveness of each module of the HL-FER separately. Weighted average recognition accuracy of 98.7% across three different datasets, using three classifiers, indicates the success of employing the HL-FER for human FER. PMID:24316568

  13. Expression of the endocannabinoid system in fibroblasts and myofascial tissues.

    PubMed

    McPartland, John M

    2008-04-01

    The endocannabinoid (eCB) system, like the better-known endorphin system, consists of cell membrane receptors, endogenous ligands and ligand-metabolizing enzymes. Two cannabinoid receptors are known: CB(1) is principally located in the nervous system, whereas CB(2) is primarily associated with the immune system. Two eCB ligands, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), are mimicked by cannabis plant compounds. The first purpose of this paper was to review the eCB system in detail, highlighting aspects of interest to bodyworkers, especially eCB modulation of pain and inflammation. Evidence suggests the eCB system may help resolve myofascial trigger points and relieve symptoms of fibromyalgia. However, expression of the eCB system in myofascial tissues has not been established. The second purpose of this paper was to investigate the eCB system in fibroblasts and other fascia-related cells. The investigation used a bioinformatics approach, obtaining microarray data via the GEO database (www.ncbi.nlm.nih.gov/geo/). GEO data mining revealed that fibroblasts, myofibroblasts, chondrocytes and synoviocytes expressed CB(1), CB(2) and eCB ligand-metabolizing enzymes. Fibroblast CB(1) levels nearly equalled levels expressed by adipocytes. CB(1) levels upregulated after exposure to inflammatory cytokines and equiaxial stretching of fibroblasts. The eCB system affects fibroblast remodeling through lipid rafts associated with focal adhesions and dampens cartilage destruction by decreasing fibroblast-secreted metalloproteinase enzymes. In conclusion, the eCB system helps shape biodynamic embryological development, diminishes nociception and pain, reduces inflammation in myofascial tissues and plays a role in fascial reorganization. Practitioners wield several tools that upregulate eCB activity, including myofascial manipulation, diet and lifestyle modifications, and pharmaceutical approaches.

  14. Development of an efficient expression system for Flavobacterium strains

    PubMed Central

    Chen, Shicheng; Kaufman, Michael G.; Bagdasarian, Michael; Bates, Adam K.; Walker, Edward D.

    2014-01-01

    Strong promoters were isolated from Flavobacterium johnsoniae in a promoter-trap vector incorporating a gfp reporter system, and were used to express fluorescent protein markers (including GFP, YFP, mOrange and mStrawberry) and insecticidal protein genes in Flavobacterium strains. Sequence analysis of trapped DNA fragments showed conserved Bacteroidetes promoter motifs (TTG-N19-TAnnTTTG) located upstream of putative open reading frames. Plasmids harboring these genomic DNA fragments from F. johnsoniae promoted strong production of fluorescent proteins in Flavobacterium hibernum but not in E. coli. The most potent promoter (PompA) identified in this work was cloned upstream of genes encoding fluorescent proteins, and these were co-expressed in Flavobacterium strains. The p42 and p51 genes (binary toxins from Bacillus sphaericus) when translationally fused to the 3’-end of gfp showed strong expression. Flavobacteria expressing these genes exhibited toxicity against larvae of the mosquitoes Culex quinquefasciatus, Anopheles gambiae, and Ochlerotatus triseriatus. However, transformants with the transcriptional fusion construct between cry11A with p20 from Bacillus thuringiensis did not express Cry11A protein indicating that constitutive expression of cry11A may be problematic in Flavobacterium. PMID:20206244

  15. Methods for high yield production of terpenes

    DOEpatents

    Kutchan, Toni; Higashi, Yasuhiro; Feng, Xiaohong

    2017-01-03

    Provided are enhanced high yield production systems for producing terpenes in plants via the expression of fusion proteins comprising various combinations of geranyl diphosphate synthase large and small subunits and limonene synthases. Also provided are engineered oilseed plants that accumulate monoterpene and sesquiterpene hydrocarbons in their seeds, as well as methods for producing such plants, providing a system for rapidly engineering oilseed crop production platforms for terpene-based biofuels.

  16. Closing system-wide yield gaps to increase food production and mitigate GHGs among mixed crop-livestock smallholders in Sub-Saharan Africa.

    PubMed

    Henderson, B; Godde, C; Medina-Hidalgo, D; van Wijk, M; Silvestri, S; Douxchamps, S; Stephenson, E; Power, B; Rigolot, C; Cacho, O; Herrero, M

    2016-03-01

    In this study we estimate yield gaps for mixed crop-livestock smallholder farmers in seven Sub-Saharan African sites covering six countries (Kenya, Tanzania, Uganda, Ethiopia, Senegal and Burkina Faso). We also assess their potential to increase food production and reduce the GHG emission intensity of their products, as a result of closing these yield gaps. We use stochastic frontier analysis to construct separate production frontiers for each site, based on 2012 survey data prepared by the International Livestock Research Institute for the Climate Change, Agriculture and Food Security program. Instead of relying on theoretically optimal yields-a common approach in yield gap assessments-our yield gaps are based on observed differences in technical efficiency among farms within each site. Sizeable yield gaps were estimated to be present in all of the sites. Expressed as potential percentage increases in outputs, the average site-based yield gaps ranged from 28 to 167% for livestock products and from 16 to 209% for crop products. The emission intensities of both livestock and crop products registered substantial falls as a consequence of closing yield gaps. The relationships between farm attributes and technical efficiency were also assessed to help inform policy makers about where best to target capacity building efforts. We found a strong and statistically significant relationship between market participation and performance across most sites. We also identified an efficiency dividend associated with the closer integration of crop and livestock enterprises. Overall, this study reveals that there are large yield gaps and that substantial benefits for food production and environmental performance are possible through closing these gaps, without the need for new technology.

  17. Expression and characterization of a Talaromyces marneffei active phospholipase B expressed in a Pichia pastoris expression system

    PubMed Central

    He, Yan; Li, Linghua; Hu, Fengyu; Chen, Wanshan; Lei, Huali; Chen, Xiejie; Cai, Weiping; Tang, Xiaoping

    2016-01-01

    Phospholipase B is a virulence factor for several clinically important pathogenic fungi, including Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus, but its role in the thermally dimorphic fungus Talaromyces marneffei remains unclear. Here, we provide the first report of the expression of a novel phospholipase gene, designated TmPlb1, from T. marneffei in the eukaryotic expression system of Pichia pastoris GS115. Sensitive real-time quantitative reverse-transcription PCR (qRT-PCR) demonstrated that the expression of TmPlb1 increased 1.85-fold in the yeast phase compared with the mycelial phase. TmPlb1 contains an open reading frame (ORF) of 732 bp that encodes a protein of 243 amino acids. The conserved serine, aspartate and histidine catalytic triad and the G-X-S-X-G domain of TmPLB1 provide the structural basis for its molecular activity. The ORF of TmPlb1 was successfully cloned into a pPIC9K vector containing an α-mating factor secretion signal that allowed the secretory expression of TmPLB1 in P. pastoris. The heterologous protein expression began 12 h after methanol induction and peaked at 96 h. Through analysis with SDS–polyacrylamide gel electrophoresis (SDS-PAGE), western blotting and mass spectrometry, we confirmed that TmPLB1 was successfully expressed. Through Ni-affinity chromatography, TmPLB1 was highly purified, and its concentration reached 240.4 mg/L of culture medium. With specific substrates, the phospholipase A1 and phospholipase A2 activities of TmPLB1 were calculated to be 5.96 and 1.59 U/mg, respectively. The high purity and activity of the TmPLB1 obtained here lay a solid foundation for further investigation. PMID:27876784

  18. The effect of lactation number, stage, length, and milking frequency on milk yield in Korean Holstein dairy cows using automatic milking system

    PubMed Central

    Vijayakumar, Mayakrishnan; Park, Ji Hoo; Ki, Kwang Seok; Lim, Dong Hyun; Kim, Sang Bum; Park, Seong Min; Jeong, Ha Yeon; Park, Beom Young; Kim, Tae Il

    2017-01-01

    Objective The aim of the current study was to describe the relationship between milk yield and lactation number, stage, length and milking frequency in Korean Holstein dairy cows using an automatic milking system (AMS). Methods The original data set consisted of observations from April to October 2016 of 780 Holstein cows, with a total of 10,751 milkings. Each time a cow was milked by an AMS during the 24 h, the AMS management system recorded identification numbers of the AMS unit, the cow being milking, date and time of the milking, and milk yield (kg) as measured by the milk meters installed on each AMS unit, date and time of the lactation, lactation stage, milking frequency (NoM). Lactation stage is defined as the number of days milking per cows per lactation. Milk yield was calculated per udder quarter in the AMS and was added to 1 record per cow and trait for each milking. Milking frequency was measured the number of milkings per cow per 24 hour. Results From the study results, a significant relationship was found between the milk yield and lactation number (p<0.001), with the maximum milk yield occurring in the third lactation cows. We recorded the highest milk yield, in a greater lactation length period of early stage (55 to 90 days) at a 4× milking frequency/d, and the lowest milk yield was observed in the later stage (>201 days) of cows. Also, milking frequency had a significant influence on milk yield (p<0.001) in Korean Holstein cows using AMS. Conclusion Detailed knowledge of these factors such as lactation number, stage, length, and milking frequency associated with increasing milk yield using AMS will help guide future recommendations to producers for maximizing milk yield in Korean Dairy industries. PMID:28423887

  19. Yield and economic performance of organic and conventional cotton-based farming systems--results from a field trial in India.

    PubMed

    Forster, Dionys; Andres, Christian; Verma, Rajeev; Zundel, Christine; Messmer, Monika M; Mäder, Paul

    2013-01-01

    The debate on the relative benefits of conventional and organic farming systems has in recent time gained significant interest. So far, global agricultural development has focused on increased productivity rather than on a holistic natural resource management for food security. Thus, developing more sustainable farming practices on a large scale is of utmost importance. However, information concerning the performance of farming systems under organic and conventional management in tropical and subtropical regions is scarce. This study presents agronomic and economic data from the conversion phase (2007-2010) of a farming systems comparison trial on a Vertisol soil in Madhya Pradesh, central India. A cotton-soybean-wheat crop rotation under biodynamic, organic and conventional (with and without Bt cotton) management was investigated. We observed a significant yield gap between organic and conventional farming systems in the 1(st) crop cycle (cycle 1: 2007-2008) for cotton (-29%) and wheat (-27%), whereas in the 2(nd) crop cycle (cycle 2: 2009-2010) cotton and wheat yields were similar in all farming systems due to lower yields in the conventional systems. In contrast, organic soybean (a nitrogen fixing leguminous plant) yields were marginally lower than conventional yields (-1% in cycle 1, -11% in cycle 2). Averaged across all crops, conventional farming systems achieved significantly higher gross margins in cycle 1 (+29%), whereas in cycle 2 gross margins in organic farming systems were significantly higher (+25%) due to lower variable production costs but similar yields. Soybean gross margin was significantly higher in the organic system (+11%) across the four harvest years compared to the conventional systems. Our results suggest that organic soybean production is a viable option for smallholder farmers under the prevailing semi-arid conditions in India. Future research needs to elucidate the long-term productivity and profitability, particularly of cotton and

  20. Yield and Economic Performance of Organic and Conventional Cotton-Based Farming Systems – Results from a Field Trial in India

    PubMed Central

    Forster, Dionys; Andres, Christian; Verma, Rajeev; Zundel, Christine; Messmer, Monika M.; Mäder, Paul

    2013-01-01

    The debate on the relative benefits of conventional and organic farming systems has in recent time gained significant interest. So far, global agricultural development has focused on increased productivity rather than on a holistic natural resource management for food security. Thus, developing more sustainable farming practices on a large scale is of utmost importance. However, information concerning the performance of farming systems under organic and conventional management in tropical and subtropical regions is scarce. This study presents agronomic and economic data from the conversion phase (2007–2010) of a farming systems comparison trial on a Vertisol soil in Madhya Pradesh, central India. A cotton-soybean-wheat crop rotation under biodynamic, organic and conventional (with and without Bt cotton) management was investigated. We observed a significant yield gap between organic and conventional farming systems in the 1st crop cycle (cycle 1: 2007–2008) for cotton (−29%) and wheat (−27%), whereas in the 2nd crop cycle (cycle 2: 2009–2010) cotton and wheat yields were similar in all farming systems due to lower yields in the conventional systems. In contrast, organic soybean (a nitrogen fixing leguminous plant) yields were marginally lower than conventional yields (−1% in cycle 1, −11% in cycle 2). Averaged across all crops, conventional farming systems achieved significantly higher gross margins in cycle 1 (+29%), whereas in cycle 2 gross margins in organic farming systems were significantly higher (+25%) due to lower variable production costs but similar yields. Soybean gross margin was significantly higher in the organic system (+11%) across the four harvest years compared to the conventional systems. Our results suggest that organic soybean production is a viable option for smallholder farmers under the prevailing semi-arid conditions in India. Future research needs to elucidate the long-term productivity and profitability, particularly of

  1. Crop yield and light/energy efficiency in a closed ecological system: Laboratory Biosphere experiments with wheat and sweet potato.

    PubMed

    Nelson, M; Dempster, W F; Silverstone, S; Alling, A; Allen, J P; van Thillo, M

    2005-01-01

    greenhouse trials at comparable light levels, and the best portion of the crop at 0.22 g mol-1 was in-between those values. Sweet potato production was overall close to 50% higher than trials using hydroponic methods with TU-82-155 at NASA JSC. Compared to projected yields for the Mars on Earth life support system, these wheat yields were about 15% higher, and the sweet potato yields averaged over 80% higher.

  2. Crop yield and light/energy efficiency in a closed ecological system: Laboratory Biosphere experiments with wheat and sweet potato

    NASA Astrophysics Data System (ADS)

    Nelson, M.; Dempster, W. F.; Silverstone, S.; Alling, A.; Allen, J. P.; van Thillo, M.

    somewhat below greenhouse trials at comparable light levels, and the best portion of the crop at 0.22 g mol -1 was in-between those values. Sweet potato production was overall close to 50% higher than trials using hydroponic methods with TU-82-155 at NASA JSC. Compared to projected yields for the Mars on Earth life support system, these wheat yields were about 15% higher, and the sweet potato yields averaged over 80% higher.

  3. High yield expression of an AHL-lactonase from Bacillus sp. B546 in Pichia pastoris and its application to reduce Aeromonas hydrophila mortality in aquaculture

    PubMed Central

    2010-01-01

    Background Aeromonas hydrophila is a serious pathogen and can cause hemorrhagic septicemia in fish. To control this disease, antibiotics and chemicals are widely used which can consequently result in "superbugs" and chemical accumulation in the food chain. Though vaccine against A. hydrophila is available, its use is limited due to multiple serotypes of this pathogen and problems of safety and efficacy. Another problem with vaccination is the ability to apply it to small fish especially in high numbers. In this study, we tried a new way to attenuate the A. hydrophila infection by using a quorum quenching strategy with a recombinant AHL-lactonase expressed in Pichia pastoris. Results The AHL-lactonase (AiiAB546) from Bacillus sp. B546 was produced extracellularly in P. pastoris with a yield of 3,558.4 ± 81.3 U/mL in a 3.7-L fermenter when using 3-oxo-C8-HSL as the substrate. After purification with a HiTrap Q Sepharose column, the recombinant homogenous protein showed a band of 33.6 kDa on SDS-PAGE, higher than the calculated molecular mass (28.14 kDa). Deglycosylation of AiiAB546 with Endo H confirmed the occurrence of N-glycosylation. The purified recombinant AiiAB546 showed optimal activity at pH 8.0 and 20°C, exhibited excellent stability at pH 8.0-12.0 and thermal stability at 70°C, was firstly confirmed to be significantly protease-resistant, and had wide substrate specificity. In application test, when co-injected with A. hydrophila in common carp, recombinant AiiAB546 decreased the mortality rate and delayed the mortality time of fish. Conclusions Our results not only indicate the possibility of mass-production of AHL-lactonase at low cost, but also open up a promising foreground of application of AHL-lactonase in fish to control A. hydrophila disease by regulating its virulence. To our knowledge, this is the first report on heterologous expression of AHL-lactonase in P. pastoris and attenuating A. hydrophila virulence by co-injection with AHL

  4. Investigating the implications of meteorological indicators of seasonal rainfall performance on maize yield in a rain-fed agricultural system: case study of Mt. Darwin District in Zimbabwe

    NASA Astrophysics Data System (ADS)

    Mushore, Terence; Manatsa, Desmond; Pedzisai, Ezra; Muzenda-Mudavanhu, Chipo; Mushore, Washington; Kudzotsa, Innocent

    2017-08-01

    The study focuses on the impacts of climate variability and change on maize yield in Mt. Darwin District. The rainfall and temperature data for the period under study that is from 1992 to 2012 were obtained from Meteorological Services Department of Zimbabwe at daily resolution while crop yield data were obtained from Department of Agricultural, Technical and Extension Services (AGRITEX) and Zimbabwe Statistics Agency (ZIMSTAT) at seasonal/yearly resolution. In order to capture full rainfall seasons, a year was set to begin on 1 June and end on 31 July the next year. Yearly yield, temperature and rainfall data were used to compute time series analysis of rainfall, temperature and yield. The relationship between temperature, rainfall, quality of season (start, cessation, dry days, wet days and length) and yield was also investigated. The study also investigated the link between meteorological normal and maize yield. The study revealed that temperature is rising while rainfall is decreasing with time hence increasing risk of low maize yield in Mt. Darwin. Correlation between maize yield was higher using a non-linear ( R 2 = 0.630) than a linear regression model ( R 2 = 0.173). There was a very high correlation between maize yield and number of dry days ( R = -0.905) as well as between maize yield and length of season ( R = 0.777). We also observed a strong correlation between percentage normal rainfall and percentage normal maize yield ( R 2 = 0.753). This was also agreed between rainfall tessiles and maize yield tessiles as 50 % of the seasons had normal and above normal rainfall coinciding with normal and above normal maize yield. Of the 21 seasons considered, only one season had above normal rainfall while maize yield was below normal. The study concluded that there is a strong association between meteorological normal and maize yield in a rain-fed agricultural system. Climate information remains crucial to agricultural productivity hence the need to train farmers

  5. Investigating the implications of meteorological indicators of seasonal rainfall performance on maize yield in a rain-fed agricultural system: case study of Mt. Darwin District in Zimbabwe

    NASA Astrophysics Data System (ADS)

    Mushore, Terence; Manatsa, Desmond; Pedzisai, Ezra; Muzenda-Mudavanhu, Chipo; Mushore, Washington; Kudzotsa, Innocent

    2016-06-01

    The study focuses on the impacts of climate variability and change on maize yield in Mt. Darwin District. The rainfall and temperature data for the period under study that is from 1992 to 2012 were obtained from Meteorological Services Department of Zimbabwe at daily resolution while crop yield data were obtained from Department of Agricultural, Technical and Extension Services (AGRITEX) and Zimbabwe Statistics Agency (ZIMSTAT) at seasonal/yearly resolution. In order to capture full rainfall seasons, a year was set to begin on 1 June and end on 31 July the next year. Yearly yield, temperature and rainfall data were used to compute time series analysis of rainfall, temperature and yield. The relationship between temperature, rainfall, quality of season (start, cessation, dry days, wet days and length) and yield was also investigated. The study also investigated the link between meteorological normal and maize yield. The study revealed that temperature is rising while rainfall is decreasing with time hence increasing risk of low maize yield in Mt. Darwin. Correlation between maize yield was higher using a non-linear (R 2 = 0.630) than a linear regression model (R 2 = 0.173). There was a very high correlation between maize yield and number of dry days (R = -0.905) as well as between maize yield and length of season (R = 0.777). We also observed a strong correlation between percentage normal rainfall and percentage normal maize yield (R 2 = 0.753). This was also agreed between rainfall tessiles and maize yield tessiles as 50 % of the seasons had normal and above normal rainfall coinciding with normal and above normal maize yield. Of the 21 seasons considered, only one season had above normal rainfall while maize yield was below normal. The study concluded that there is a strong association between meteorological normal and maize yield in a rain-fed agricultural system. Climate information remains crucial to agricultural productivity hence the need to train farmers to

  6. Development of a System for Automatic Facial Expression Analysis

    NASA Astrophysics Data System (ADS)

    Diago, Luis A.; Kitaoka, Tetsuko; Hagiwara, Ichiro

    Automatic recognition of facial expressions can be an important component of natural human-machine interactions. While a lot of samples are desirable for estimating more accurately the feelings of a person (e.g. likeness) about a machine interface, in real world situation, only a small number of samples must be obtained because the high cost in collecting emotions from observed person. This paper proposes a system that solves this problem conforming to individual differences. A new method is developed for facial expression classification based on the combination of Holographic Neural Networks (HNN) and Type-2 Fuzzy Logic. For the recognition of emotions induced by facial expressions, compared with former HNN and Support Vector Machines (SVM) classifiers, proposed method achieved the best generalization performance using less learning time than SVM classifiers.

  7. High-level protein expression following single and dual gene cloning of infectious bronchitis virus N and S genes using baculovirus systems.

    PubMed

    Abdel-Moneim, Ahmed S; Giesow, Katrin; Keil, Günther M

    2014-03-01

    Baculovirus is an efficient system for the gene expression that can be used for gene transfer to both insect and different vertebrate hosts. The nucleocapsid gene (N) of the infectious bronchitis virus was cloned in a baculovirus expression system for insect cell expression. Dual expression vectors containing IBV N and spike (S) proteins of the avian infectious bronchitis virus were engineered under the control of human and murine cytomegalovirus immediate-early enhancer/promoter elements in combination with the baculoviral polyhedrin and p10 promoters for simultaneous expression in both vertebrate and insect cells. Transduction of the N gene in the insect Sf9 cells revealed a high level of protein expression. The expressed protein, used in ELISA, effectively detected chicken anti-IBV antibodies with high specificity. Transduction of mammalian and avian cells with BacMam viruses revealed that dual expression cassettes yielded high levels of protein from both transcription units.

  8. Relating Perturbation Magnitude to Temporal Gene Expression in Biological Systems

    SciTech Connect

    Callister, Stephen J.; Parnell, John J.; Pfrender, Michael E.; Hashsham, Syed

    2009-03-19

    A method to quantitatively relate stress to response at the level of gene expression is described using Saccharomyces cerevisiae as a model organism. Stress was defined as the magnitude of perturbation and strain was defined as the magnitude of cumulative response in terms of gene expression. Expression patterns of sixty genes previously reported to be significantly impacted by osmotic shock or belonging to the high-osmotic glycerol, glycerolipid metabolism, and glycolysis pathways were determined following perturbations of increasing sodium chloride concentrations (0, 0.5, 0.7, 1.0, 1.5, and 1.4 M). Expression of these genes was quantified temporally using reverse transcriptase real time polymerase chain reaction. The magnitude of cumulative response was obtained by calculating the total moment of area of the temporal response envelope for all the 60 genes, either together or for the set of genes related to each pathway. A non-linear relationship between stress and response was observed for the range of stress studied. This study examines a quantitative approach to quantify the strain at the level of gene expression to relate stress to strain in biological systems. The approach should be generally applicable to quantitatively evaluate the response of organisms to environmental change.

  9. Proteoglycan and collagen expression during human air conducting system development

    PubMed Central

    Godoy-Guzmán, C.; San Martin, S.; Pereda, J.

    2012-01-01

    The lung is formed from a bud that grows and divides in a dichotomous way. A bud is a new growth center which is determined by epithelial-mesenchymal interactions where proteins of the extracellular matrix (ECM) might be involved. To understand this protein participation during human lung development, we examined the expression and distribution of proteoglycans in relation to the different types of collagens during the period in which the air conducting system is installed. Using light microscopy and immunohistochemistry we evaluate the expression of collagens (I, III and VI) and proteoglycans (decorin, biglycan and lumican) between 8 to 10 weeks post fertilization and 11 to 14 weeks of gestational age of human embryo and fetus lungs. We show that decorin, lumican and all the collagen types investigated were expressed at the epithelium-mesenchymal interface, forming a sleeve around the bronchiolar ducts. In addition, biglycan was expressed in both the endothelial cells and the smooth muscle of the blood vessels. Thus, the similar distribution pattern of collagen and proteoglycans in the early developmental stages of the human lung may be closely related to the process of dichotomous division of the bronchial tree. This study provides a new insight concerning the participation of collagens and proteoglycans in the epithelial-mesenchymal interface during the period in which the air conducting system is installed in the human fetal lung. PMID:23027345

  10. Dynamic Visualization of Co-expression in Systems Genetics Data

    SciTech Connect

    New, Joshua Ryan; Huang, Jian; Chesler, Elissa J

    2008-01-01

    Biologists hope to address grand scientific challenges by exploring the abundance of data made available through modern microarray technology and other high-throughput techniques. The impact of this data, however, is limited unless researchers can effectively assimilate such complex information and integrate it into their daily research; interactive visualization tools are called for to support the effort. Specifically, typical studies of gene co-expression require novel visualization tools that enable the dynamic formulation and fine-tuning of hypotheses to aid the process of evaluating sensitivity of key parameters. These tools should allow biologists to develop an intuitive understanding of the structure of biological networks and discover genes which reside in critical positions in networks and pathways. By using a graph as a universal data representation of correlation in gene expression data, our novel visualization tool employs several techniques that when used in an integrated manner provide innovative analytical capabilities. Our tool for interacting with gene co-expression data integrates techniques such as: graph layout, qualitative subgraph extraction through a novel 2D user interface, quantitative subgraph extraction using graph-theoretic algorithms or by querying an optimized b-tree, dynamic level-of-detail graph abstraction, and template-based fuzzy classification using neural networks. We demonstrate our system using a real-world workflow from a large-scale, systems genetics study of mammalian gene co-expression.

  11. An amplified promoter system for targeted expression of calcium indicator proteins in the cerebellar cortex

    PubMed Central

    Kuhn, Bernd; Ozden, Ilker; Lampi, Yulia; Hasan, Mazahir T.; Wang, Samuel S.-H.

    2012-01-01

    Recording of identified neuronal network activity using genetically encoded calcium indicators (GECIs) requires labeling that is cell type-specific and bright enough for the detection of functional signals. However, specificity and strong expression are often not achievable using the same promoter. Here we present a combinatorial approach for targeted expression and single-cell-level quantification in which a weak promoter is used to drive trans-amplification under a strong general promoter. We demonstrated this approach using recombinant adeno-associated viruses (rAAVs) to deliver the sequence of the GECI D3cpv in the mouse cerebellar cortex. Direct expression under the human synapsin promoter (hSYN) led to high levels of expression (50–100 μM) in five interneuron types of the cerebellar cortex but not in Purkinje cells (PCs) (≤10 μM), yielding sufficient contrast to allow functional signals to be recorded from somata and processes in awake animals using two-photon microscopy. When the hSYN promoter was used to drive expression of the tetracycline transactivator (tTA), a second rAAV containing the bidirectional TET promoter (Ptetbi) could drive strong D3cpv expression in PCs (10–300 μM), enough to allow reliable complex spike detection in the dendritic arbor. An amplified approach should be of use in monitoring neural processing in selected cell types and boosting expression of optogenetic probes. Additionally, we overcome cell toxicity associated with rAAV injection and/or local GECI overexpression by combining the virus injection with systemic pre-injection of hyperosmotic D-mannitol, and by this double the time window for functional imaging. PMID:22866030

  12. Improved Production Efficiency of Virus-Like Particles by the Baculovirus Expression Vector System

    PubMed Central

    Bárcena, Juan; Nuñez, Maria del Carmen; Martínez-Alonso, Diego; Dudognon, Benoit; Guijarro, Eva; Escribano, José M.

    2015-01-01

    Vaccines based on virus-like particles (VLPs) have proven effective in humans and animals. In this regard, the baculovirus expression vector system (BEVS) is one of the technologies of choice to generate such highly immunogenic vaccines. The extended use of these vaccines for human and animal populations is constrained because of high production costs, therefore a significant improvement in productivity is crucial to ensure their commercial viability. Here we describe the use of the previously described baculovirus expression cassette, called TB, to model the production of two VLP-forming vaccine antigens in insect cells. Capsid proteins from porcine circovirus type 2 (PCV2 Cap) and from the calicivirus that causes rabbit hemorrhagic disease (RHDV VP60) were expressed in insect cells using baculoviruses genetically engineered with the TB expression cassette. Productivity was compared to that obtained using standard counterpart vectors expressing the same proteins under the control of the polyhedrin promoter. Our results demonstrate that the use of the TB expression cassette increased the production yields of these vaccine antigens by around 300% with respect to the standard vectors. The recombinant proteins produced by TB-modified vectors were fully functional, forming VLPs identical in size and shape to those generated by the standard baculoviruses, as determined by electron microscopy analysis. The use of the TB expression cassette implies a simple modification of the baculovirus vectors that significantly improves the cost efficiency of VLP-based vaccine production, thereby facilitating the commercial viability and broad application of these vaccines for human and animal health. PMID:26458221

  13. Functional protein expression from a DNA based wheat germ cell-free system.

    PubMed

    Zhao, Kate Qin; Hurst, Robin; Slater, Michael R; Bulleit, Robert F

    2007-12-01

    Wheat germ based eukaryotic cell-free systems have been shown to be applicable for both functional and structural analyses of proteins. However, the existing methods might require specialized instrumentation and/or a separate mRNA synthesis step. We have developed a DNA based, highly productive, coupled transcription/translation wheat germ cell-free system that incorporates the normally separate mRNA synthesis step and does not require specialized instrumentation. Using a small-volume batch reaction with fluorescence labeling, DNA templates predicted to encode proteins could be quickly screened for their ability to direct the expression of proteins of the appropriate size. Protein yield can be increased as much as 2 to 4-fold in this system using a dialysis reaction, reaching approximately 200-440 microg/ml in 10-20 h. Furthermore, enzyme activities can be assayed directly in the extract without further purification. Simple purification with affinity tags can be achieved in one-step and with minor modifications, efficient SeMet and [U-15N] labeling of >95% can be accomplished in this system. Thus, this efficient cell-free expression system can facilitate both functional and structural proteomics.

  14. A Genome-Wide Screen Reveals that the Vibrio cholerae Phosphoenolpyruvate Phosphotransferase System Modulates Virulence Gene Expression

    PubMed Central

    Millet, Yves A.; Chao, Michael C.; Sasabe, Jumpei; Davis, Brigid M.

    2015-01-01

    Diverse environmental stimuli and a complex network of regulatory factors are known to modulate expression of Vibrio cholerae's principal virulence factors. However, there is relatively little known about how metabolic factors impinge upon the pathogen's well-characterized cascade of transcription factors that induce expression of cholera toxin and the toxin-coregulated pilus (TCP). Here, we used a transposon insertion site (TIS) sequencing-based strategy to identify new factors required for expression of tcpA, which encodes the major subunit of TCP, the organism's chief intestinal colonization factor. Besides identifying most of the genes known to modulate tcpA expression, the screen yielded ptsI and ptsH, which encode the enzyme I (EI) and Hpr components of the V. cholerae phosphoenolpyruvate phosphotransferase system (PTS). In addition to reduced expression of TcpA, strains lacking EI, Hpr, or the associated EIIAGlc protein produced less cholera toxin (CT) and had a diminished capacity to colonize the infant mouse intestine. The PTS modulates virulence gene expression by regulating expression of tcpPH and aphAB, which themselves control expression of toxT, the central activator of virulence gene expression. One mechanism by which PTS promotes virulence gene expression appears to be by modulating the amounts of intracellular cyclic AMP (cAMP). Our findings reveal that the V. cholerae PTS is an additional modulator of the ToxT regulon and demonstrate the potency of loss-of-function TIS sequencing screens for defining regulatory networks. PMID:26056384

  15. Closing system-wide yield gaps to increase food production and mitigate GHGs among mixed crop–livestock smallholders in Sub-Saharan Africa

    PubMed Central

    Henderson, B.; Godde, C.; Medina-Hidalgo, D.; van Wijk, M.; Silvestri, S.; Douxchamps, S.; Stephenson, E.; Power, B.; Rigolot, C.; Cacho, O.; Herrero, M.

    2016-01-01

    In this study we estimate yield gaps for mixed crop–livestock smallholder farmers in seven Sub-Saharan African sites covering six countries (Kenya, Tanzania, Uganda, Ethiopia, Senegal and Burkina Faso). We also assess their potential to increase food production and reduce the GHG emission intensity of their products, as a result of closing these yield gaps. We use stochastic frontier analysis to construct separate production frontiers for each site, based on 2012 survey data prepared by the International Livestock Research Institute for the Climate Change, Agriculture and Food Security program. Instead of relying on theoretically optimal yields—a common approach in yield gap assessments—our yield gaps are based on observed differences in technical efficiency among farms within each site. Sizeable yield gaps were estimated to be present in all of the sites. Expressed as potential percentage increases in outputs, the average site-based yield gaps ranged from 28 to 167% for livestock products and from 16 to 209% for crop products. The emission intensities of both livestock and crop products registered substantial falls as a consequence of closing yield gaps. The relationships between farm attributes and technical efficiency were also assessed to help inform policy makers about where best to target capacity building efforts. We found a strong and statistically significant relationship between market participation and performance across most sites. We also identified an efficiency dividend associated with the closer integration of crop and livestock enterprises. Overall, this study reveals that there are large yield gaps and that substantial benefits for food production and environmental performance are possible through closing these gaps, without the need for new technology. PMID:26941474

  16. Severe Traumatic Head Injury Affects Systemic Cytokine Expression

    PubMed Central

    LaPar, Damien J; Rosenberger, Laura H; Walters, Dustin M; Hedrick, Traci L; Swenson, Brian R; Young, Jeffrey S; Dossett, Lesly A; May, Addison K; Sawyer, Robert G

    2012-01-01

    Background The neuroimmunologic effect of traumatic head injury remains ill-defined. This study aimed to characterize systemic cytokine profiles among traumatically injured patients to assess the effect of traumatic head injury on the systemic inflammatory response. Study Design Over five years, 1,022 patients were evaluated from a multi-institutional trauma immunomodulatory database (TIMD). Patients were stratified by presence of severe head injury (SHI, Head ISS ≥ 4, n=335) versus non-severe head injury (NHI, Head ISS ≤ 3, n=687). Systemic cytokine expression was quantified by ELISA within 72 hours of admission. Patient factors, outcomes, and cytokine profiles were compared by univariate analyses. Results SHI patients were more severely injured with higher mortality despite similar ICU infection and ventilator associated pneumonia (VAP) rates. Expression of early pro-inflammatory cytokines, IL-6 (p<0.001) and tumor necrosis factor (TNF)-α (p=0.02), were higher among NHI patients, while expression of immunomodulatory cytokines, interferon-γ (p=0.01) and IL-12 (p=0.003), was higher in SHI patients. High TNF-α levels in NHI patients were associated with mortality (p=0.01), increased mechanical ventilation (p=0.02), and development of VAP (p=0.01). Alternatively, among SHI patients, high IL-2 levels were associated with survival, decreased mechanical ventilation, and absence of VAP. Conclusions The presence of severe traumatic head injury significantly alters systemic cytokine expression and exerts an immunomodulatory effect. Early recognition of these profiles may allow for targeted intervention to reduce patient morbidity and mortality. PMID:22342787

  17. High-yield expression and purification of soluble forms of the anti-apoptotic Bcl-x(L) and Bcl-2 as TolAIII-fusion proteins.

    PubMed

    Nedelkina, Svetlana; Gokce, Isa; Ridley, Helen; Weckerle, Céline; Magnin, Thierry; Vallette, François; Pattus, Franc; Lakey, Jeremy H; Bechinger, Burkhard

    2008-08-01

    A method is presented to produce large amounts of Bcl-2 and Bcl-x(L), two anti-apoptotic proteins of considerable biomedical interest. Expression constructs were prepared in which the Escherichia coli protein TolAIII, known to promote over expression of soluble product, was added to the N-terminus of Bcl-2 or Bcl-x(L) proteins, which had their C-terminal hydrophobic anchors deleted. Here the expression of these TolAIII-fusion constructs, followed by a two-step metal-affinity based purification protocol is described. The method delivers at least 20 and 10mg of more than 90% pure TolAIII-Bcl-x(L)DeltaC and TolAIII-Bcl-2(2)DeltaC proteins, respectively, per liter of E. coli cell culture. The proteins are released by proteolysis with thrombin providing > 12 mg of Bcl-x(L)DeltaC or > 6 mg of Bcl-2(2)DeltaC per liter of E. coli cell culture with a purity of more than 95%. Whereas Bcl-x(L)DeltaC is soluble both before and after TolAIII removal, Triton X-100 can significantly increase the extraction of TolAIII- Bcl-2(2)DeltaC from the bacterial cells and its subsequent solubility. Far-UV CD spectroscopy demonstrated that they both have an alpha-helical structure. Fluorescence spectroscopy was used to quantitatively analyze the binding of the respiratory inhibitor antimycin A to recombinant Bcl-2 and Bcl-x(L) proteins as well as the displacement of this ligand from the hydrophobic pocket with BH3 Bad-derived peptide. Purified Bcl-x(L)DeltaC and Bcl-2(2)DeltaC both protect isolated mitochondria from Bax-induced release of cytochrome c. The ensemble of data shows that the expressed proteins are correctly folded and functional. Therefore, the TolAIII-fusion system provides a convenient tool for functional characterization and structural studies of anti-apoptotic proteins.

  18. Expression Systems and Species Used for Transgenic Animal Bioreactors

    PubMed Central

    Wang, Yanli; Zhao, Sihai; Bai, Liang; Fan, Jianglin; Liu, Enqi

    2013-01-01

    Transgenic animal bioreactors can produce therapeutic proteins with high value for pharmaceutical use. In this paper, we compared different systems capable of producing therapeutic proteins (bacteria, mammalian cells, transgenic plants, and transgenic animals) and found that transgenic animals were potentially ideal bioreactors for the synthesis of pharmaceutical protein complexes. Compared with other transgenic animal expression systems (egg white, blood, urine, seminal plasma, and silkworm cocoon), the mammary glands of transgenic animals have enormous potential. Compared with other mammalian species (pig, goat, sheep, and cow) that are currently being studied as bioreactors, rabbits offer many advantages: high fertility, easy generation of transgenic founders and offspring, insensitivity to prion diseases, relatively high milk production, and no transmission of severe diseases to humans. Noticeably, for a small- or medium-sized facility, the rabbit system is ideal to produce up to 50 kg of protein per year, considering both economical and hygienic aspects; rabbits are attractive candidates for the mammary-gland-specific expression of recombinant proteins. We also reviewed recombinant proteins that have been produced by targeted expression in the mammary glands of rabbits and discussed the limitations of transgenic animal bioreactors. PMID:23586046

  19. Expression systems and species used for transgenic animal bioreactors.

    PubMed

    Wang, Yanli; Zhao, Sihai; Bai, Liang; Fan, Jianglin; Liu, Enqi

    2013-01-01

    Transgenic animal bioreactors can produce therapeutic proteins with high value for pharmaceutical use. In this paper, we compared different systems capable of producing therapeutic proteins (bacteria, mammalian cells, transgenic plants, and transgenic animals) and found that transgenic animals were potentially ideal bioreactors for the synthesis of pharmaceutical protein complexes. Compared with other transgenic animal expression systems (egg white, blood, urine, seminal plasma, and silkworm cocoon), the mammary glands of transgenic animals have enormous potential. Compared with other mammalian species (pig, goat, sheep, and cow) that are currently being studied as bioreactors, rabbits offer many advantages: high fertility, easy generation of transgenic founders and offspring, insensitivity to prion diseases, relatively high milk production, and no transmission of severe diseases to humans. Noticeably, for a small- or medium-sized facility, the rabbit system is ideal to produce up to 50 kg of protein per year, considering both economical and hygienic aspects; rabbits are attractive candidates for the mammary-gland-specific expression of recombinant proteins. We also reviewed recombinant proteins that have been produced by targeted expression in the mammary glands of rabbits and discussed the limitations of transgenic animal bioreactors.

  20. Cloning and Expression of Recombinant Human Endostatin in Periplasm of Escherichia coli Expression System

    PubMed Central

    Mohajeri, Abbas; Pilehvar-Soltanahmadi, Yones; Pourhassan-Moghaddam, Mohammad; Abdolalizadeh, Jalal; Karimi, Pouran; Zarghami, Nosratollah

    2016-01-01

    Purpose: Recombinant human endostatin (rhEs) is an angiogenesis inhibitor which is used as a specific drug in the treatment of non-small-cell lung cancer. In the current research, we developed an efficient method for expressing soluble form of the rhEs protein in the periplasmic space of Escherichia coli via fusing with pelB signal peptide. Methods: The human endostatin (hEs) gene was amplified using synthetic (hEs) gene as a template; then, cloned and expressed under T7 lac promoter. IPTG was used as an inducer for rhEs expression. Next, the osmotic shock was used to extraction of protein from the periplasmic space. The presence of rhEs in the periplasmic space was approved by SDS-PAGE and Western blotting. Results: The results show the applicability of pelB fusion protein system usage for secreting rhEs in the periplasm of E. coli in the laboratory scale. The rhEs represents approximately 35 % (0.83mg/l) of the total cell protein. Conclusion: The present study apparently is the first report of codon-optimized rhEs expression as a fusion with pelB signal peptide. The results presented the successful secretion of soluble rhEs to the periplasmic space. PMID:27478780

  1. Control of gene expression by CRISPR-Cas systems.

    PubMed

    Bikard, David; Marraffini, Luciano A

    2013-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) loci and their associated cas (CRISPR-associated) genes provide adaptive immunity against viruses (phages) and other mobile genetic elements in bacteria and archaea. While most of the early work has largely been dominated by examples of CRISPR-Cas systems directing the cleavage of phage or plasmid DNA, recent studies have revealed a more complex landscape where CRISPR-Cas loci might be involved in gene regulation. In this review, we summarize the role of these loci in the regulation of gene expression as well as the recent development of synthetic gene regulation using engineered CRISPR-Cas systems.

  2. Control of gene expression by CRISPR-Cas systems

    PubMed Central

    2013-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) loci and their associated cas (CRISPR-associated) genes provide adaptive immunity against viruses (phages) and other mobile genetic elements in bacteria and archaea. While most of the early work has largely been dominated by examples of CRISPR-Cas systems directing the cleavage of phage or plasmid DNA, recent studies have revealed a more complex landscape where CRISPR-Cas loci might be involved in gene regulation. In this review, we summarize the role of these loci in the regulation of gene expression as well as the recent development of synthetic gene regulation using engineered CRISPR-Cas systems. PMID:24273648

  3. Periplasmic production via the pET expression system of soluble, bioactive human growth hormone.

    PubMed

    Sockolosky, Jonathan T; Szoka, Francis C

    2013-02-01

    A pET based expression system for the production of recombinant human growth hormone (hGH) directed to the Escherichia coli periplasmic space was developed. The pET22b plasmid was used as a template for creating vectors that encode hGH fused to either a pelB or ompA secretion signal under control of the strong bacteriophage T7 promoter. The pelB- and ompA-hGH constructs expressed in BL21 (λDE3)-RIPL E. coli are secreted into the periplasm which facilitates isolation of soluble hGH by selective disruption of the outer membrane. A carboxy-terminal poly-histidine tag enabled purification by Ni(2+) affinity chromatography with an average yield of 1.4 mg/L culture of purified hGH, independent of secretion signal. Purified pelB- and ompA-hGH are monomeric based on size exclusion chromatography with an intact mass corresponding to mature hGH indicating proper cleavage of the signal peptide and folding in the periplasm. Both pelB- and ompA-hGH bind the hGH receptor with high affinity and potently stimulate Nb2 cell growth. These results demonstrate that the pET expression system is suitable for the rapid and simple isolation of bioactive, soluble hGH from E. coli.

  4. Effects of reduced nitrogen inputs on crop yield and nitrogen use efficiency in a long-term maize-soybean relay strip intercropping system.

    PubMed

    Chen, Ping; Du, Qing; Liu, Xiaoming; Zhou, Li; Hussain, Sajad; Lei, Lu; Song, Chun; Wang, Xiaochun; Liu, Weiguo; Yang, Feng; Shu, Kai; Liu, Jiang; Du, Junbo; Yang, Wenyu; Yong, Taiwen

    2017-01-01

    The blind pursuit of high yields via increased fertilizer inputs increases the environmental costs. Relay intercropping has advantages for yield, but a strategy for N management is urgently required to decrease N inputs without yield loss in maize-soybean relay intercropping systems (IMS). Experiments were conducted with three levels of N and three planting patterns, and dry matter accumulation, nitrogen uptake, nitrogen use efficiency (NUE), competition ratio (CR), system productivity index (SPI), land equivalent ratio (LER), and crop root distribution were investigated. Our results showed that the CR of soybean was greater than 1, and that the change in root distribution in space and time resulted in an interspecific facilitation in IMS. The maximum yield of maize under monoculture maize (MM) occurred with conventional nitrogen (CN), whereas under IMS, the maximum yield occurred with reduced nitrogen (RN). The yield of monoculture soybean (MS) and of soybean in IMS both reached a maximum under RN. The LER of IMS varied from 1.85 to 2.36, and the SPI peaked under RN. Additionally, the NUE of IMS increased by 103.7% under RN compared with that under CN. In conclusion, the separation of the root ecological niche contributed to a positive interspecific facilitation, which increased the land productivity. Thus, maize-soybean relay intercropping with reduced N input provides a very useful approach to increase land productivity and avert environmental pollution.

  5. The expression of a mountain cedar allergen comparing plant-viral apoplastic and yeast expression systems.

    PubMed

    Moehnke, Marcie H; Midoro-Horiuti, Terumi; Goldblum, Randall M; Kearney, Christopher M

    2008-07-01

    Jun a 3, a major allergenic protein in mountain cedar pollen, causes seasonal allergic rhinitis in hypersensitive individuals. Recombinant Jun a 3 was expressed in Nicotiana benthamiana interstitial fluid (300 microg/g leaf material) and Pichia pastoris (100 microg/ml media). Polyclonal anti-Jun a 3 and IgE antibodies from the sera of allergic patients both reacted with the recombinant protein. Of the two systems, recombinant protein from the plant apoplast contained fewer contaminating proteins. This method allows for a more convenient and inexpensive expression of the recombinant allergen, which will allow for further structural studies and may prove useful in diagnostic and/or immunotherapeutic strategies for cedar allergy.

  6. Renin-angiotensin system gene expression and neurodegenerative diseases.

    PubMed

    Goldstein, Benjamin; Speth, Robert C; Trivedi, Malav

    2016-07-01

    Single nucleotide polymorphisms and altered gene expression of components of the renin-angiotensin system (RAS) are associated with neurodegenerative diseases. Drugs that interact with the RAS have been shown to affect the course of neurodegenerative disease, suggesting that abnormalities in the RAS may contribute to neurodegenerative disease. A meta-analysis of genome-wide association studies and gene expression data for 14 RAS-related proteins was carried out for five neurodegenerative diseases: Alzheimer's disease, Parkinson's disease, narcolepsy, amyotrophic lateral sclerosis and multiple sclerosis. No single nucleotide polymorphisms in any of the 14 RAS-related protein genes were significantly associated with the five neurodegenerative diseases investigated. There was an inverse association between expression of ATP6AP2, which encodes the (pro)renin receptor, and multiple sclerosis, Alzheimer's disease and Parkinson's disease. An association of AGTR, which encodes the AT1 angiotensin II receptor, and Parkinson's disease and Alzheimer's disease was also observed. To date, no single nucleotide polymorphisms in components of the RAS can be definitively linked to the neurodegenerative diseases evaluated in this study. However, altered gene expression of several components of the RAS is associated with several neurodegenerative diseases, which may indicate that the RAS contributes to the pathology of these diseases. © The Author(s) 2016.

  7. Annual Variation in Flowering Phenology, Pollination, Mating System, and Pollen Yield in Two Natural Populations of Schima wallichii (DC.) Korth

    PubMed Central

    Khanduri, Vinod Prasad; Sharma, C. M.; Kumar, K. S.; Ghildiyal, S. K.

    2013-01-01

    Background. Schima wallichii is a highly valuable tree of tropical forest in north-east Himalaya region that grows naturally in a wide range of altitudes between 750 and 2400 m asl with varying environments. Flowering phenology of tropical tree species at population level is generally ignored and therefore a detailed knowledge of flowering and fruiting patterns of important multipurpose tree species is critical to the successful management of forest genetic resources. Materials and Methods. The study was conducted at two different altitudes (i.e., 750 m and 900 m asl) in the tropical semideciduous forest of north-east Himalaya. The floral phenology including flowering synchrony in the populations, anthesis, anther dehiscence, stigma receptivity, pollinators visitation frequency, and mating system including index of self-incompatibility were worked out in Schima wallichii according to the ear-marked standard methods given by various scientists for each parameter. Results. The flowering period in Schima wallichii varied from 33 to 42 days with mean synchrony of 0.54 to 0.68 between the populations. The stigma was receptive up to 2.5 days only and showed slightly protandrous type of dichogamy. Average pollen production ranged between 6.90 × 107 pollen per tree in 2007 and 15.49 × 108 pollen per tree in 2011. A three-year masting cycle was noticed in this species. The frequency of visitation of honey bees was fairly high (5.2 ± 1.12 visits/flower/hour) as compared to other pollinators. The hand pollination revealed maximum fruit (74.2 ± 5.72%) and seed (70.8 ± 7.46%) settings. Conclusions. The variation in flowering phenology and pollen yield individually and annually along with temporal separation in anther dehiscence and pollinator's visitation cause pollen limited reproduction, which ultimately influences the reproductive success in Schima wallichii. PMID:24501577

  8. Engineering of a wheat germ expression system to provide compatibility with a high throughput pET-based cloning platform.

    PubMed

    Zhao, Li; Zhao, Kate Q; Hurst, Robin; Slater, Michael R; Acton, Thomas B; Swapna, G V T; Shastry, Ritu; Kornhaber, Gregory J; Montelione, Gaetano T

    2010-09-01

    Wheat germ cell-free methods provide an important approach for the production of eukaryotic proteins. We have developed a protein expression vector for the TNT((R)) SP6 High-Yield Wheat Germ Cell-Free (TNT WGCF) expression system (Promega) that is also compatible with our T7-based Escherichia coli intracellular expression vector pET15_NESG. This allows cloning of the same PCR product into either one of several pET_NESG vectors and this modified WGCF vector (pWGHisAmp) by In-Fusion LIC cloning (Zhu et al. in Biotechniques 43:354-359, 2007). Integration of these two vector systems allowed us to explore the efficacy of the TNT WGCF system by comparing the expression and solubility characteristics of 59 human protein constructs in both WGCF and pET15_NESG E. coli intracellular expression. While only 30% of these human proteins could be produced in soluble form using the pET15_NESG based system, some 70% could be produced in soluble form using the TNT WGCF system. This high success rate underscores the importance of eukaryotic expression host systems like the TNT WGCF system for eukaryotic protein production in a structural genomics sample production pipeline. To further demonstrate the value of this WGCF system in producing protein suitable for structural studies, we scaled up, purified, and analyzed by 2D NMR two (15)N-, (13)C-enriched human proteins. The results of this study indicate that the TNT WGCF system is a successful salvage pathway for producing samples of difficult-to-express small human proteins for NMR studies, providing an important complementary pathway for eukaryotic sample production in the NESG NMR structure production pipeline.

  9. Analytic approaches to stochastic gene expression in multicellular systems.

    PubMed

    Boettiger, Alistair Nicol

    2013-12-17

    Deterministic thermodynamic models of the complex systems, which control gene expression in metazoa, are helping researchers identify fundamental themes in the regulation of transcription. However, quantitative single cell studies are increasingly identifying regulatory mechanisms that control variability in expression. Such behaviors cannot be captured by deterministic models and are poorly suited to contemporary stochastic approaches that rely on continuum approximations, such as Langevin methods. Fortunately, theoretical advances in the modeling of transcription have assembled some general results that can be readily applied to systems being explored only through a deterministic approach. Here, I review some of the recent experimental evidence for the importance of genetically regulating stochastic effects during embryonic development and discuss key results from Markov theory that can be used to model this regulation. I then discuss several pairs of regulatory mechanisms recently investigated through a Markov approach. In each case, a deterministic treatment predicts no difference between the mechanisms, but the statistical treatment reveals the potential for substantially different distributions of transcriptional activity. In this light, features of gene regulation that seemed needlessly complex evolutionary baggage may be appreciated for their key contributions to reliability and precision of gene expression.

  10. From genetic variability to phenotypic expression of blood group systems.

    PubMed

    Raud, L; Férec, C; Fichou, Y

    2017-06-29

    More than 300 red blood cell (RBC) antigens belonging to 36 blood group systems have been officially reported in humans by the International Society of Blood Transfusion (ISBT). Phenotypic variability is directly linked to the expression of the 41 blood group genes. The Rh blood group system, which is composed of 54 antigens, is the most complex and polymorphic system. Many rare genetic variants within the RH (RHD and RHCE) genes, involving various mutational mechanisms (single-nucleotide substitutions, short insertions/deletions, rearrangements, large deletions), have been reported in the literature and reference databases. Expression of the variants induces variable clinical outcomes depending on their nature and impact on antigen structure. Their respective molecular and cellular effects remain however poorly studied. Biological resources to conduct this research are also barely available. We have paid a specific attention to three different classes of single-nucleotide subs