Science.gov

Sample records for domesticated piggybac transposase

  1. PGBD5: a neural-specific intron-containing piggyBac transposase domesticated over 500 million years ago and conserved from cephalochordates to humans

    PubMed Central

    2013-01-01

    Background piggyBac domain (PGBD) transposons are found in organisms ranging from fungi to humans. Three domesticated piggyBac elements have been described. In the ciliates Paramecium tetraurelia and Tetrahymena thermophila, homologs known as piggyMacs excise internal eliminated sequences from germline micronuclear DNA during regeneration of the new somatic macronucleus. In primates, a PGBD3 element inserted into the Cockayne syndrome group B (CSB) gene over 43 Mya serves as an alternative 3′ terminal exon, enabling the CSB gene to generate both full length CSB and a conserved CSB-PGBD3 fusion protein that joins an N-terminal CSB domain to the C-terminal transposase domain. Results We describe a fourth domesticated piggyBac element called PGBD5. We show that i) PGBD5 was first domesticated in the common ancestor of the cephalochordate Branchiostoma floridae (aka lancelet or amphioxus) and vertebrates, and is conserved in all vertebrates including lamprey but cannot be found in more basal urochordates, hemichordates, or echinoderms; ii) the lancelet, lamprey, and human PGBD5 genes are syntenic and orthologous; iii) no potentially mobile ancestral PGBD5 elements can be identified in other more deeply rooted organisms; iv) although derived from an IS4-related transposase of the RNase H clan, PGBD5 protein is unlikely to retain enzymatic activity because the catalytic DDD(D) motif is not conserved; v) PGBD5 is preferentially expressed in certain granule cell lineages of the brain and in the central nervous system based on available mouse and human in situ hybridization data, and the tissue-specificity of documented mammalian EST and mRNA clones; vi) the human PGBD5 promoter and gene region is rich in bound regulatory factors including the neuron-restrictive silencer factors NRSF/REST and CoREST, as well as SIN3, KAP1, STAT3, and CTCF; and vii) despite preferential localization within the nucleus, PGBD5 protein is unlikely to bind DNA or chromatin as neither DNase I

  2. Hyperactive self-inactivating piggyBac for transposase-enhanced pronuclear microinjection transgenesis.

    PubMed

    Marh, Joel; Stoytcheva, Zoia; Urschitz, Johann; Sugawara, Atsushi; Yamashiro, Hideaki; Owens, Jesse B; Stoytchev, Ilko; Pelczar, Pawel; Yanagimachi, Ryuzo; Moisyadi, Stefan

    2012-11-20

    We have developed a unique method for mouse transgenesis. The transposase-enhanced pronuclear microinjection (PNI) technique described herein uses the hyperactive piggyBac transposase to insert a large transgene into the mouse genome. This procedure increased transgene integration efficiency by fivefold compared with conventional PNI or intracytoplasmic sperm injection-mediated transgenesis. Our data indicate that the transposase-enhanced PNI technique additionally requires fewer embryos to be microinjected than traditional methods to obtain transgenic animals. This transposase-mediated approach is also very efficient for single-cell embryo cytoplasmic injections, offering an easy-to-implement transgenesis method to the scientific community. PMID:23093669

  3. DNA transposition by protein transduction of the piggyBac transposase from lentiviral Gag precursors

    PubMed Central

    Cai, Yujia; Bak, Rasmus O.; Krogh, Louise Bechmann; Staunstrup, Nicklas H.; Moldt, Brian; Corydon, Thomas J.; Schrøder, Lisbeth Dahl; Mikkelsen, Jacob Giehm

    2014-01-01

    DNA transposon-based vectors have emerged as gene vehicles with a wide biomedical and therapeutic potential. So far, genomic insertion of such vectors has relied on the co-delivery of genetic material encoding the gene-inserting transposase protein, raising concerns related to persistent expression, insertional mutagenesis and cytotoxicity. This report describes potent DNA transposition achieved by direct delivery of transposase protein. By adapting integrase-deficient lentiviral particles (LPs) as carriers of the hyperactive piggyBac transposase protein (hyPBase), we demonstrate rates of DNA transposition that are comparable with the efficiency of a conventional plasmid-based strategy. Embedded in the Gag polypeptide, hyPBase is robustly incorporated into LPs and liberated from the viral proteins by the viral protease during particle maturation. We demonstrate lentiviral co-delivery of the transposase protein and vector RNA carrying the transposon sequence, allowing robust DNA transposition in a variety of cell types. Importantly, this novel delivery method facilitates a balanced cellular uptake of hyPBase, as shown by confocal microscopy, and allows high-efficiency production of clones harboring a single transposon insertion. Our findings establish engineered LPs as a new tool for transposase delivery. We believe that protein transduction methods will increase applicability and safety of DNA transposon-based vector technologies. PMID:24270790

  4. Chimeric piggyBac transposases for genomic targeting in human cells.

    PubMed

    Owens, Jesse B; Urschitz, Johann; Stoytchev, Ilko; Dang, Nong C; Stoytcheva, Zoia; Belcaid, Mahdi; Maragathavally, Kommineni J; Coates, Craig J; Segal, David J; Moisyadi, Stefan

    2012-08-01

    Integrating vectors such as viruses and transposons insert transgenes semi-randomly and can potentially disrupt or deregulate genes. For these techniques to be of therapeutic value, a method for controlling the precise location of insertion is required. The piggyBac (PB) transposase is an efficient gene transfer vector active in a variety of cell types and proven to be amenable to modification. Here we present the design and validation of chimeric PB proteins fused to the Gal4 DNA binding domain with the ability to target transgenes to pre-determined sites. Upstream activating sequence (UAS) Gal4 recognition sites harbored on recipient plasmids were preferentially targeted by the chimeric Gal4-PB transposase in human cells. To analyze the ability of these PB fusion proteins to target chromosomal locations, UAS sites were randomly integrated throughout the genome using the Sleeping Beauty transposon. Both N- and C-terminal Gal4-PB fusion proteins but not native PB were capable of targeting transposition nearby these introduced sites. A genome-wide integration analysis revealed the ability of our fusion constructs to bias 24% of integrations near endogenous Gal4 recognition sequences. This work provides a powerful approach to enhance the properties of the PB system for applications such as genetic engineering and gene therapy. PMID:22492708

  5. Chimeric piggyBac transposases for genomic targeting in human cells

    PubMed Central

    Owens, Jesse B.; Urschitz, Johann; Stoytchev, Ilko; Dang, Nong C.; Stoytcheva, Zoia; Belcaid, Mahdi; Maragathavally, Kommineni J.; Coates, Craig J.; Segal, David J.; Moisyadi, Stefan

    2012-01-01

    Integrating vectors such as viruses and transposons insert transgenes semi-randomly and can potentially disrupt or deregulate genes. For these techniques to be of therapeutic value, a method for controlling the precise location of insertion is required. The piggyBac (PB) transposase is an efficient gene transfer vector active in a variety of cell types and proven to be amenable to modification. Here we present the design and validation of chimeric PB proteins fused to the Gal4 DNA binding domain with the ability to target transgenes to pre-determined sites. Upstream activating sequence (UAS) Gal4 recognition sites harbored on recipient plasmids were preferentially targeted by the chimeric Gal4–PB transposase in human cells. To analyze the ability of these PB fusion proteins to target chromosomal locations, UAS sites were randomly integrated throughout the genome using the Sleeping Beauty transposon. Both N- and C-terminal Gal4-PB fusion proteins but not native PB were capable of targeting transposition nearby these introduced sites. A genome-wide integration analysis revealed the ability of our fusion constructs to bias 24% of integrations near endogenous Gal4 recognition sequences. This work provides a powerful approach to enhance the properties of the PB system for applications such as genetic engineering and gene therapy. PMID:22492708

  6. Tracking and transforming neocortical progenitors by CRISPR/Cas9 gene targeting and piggyBac transposase lineage labeling.

    PubMed

    Chen, Fuyi; Rosiene, Joel; Che, Alicia; Becker, Albert; LoTurco, Joseph

    2015-10-15

    The ability to induce targeted mutations in somatic cells in developing organisms and then track the fates of those cells is a powerful approach both for studying neural development and for modeling human disease. The CRISPR/Cas9 system allows for such targeted mutagenesis, and we therefore tested it in combination with a piggyBac transposase lineage labeling system to track the development of neocortical neural progenitors with targeted mutations in genes linked to neurodevelopmental disruptions and tumor formation. We show that sgRNAs designed to target PTEN successfully decreased PTEN expression, and led to neuronal hypertrophy and altered neuronal excitability. Targeting NF1, by contrast, caused increased astrocytogenesis at the expense of neurogenesis, and combined targeting of three tumor suppressors (PTEN, NF1 and P53) resulted in formation of glioblastoma tumors. Our results demonstrate that CRISPR/Cas9 combined with piggyBac transposase lineage labeling can produce unique models of neurodevelopmental disruption and tumors caused by somatic mutation in neural progenitors. PMID:26400094

  7. A Nucleolus-Predominant piggyBac Transposase, NP-mPB, Mediates Elevated Transposition Efficiency in Mammalian Cells

    PubMed Central

    Ku, Amy T.; Fan, Hsiang-Hsuan; Lee, Tung-Lung; Huang, Yung-Hsin; Yang, Tsung-Lin; Su, I-Chang; Yu, I-Shing; Lin, Shu-Wha; Chien, Chung-Liang; Ho, Hong-Nerng; Chen, You-Tzung

    2014-01-01

    PiggyBac is a prevalent transposon system used to deliver transgenes and functionally explore the mammalian untouched genomic territory. The important features of piggyBac transposon are the relatively low insertion site preference and the ability of seamless removal from genome, which allow its potential uses in functional genomics and regenerative medicine. Efforts to increase its transposition efficiency in mammals were made through engineering the corresponding transposase (PBase) codon usage to enhance its expression level and through screening for mutant PBase variants with increased enzyme activity. To improve the safety for its potential use in regenerative medicine applications, site-specific transposition was achieved by using engineered zinc finger- and Gal4-fused PBases. An excision-prone PBase variant has also been successfully developed. Here we describe the construction of a nucleolus-predominant PBase, NP-mPB, by adding a nucleolus-predominant (NP) signal peptide from HIV-1 TAT protein to a mammalian codon-optimized PBase (mPB). Although there is a predominant fraction of the NP-mPB-tGFP fusion proteins concentrated in the nucleoli, an insertion site preference toward nucleolar organizer regions is not detected. Instead a 3–4 fold increase in piggyBac transposition efficiency is reproducibly observed in mouse and human cells. PMID:24586748

  8. A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells.

    PubMed

    Hong, Jin-Bon; Chou, Fu-Ju; Ku, Amy T; Fan, Hsiang-Hsuan; Lee, Tung-Lung; Huang, Yung-Hsin; Yang, Tsung-Lin; Su, I-Chang; Yu, I-Shing; Lin, Shu-Wha; Chien, Chung-Liang; Ho, Hong-Nerng; Chen, You-Tzung

    2014-01-01

    PiggyBac is a prevalent transposon system used to deliver transgenes and functionally explore the mammalian untouched genomic territory. The important features of piggyBac transposon are the relatively low insertion site preference and the ability of seamless removal from genome, which allow its potential uses in functional genomics and regenerative medicine. Efforts to increase its transposition efficiency in mammals were made through engineering the corresponding transposase (PBase) codon usage to enhance its expression level and through screening for mutant PBase variants with increased enzyme activity. To improve the safety for its potential use in regenerative medicine applications, site-specific transposition was achieved by using engineered zinc finger- and Gal4-fused PBases. An excision-prone PBase variant has also been successfully developed. Here we describe the construction of a nucleolus-predominant PBase, NP-mPB, by adding a nucleolus-predominant (NP) signal peptide from HIV-1 TAT protein to a mammalian codon-optimized PBase (mPB). Although there is a predominant fraction of the NP-mPB-tGFP fusion proteins concentrated in the nucleoli, an insertion site preference toward nucleolar organizer regions is not detected. Instead a 3-4 fold increase in piggyBac transposition efficiency is reproducibly observed in mouse and human cells.

  9. Generation of transgenic pigs by cytoplasmic injection of piggyBac transposase-based pmGENIE-3 plasmids.

    PubMed

    Li, Zicong; Zeng, Fang; Meng, Fanming; Xu, Zhiqian; Zhang, Xianwei; Huang, Xiaoling; Tang, Fei; Gao, Wenchao; Shi, Junsong; He, Xiaoyan; Liu, Dewu; Wang, Chong; Urschitz, Johann; Moisyadi, Stefan; Wu, Zhenfang

    2014-05-01

    The process of transgenesis involves the introduction of a foreign gene, the transgene, into the genome of an animal. Gene transfer by pronuclear microinjection (PNI) is the predominant method used to produce transgenic animals. However, this technique does not always result in germline transgenic offspring and has a low success rate for livestock. Alternate approaches, such as somatic cell nuclear transfer using transgenic fibroblasts, do not show an increase in efficiency compared to PNI, while viral-based transgenesis is hampered by issues regarding transgene size and biosafety considerations. We have recently described highly successful transgenesis experiments with mice using a piggyBac transposase-based vector, pmhyGENIE-3. This construct, a single and self-inactivating plasmid, contains all the transpositional elements necessary for successful gene transfer. In this series of experiments, our laboratories have implemented cytoplasmic injection (CTI) of pmGENIE-3 for transgene delivery into in vivo-fertilized pig zygotes. More than 8.00% of the injected embryos developed into transgenic animals containing monogenic and often single transgenes in their genome. However, the CTI technique was unsuccessful during the injection of in vitro-fertilized pig zygotes. In summary, here we have described a method that is not only easy to implement, but also demonstrated the highest efficiency rate for nonviral livestock transgenesis.

  10. piggyBac transposon/transposase system to generate CD19-specific T cells for the treatment of B-lineage malignancies.

    PubMed

    Manuri, Pallavi V Raja; Wilson, Matthew H; Maiti, Sourindra N; Mi, Tiejuan; Singh, Harjeet; Olivares, Simon; Dawson, Margaret J; Huls, Helen; Lee, Dean A; Rao, Pulivarthi H; Kaminski, Joseph M; Nakazawa, Yozo; Gottschalk, Stephen; Kebriaei, Partow; Shpall, Elizabeth J; Champlin, Richard E; Cooper, Laurence J N

    2010-04-01

    Nonviral integrating vectors can be used for expression of therapeutic genes. piggyBac (PB), a transposon/transposase system, has been used to efficiently generate induced pluripotent stems cells from somatic cells, without genetic alteration. In this paper, we apply PB transposition to express a chimeric antigen receptor (CAR) in primary human T cells. We demonstrate that T cells electroporated to introduce the PB transposon and transposase stably express CD19-specific CAR and when cultured on CD19(+) artificial antigen-presenting cells, numerically expand in a CAR-dependent manner, display a phenotype associated with both memory and effector T cell populations, and exhibit CD19-dependent killing of tumor targets. Integration of the PB transposon expressing CAR was not associated with genotoxicity, based on chromosome analysis. PB transposition for generating human T cells with redirected specificity to a desired target such as CD19 is a new genetic approach with therapeutic implications. PMID:19905893

  11. Domesticated transposase Kat1 and its fossil imprints induce sexual differentiation in yeast

    PubMed Central

    Rajaei, Naghmeh; Chiruvella, Kishore K.; Lin, Feng; Åström, Stefan U.

    2014-01-01

    Transposable elements (TEs) have had a major influence on shaping both prokaryotic and eukaryotic genomes, largely through stochastic events following random or near-random insertions. In the mammalian immune system, the recombination activation genes1/2 (Rag1/2) recombinase has evolved from a transposase gene, demonstrating that TEs can be domesticated by the host. In this study, we uncovered a domesticated transposase, Kluyveromyces lactis hobo/Activator/Tam3 (hAT) transposase 1 (Kat1), operating at the fossil imprints of an ancient transposon, that catalyzes the differentiation of cell type. Kat1 induces mating-type switching from mating type a (MATa) to MATα in the yeast K. lactis. Kat1 activates switching by introducing two hairpin-capped DNA double-strand breaks (DSBs) in the MATa1–MATa2 intergenic region, as we demonstrate both in vivo and in vitro. The DSBs stimulate homologous recombination with the cryptic hidden MAT left alpha (HMLα) locus resulting in a switch of the cell type. The sites where Kat1 acts in the MATa locus most likely are ancient remnants of terminal inverted repeats from a long-lost TE. The KAT1 gene is annotated as a pseudogene because it contains two overlapping ORFs. We demonstrate that translation of full-length Kat1 requires a programmed −1 frameshift. The frameshift limited Kat1 activity, because restoring the zero frame causes switching to the MATα genotype. Kat1 also was transcriptionally activated by nutrient limitation via the transcription factor mating type switch 1 (Mts1). A phylogenetic analysis indicated that KAT1 was domesticated specifically in the Kluyveromyces clade of the budding yeasts. We conclude that Kat1 is a highly regulated transposase-derived endonuclease vital for sexual differentiation. PMID:25313032

  12. Functional characterization of sugarcane mustang domesticated transposases and comparative diversity in sugarcane, rice, maize and sorghum

    PubMed Central

    Kajihara, Daniela; de Godoy, Fabiana; Hamaji, Thais Alves; Blanco, Silvia Regina; Van Sluys, Marie-Anne; Rossi, Magdalena

    2012-01-01

    Transposable elements (TEs) account for a large portion of plant genomes, particularly in grasses, in which they correspond to 50%–80% of the genomic content. TEs have recently been shown to be a source of new genes and new regulatory networks. The most striking contribution of TEs is referred as “molecular domestication”, by which the element coding sequence loses its movement capacity and acquires cellular function. Recently, domesticated transposases known as mustang and derived from the Mutator element have been described in sugarcane. In order to improve our understanding of the function of these proteins, we identified mustang genes from Sorghum bicolor and Zea mays and performed a phenetic analysis to assess the diversity and evolutionary history of this gene family. This analysis identified orthologous groups and showed that mustang genes are highly conserved in grass genomes. We also explored the transcriptional activity of sugarcane mustang genes in heterologous and homologous systems. These genes were found to be ubiquitously transcribed, with shoot apical meristem having the highest expression levels, and were downregulated by phytohormones. Together, these findings suggest the possible involvement of mustang proteins in the maintenance of hormonal homeostasis. PMID:23055803

  13. Biochemical Characterization of Kat1: a Domesticated hAT-Transposase that Induces DNA Hairpin Formation and MAT-Switching

    PubMed Central

    Chiruvella, Kishore K.; Rajaei, Naghmeh; Jonna, Venkateswara Rao; Hofer, Anders; Åström, Stefan U.

    2016-01-01

    Kluyveromyces lactis hAT-transposase 1 (Kat1) generates hairpin-capped DNA double strand breaks leading to MAT-switching (MATa to MATα). Using purified Kat1, we demonstrate the importance of terminal inverted repeats and subterminal repeats for its endonuclease activity. Kat1 promoted joining of the transposon end into a target DNA molecule in vitro, a biochemical feature that ties Kat1 to transposases. Gas-phase Electrophoretic Mobility Macromolecule analysis revealed that Kat1 can form hexamers when complexed with DNA. Kat1 point mutants were generated in conserved positions to explore structure-function relationships. Mutants of predicted catalytic residues abolished both DNA cleavage and strand-transfer. Interestingly, W576A predicted to be impaired for hairpin formation, was active for DNA cleavage and supported wild type levels of mating-type switching. In contrast, the conserved CXXH motif was critical for hairpin formation because Kat1 C402A/H405A completely blocked hairpinning and switching, but still generated nicks in the DNA. Mutations in the BED zinc-finger domain (C130A/C133A) resulted in an unspecific nuclease activity, presumably due to nonspecific DNA interaction. Kat1 mutants that were defective for cleavage in vitro were also defective for mating-type switching. Collectively, this study reveals Kat1 sharing extensive biochemical similarities with cut and paste transposons despite being domesticated and evolutionary diverged from active transposons. PMID:26902909

  14. Excision of the piggyBac transposable element in maize cells is a precise event

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The piggyBac transposable element (TE) from the moth Trichoplusia ni encodes a ‘cut and paste’ DNA transposase that has been used to transform a number of insects, as well as planaria, mammalian cells, and mice. The wild type and a mutated piggyBac TE excised from a DNA vector in transient assays u...

  15. Post-integration stability of piggyBac in Aedes aegypti.

    PubMed

    Sethuraman, Nagaraja; Fraser, Malcolm J; Eggleston, Paul; O'Brochta, David A

    2007-09-01

    The post-integration activity of piggyBac transposable element gene vectors in Aedes aegypti mosquitoes was tested under a variety of conditions. The embryos from five independent transgenic lines of Ae. aegypti, each with a single integrated non-autonomous piggyBac transposable element gene vector, were injected with plasmids containing the piggyBac transposase open-reading frame under the regulatory control of the Drosophila melanogaster hsp70 promoter. No evidence for somatic remobilization was detected in the subsequent adults whereas somatic remobilization was readily detected when similar lines of transgenic D. melanogaster were injected with the same piggyBac transposase-expressing plasmid. Ae. aegypti heterozygotes of piggyBac reporter-containing transgenes and piggyBac transposase-expressing transgenes showed no evidence of somatic and germ-line remobilization based on phenotypic and molecular detection methods. The post-integration mobility properties of piggyBac in Ae. aegypti enhance the utility of this gene vector for certain applications, particularly those where any level of vector remobilization is unacceptable. PMID:17681233

  16. Kicking against the PRCs – A Domesticated Transposase Antagonises Silencing Mediated by Polycomb Group Proteins and Is an Accessory Component of Polycomb Repressive Complex 2

    PubMed Central

    Perera, Pumi; Mora-García, Santiago; de Leau, Erica; Thornton, Harry; de Alves, Flavia Lima; Rapsilber, Juri; Yang, Suxin; James, Geo Velikkakam; Schneeberger, Korbinian; Finnegan, E. Jean; Turck, Franziska; Goodrich, Justin

    2015-01-01

    G silencing by blocking the interaction of the core PRC2 with accessory components that promote its HMTase activity or its role in inhibiting transcription. ALP1 is the first example of a domesticated transposase acquiring a novel function as a PcG component. The antagonistic interaction of a modified transposase with the PcG machinery is novel and may have arisen as a means for the cognate transposon to evade host surveillance or for the host to exploit features of the transposition machinery beneficial for epigenetic regulation of gene activity. PMID:26642436

  17. Genome-wide target profiling of piggyBac and Tol2 in HEK 293: pros and cons for gene discovery and gene therapy

    PubMed Central

    2011-01-01

    Background DNA transposons have emerged as indispensible tools for manipulating vertebrate genomes with applications ranging from insertional mutagenesis and transgenesis to gene therapy. To fully explore the potential of two highly active DNA transposons, piggyBac and Tol2, as mammalian genetic tools, we have conducted a side-by-side comparison of the two transposon systems in the same setting to evaluate their advantages and disadvantages for use in gene therapy and gene discovery. Results We have observed that (1) the Tol2 transposase (but not piggyBac) is highly sensitive to molecular engineering; (2) the piggyBac donor with only the 40 bp 3'-and 67 bp 5'-terminal repeat domain is sufficient for effective transposition; and (3) a small amount of piggyBac transposases results in robust transposition suggesting the piggyBac transpospase is highly active. Performing genome-wide target profiling on data sets obtained by retrieving chromosomal targeting sequences from individual clones, we have identified several piggyBac and Tol2 hotspots and observed that (4) piggyBac and Tol2 display a clear difference in targeting preferences in the human genome. Finally, we have observed that (5) only sites with a particular sequence context can be targeted by either piggyBac or Tol2. Conclusions The non-overlapping targeting preference of piggyBac and Tol2 makes them complementary research tools for manipulating mammalian genomes. PiggyBac is the most promising transposon-based vector system for achieving site-specific targeting of therapeutic genes due to the flexibility of its transposase for being molecularly engineered. Insights from this study will provide a basis for engineering piggyBac transposases to achieve site-specific therapeutic gene targeting. PMID:21447194

  18. Transgene-free production of pluripotent stem cells using piggyBac transposons.

    PubMed

    Woltjen, Knut; Hämäläinen, Riikka; Kibschull, Mark; Mileikovsky, Maria; Nagy, Andras

    2011-01-01

    Reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) allows the derivation of -personalized stem cells. Transposon transgenesis is a novel and viable alternative to viral transduction methods for the delivery of reprogramming factors (Oct4, Sox2, Klf4, c-Myc) to somatic cells. Since transposons can be introduced as naked DNA using common plasmid transfection protocols, they provide a safer alternative to viral methods. piggyBac transposons are host-factor independent and integrate stably into the target genome, yet benefit from the unique characteristic of seamless removal mediated by transient expression of piggyBac transposase. Thus, piggyBac transposition provides an effective means to generate human, transgene-free iPSCs. The protocol describes the production of iPSCs from human embryonic fibroblasts, delivering reprogramming factors via plasmid transfection and piggyBac transposition.

  19. Precise marker excision system using an animal-derived piggyBac transposon in plants.

    PubMed

    Nishizawa-Yokoi, Ayako; Endo, Masaki; Osakabe, Keishi; Saika, Hiroaki; Toki, Seiichi

    2014-02-01

    Accurate and effective positive marker excision is indispensable for the introduction of desired mutations into the plant genome via gene targeting (GT) using a positive/negative counter selection system. In mammals, the moth-derived piggyBac transposon system has been exploited successfully to eliminate a selectable marker from a GT locus without leaving a footprint. Here, we present evidence that the piggyBac transposon also functions in plant cells. To demonstrate the use of the piggyBac transposon for effective marker excision in plants, we designed a transposition assay system that allows the piggyBac transposition to be visualized as emerald luciferase (Eluc) luminescence in rice cells. The Eluc signal derived from piggyBac excision was observed in hyperactive piggyBac transposase-expressing rice calli. Polymerase chain reaction, Southern blot analyses and sequencing revealed the efficient and precise transposition of piggyBac in these calli. Furthermore, we have demonstrated the excision of a selection marker from a reporter locus in T0 plants without concomitant re-integration of the transposon and at a high frequency (44.0% of excision events), even in the absence of negative selection. PMID:24164672

  20. Precise marker excision system using an animal-derived piggyBac transposon in plants

    PubMed Central

    Nishizawa-Yokoi, Ayako; Endo, Masaki; Osakabe, Keishi; Saika, Hiroaki; Toki, Seiichi

    2014-01-01

    Accurate and effective positive marker excision is indispensable for the introduction of desired mutations into the plant genome via gene targeting (GT) using a positive/negative counter selection system. In mammals, the moth-derived piggyBac transposon system has been exploited successfully to eliminate a selectable marker from a GT locus without leaving a footprint. Here, we present evidence that the piggyBac transposon also functions in plant cells. To demonstrate the use of the piggyBac transposon for effective marker excision in plants, we designed a transposition assay system that allows the piggyBac transposition to be visualized as emerald luciferase (Eluc) luminescence in rice cells. The Eluc signal derived from piggyBac excision was observed in hyperactive piggyBac transposase-expressing rice calli. Polymerase chain reaction, Southern blot analyses and sequencing revealed the efficient and precise transposition of piggyBac in these calli. Furthermore, we have demonstrated the excision of a selection marker from a reporter locus in T0 plants without concomitant re-integration of the transposon and at a high frequency (44.0% of excision events), even in the absence of negative selection. PMID:24164672

  1. piggyBac is a flexible and highly active transposon as compared to sleeping beauty, Tol2, and Mos1 in mammalian cells.

    PubMed

    Wu, Sareina Chiung-Yuan; Meir, Yaa-Jyuhn James; Coates, Craig J; Handler, Alfred M; Pelczar, Pawel; Moisyadi, Stefan; Kaminski, Joseph M

    2006-10-10

    A nonviral vector for highly efficient site-specific integration would be desirable for many applications in transgenesis, including gene therapy. In this study we directly compared the genomic integration efficiencies of piggyBac, hyperactive Sleeping Beauty (SB11), Tol2, and Mos1 in four mammalian cell lines. piggyBac demonstrated significantly higher transposition activity in all cell lines whereas Mos1 had no activity. Furthermore, piggyBac transposase coupled to the GAL4 DNA-binding domain retains transposition activity whereas similarly manipulated gene products of Tol2 and SB11 were inactive. The high transposition activity of piggyBac and the flexibility for molecular modification of its transposase suggest the possibility of using it routinely for mammalian transgenesis. PMID:17005721

  2. Hybrid Nonviral/Viral Vector Systems for Improved piggyBac DNA Transposon In Vivo Delivery

    PubMed Central

    Cooney, Ashley L; Singh, Brajesh K; Sinn, Patrick L

    2015-01-01

    The DNA transposon piggyBac is a potential therapeutic agent for multiple genetic diseases such as cystic fibrosis (CF). Recombinant piggyBac transposon and transposase are typically codelivered by plasmid transfection; however, plasmid delivery is inefficient in somatic cells in vivo and is a barrier to the therapeutic application of transposon-based vector systems. Here, we investigate the potential for hybrid piggyBac/viral vectors to transduce cells and support transposase-mediated genomic integration of the transposon. We tested both adenovirus (Ad) and adeno-associated virus (AAV) as transposon delivery vehicles. An Ad vector expressing hyperactive insect piggyBac transposase (iPB7) was codelivered. We show transposase-dependent transposition activity and mapped integrations in mammalian cells in vitro and in vivo from each viral vector platform. We also demonstrate efficient and persistent transgene expression following nasal delivery of piggyBac/viral vectors to mice. Furthermore, using piggyBac/Ad expressing Cystic Fibrosis transmembrane Conductance Regulator (CFTR), we show persistent correction of chloride current in well-differentiated primary cultures of human airway epithelial cells derived from CF patients. Combining the emerging technologies of DNA transposon-based vectors with well-studied adenoviral and AAV delivery provides new tools for in vivo gene transfer and presents an exciting opportunity to increase the delivery efficiency for therapeutic genes such as CFTR. PMID:25557623

  3. Vast potential for using the piggyBac transposon to engineer transgenic plants at specific genomic locations.

    PubMed

    Johnson, Eric T; Owens, Jesse B; Moisyadi, Stefan

    2016-01-01

    The acceptance of bioengineered plants by some nations is hampered by a number of factors, including the random insertion of a transgene into the host genome. Emerging technologies, such as site-specific nucleases, are enabling plant scientists to promote recombination or mutations at specific plant loci. Off target activity of these nucleases may limit widespread use. Insertion of transgenes by transposases engineered with a specific DNA binding domain has been accomplished in a number of organisms, but not in plants. The piggyBac transposon system, originally isolated from an insect, has been utilized to transform a variety of organisms. The piggyBac transposase is amendable to structural modifications, and was able to insert a transgene at a specific human locus through fusion of a DNA binding domain to its N-terminus. Recent developments demonstrating the activity of piggyBac transposase in plants is an important first step toward the potential use of engineered versions of piggyBac transposase for site-specific transgene insertion in plants. PMID:26930269

  4. piggyBac

    PubMed Central

    Lok, James

    2013-01-01

    In addition to their natural role in eukaryotic genome evolution, transposons can be powerful tools for functional genomics in diverse taxa. The piggyBac transposon has been applied as such in eukaryotic parasites, both protozoa and helminths, and in several important vector mosquitoes. piggyBac is advantageous for functional genomics because of its ability to transduce a wide range of taxa, its capacity to integrate large DNA ‘cargoes’ relative to other mobile genetic elements, its propensity to target transcriptional units and its ability to re-mobilize without leaving a pattern of non-excised sequences or ‘footprint’ in the genome. We recently demonstrated that piggyBac can integrate transgenes into the genome of the parasitic nematode Strongyloides ratti, an important model for parasitic nematode biology and a close relative of the significant human pathogen S. stercoralis. Unlike transgenes encoded in conventional plasmid vectors, which we assume are assembled into multi-copy episomal arrays as they are in Caenorhabditis elegans, transgenes integrated via piggyBac are not only stably inherited in S. ratti, they are also continuously expressed. This has allowed derivation of the first stable transgene expressing lines in any parasitic nematode, a significant advance in the development of functional genomic tools for these important pathogens. PMID:23914309

  5. Optimization of the piggyBac Transposon Using mRNA and Insulators: Toward a More Reliable Gene Delivery System

    PubMed Central

    Bire, Solenne; Ley, Déborah; Casteret, Sophie; Mermod, Nicolas; Bigot, Yves; Rouleux-Bonnin, Florence

    2013-01-01

    Integrating and expressing stably a transgene into the cellular genome remain major challenges for gene-based therapies and for bioproduction purposes. While transposon vectors mediate efficient transgene integration, expression may be limited by epigenetic silencing, and persistent transposase expression may mediate multiple transposition cycles. Here, we evaluated the delivery of the piggyBac transposase messenger RNA combined with genetically insulated transposons to isolate the transgene from neighboring regulatory elements and stabilize expression. A comparison of piggyBac transposase expression from messenger RNA and DNA vectors was carried out in terms of expression levels, transposition efficiency, transgene expression and genotoxic effects, in order to calibrate and secure the transposition-based delivery system. Messenger RNA reduced the persistence of the transposase to a narrow window, thus decreasing side effects such as superfluous genomic DNA cleavage. Both the CTF/NF1 and the D4Z4 insulators were found to mediate more efficient expression from a few transposition events. We conclude that the use of engineered piggyBac transposase mRNA and insulated transposons offer promising ways of improving the quality of the integration process and sustaining the expression of transposon vectors. PMID:24312663

  6. Generation of an inducible and optimized piggyBac transposon system†

    PubMed Central

    Cadiñanos, Juan; Bradley, Allan

    2007-01-01

    Genomic studies in the mouse have been slowed by the lack of transposon-mediated mutagenesis. However, since the resurrection of Sleeping Beauty (SB), the possibility of performing forward genetics in mice has been reinforced. Recently, piggyBac (PB), a functional transposon from insects, was also described to work in mammals. As the activity of PB is higher than that of SB11 and SB12, two hyperactive SB transposases, we have characterized and improved the PB system in mouse ES cells. We have generated a mouse codon-optimized version of the PB transposase coding sequence (CDS) which provides transposition levels greater than the original. We have also found that the promoter sequence predicted in the 5′-terminal repeat of the PB transposon is active in the mammalian context. Finally, we have engineered inducible versions of the optimized piggyBac transposase fused with ERT2. One of them, when induced, provides higher levels of transposition than the native piggyBac CDS, whereas in the absence of induction its activity is indistinguishable from background. We expect that these tools, adaptable to perform mouse-germline mutagenesis, will facilitate the identification of genes involved in pathological and physiological processes, such as cancer or ES cell differentiation. PMID:17576687

  7. Hyperactive PiggyBac Transposons for Sustained and Robust Liver-targeted Gene Therapy

    PubMed Central

    Di Matteo, Mario; Samara-Kuko, Emira; Ward, Natalie J; Waddingon, Simon N; McVey, John H; Chuah, Marinee KL; VandenDriessche, Thierry

    2014-01-01

    The development of robust nonviral vectors could facilitate clinical gene therapy applications and may overcome some of the immune complications of viral vectors. Nevertheless, most nonviral gene deliver approaches typically yield only transient and/or low gene expression. To address these caveats, we have explored piggyBac transposons to correct hemophilia B by liver-directed factor IX (FIX) gene therapy in hemophilic mice. To achieve this, we combined the use of: (i) a hyperactive codon-optimized piggyBac transposase, (ii) a computationally enhanced liver-specific promoter, (iii) a hyperfunctional codon-optimized FIX transgene (FIX R338L Padua), and (iv) a modification of the transposon terminal repeats. This combination strategy resulted in a robust 400-fold improvement in vector performance in hepatocytes, yielding stable supraphysiologic human FIX activity (>1 year). Liver-specific expression resulted in the induction of FIX-specific immune tolerance. Remarkably, only very low transposon/transposase doses were required to cure the bleeding diathesis. Similarly, PB transposons could be used to express supraphysiologic factor VIII levels using low transposon/transposase doses. PB transposition did not induce tumors in a sensitive hepatocellular carcinoma-prone mouse model. These results underscore the potency and relative safety of the latest generation PB transposons, which constitutes a versatile platform for stable and robust secretion of therapeutic proteins. PMID:25034357

  8. Hyperactive piggyBac transposons for sustained and robust liver-targeted gene therapy.

    PubMed

    Di Matteo, Mario; Samara-Kuko, Emira; Ward, Natalie J; Waddington, Simon N; Waddingon, Simon N; McVey, John H; Chuah, Marinee K L; VandenDriessche, Thierry

    2014-09-01

    The development of robust nonviral vectors could facilitate clinical gene therapy applications and may overcome some of the immune complications of viral vectors. Nevertheless, most nonviral gene deliver approaches typically yield only transient and/or low gene expression. To address these caveats, we have explored piggyBac transposons to correct hemophilia B by liver-directed factor IX (FIX) gene therapy in hemophilic mice. To achieve this, we combined the use of: (i) a hyperactive codon-optimized piggyBac transposase, (ii) a computationally enhanced liver-specific promoter, (iii) a hyperfunctional codon-optimized FIX transgene (FIX R338L Padua), and (iv) a modification of the transposon terminal repeats. This combination strategy resulted in a robust 400-fold improvement in vector performance in hepatocytes, yielding stable supraphysiologic human FIX activity (>1 year). Liver-specific expression resulted in the induction of FIX-specific immune tolerance. Remarkably, only very low transposon/transposase doses were required to cure the bleeding diathesis. Similarly, PB transposons could be used to express supraphysiologic factor VIII levels using low transposon/transposase doses. PB transposition did not induce tumors in a sensitive hepatocellular carcinoma-prone mouse model. These results underscore the potency and relative safety of the latest generation PB transposons, which constitutes a versatile platform for stable and robust secretion of therapeutic proteins.

  9. Sustained high level transgene expression in mammalian cells mediated by the optimized piggyBac transposon system

    PubMed Central

    Chen, Xiang; Cui, Jing; Yan, Zhengjian; Zhang, Hongmei; Chen, Xian; Wang, Ning; Shah, Palak; Deng, Fang; Zhao, Chen; Geng, Nisha; Li, Melissa; Denduluri, Sahitya K.; Haydon, Rex C.; Luu, Hue H.; Reid, Russell R.; He, Tong-Chuan

    2015-01-01

    Sustained, high level transgene expression in mammalian cells, especially stem cells, may be desired in many cases for studying gene functions. Traditionally, stable transgene expression has been accomplished by using retroviral or lentiviral vectors. However, such viral vector-mediated transgene expression is often at low levels and can be reduced over time due to low copy numbers and/or chromatin remodeling repression. The piggyBac transposon has emerged as a promising non-viral vector system for efficient gene transfer into mammalian cells. Despite its inherent advantages over lentiviral and retroviral systems, piggyBac system has not been widely used, at least in part due to the limited availability of piggyBac vectors with manipulation flexibilities. Here, we seek to optimize piggyBac-mediated transgene expression and generate a more efficient, user-friendly piggyBac system. By engineering a panel of versatile piggyBac vectors and constructing recombinant adenoviruses expressing piggyBac transposase (PBase), we demonstrate that adenovirus-mediated PBase expression significantly enhances the integration efficiency and expression level of transgenes in mesenchymal stem cells and osteosarcoma cells, compared to that obtained from co-transfection of the CMV-PBase plasmid. We further determine the drug selection timeline to achieve optimal stable transgene expression. Moreover, we demonstrate that the transgene copy number of piggyBac-mediated integration is approximately 10 times higher than that mediated by retroviral vectors. Using the engineered tandem expression vector, we show that three transgenes can be simultaneously expressed in a single vector with high efficiency. Thus, these results strongly suggest that the optimized piggyBac system is a valuable tool for making stable cell lines with sustained, high transgene expression. PMID:25815368

  10. [The improvement and application of piggyBac transposon system in mammals].

    PubMed

    Qian, Qiujie; Che, Jiaqian; Ye, Lupeng; Zhong, Boxiong

    2014-10-01

    The piggyBac (PB) transposon system is a useful genomic engineering tool due to its high transposition efficiency, precise excision, semi-random insertion and large cargo capacity. But, it still needs to further improve the transgenic efficiency and reduce the risk of endogenous disruption caused by the random insertion of exogenous gene, especially in transgenic experiments of individual mammals. In recent studies, the PB transposase is fused with a DNA binding protein as a chimeric protein, which can guide the transposon to pre-designed loci. Besides, PB transposases obtained by mutagenesis have dramatically enhanced transposition activity and generated a novel function which is excision competent and integration defective. Furthermore, PB transposon system can carry large exogenous DNA fragments up to 207 kb when combining with the bacterial artificial chromosome vector. So far, these modified transposon systems have been widely applied in genome studies, gene therapy and induced pluripotent stem cells (iPS cells). In this study, we review the latest studies on piggyBac transposon system and its application prospect.

  11. Generation of transgene-free induced pluripotent mouse stem cells by the piggyBac transposon

    PubMed Central

    Yusa, Kosuke; Rad, Roland; Takeda, Junji; Bradley, Allan

    2009-01-01

    Induced pluripotent stem cells (iPSCs) have been generated from somatic cells by transgenic expression of Oct4, Sox2, Klf4, and cMyc. A major difficulty in the application of this technology for regenerative medicine, however, is the delivery of reprogramming factors. Whereas retroviral transduction increases the risk of tumorigenicity, transient expression methods have considerably lower reprogramming efficiencies. Here we show a highly efficient piggyBac transposon-based approach to generate integration-free iPSCs. Transposons carrying 2A peptide-linked reprogramming factors induced reprogramming of mouse embryonic fibroblasts with equivalent efficiencies to retroviral transduction. Transposons were removed from these primary iPSCs by re-expressing transposase. Transgene-free iPSCs could be easily identified by HSVtk-FIAU selection. piggyBac excises without a footprint, leaving the iPSC genome without any genetic alteration. iPSCs fulfilled all criteria of pluripotency, such as expression of embryonic stem cell-specific markers, teratoma formation and contribution to chimeras. piggyBac transposon-based reprogramming may be used to generate therapeutically applicable iPSCs. PMID:19337237

  12. In Vitro Synthesis, Delivery, and Bioavailability of Exogenous mRNA in Gene Transfer Mediated by PiggyBac Transposition.

    PubMed

    Bire, Solenne; Ishac, Nicole; Rouleux-Bonnin, Florence

    2016-01-01

    Nowadays, nonviral gene transfer is currently of great importance for introducing exogenous genes into genomes and for ensuring that transgene expression is suitable for therapeutic and bioproduction purposes. The piggyBac transposon-based system is particularly interesting since it is easy to engineer and has a large cargo capacity, up to 100 kb. In its setup, the system requires only the piggyBac transposase protein and the transgene delineated by the two piggyBac-specific inverted terminal repeats. Usually the source of transposase is carried by a DNA plasmid. However, the principal drawback of this method is the lasting presence of the transposase, due to episomal persistence or possible integration of the transposase gene vector into the cell's genome. This can lead to genotoxic effects such as multiple genomic integration events and remobilization of the transposon vector once it has been integrated. One alternative to improve the safety of the system is to deliver the transposase as in vitro-synthesized messenger RNA in order to define a very narrow expression window during which a one-shot transposition process would occur. Issues that can be encountered when working on mRNA cell transfer are related to the quality of the synthetic mRNA, the system used to introduce mRNA into the cells and the bioavailability of the mRNA molecules. Here we describe a method to produce mRNA, verify its quality, determine which transfecting reagents can be used and how this mRNA is available to promote the transposition process in HeLa cells. Additionally, we illustrate this method in stromal mesenchymal cell lines in order to support hematopoiesis. PMID:27236801

  13. A piggyBac transposon gene trap for the analysis of gene expression and function in Drosophila.

    PubMed Central

    Bonin, Christopher P; Mann, Richard S

    2004-01-01

    P-element-based gene and enhancer trap strategies have provided a wealth of information on the expression and function of genes in Drosophila melanogaster. Here we present a new vector that utilizes the simple insertion requirements of the piggyBac transposon, coupled to a splice acceptor (SA) site fused to the sequence encoding enhanced green fluorescent protein (EGFP) and a transcriptional terminator. Mobilization of the piggyBac splice site gene trap vector (PBss) was accomplished by heat-shock-induced expression of piggyBac transposase (PBase). We show that insertion of PBss into genes leads to fusions between the gene's mRNA and the PBss-encoded EGFP transcripts. As heterozygotes, these fusions report the normal pattern of expression of the trapped gene. As homozygotes, these fusions can inactivate the gene and lead to lethality. Molecular characterization of PBss insertion events shows that they are single copy, that they always occur at TTAA sequences, and that splicing utilizes the engineered splice site in PBss. In those instances where protein-EGFP fusions are predicted to occur, the subcellular localization of the wild-type protein can be inferred from the localization of the EGFP fusion protein. These experiments highlight the utility of the PBss system for expanding the functional genomics tools that are available in Drosophila. PMID:15342518

  14. Mobility of the piggyBac transposon in embryos of the vectors of Dengue fever (Aedes albopictus) and La Crosse encephalitis (Ae. triseriatus).

    PubMed

    Lobo, N; Li, X; Hua-Van, A; Fraser, M J

    2001-03-01

    The re-emergence of arboviral diseases such as Dengue Fever and La Crosse encephalitis is primarily due to the failure of insect vector control strategies. The development of a procedure capable of producing stable germ-line transformants in the insect vectors of these diseases would bridge the gap between gene expression systems being developed to curb vector transmission and the identification of important genes and regulatory sequences and their reintroduction back into the insect genome in the form of vector control strategies. The transposable element piggyBac is capable of transposition in a variety of insect species, and could serve as a versatile insect transformation vector. Using plasmid-based excision and transposition assays, we report that this short-ITR transposon undergoes precise, transposase-dependent excision and transposition in embryos of Aedes albopictus and Aedes triseriatus, the vectors of Dengue fever and LaCrosse encephalitis, respectively. These assays allow us easily and rapidly to confirm and assess the potential utility of piggyBac as a gene transfer tool in a given species. piggyBac is an exceptionally mobile and versatile genetic transformation vector, comparable to other transposons currently in use for the transformation of insects. The mobility of the piggyBac element seen in both Ae. albopictus and Ae. triseriatus is further evidence that it can be employed as a germ-line vector in important insect disease vectors.

  15. Ultrasound directs a transposase system for durable hepatic gene delivery in mice

    PubMed Central

    Anderson, Cynthia D; Urschitz, Johann; Khemmani, Mark; Owens, Jesse B; Moisyadi, Stefan

    2013-01-01

    Our aim was to evaluate the delivery of transposase-based vectors by Ultrasound Targeted Microbubble Destruction (UTMD) in mice. DNA vectors were attached to cationic lipid microbubbles (1-3 μm in diameter), injected intravenously, and delivered to the liver by destruction of the carrier bubbles with ultrasound in burst mode at a 1.0 MHz, 20 μs pulse duration, 10 Hz pulse repetition frequency, and acoustic peak negative pressure of ~1.3 MPa. We evaluated the expression and genomic integration of conventional (pcDNA3) and piggyBac transposase-based (pmGENIE) reporter vectors. In vivo, we observed UTMD-mediated liver-specific expression of pmGENIE for an average of 24 days, compared to 4 days with pcDNA3. Reporter expression was predominately located in proximity to blood vessels initially, while expression after three days was more evenly distributed through the parenchyma of the liver. We confirmed random genomic integration for pmGENIE in vitro, however, integration events for pmGENIE in vivo were targeted to specific areas of chromosome 14. Our results suggest that a combination of UTMD with nonviral DNA transposase vectors can mediate weeks of hepatic-specific gene transfer in vivo, and analyses performed by non-restrictive linear amplification-mediated (nrLAM) PCR, cloning, and sequencing identify an unexpected tropism for integration within a specific sequence on chromosome 14 in mice. UTMD delivery of transgenes may be useful for the treatment of hepatic gene deficiency disorders. PMID:24035623

  16. Structural Determinants of Sleeping Beauty Transposase Activity.

    PubMed

    Abrusán, György; Yant, Stephen R; Szilágyi, András; Marsh, Joseph A; Mátés, Lajos; Izsvák, Zsuzsanna; Barabás, Orsolya; Ivics, Zoltán

    2016-08-01

    Transposases are important tools in genome engineering, and there is considerable interest in engineering more efficient ones. Here, we seek to understand the factors determining their activity using the Sleeping Beauty transposase. Recent work suggests that protein coevolutionary information can be used to classify groups of physically connected, coevolving residues into elements called "sectors", which have proven useful for understanding the folding, allosteric interactions, and enzymatic activity of proteins. Using extensive mutagenesis data, protein modeling and analysis of folding energies, we show that (i) The Sleeping Beauty transposase contains two sectors, which span across conserved domains, and are enriched in DNA-binding residues, indicating that the DNA binding and endonuclease functions of the transposase coevolve; (ii) Sector residues are highly sensitive to mutations, and most mutations of these residues strongly reduce transposition rate; (iii) Mutations with a strong effect on free energy of folding in the DDE domain of the transposase significantly reduce transposition rate. (iv) Mutations that influence DNA and protein-protein interactions generally reduce transposition rate, although most hyperactive mutants are also located on the protein surface, including residues with protein-protein interactions. This suggests that hyperactivity results from the modification of protein interactions, rather than the stabilization of protein fold.

  17. Structural Determinants of Sleeping Beauty Transposase Activity.

    PubMed

    Abrusán, György; Yant, Stephen R; Szilágyi, András; Marsh, Joseph A; Mátés, Lajos; Izsvák, Zsuzsanna; Barabás, Orsolya; Ivics, Zoltán

    2016-08-01

    Transposases are important tools in genome engineering, and there is considerable interest in engineering more efficient ones. Here, we seek to understand the factors determining their activity using the Sleeping Beauty transposase. Recent work suggests that protein coevolutionary information can be used to classify groups of physically connected, coevolving residues into elements called "sectors", which have proven useful for understanding the folding, allosteric interactions, and enzymatic activity of proteins. Using extensive mutagenesis data, protein modeling and analysis of folding energies, we show that (i) The Sleeping Beauty transposase contains two sectors, which span across conserved domains, and are enriched in DNA-binding residues, indicating that the DNA binding and endonuclease functions of the transposase coevolve; (ii) Sector residues are highly sensitive to mutations, and most mutations of these residues strongly reduce transposition rate; (iii) Mutations with a strong effect on free energy of folding in the DDE domain of the transposase significantly reduce transposition rate. (iv) Mutations that influence DNA and protein-protein interactions generally reduce transposition rate, although most hyperactive mutants are also located on the protein surface, including residues with protein-protein interactions. This suggests that hyperactivity results from the modification of protein interactions, rather than the stabilization of protein fold. PMID:27401040

  18. Metaproteomics reveals abundant transposase expression in mutualistic endosymbionts

    SciTech Connect

    Kleiner, Manuel; Young, Jacque C; Shah, Manesh B; Verberkmoes, Nathan C; Dubilier, Nicole

    2013-01-01

    Transposases, enzymes that catalyze the movement of mobile genetic elements, are the most abundant genes in nature. While many bacteria encode an abundance of transposases in their genomes, the current paradigm is that transposase gene expression is tightly regulated and generally low due to its severe mutagenic effects. In the current study, we detected the highest number of transposase proteins ever reported in bacteria, in symbionts of the gutless marine worm Olavius algarvensis using metaproteomics. At least 26 different transposases from 12 different families were detected and genomic and proteomic analyses suggest many of these are active. This high expression of transposases indicates that the mechanisms for their tight regulation have been disabled or destroyed. Based on recent studies on other symbionts and pathogens that showed high transposase transcription, we speculate that abundant transposase expression might be common in symbionts and pathogens.

  19. Suicidal autointegration of sleeping beauty and piggyBac transposons in eukaryotic cells.

    PubMed

    Wang, Yongming; Wang, Jichang; Devaraj, Anatharam; Singh, Manvendra; Jimenez Orgaz, Ana; Chen, Jia-Xuan; Selbach, Matthias; Ivics, Zoltán; Izsvák, Zsuzsanna

    2014-03-01

    Transposons are discrete segments of DNA that have the distinctive ability to move and replicate within genomes across the tree of life. 'Cut and paste' DNA transposition involves excision from a donor locus and reintegration into a new locus in the genome. We studied molecular events following the excision steps of two eukaryotic DNA transposons, Sleeping Beauty (SB) and piggyBac (PB) that are widely used for genome manipulation in vertebrate species. SB originates from fish and PB from insects; thus, by introducing these transposons to human cells we aimed to monitor the process of establishing a transposon-host relationship in a naïve cellular environment. Similarly to retroviruses, neither SB nor PB is capable of self-avoidance because a significant portion of the excised transposons integrated back into its own genome in a suicidal process called autointegration. Barrier-to-autointegration factor (BANF1), a cellular co-factor of certain retroviruses, inhibited transposon autointegration, and was detected in higher-order protein complexes containing the SB transposase. Increasing size sensitized transposition for autointegration, consistent with elevated vulnerability of larger transposons. Both SB and PB were affected similarly by the size of the transposon in three different assays: excision, autointegration and productive transposition. Prior to reintegration, SB is completely separated from the donor molecule and followed an unbiased autointegration pattern, not associated with local hopping. Self-disruptive autointegration occurred at similar frequency for both transposons, while aberrant, pseudo-transposition events were more frequently observed for PB.

  20. piggyBac transposons expressing full-length human dystrophin enable genetic correction of dystrophic mesoangioblasts.

    PubMed

    Loperfido, Mariana; Jarmin, Susan; Dastidar, Sumitava; Di Matteo, Mario; Perini, Ilaria; Moore, Marc; Nair, Nisha; Samara-Kuko, Ermira; Athanasopoulos, Takis; Tedesco, Francesco Saverio; Dickson, George; Sampaolesi, Maurilio; VandenDriessche, Thierry; Chuah, Marinee K

    2016-01-29

    Duchenne muscular dystrophy (DMD) is a genetic neuromuscular disorder caused by the absence of dystrophin. We developed a novel gene therapy approach based on the use of the piggyBac (PB) transposon system to deliver the coding DNA sequence (CDS) of either full-length human dystrophin (DYS: 11.1 kb) or truncated microdystrophins (MD1: 3.6 kb; MD2: 4 kb). PB transposons encoding microdystrophins were transfected in C2C12 myoblasts, yielding 65±2% MD1 and 66±2% MD2 expression in differentiated multinucleated myotubes. A hyperactive PB (hyPB) transposase was then deployed to enable transposition of the large-size PB transposon (17 kb) encoding the full-length DYS and green fluorescence protein (GFP). Stable GFP expression attaining 78±3% could be achieved in the C2C12 myoblasts that had undergone transposition. Western blot analysis demonstrated expression of the full-length human DYS protein in myotubes. Subsequently, dystrophic mesoangioblasts from a Golden Retriever muscular dystrophy dog were transfected with the large-size PB transposon resulting in 50±5% GFP-expressing cells after stable transposition. This was consistent with correction of the differentiated dystrophic mesoangioblasts following expression of full-length human DYS. These results pave the way toward a novel non-viral gene therapy approach for DMD using PB transposons underscoring their potential to deliver large therapeutic genes.

  1. piggyBac transposons expressing full-length human dystrophin enable genetic correction of dystrophic mesoangioblasts

    PubMed Central

    Loperfido, Mariana; Jarmin, Susan; Dastidar, Sumitava; Di Matteo, Mario; Perini, Ilaria; Moore, Marc; Nair, Nisha; Samara-Kuko, Ermira; Athanasopoulos, Takis; Tedesco, Francesco Saverio; Dickson, George; Sampaolesi, Maurilio; VandenDriessche, Thierry; Chuah, Marinee K.

    2016-01-01

    Duchenne muscular dystrophy (DMD) is a genetic neuromuscular disorder caused by the absence of dystrophin. We developed a novel gene therapy approach based on the use of the piggyBac (PB) transposon system to deliver the coding DNA sequence (CDS) of either full-length human dystrophin (DYS: 11.1 kb) or truncated microdystrophins (MD1: 3.6 kb; MD2: 4 kb). PB transposons encoding microdystrophins were transfected in C2C12 myoblasts, yielding 65±2% MD1 and 66±2% MD2 expression in differentiated multinucleated myotubes. A hyperactive PB (hyPB) transposase was then deployed to enable transposition of the large-size PB transposon (17 kb) encoding the full-length DYS and green fluorescence protein (GFP). Stable GFP expression attaining 78±3% could be achieved in the C2C12 myoblasts that had undergone transposition. Western blot analysis demonstrated expression of the full-length human DYS protein in myotubes. Subsequently, dystrophic mesoangioblasts from a Golden Retriever muscular dystrophy dog were transfected with the large-size PB transposon resulting in 50±5% GFP-expressing cells after stable transposition. This was consistent with correction of the differentiated dystrophic mesoangioblasts following expression of full-length human DYS. These results pave the way toward a novel non-viral gene therapy approach for DMD using PB transposons underscoring their potential to deliver large therapeutic genes. PMID:26682797

  2. Suicidal Autointegration of Sleeping Beauty and piggyBac Transposons in Eukaryotic Cells

    PubMed Central

    Devaraj, Anatharam; Singh, Manvendra; Jimenez Orgaz, Ana; Chen, Jia-Xuan; Selbach, Matthias; Ivics, Zoltán; Izsvák, Zsuzsanna

    2014-01-01

    Transposons are discrete segments of DNA that have the distinctive ability to move and replicate within genomes across the tree of life. ‘Cut and paste’ DNA transposition involves excision from a donor locus and reintegration into a new locus in the genome. We studied molecular events following the excision steps of two eukaryotic DNA transposons, Sleeping Beauty (SB) and piggyBac (PB) that are widely used for genome manipulation in vertebrate species. SB originates from fish and PB from insects; thus, by introducing these transposons to human cells we aimed to monitor the process of establishing a transposon-host relationship in a naïve cellular environment. Similarly to retroviruses, neither SB nor PB is capable of self-avoidance because a significant portion of the excised transposons integrated back into its own genome in a suicidal process called autointegration. Barrier-to-autointegration factor (BANF1), a cellular co-factor of certain retroviruses, inhibited transposon autointegration, and was detected in higher-order protein complexes containing the SB transposase. Increasing size sensitized transposition for autointegration, consistent with elevated vulnerability of larger transposons. Both SB and PB were affected similarly by the size of the transposon in three different assays: excision, autointegration and productive transposition. Prior to reintegration, SB is completely separated from the donor molecule and followed an unbiased autointegration pattern, not associated with local hopping. Self-disruptive autointegration occurred at similar frequency for both transposons, while aberrant, pseudo-transposition events were more frequently observed for PB. PMID:24625543

  3. Excision Efficiency Is Not Strongly Coupled to Transgenic Rate: Cell Type-Dependent Transposition Efficiency of Sleeping Beauty and piggyBac DNA Transposons

    PubMed Central

    Kolacsek, Orsolya; Erdei, Zsuzsa; Apáti, Ágota; Sándor, Sára; Izsvák, Zsuzsanna; Ivics, Zoltán; Sarkadi, Balázs

    2014-01-01

    Abstract The Sleeping Beauty (SB) and piggyBac (PB) DNA transposons represent an emerging new gene delivery technology, potentially suitable for human gene therapy applications. Previous studies pointed to important differences between these transposon systems, depending on the cell types examined and the methodologies applied. However, efficiencies cannot always be compared because of differences in applications. In addition, “overproduction inhibition,” a phenomenon believed to be a characteristic of DNA transposons, can remarkably reduce the overall transgenic rate, emphasizing the importance of transposase dose applied. Therefore, because of lack of comprehensive analysis, researchers are forced to optimize the technology for their own “in-house” platforms. In this study, we investigated the transposition of several SB (SB11, SB32, SB100X) and PB (mPB and hyPB) variants in various cell types at three levels: comparing the excision efficiency of the reaction by real-time PCR, testing the overall transgenic rate by detecting cells with stable integrations, and determining the average copy number when using different transposon systems and conditions. We concluded that high excision activity is not always followed by a higher transgenic rate, as exemplified by the hyperactive transposases, indicating that the excision and the integration steps of transposition are not strongly coupled as previously thought. In general, all levels of transposition show remarkable differences depending on the transposase used and cell lines examined, being the least efficient in human embryonic stem cells (hESCs). In spite of the comparably low activity in those special cell types, the hyperactive SB100X and hyPB systems could be used in hESCs with similar transgenic efficiency and with reasonably low (2–3) transgene copy numbers, indicating their potential applicability for gene therapy purposes in the future. PMID:25045962

  4. TAL effectors mediate high-efficiency transposition of the piggyBac transposon in silkworm Bombyx mori L.

    PubMed

    Ye, Lupeng; You, Zhengying; Qian, Qiujie; Zhang, Yuyu; Che, Jiaqian; Song, Jia; Zhong, Boxiong

    2015-11-26

    The piggyBac (PB) transposon is one of the most useful transposable elements, and has been successfully used for genetic manipulation in more than a dozen species. However, the efficiency of PB-mediated transposition is still insufficient for many purposes. Here, we present a strategy to enhance transposition efficiency using a fusion of transcription activator-like effector (TALE) and the PB transposase (PBase). The results demonstrate that the TALE-PBase fusion protein which is engineered in this study can produce a significantly improved stable transposition efficiency of up to 63.9%, which is at least 7 times higher than the current transposition efficiency in silkworm. Moreover, the average number of transgene-positive individuals increased up to 5.7-fold, with each positive brood containing an average of 18.1 transgenic silkworms. Finally, we demonstrate that TALE-PBase fusion-mediated PB transposition presents a new insertional preference compared with original insertional preference. This method shows a great potential and value for insertional therapy of many genetic diseases. In conclusion, this new and powerful transposition technology will efficiently promote genetic manipulation studies in both invertebrates and vertebrates.

  5. PiggyBac transposon-based polyadenylation-signal trap for genome-wide mutagenesis in mice

    PubMed Central

    Li, Limei; Liu, Peng; Sun, Liangliang; Bin Zhou; Fei, Jian

    2016-01-01

    We designed a new type of polyadenylation-signal (PAS) trap vector system in living mice, the piggyBac (PB) (PAS-trapping (EGFP)) gene trapping vector, which takes advantage of the efficient transposition ability of PB and efficient gene trap and insertional mutagenesis of PAS-trapping. The reporter gene of PB(PAS-trapping (EGFP)) is an EGFP gene with its own promoter, but lacking a poly(A) signal. Transgenic mouse lines carrying PB(PAS-trapping (EGFP)) and protamine 1 (Prm1) promoter-driven PB transposase transgenes (Prm1-PBase) were generated by microinjection. Male mice doubly positive for PB(PAS-trapping (EGFP)) and Prm1-PBase were crossed with WT females, generating offspring with various insertion mutations. We found that 44.8% (26/58) of pups were transposon-positive progenies. New transposon integrations comprised 26.9% (7/26) of the transposon-positive progenies. We found that 100% (5/5) of the EGFP fluorescence-positive mice had new trap insertions mediated by a PB transposon in transcriptional units. The direction of the EGFP gene in the vector was consistent with the direction of the endogenous gene reading frame. Furthermore, mice that were EGFP-PCR positive, but EGFP fluorescent negative, did not show successful gene trapping. Thus, the novel PB(PAS-trapping (EGFP)) system is an efficient genome-wide gene-trap mutagenesis in mice. PMID:27292714

  6. NMR structural analysis of Sleeping Beauty transposase binding to DNA

    PubMed Central

    E Carpentier, Claire; Schreifels, Jeffrey M; Aronovich, Elena L; Carlson, Daniel F; Hackett, Perry B; Nesmelova, Irina V

    2014-01-01

    The Sleeping Beauty (SB) transposon is the most widely used DNA transposon in genetic applications and is the only DNA transposon thus far in clinical trials for human gene therapy. In the absence of atomic level structural information, the development of SB transposon relied primarily on the biochemical and genetic homology data. While these studies were successful and have yielded hyperactive transposases, structural information is needed to gain a mechanistic understanding of transposase activity and guides to further improvement. We have initiated a structural study of SB transposase using Nuclear Magnetic Resonance (NMR) and Circular Dichroism (CD) spectroscopy to investigate the properties of the DNA-binding domain of SB transposase in solution. We show that at physiologic salt concentrations, the SB DNA-binding domain remains mostly unstructured but its N-terminal PAI subdomain forms a compact, three-helical structure with a helix-turn-helix motif at higher concentrations of NaCl. Furthermore, we show that the full-length SB DNA-binding domain associates differently with inner and outer binding sites of the transposon DNA. We also show that the PAI subdomain of SB DNA-binding domain has a dominant role in transposase's attachment to the inverted terminal repeats of the transposon DNA. Overall, our data validate several earlier predictions and provide new insights on how SB transposase recognizes transposon DNA. PMID:24243759

  7. Identification of nuclear localization signal within goldfish Tgf2 transposase.

    PubMed

    Shen, Xiao-Dan; Hou, Fei; Chen, Jie; Jiang, Xia-Yun; Zou, Shu-Ming

    2016-11-15

    The structure of goldfish (Carassius auratus) Tgf2 transposase is still poorly understood, although it can mediate efficient gene transfer in teleost fish. We hypothesized the existence of a nuclear localization signal (NLS) within Tgf2 transposase to assist transport into the nucleus. To explore this, 15 consecutive amino acid residues (656-670 aa) within the C-terminus of Tgf2 transposase were predicted in silico to be a NLS domain. The pEGFP-C1-Tgf2TP(△31C) plasmid encoding the NLS-domain-deleted Tgf2 transposase fused to EGFP was constructed, and transfected into 293T cells. After transfection with pEGFP-C1-Tgf2TP(△31C), EGFP was not detected in the nucleus alone, while 67.0% of cells expressed EGFP only in the cytoplasm. In contrast, after transfection with control plasmids containing C- or N-terminal truncated Tgf2 transposases with an intact NLS domain, EGFP was not detected in the cytoplasm alone, while approximately 40% of cells expressed EGFP only in the nucleus, and the remaining 60% expressed EGFP in both the nucleus and cytoplasm. Our results demonstrated that loss of the NLS domain results in expression in the cytoplasm but not in the nucleus. These findings suggest that 15 aa residues located from 656 to 670 aa within the C-terminus of Tgf2 transposase can function as a NLS to assist the transfer of the transposase into the nucleus where it mediates DNA transposition. PMID:27468946

  8. Sperm-mediated transgenesis in chicken using a PiggyBac transposon system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Towards development of transgenic chickens without the use of viral vectors, factors affecting sperm mediated gene transfer (SMGT) using a piggyBac vector are being studied. The piggyBac pPBCAG-LacZ contains 13bp terminal inverted repeats flanking a LacZ gene driven by the CAG promoter. A helper pla...

  9. A non-autonomous insect piggyBac trasposable element is mobile in tobacco

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The piggyBac transposable element, originally isolated from a virus in an insect cell line, is a valuable molecular tool for transgenesis and mutagenesis of invertebrates. For heterologous transgenesis in a variety of mammals, transfer of the piggyBac transposable element from an ectopic plasmid onl...

  10. DNA binding activities of the Caenorhabditis elegans Tc3 transposase.

    PubMed Central

    Colloms, S D; van Luenen, H G; Plasterk, R H

    1994-01-01

    Tc3 is a member of the Tc1/mariner family of transposable elements. All these elements have terminal inverted repeats, encode related transposases and insert exclusively into TA dinucleotides. We have studied the DNA binding properties of Tc3 transposase and found that an N-terminal domain of 65 amino acids binds specifically to two regions within the 462 bp Tc3 inverted repeat; one region is located at the end of the inverted repeat, the other is located approximately 180 bp from the end. Methylation interference experiments indicate that this N-terminal DNA binding domain of the Tc3 transposase interacts with nucleotides on one face of the DNA helix over adjacent major and minor grooves. Images PMID:7838706

  11. Purification, crystallization and preliminary crystallographic analysis of the Hermes transposase

    SciTech Connect

    Perez, Zhanita N.; Musingarimi, Primrose; Craig, Nancy L.; Dyda, Fred; Hickman, Alison Burgess

    2005-06-01

    Upon purification, an N-terminally deleted version of the Hermes transposase exists in solution as a mixture of two species that are approximately hexameric and dimeric. Crystals have been obtained of the smaller species that diffract to 2.1 Å resolution. DNA transposition is the movement of a defined segment of DNA from one location to another. Although the enzymes that catalyze transposition in bacterial systems have been well characterized, much less is known about the families of transposase enzymes that function in higher organisms. Active transposons have been identified in many insect species, providing tools for gene identification and offering the possibility of altering the genotypes of natural insect populations. One of these active transposons is Hermes, a 2749-base-pair element from Musca domestica that encodes its own transposase. An N-terminally deleted version of the Hermes transposase (residues 79–612) has been overexpressed and purified, and crystals that diffract to 2.1 Å resolution have been obtained at 277 K by the hanging-drop method.

  12. [Construction of a general piggyBac transposon inducible cell immortalization vector and verification of its basic properties].

    PubMed

    Huang, Hui; Hu, Guangdong; Kang, Jian; Qing, Suzhu; Zhang, Yong

    2014-08-01

    In order to construct generally efficient cell immortalization vector, pTP-hTERT, we modified the traditional piggyBac (PB) transposon using artificial synthesis, PCR and enzyme digestion. The modified vector contained the necessary transposon elements, a PB transposase expression cassette, a co-expression selectable element and a human telomerase reverse transcriptase (hTERT) expression cassette. The co-expression selectable element had two markers, enhanced green fluorescent protein (EGFP) gene and puromycin-resistance (Puro) gene, linked by porcine teschovirus-1 2A peptide (P2A). To validate the functionality of vector elements, we transfected pTP-hTERT into HEK293 cell, selected the positive cell clones and then conducted RT-PCR, Western blotting (WB) and Tail-PCR, methylene blue staining and statistic analysis on selected cells. The results of sequencing and cell culture show that the pTP-hTERT was constructed successfully and the positive cell could be selected by puromycin. The WB results, P2A cutting EGFP and Puro fusion protein with high efficiency, reflected the selectable element worked. The sequencing result of Tail-PCR confirmed the vector integrated into the genome through transposition. The results of methylene blue staining and statistic analysis indicated the clone of positive cells triggered by pTP-hTERT significantly increased (P < 0.01) compared with control group. The construction of pTP-hTERT provides an efficient tool for establishing immortalized cell lines and a demonstration for building other eukaryotic plasmids.

  13. Sleeping Beauty transposase structure allows rational design of hyperactive variants for genetic engineering

    PubMed Central

    Voigt, Franka; Wiedemann, Lisa; Zuliani, Cecilia; Querques, Irma; Sebe, Attila; Mátés, Lajos; Izsvák, Zsuzsanna; Ivics, Zoltán; Barabas, Orsolya

    2016-01-01

    Sleeping Beauty (SB) is a prominent Tc1/mariner superfamily DNA transposon that provides a popular genome engineering tool in a broad range of organisms. It is mobilized by a transposase enzyme that catalyses DNA cleavage and integration at short specific sequences at the transposon ends. To facilitate SB's applications, here we determine the crystal structure of the transposase catalytic domain and use it to model the SB transposase/transposon end/target DNA complex. Together with biochemical and cell-based transposition assays, our structure reveals mechanistic insights into SB transposition and rationalizes previous hyperactive transposase mutations. Moreover, our data enables us to design two additional hyperactive transposase variants. Our work provides a useful resource and proof-of-concept for structure-based engineering of tailored SB transposases. PMID:27025571

  14. Sleeping Beauty transposase structure allows rational design of hyperactive variants for genetic engineering.

    PubMed

    Voigt, Franka; Wiedemann, Lisa; Zuliani, Cecilia; Querques, Irma; Sebe, Attila; Mátés, Lajos; Izsvák, Zsuzsanna; Ivics, Zoltán; Barabas, Orsolya

    2016-03-30

    Sleeping Beauty (SB) is a prominent Tc1/mariner superfamily DNA transposon that provides a popular genome engineering tool in a broad range of organisms. It is mobilized by a transposase enzyme that catalyses DNA cleavage and integration at short specific sequences at the transposon ends. To facilitate SB's applications, here we determine the crystal structure of the transposase catalytic domain and use it to model the SB transposase/transposon end/target DNA complex. Together with biochemical and cell-based transposition assays, our structure reveals mechanistic insights into SB transposition and rationalizes previous hyperactive transposase mutations. Moreover, our data enables us to design two additional hyperactive transposase variants. Our work provides a useful resource and proof-of-concept for structure-based engineering of tailored SB transposases.

  15. Sleeping Beauty transposase structure allows rational design of hyperactive variants for genetic engineering.

    PubMed

    Voigt, Franka; Wiedemann, Lisa; Zuliani, Cecilia; Querques, Irma; Sebe, Attila; Mátés, Lajos; Izsvák, Zsuzsanna; Ivics, Zoltán; Barabas, Orsolya

    2016-01-01

    Sleeping Beauty (SB) is a prominent Tc1/mariner superfamily DNA transposon that provides a popular genome engineering tool in a broad range of organisms. It is mobilized by a transposase enzyme that catalyses DNA cleavage and integration at short specific sequences at the transposon ends. To facilitate SB's applications, here we determine the crystal structure of the transposase catalytic domain and use it to model the SB transposase/transposon end/target DNA complex. Together with biochemical and cell-based transposition assays, our structure reveals mechanistic insights into SB transposition and rationalizes previous hyperactive transposase mutations. Moreover, our data enables us to design two additional hyperactive transposase variants. Our work provides a useful resource and proof-of-concept for structure-based engineering of tailored SB transposases. PMID:27025571

  16. Germ-line transformation of the Queensland fruit fly, Bactrocera tryoni, using a piggyBac vector in the presence of endogenous piggyBac elements

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report the stable genetic transformation of the Queensland fruit fly Bactrocera tryoni using a piggyBac vector marked with either the fluorescent protein DsRed or EGFP.A transformation frequency of 5–10% was obtained.Inheritance of the transgenes has remained stable over eight generations despite...

  17. DNA repair by the cryptic endonuclease activity of Mu transposase.

    PubMed

    Choi, Wonyoung; Harshey, Rasika M

    2010-06-01

    Phage Mu transposes by two distinct pathways depending on the specific stage of its life cycle. A common strand transfer intermediate is resolved differentially in the two pathways. During lytic growth, the intermediate is resolved by replication of Mu initiated within the flanking target DNA; during integration of infecting Mu, it is resolved without replication, by removal and repair of DNA from a previous host that is still attached to the ends of the incoming Mu genome. We have discovered that the cryptic endonuclease activity reported for the isolated C-terminal domain of the transposase MuA [Wu Z, Chaconas G (1995) A novel DNA binding and nuclease activity in domain III of Mu transposase: Evidence for a catalytic region involved in donor cleavage. EMBO J 14:3835-3843], which is not observed in the full-length protein or in the assembled transpososome in vitro, is required in vivo for removal of the attached host DNA or "5'flap" after the infecting Mu genome has integrated into the E. coli chromosome. Efficient flap removal also requires the host protein ClpX, which is known to interact with the C-terminus of MuA to remodel the transpososome for replication. We hypothesize that ClpX constitutes part of a highly regulated mechanism that unmasks the cryptic nuclease activity of MuA specifically in the repair pathway. PMID:20167799

  18. MuA Transposase: Organizing Two Reactions With One Active Site

    NASA Astrophysics Data System (ADS)

    Goldhaber-Gordon, Ilana; Baker, Tania A.

    2002-03-01

    Transposases are unusual proteins, performing at least two types of chemical reaction with a single active site. In the first reaction, the transposase cleaves the 3' end of a transposon DNA sequence. In the second reaction, the transposase transfers the cleaved 3' end into a new DNA molecule called the target. The final product is the transposon DNA attached by its 3' strands to the target DNA. The two reactions, DNA cleavage and DNA strand transfer, are chemically very similar, raising the question of how transposases avoid unintentionally cleaving the target. We have now observed that at least one transposase, the MuA protein, can cleave a target DNA --- when in the presence of a transposon DNA with a special (dideoxy nucleotide) termination. By exploring the protein's ability to cleave the target DNA under these unusual circumstances, we are learning how it prevents target cleavage when working with more standard DNA substrates.

  19. The piggyBac transposon displays local and distant reintegration preferences and can cause mutations at noncanonical integration sites.

    PubMed

    Li, Meng Amy; Pettitt, Stephen J; Eckert, Sabine; Ning, Zemin; Rice, Stephen; Cadiñanos, Juan; Yusa, Kosuke; Conte, Nathalie; Bradley, Allan

    2013-04-01

    The DNA transposon piggyBac is widely used as a tool in mammalian experimental systems for transgenesis, mutagenesis, and genome engineering. We have characterized genome-wide insertion site preferences of piggyBac by sequencing a large set of integration sites arising from transposition from two separate genomic loci and a plasmid donor in mouse embryonic stem cells. We found that piggyBac preferentially integrates locally to the excision site when mobilized from a chromosomal location and identified other nonlocal regions of the genome with elevated insertion frequencies. piggyBac insertions were associated with expressed genes and markers of open chromatin structure and were excluded from heterochromatin. At the nucleotide level, piggyBac prefers to insert into TA-rich regions within a broader GC-rich context. We also found that piggyBac can insert into sites other than its known TTAA insertion site at a low frequency (2%). Such insertions introduce mismatches that are repaired with signatures of host cell repair pathways. Transposons could be mobilized from plasmids with the observed noncanonical flanking regions, indicating that piggyBac could generate point mutations in the genome. PMID:23358416

  20. Insect transformation with piggyBac: getting the number of injections just right

    PubMed Central

    Morrison, N. I.; Shimeld, S. M.

    2016-01-01

    Abstract The insertion of exogenous genetic cargo into insects using transposable elements is a powerful research tool with potential applications in meeting food security and public health challenges facing humanity. piggyBac is the transposable element most commonly utilized for insect germline transformation. The described efficiency of this process is variable in the published literature, and a comprehensive review of transformation efficiency in insects is lacking. This study compared and contrasted all available published data with a comprehensive data set provided by a biotechnology group specializing in insect transformation. Based on analysis of these data, with particular focus on the more complete observational data from the biotechnology group, we designed a decision tool to aid researchers' decision‐making when using piggyBac to transform insects by microinjection. A combination of statistical techniques was used to define appropriate summary statistics of piggyBac transformation efficiency by species and insect order. Publication bias was assessed by comparing the data sets. The bias was assessed using strategies co‐opted from the medical literature. The work culminated in building the Goldilocks decision tool, a Markov‐Chain Monte‐Carlo simulation operated via a graphical interface and providing guidance on best practice for those seeking to transform insects using piggyBac. PMID:27027400

  1. Insect transformation with piggyBac: getting the number of injections just right.

    PubMed

    Gregory, M; Alphey, L; Morrison, N I; Shimeld, S M

    2016-06-01

    The insertion of exogenous genetic cargo into insects using transposable elements is a powerful research tool with potential applications in meeting food security and public health challenges facing humanity. piggyBac is the transposable element most commonly utilized for insect germline transformation. The described efficiency of this process is variable in the published literature, and a comprehensive review of transformation efficiency in insects is lacking. This study compared and contrasted all available published data with a comprehensive data set provided by a biotechnology group specializing in insect transformation. Based on analysis of these data, with particular focus on the more complete observational data from the biotechnology group, we designed a decision tool to aid researchers' decision-making when using piggyBac to transform insects by microinjection. A combination of statistical techniques was used to define appropriate summary statistics of piggyBac transformation efficiency by species and insect order. Publication bias was assessed by comparing the data sets. The bias was assessed using strategies co-opted from the medical literature. The work culminated in building the Goldilocks decision tool, a Markov-Chain Monte-Carlo simulation operated via a graphical interface and providing guidance on best practice for those seeking to transform insects using piggyBac. PMID:27027400

  2. Insect transformation with piggyBac: getting the number of injections just right.

    PubMed

    Gregory, M; Alphey, L; Morrison, N I; Shimeld, S M

    2016-06-01

    The insertion of exogenous genetic cargo into insects using transposable elements is a powerful research tool with potential applications in meeting food security and public health challenges facing humanity. piggyBac is the transposable element most commonly utilized for insect germline transformation. The described efficiency of this process is variable in the published literature, and a comprehensive review of transformation efficiency in insects is lacking. This study compared and contrasted all available published data with a comprehensive data set provided by a biotechnology group specializing in insect transformation. Based on analysis of these data, with particular focus on the more complete observational data from the biotechnology group, we designed a decision tool to aid researchers' decision-making when using piggyBac to transform insects by microinjection. A combination of statistical techniques was used to define appropriate summary statistics of piggyBac transformation efficiency by species and insect order. Publication bias was assessed by comparing the data sets. The bias was assessed using strategies co-opted from the medical literature. The work culminated in building the Goldilocks decision tool, a Markov-Chain Monte-Carlo simulation operated via a graphical interface and providing guidance on best practice for those seeking to transform insects using piggyBac.

  3. Genetic Transformation of the Codling Moth, Cydia pomonella L., with piggyBac EGFP

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genetic transformation of the codling moth, Cydia pomonella, was accomplished through embryo microinjection with a plasmid-based piggyBac vector containing the enhanced green fluorescent protein (EGFP) gene. Sequencing of the flanking regions around the inserted construct results in identification o...

  4. Sperm-mediated transgenesis in chicken using a PiggyBac transposon system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sperm-mediated transgenesis in chicken using a PiggyBac transposon system Emmanuel Quansah1,2, Julie Long2, David Donovan2, Stephen Becker2, Bhanu Telugu2, Juli Frey2, Nigel Urwin1 1,Charles Sturt University, Graham Center of Agricultural Innovation, Wagga Wagga. Australia and 2Beltsville Agricultu...

  5. Avoiding cytotoxicity of transposases by dose-controlled mRNA delivery

    PubMed Central

    Galla, Melanie; Schambach, Axel; Falk, Christine S.; Maetzig, Tobias; Kuehle, Johannes; Lange, Kathrin; Zychlinski, Daniela; Heinz, Niels; Brugman, Martijn H.; Göhring, Gudrun; Izsvák, Zsuzsanna; Ivics, Zoltán; Baum, Christopher

    2011-01-01

    The Sleeping Beauty (SB) transposase and its newly developed hyperactive variant, SB100X, are of increasing interest for genome modification in experimental models and gene therapy. The potential cytotoxicity of transposases requires careful assessment, considering that residual integration events of transposase expression vectors delivered by physicochemical transfection or episomal retroviral vectors may lead to permanent transposase expression and resulting uncontrollable transposition. Comparing retrovirus-based approaches for delivery of mRNA, episomal DNA or integrating DNA, we found that conventional SB transposase, SB100X and a newly developed codon-optimized SB100Xo may trigger premitotic arrest and apoptosis. Cell stress induced by continued SB overexpression was self-limiting due to the induction of cell death, which occurred even in the absence of a co-transfected transposable element. The cytotoxic effects of SB transposase were strictly dose dependent and heralded by induction of p53 and c-Jun. Inactivating mutations in SB’s catalytic domain could not abrogate cytotoxicity, suggesting a mechanism independent of DNA cleavage activity. An improved approach of retrovirus particle-mediated mRNA transfer allowed transient and dose-controlled expression of SB100X, supported efficient transposition and prevented cytotoxicity. Transposase-mediated gene transfer can thus be tuned to maintain high efficiency in the absence of overt cell damage. PMID:21609958

  6. Genome-wide Profiling Reveals Remarkable Parallels Between Insertion Site Selection Properties of the MLV Retrovirus and the piggyBac Transposon in Primary Human CD4+ T Cells

    PubMed Central

    Gogol-Döring, Andreas; Ammar, Ismahen; Gupta, Saumyashree; Bunse, Mario; Miskey, Csaba; Chen, Wei; Uckert, Wolfgang; Schulz, Thomas F; Izsvák, Zsuzsanna; Ivics, Zoltán

    2016-01-01

    The inherent risks associated with vector insertion in gene therapy need to be carefully assessed. We analyzed the genome-wide distributions of Sleeping Beauty (SB) and piggyBac (PB) transposon insertions as well as MLV retrovirus and HIV lentivirus insertions in human CD4+ T cells with respect to a panel of 40 chromatin states. The distribution of SB transposon insertions displayed the least deviation from random, while the PB transposon and the MLV retrovirus showed unexpected parallels across all chromatin states. Both MLV and PB insertions are enriched at transcriptional start sites (TSSs) and co-localize with BRD4-associated sites. We demonstrate physical interaction between the PB transposase and bromodomain and extraterminal domain proteins (including BRD4), suggesting convergent evolution of a tethering mechanism that directs integrating genetic elements into TSSs. We detect unequal biases across the four systems with respect to targeting genes whose deregulation has been previously linked to serious adverse events in gene therapy clinical trials. The SB transposon has the highest theoretical chance of targeting a safe harbor locus in the human genome. The data underscore the significance of vector choice to reduce the mutagenic load on cells in clinical applications. PMID:26755332

  7. Transposase interaction with the β sliding clamp: effects on insertion sequence proliferation and transposition rate

    PubMed Central

    Díaz-Maldonado, Héctor; Gómez, Manuel J.; Moreno-Paz, Mercedes; San Martín-Úriz, Patxi; Amils, Ricardo; Parro, Víctor; López de Saro, Francisco J.

    2015-01-01

    Insertion sequences (ISs) are ubiquitous and abundant mobile genetic elements in prokaryotic genomes. ISs often encode only one protein, the transposase, which catalyzes their transposition. Recent studies have shown that transposases of many different IS families interact with the β sliding clamp, a DNA replication factor of the host. However, it was unclear to what extent this interaction limits or favors the ability of ISs to colonize a chromosome from a phylogenetically-distant organism, or if the strength of this interaction affects the transposition rate. Here we describe the proliferation of a member of the IS1634 family in Acidiphilium over ~600 generations of cultured growth. We demonstrate that the purified transposase binds to the β sliding clamp of Acidiphilium, Leptospirillum and E. coli. Further, we also demonstrate that the Acidiphilium IS1634 transposase binds to the archaeal sliding clamp (PCNA) from Methanosarcina, and that the transposase encoded by Methanosarcina IS1634 binds to Acidiphilium β. Finally, we demonstrate that increasing the strength of the interaction between β and transposase results in a higher transposition rate in vivo. Our results suggest that the interaction could determine the potential of ISs to be mobilized in bacterial populations and also their ability to proliferate within chromosomes. PMID:26306550

  8. Crystal Structure of the Human Hsmar1-Derived Transposase Domain in the DNA Repair Enzyme Metnase

    SciTech Connect

    Goodwin, Kristie D.; He, Hongzhen; Imasaki, Tsuyoshi; Lee, Suk-Hee; Georgiadis, Millie M.

    2010-08-12

    Although the human genome is littered with sequences derived from the Hsmar1 transposon, the only intact Hsmar1 transposase gene exists within a chimeric SET-transposase fusion protein referred to as Metnase or SETMAR. Metnase retains many of the transposase activities including terminal inverted repeat (TIR) specific DNA-binding activity, DNA cleavage activity, albeit uncoupled from TIR-specific binding, and the ability to form a synaptic complex. However, Metnase has evolved as a DNA repair protein that is specifically involved in nonhomologous end joining. Here, we present two crystal structures of the transposase catalytic domain of Metnase revealing a dimeric enzyme with unusual active site plasticity that may be involved in modulating metal binding. We show through characterization of a dimerization mutant, F460K, that the dimeric form of the enzyme is required for its DNA cleavage, DNA-binding, and nonhomologous end joining activities. Of significance is the conservation of F460 along with residues that we propose may be involved in the modulation of metal binding in both the predicted ancestral Hsmar1 transposase sequence as well as in the modern enzyme. The Metnase transposase has been remarkably conserved through evolution; however, there is a clustering of substitutions located in alpha helices 4 and 5 within the putative DNA-binding site, consistent with loss of transposition specific DNA cleavage activity and acquisition of DNA repair specific cleavage activity.

  9. A Transposon and Transposase System for Human Application

    PubMed Central

    Hackett, Perry B; Largaespada, David A; Cooper, Laurence JN

    2010-01-01

    The stable introduction of therapeutic transgenes into human cells can be accomplished using viral and nonviral approaches. Transduction with clinical-grade recombinant viruses offers the potential of efficient gene transfer into primary cells and has a record of therapeutic successes. However, widespread application for gene therapy using viruses can be limited by their initially high cost of manufacture at a limited number of production facilities as well as a propensity for nonrandom patterns of integration. The ex vivo application of transposon-mediated gene transfer now offers an alternative to the use of viral vectors. Clinical-grade DNA plasmids can be prepared at much reduced cost and with lower immunogenicity, and the integration efficiency can be improved by the transient coexpression of a hyperactive transposase. This has facilitated the design of human trials using the Sleeping Beauty (SB) transposon system to introduce a chimeric antigen receptor (CAR) to redirect the specificity of human T cells. This review examines the rationale and safety implications of application of the SB system to genetically modify T cells to be manufactured in compliance with current good manufacturing practice (cGMP) for phase I/II trials. PMID:20104209

  10. Mobilization of giant piggyBac transposons in the mouse genome

    PubMed Central

    Li, Meng Amy; Turner, Daniel J.; Ning, Zemin; Yusa, Kosuke; Liang, Qi; Eckert, Sabine; Rad, Lena; Fitzgerald, Tomas W.; Craig, Nancy L.; Bradley, Allan

    2011-01-01

    The development of technologies that allow the stable delivery of large genomic DNA fragments in mammalian systems is important for genetic studies as well as for applications in gene therapy. DNA transposons have emerged as flexible and efficient molecular vehicles to mediate stable cargo transfer. However, the ability to carry DNA fragments >10 kb is limited in most DNA transposons. Here, we show that the DNA transposon piggyBac can mobilize 100-kb DNA fragments in mouse embryonic stem (ES) cells, making it the only known transposon with such a large cargo capacity. The integrity of the cargo is maintained during transposition, the copy number can be controlled and the inserted giant transposons express the genomic cargo. Furthermore, these 100-kb transposons can also be excised from the genome without leaving a footprint. The development of piggyBac as a large cargo vector will facilitate a wider range of genetic and genomic applications. PMID:21948799

  11. Transcriptionally regulated and nontoxic delivery of the hyperactive Sleeping Beauty Transposase.

    PubMed

    Cocchiarella, Fabienne; Latella, Maria Carmela; Basile, Valentina; Miselli, Francesca; Galla, Melanie; Imbriano, Carol; Recchia, Alessandra

    2016-01-01

    The Sleeping Beauty (SB) transposase and, in particular, its hyperactive variant SB100X raises increasing interest for gene therapy application, including genome modification and, more recently, induced pluripotent stem cells (iPS) reprogramming. The documented cytotoxicity of the transposase, when constitutively expressed by an integrating retroviral vector (iRV), has been circumvented by the transient delivery of SB100X using retroviral mRNA transfer. In this study, we developed an alternative, safe, and efficient transposase delivery system based on a tetracycline-ON regulated expression cassette and the rtTA2(S)-M2 transactivator gene transiently delivered by integration-defective lentiviral vectors (IDLVs). Compared with iRV-mediated delivery, expression of tetracycline-induced SB100X delivered by an IDLV results in more efficient integration of a GFP transposon and reduced toxicity. Tightly regulated expression and reactivation of the transposase was achieved in HeLa cells as wells as in human primary keratinocytes. Based on these properties, the regulated transposase-IDLV vectors may represent a valuable tool for genetic engineering and therapeutic gene transfer.

  12. Transcriptionally regulated and nontoxic delivery of the hyperactive Sleeping Beauty Transposase.

    PubMed

    Cocchiarella, Fabienne; Latella, Maria Carmela; Basile, Valentina; Miselli, Francesca; Galla, Melanie; Imbriano, Carol; Recchia, Alessandra

    2016-01-01

    The Sleeping Beauty (SB) transposase and, in particular, its hyperactive variant SB100X raises increasing interest for gene therapy application, including genome modification and, more recently, induced pluripotent stem cells (iPS) reprogramming. The documented cytotoxicity of the transposase, when constitutively expressed by an integrating retroviral vector (iRV), has been circumvented by the transient delivery of SB100X using retroviral mRNA transfer. In this study, we developed an alternative, safe, and efficient transposase delivery system based on a tetracycline-ON regulated expression cassette and the rtTA2(S)-M2 transactivator gene transiently delivered by integration-defective lentiviral vectors (IDLVs). Compared with iRV-mediated delivery, expression of tetracycline-induced SB100X delivered by an IDLV results in more efficient integration of a GFP transposon and reduced toxicity. Tightly regulated expression and reactivation of the transposase was achieved in HeLa cells as wells as in human primary keratinocytes. Based on these properties, the regulated transposase-IDLV vectors may represent a valuable tool for genetic engineering and therapeutic gene transfer. PMID:27574698

  13. Regulation of the transposase of Tn4652 by the transposon-encoded protein TnpC.

    PubMed

    Hõrak, R; Kivisaar, M

    1999-10-01

    Transposition is a DNA reorganization reaction potentially deleterious for the host. The frequency of transposition is limited by the amount of transposase. Therefore, strict regulation of a transposase is required to keep control over the destructive multiplication of the mobile element. We have shown previously that the expression of the transposase (tnpA) of the Pseudomonas putida PaW85 transposon Tn4652 is positively affected by integration host factor. Here, we present evidence that the amount of the transposase of Tn4652 in P. putida cells is controlled by the transposon-encoded protein (TnpC). Sequence analysis of the 120-amino-acid-long TnpC, coded just downstream of the tnpA gene, showed that it has remarkable similarity to the putative polypeptide encoded by the mercury resistance transposon Tn5041. As determined by quantitative Western blot analysis, the abundance of TnpA was reduced up to 10-fold in the intact tnpC background. In vivo experiments using transcriptional and translational fusions of the tnpA gene and the reporter gene gusA indicated that TnpC operates in the regulation of the transposase of Tn4652 at the post-transcriptional level. PMID:10515920

  14. Transcriptionally regulated and nontoxic delivery of the hyperactive Sleeping Beauty Transposase

    PubMed Central

    Cocchiarella, Fabienne; Latella, Maria Carmela; Basile, Valentina; Miselli, Francesca; Galla, Melanie; Imbriano, Carol; Recchia, Alessandra

    2016-01-01

    The Sleeping Beauty (SB) transposase and, in particular, its hyperactive variant SB100X raises increasing interest for gene therapy application, including genome modification and, more recently, induced pluripotent stem cells (iPS) reprogramming. The documented cytotoxicity of the transposase, when constitutively expressed by an integrating retroviral vector (iRV), has been circumvented by the transient delivery of SB100X using retroviral mRNA transfer. In this study, we developed an alternative, safe, and efficient transposase delivery system based on a tetracycline-ON regulated expression cassette and the rtTA2S-M2 transactivator gene transiently delivered by integration-defective lentiviral vectors (IDLVs). Compared with iRV-mediated delivery, expression of tetracycline-induced SB100X delivered by an IDLV results in more efficient integration of a GFP transposon and reduced toxicity. Tightly regulated expression and reactivation of the transposase was achieved in HeLa cells as wells as in human primary keratinocytes. Based on these properties, the regulated transposase-IDLV vectors may represent a valuable tool for genetic engineering and therapeutic gene transfer. PMID:27574698

  15. ATP-dependent specific binding of Tn3 transposase to Tn3 inverted repeats

    NASA Astrophysics Data System (ADS)

    Wishart, W. L.; Broach, J. R.; Ohtsubo, E.

    1985-04-01

    Transposons are discrete segments of DNA which are capable of moving from one site in a genome to many different sites1,2. Tn3 is a prokaryotic transposon which is 4,957 base pairs (bp) long and encodes a transposase protein which is essential for transposition3-7. We report here a simple method for purifying Tn3 transposase and demonstrate that the transposase protein binds specifically to the ends of the Tn3 transposon in an ATP-dependent manner. The transposase protein binds to linear double-stranded DNA both nonspecifically and specifically; the nonspecific DNA binding activity is sensitive to challenge with heparin. Site-specific DNA binding to the ends (inverted repeats) of Tn3 is observed only when binding is performed in the presence of ATP; this ATP-dependent site-specific DNA binding activity is resistant to heparin challenge. Our results indicate that ATP qualitatively alters the DNA binding activity of the transposase protein so that the protein is able to bind specifically to the ends of the Tn3 transposon.

  16. Active Site Sharing and Subterminal Hairpin Recognition in a New Class of DNA Transposases

    SciTech Connect

    Ronning, Donald R.; Guynet, Catherine; Ton-Hoang, Bao; Perez, Zhanita N.; Ghirlando, Rodolfo; Chandler, Michael; Dyda, Fred

    2010-07-20

    Many bacteria harbor simple transposable elements termed insertion sequences (IS). In Helicobacter pylori, the chimeric IS605 family elements are particularly interesting due to their proximity to genes encoding gastric epithelial invasion factors. Protein sequences of IS605 transposases do not bear the hallmarks of other well-characterized transposases. We have solved the crystal structure of full-length transposase (TnpA) of a representative member, ISHp608. Structurally, TnpA does not resemble any characterized transposase; rather, it is related to rolling circle replication (RCR) proteins. Consistent with RCR, Mg{sup 2+} and a conserved tyrosine, Tyr127, are essential for DNA nicking and the formation of a covalent intermediate between TnpA and DNA. TnpA is dimeric, contains two shared active sites, and binds two DNA stem loops representing the conserved inverted repeats near each end of ISHp608. The cocrystal structure with stem-loop DNA illustrates how this family of transposases specifically recognizes and pairs ends, necessary steps during transposition.

  17. Germline transformation of Aedes fluviatilis (Diptera:Culicidae) with the piggyBac transposable element.

    PubMed

    Rodrigues, Flávia Guimarães; Oliveira, Sabrina Barbosa; Rocha, Bruno Coelho; Moreira, Luciano Andrade

    2006-11-01

    The technique to generate transgenic mosquitoes requires adaptation for each target species because of aspects related to species biology, sensitivity to manipulation and rearing conditions. Here we tested different parameters on the microinjection procedure in order to obtain a transgenic Neotropical mosquito species. By using a transposon-based strategy we were able to successfully transform Aedes fluviatilis (Lutz), which can be used as an avian malaria model. These results demonstrate the usefulness of the piggyBac transposable element as a transformation vector for Neotropical mosquito species and opens up new research frontiers for South American mosquito vectors. PMID:17160283

  18. Goldfish transposase Tgf2 presumably from recent horizontal transfer is active.

    PubMed

    Jiang, Xia-Yun; Du, Xue-Di; Tian, Yu-Mei; Shen, Rui-Jie; Sun, Cheng-Fei; Zou, Shu-Ming

    2012-07-01

    Hobo/Activator/Tam3 (hAT) superfamily transposons occur in plants and animals and play a role in genomic evolution. Certain hAT transposons are active and have been developed as incisive genetic tools. Active vertebrate elements are rarely discovered; however, Tgf2 transposon was recently discovered in goldfish (Carassius auratus). Here, we found that the endogenous Tgf2 element can transpose in goldfish genome. Seven different goldfish mRNA transcripts, encoding three lengths of Tgf2 transposase, were identified. Tgf2 transposase mRNA was detected in goldfish embryos, mainly in epithelial cells; levels were high in ovaries and mature eggs and in all adult tissues tested. Endogenous Tgf2 transposase mRNA is active in mature eggs and can mediate high rates of transposition (>30%) when injected with donor plasmids harboring a Tgf2 cis-element. When donor plasmid was coinjected with capped Tgf2 transposase mRNA, the insertion rate reached >90% at 1 yr. Nonautonomous copies of the Tgf2 transposon with large-fragment deletions and low levels of point mutations were also detected in common goldfish. Phylogenetic analysis indicates the taxonomic distribution of Tgf2 in goldfish is not due to vertical inheritance. We propose that the goldfish Tgf2 transposon originated by recent horizontal transfer and maintains a highly native activity.

  19. piggyBac transposon-mediated cellular transgenesis in mammalian forebrain by in utero electroporation.

    PubMed

    Chen, Fuyi; Maher, Brady J; LoTurco, Joseph J

    2014-07-01

    In utero electroporation (IUE) is an effective transfection method for delivering plasmid DNA into neural progenitor cells and neurons of mammalian neocortex in vivo. Although IUE is effective at delivering multiple DNA plasmids into populations of cells, unfortunately plasmids delivered into neural progenitor cells remain largely episomal and often get inactivated or lost after cell division. This results in a form of "birthdate" labeling in which only the cell types that do not undergo a second cell division continue to express the transfected plasmids. This limits the application of IUE with standard plasmids and precludes its use in experiments where manipulating or labeling the complete cell lineage of a progenitor is desired. To circumvent this episomal loss of plasmid in IUE, we have used a binary piggyBac transposon system to induce nonviral genomic integration of transgenes. These transgenes do not appear to inactivate after cell division, and this results in stable somatic cellular transgenesis of neurons and glia. Like standard IUE, the system can be used with multiple combinations of plasmids to achieve multicolor labeling and both loss-of-function and gain-of-function manipulations. In this protocol, we describe the method for delivering a binary piggyBac transposon plasmid system by IUE.

  20. piggyBac transposon-mediated cellular transgenesis in mammalian forebrain by in utero electroporation.

    PubMed

    Chen, Fuyi; Maher, Brady J; LoTurco, Joseph J

    2014-07-01

    In utero electroporation (IUE) is an effective transfection method for delivering plasmid DNA into neural progenitor cells and neurons of mammalian neocortex in vivo. Although IUE is effective at delivering multiple DNA plasmids into populations of cells, unfortunately plasmids delivered into neural progenitor cells remain largely episomal and often get inactivated or lost after cell division. This results in a form of "birthdate" labeling in which only the cell types that do not undergo a second cell division continue to express the transfected plasmids. This limits the application of IUE with standard plasmids and precludes its use in experiments where manipulating or labeling the complete cell lineage of a progenitor is desired. To circumvent this episomal loss of plasmid in IUE, we have used a binary piggyBac transposon system to induce nonviral genomic integration of transgenes. These transgenes do not appear to inactivate after cell division, and this results in stable somatic cellular transgenesis of neurons and glia. Like standard IUE, the system can be used with multiple combinations of plasmids to achieve multicolor labeling and both loss-of-function and gain-of-function manipulations. In this protocol, we describe the method for delivering a binary piggyBac transposon plasmid system by IUE. PMID:24987137

  1. The diversity of prokaryotic DDE transposases of the mutator superfamily, insertion specificity, and association with conjugation machineries.

    PubMed

    Guérillot, Romain; Siguier, Patricia; Gourbeyre, Edith; Chandler, Michael; Glaser, Philippe

    2014-02-01

    Transposable elements (TEs) are major components of both prokaryotic and eukaryotic genomes and play a significant role in their evolution. In this study, we have identified new prokaryotic DDE transposase families related to the eukaryotic Mutator-like transposases. These genes were retrieved by cascade PSI-Blast using as initial query the transposase of the streptococcal integrative and conjugative element (ICE) TnGBS2. By combining secondary structure predictions and protein sequence alignments, we predicted the DDE catalytic triad and the DNA-binding domain recognizing the terminal inverted repeats. Furthermore, we systematically characterized the organization and the insertion specificity of the TEs relying on these prokaryotic Mutator-like transposases (p-MULT) for their mobility. Strikingly, two distant TE families target their integration upstream σA dependent promoters. This allowed us to identify a transposase sequence signature associated with this unique insertion specificity and to show that the dissymmetry between the two inverted repeats is responsible for the orientation of the insertion. Surprisingly, while DDE transposases are generally associated with small and simple transposons such as insertion sequences (ISs), p-MULT encoding TEs show an unprecedented diversity with several families of IS, transposons, and ICEs ranging in size from 1.1 to 52 kb.

  2. Messenger RNA as a source of transposase for sleeping beauty transposon-mediated correction of hereditary tyrosinemia type I.

    PubMed

    Wilber, Andrew; Wangensteen, Kirk J; Chen, Yixin; Zhuo, Lijuan; Frandsen, Joel L; Bell, Jason B; Chen, Zongyu J; Ekker, Stephen C; McIvor, R Scott; Wang, Xin

    2007-07-01

    The Sleeping Beauty (SB) transposon system mediates chromosomal integration and stable gene expression when an engineered SB transposon is delivered along with transposase. One concern in the therapeutic application of the SB system is that persistent expression of transposase could result in transposon instability and genotoxicity. Here, we tested the use of transposase-encoding RNA plus transposon DNA for correction of murine fumarylacetoacetate hydrolase (FAH) deficiency. A bi-functional transposon containing both mouse FAH and firefly luciferase sequences was used to track the growth of genetically corrected liver tissue by in vivo bioluminescence imaging after delivery of DNA or RNA as a source of transposase. Supplying SB transposase in the form of RNA resulted in selective repopulation of corrected hepatocytes with stable expression of FAH and luciferase. Plasma succinylacetone and amino acid levels were normalized, suggesting normal liver metabolism of catabolized protein products. Secondary FAH-deficient animals transplanted with hepatocytes (250,000) isolated from primary treated animals survived 2-(2-nitro-4-trifluoro-methylbenzoyl)-1,3-cyclohexanedione (NTBC) withdrawal, gained weight consistently, and demonstrated stable expression of luciferase. We conclude that transposase-encoding messenger RNA (mRNA) can be used to mediate stable non-viral gene therapy, resulting in complete phenotypic correction, and is thus an effective source of recombinase activity for use in human gene therapy.

  3. Messenger RNA as a source of transposase for sleeping beauty transposon-mediated correction of hereditary tyrosinemia type I.

    PubMed

    Wilber, Andrew; Wangensteen, Kirk J; Chen, Yixin; Zhuo, Lijuan; Frandsen, Joel L; Bell, Jason B; Chen, Zongyu J; Ekker, Stephen C; McIvor, R Scott; Wang, Xin

    2007-07-01

    The Sleeping Beauty (SB) transposon system mediates chromosomal integration and stable gene expression when an engineered SB transposon is delivered along with transposase. One concern in the therapeutic application of the SB system is that persistent expression of transposase could result in transposon instability and genotoxicity. Here, we tested the use of transposase-encoding RNA plus transposon DNA for correction of murine fumarylacetoacetate hydrolase (FAH) deficiency. A bi-functional transposon containing both mouse FAH and firefly luciferase sequences was used to track the growth of genetically corrected liver tissue by in vivo bioluminescence imaging after delivery of DNA or RNA as a source of transposase. Supplying SB transposase in the form of RNA resulted in selective repopulation of corrected hepatocytes with stable expression of FAH and luciferase. Plasma succinylacetone and amino acid levels were normalized, suggesting normal liver metabolism of catabolized protein products. Secondary FAH-deficient animals transplanted with hepatocytes (250,000) isolated from primary treated animals survived 2-(2-nitro-4-trifluoro-methylbenzoyl)-1,3-cyclohexanedione (NTBC) withdrawal, gained weight consistently, and demonstrated stable expression of luciferase. We conclude that transposase-encoding messenger RNA (mRNA) can be used to mediate stable non-viral gene therapy, resulting in complete phenotypic correction, and is thus an effective source of recombinase activity for use in human gene therapy. PMID:17440442

  4. Harbinger transposons and an ancient HARBI1 gene derived from a transposase.

    PubMed

    Kapitonov, Vladimir V; Jurka, Jerzy

    2004-05-01

    In this study we report main properties of Harbinger DNA transposons identified in protists, plants, insects, worms, and vertebrates. This is the first superfamily of eukaryotic DNA transposons where all autonomous transposons, even those that are hosted by species from different kingdoms, encode two proteins: a superfamily-specific transposase and a DNA-binding protein characterized by the presence of the conserved SANT/myb/trihelix motif. The last motif is known to be important for the DNA binding by different transcription regulators. Therefore, we suggest that this protein is necessary for coordinated expression of the Harbinger transposase. Although mammalian genomes are free of recognizable remnants of Harbingers, we identified a widely expressed HARBI1 gene encoding a 350-aa protein entirely derived from a Harbinger transposase some 450-500 million years ago. The HARBI1 proteins are conserved in humans, rats, mice, cows, pigs, chickens, frogs, and various bony fish, as well as other extremely important proteins, including RAG1 and RAG2. Conserved motifs detected in the Harbinger transposases are also well preserved in the HARBI1 proteins. Therefore, the HARBI1 proteins are expected to be nucleases important for functioning of bony vertebrates. We also found that the protein most similar to HARBI1 is encoded by an autonomous Harbinger 3_DR transposon that was transpositionally active in the zebrafish genome a few million years ago. Nonautonomous transposons derived from Harbinger3_DR are characterized by a striking preference for a 17-bp target site never seen previously in any other DNA transposon. Based on this observation, we suggest that the hypothetical HARBI1 nucleases are also characterized by a strong DNA-target specificity.

  5. Alternative splicing of the maize Ac transposase transcript in transgenic sugar beet (Beta vulgaris L.).

    PubMed

    Lisson, Ralph; Hellert, Jan; Ringleb, Malte; Machens, Fabian; Kraus, Josef; Hehl, Reinhard

    2010-09-01

    The maize Activator/Dissociation (Ac/Ds) transposable element system was introduced into sugar beet. The autonomous Ac and non-autonomous Ds element excise from the T-DNA vector and integrate at novel positions in the sugar beet genome. Ac and Ds excisions generate footprints in the donor T-DNA that support the hairpin model for transposon excision. Two complete integration events into genomic sugar beet DNA were obtained by IPCR. Integration of Ac leads to an eight bp duplication, while integration of Ds in a homologue of a sugar beet flowering locus gene did not induce a duplication. The molecular structure of the target site indicates Ds integration into a double strand break. Analyses of transposase transcription using RT-PCR revealed low amounts of alternatively spliced mRNAs. The fourth intron of the transposase was found to be partially misspliced. Four different splice products were identified. In addition, the second and third exon were found to harbour two and three novel introns, respectively. These utilize each the same splice donor but several alternative splice acceptor sites. Using the SplicePredictor online tool, one of the two introns within exon two is predicted to be efficiently spliced in maize. Most interestingly, splicing of this intron together with the four major introns of Ac would generate a transposase that lacks the DNA binding domain and two of its three nuclear localization signals, but still harbours the dimerization domain.

  6. Microbial co-habitation and lateral gene transfer: what transposases can tell us

    SciTech Connect

    Hooper, Sean D.; Mavromatis, Konstantinos; Kyrpides, Nikos C.

    2009-03-01

    Determining the habitat range for various microbes is not a simple, straightforward matter, as habitats interlace, microbes move between habitats, and microbial communities change over time. In this study, we explore an approach using the history of lateral gene transfer recorded in microbial genomes to begin to answer two key questions: where have you been and who have you been with? All currently sequenced microbial genomes were surveyed to identify pairs of taxa that share a transposase that is likely to have been acquired through lateral gene transfer. A microbial interaction network including almost 800 organisms was then derived from these connections. Although the majority of the connections are between closely related organisms with the same or overlapping habitat assignments, numerous examples were found of cross-habitat and cross-phylum connections. We present a large-scale study of the distributions of transposases across phylogeny and habitat, and find a significant correlation between habitat and transposase connections. We observed cases where phylogenetic boundaries are traversed, especially when organisms share habitats; this suggests that the potential exists for genetic material to move laterally between diverse groups via bridging connections. The results presented here also suggest that the complex dynamics of microbial ecology may be traceable in the microbial genomes.

  7. Alternative splicing of the maize Ac transposase transcript in transgenic sugar beet (Beta vulgaris L.)

    PubMed Central

    Lisson, Ralph; Hellert, Jan; Ringleb, Malte; Machens, Fabian; Kraus, Josef

    2010-01-01

    The maize Activator/Dissociation (Ac/Ds) transposable element system was introduced into sugar beet. The autonomous Ac and non-autonomous Ds element excise from the T-DNA vector and integrate at novel positions in the sugar beet genome. Ac and Ds excisions generate footprints in the donor T-DNA that support the hairpin model for transposon excision. Two complete integration events into genomic sugar beet DNA were obtained by IPCR. Integration of Ac leads to an eight bp duplication, while integration of Ds in a homologue of a sugar beet flowering locus gene did not induce a duplication. The molecular structure of the target site indicates Ds integration into a double strand break. Analyses of transposase transcription using RT–PCR revealed low amounts of alternatively spliced mRNAs. The fourth intron of the transposase was found to be partially misspliced. Four different splice products were identified. In addition, the second and third exon were found to harbour two and three novel introns, respectively. These utilize each the same splice donor but several alternative splice acceptor sites. Using the SplicePredictor online tool, one of the two introns within exon two is predicted to be efficiently spliced in maize. Most interestingly, splicing of this intron together with the four major introns of Ac would generate a transposase that lacks the DNA binding domain and two of its three nuclear localization signals, but still harbours the dimerization domain. PMID:20512402

  8. Alternative splicing of the maize Ac transposase transcript in transgenic sugar beet (Beta vulgaris L.).

    PubMed

    Lisson, Ralph; Hellert, Jan; Ringleb, Malte; Machens, Fabian; Kraus, Josef; Hehl, Reinhard

    2010-09-01

    The maize Activator/Dissociation (Ac/Ds) transposable element system was introduced into sugar beet. The autonomous Ac and non-autonomous Ds element excise from the T-DNA vector and integrate at novel positions in the sugar beet genome. Ac and Ds excisions generate footprints in the donor T-DNA that support the hairpin model for transposon excision. Two complete integration events into genomic sugar beet DNA were obtained by IPCR. Integration of Ac leads to an eight bp duplication, while integration of Ds in a homologue of a sugar beet flowering locus gene did not induce a duplication. The molecular structure of the target site indicates Ds integration into a double strand break. Analyses of transposase transcription using RT-PCR revealed low amounts of alternatively spliced mRNAs. The fourth intron of the transposase was found to be partially misspliced. Four different splice products were identified. In addition, the second and third exon were found to harbour two and three novel introns, respectively. These utilize each the same splice donor but several alternative splice acceptor sites. Using the SplicePredictor online tool, one of the two introns within exon two is predicted to be efficiently spliced in maize. Most interestingly, splicing of this intron together with the four major introns of Ac would generate a transposase that lacks the DNA binding domain and two of its three nuclear localization signals, but still harbours the dimerization domain. PMID:20512402

  9. Pig transgenesis by piggyBac transposition in combination with somatic cell nuclear transfer.

    PubMed

    Wu, Zhenfang; Xu, Zhiqian; Zou, Xian; Zeng, Fang; Shi, Junsong; Liu, Dewu; Urschitz, Johann; Moisyadi, Stefan; Li, Zicong

    2013-12-01

    The production of animals by somatic cell nuclear transfer (SCNT) is inefficient, with approximately 2% of micromanipulated oocytes going to term and resulting in live births. However, it is the most commonly used method for the generation of cloned transgenic livestock as it facilitates the attainment of transgenic animals once the nuclear donor cells are stably transfected and more importantly as alternatives methods of transgenesis in farm animals have proven even less efficient. Here we describe piggyBac-mediated transposition of a transgene into porcine primary cells and use of these genetically modified cells as nuclear donors for the generation of transgenic pigs by SCNT. Gene transfer by piggyBac transposition serves to provide an alternative approach for the transfection of nuclear donor cells used in SCNT.

  10. The piggyBac Transposon-Mediated Expression of SV40 T Antigen Efficiently Immortalizes Mouse Embryonic Fibroblasts (MEFs)

    PubMed Central

    Cui, Jing; Zhang, Hongmei; Chen, Xiang; Li, Ruidong; Wu, Ningning; Chen, Xian; Wen, Sheng; Zhang, Junhui; Yin, Liangjun; Deng, Fang; Liao, Zhan; Zhang, Zhonglin; Zhang, Qian; Yan, Zhengjian; Liu, Wei; Ye, Jixing; Deng, Youlin; Wang, Zhongliang; Qiao, Min; Luu, Hue H.; Haydon, Rex C.; Shi, Lewis L.; Liang, Houjie; He, Tong-Chuan

    2014-01-01

    Mouse embryonic fibroblasts (MEFs) are mesenchymal stem cell (MSC)-like multipotent progenitor cells and can undergo self-renewal and differentiate into to multiple lineages, including bone, cartilage and adipose. Primary MEFs have limited life span in culture, which thus hampers MEFs’ basic research and translational applications. To overcome this challenge, we investigate if piggyBac transposon-mediated expression of SV40 T antigen can effectively immortalize mouse MEFs and that the immortalized MEFs can maintain long-term cell proliferation without compromising their multipotency. Using the piggyBac vector MPH86 which expresses SV40 T antigen flanked with flippase (FLP) recognition target (FRT) sites, we demonstrate that mouse embryonic fibroblasts (MEFs) can be efficiently immortalized. The immortalized MEFs (piMEFs) exhibit an enhanced proliferative activity and maintain long-term cell proliferation, which can be reversed by FLP recombinase. The piMEFs express most MEF markers and retain multipotency as they can differentiate into osteogenic, chondrogenic and adipogenic lineages upon BMP9 stimulation in vitro. Stem cell implantation studies indicate that piMEFs can form bone, cartilage and adipose tissues upon BMP9 stimulation, whereas FLP-mediated removal of SV40 T antigen diminishes the ability of piMEFs to differentiate into these lineages, possibly due to the reduced expansion of progenitor populations. Our results demonstrate that piggyBac transposon-mediated expression of SV40 T can effectively immortalize MEFs and that the reversibly immortalized piMEFs not only maintain long-term cell proliferation but also retain their multipotency. Thus, the high transposition efficiency and the potential footprint-free natures may render piggyBac transposition an effective and safe strategy to immortalize progenitor cells isolated from limited tissue supplies, which is essential for basic and translational studies. PMID:24845466

  11. A conditional piggyBac transposition system for genetic screening in mice identifies oncogenic networks in pancreatic cancer.

    PubMed

    Rad, Roland; Rad, Lena; Wang, Wei; Strong, Alexander; Ponstingl, Hannes; Bronner, Iraad F; Mayho, Matthew; Steiger, Katja; Weber, Julia; Hieber, Maren; Veltkamp, Christian; Eser, Stefan; Geumann, Ulf; Öllinger, Rupert; Zukowska, Magdalena; Barenboim, Maxim; Maresch, Roman; Cadiñanos, Juan; Friedrich, Mathias; Varela, Ignacio; Constantino-Casas, Fernando; Sarver, Aaron; Ten Hoeve, Jelle; Prosser, Haydn; Seidler, Barbara; Bauer, Judith; Heikenwälder, Mathias; Metzakopian, Emmanouil; Krug, Anne; Ehmer, Ursula; Schneider, Günter; Knösel, Thomas; Rümmele, Petra; Aust, Daniela; Grützmann, Robert; Pilarsky, Christian; Ning, Zemin; Wessels, Lodewyk; Schmid, Roland M; Quail, Michael A; Vassiliou, George; Esposito, Irene; Liu, Pentao; Saur, Dieter; Bradley, Allan

    2015-01-01

    Here we describe a conditional piggyBac transposition system in mice and report the discovery of large sets of new cancer genes through a pancreatic insertional mutagenesis screen. We identify Foxp1 as an oncogenic transcription factor that drives pancreatic cancer invasion and spread in a mouse model and correlates with lymph node metastasis in human patients with pancreatic cancer. The propensity of piggyBac for open chromatin also enabled genome-wide screening for cancer-relevant noncoding DNA, which pinpointed a Cdkn2a cis-regulatory region. Histologically, we observed different tumor subentities and discovered associated genetic events, including Fign insertions in hepatoid pancreatic cancer. Our studies demonstrate the power of genetic screening to discover cancer drivers that are difficult to identify by other approaches to cancer genome analysis, such as downstream targets of commonly mutated human cancer genes. These piggyBac resources are universally applicable in any tissue context and provide unique experimental access to the genetic complexity of cancer.

  12. A High-Capacity Adenoviral Hybrid Vector System Utilizing the Hyperactive Sleeping Beauty Transposase SB100X for Enhanced Integration.

    PubMed

    Boehme, Philip; Zhang, Wenli; Solanki, Manish; Ehrke-Schulz, Eric; Ehrhardt, Anja

    2016-07-19

    For efficient delivery of required genetic elements we utilized high-capacity adenoviral vectors in the past allowing high transgene capacities of up to 36 kb. Previously we explored the hyperactive Sleeping Beauty (SB) transposase (HSB5) for somatic integration from the high-capacity adenoviral vectors genome. To further improve this hybrid vector system we hypothesized that the previously described hyperactive SB transposase SB100X will result in significantly improved efficacies after transduction of target cells. Plasmid based delivery of the SB100X system revealed significantly increased integration efficiencies compared with the previously published hyperactive SB transposase HSB5. After optimizing experimental setups for high-capacity adenoviral vectors-based delivery of the SB100X system we observed up to eightfold and 100-fold increased integration efficiencies compared with the previously published hyperactive SB transposase HSB5 and the inactive transposase mSB, respectively. Furthermore, transposon copy numbers per cell were doubled with SB100X compared with HSB5 when using the identical multiplicity of infection. We believe that this improved hybrid vector system represents a valuable tool for achieving stabilized transgene expression in cycling cells and for treatment of numerous genetic disorders. Especially for in vivo approaches this improved adenoviral hybrid vector system will be advantageous because it may potentially allow reduction of the applied viral dose.

  13. A High-Capacity Adenoviral Hybrid Vector System Utilizing the Hyperactive Sleeping Beauty Transposase SB100X for Enhanced Integration.

    PubMed

    Boehme, Philip; Zhang, Wenli; Solanki, Manish; Ehrke-Schulz, Eric; Ehrhardt, Anja

    2016-01-01

    For efficient delivery of required genetic elements we utilized high-capacity adenoviral vectors in the past allowing high transgene capacities of up to 36 kb. Previously we explored the hyperactive Sleeping Beauty (SB) transposase (HSB5) for somatic integration from the high-capacity adenoviral vectors genome. To further improve this hybrid vector system we hypothesized that the previously described hyperactive SB transposase SB100X will result in significantly improved efficacies after transduction of target cells. Plasmid based delivery of the SB100X system revealed significantly increased integration efficiencies compared with the previously published hyperactive SB transposase HSB5. After optimizing experimental setups for high-capacity adenoviral vectors-based delivery of the SB100X system we observed up to eightfold and 100-fold increased integration efficiencies compared with the previously published hyperactive SB transposase HSB5 and the inactive transposase mSB, respectively. Furthermore, transposon copy numbers per cell were doubled with SB100X compared with HSB5 when using the identical multiplicity of infection. We believe that this improved hybrid vector system represents a valuable tool for achieving stabilized transgene expression in cycling cells and for treatment of numerous genetic disorders. Especially for in vivo approaches this improved adenoviral hybrid vector system will be advantageous because it may potentially allow reduction of the applied viral dose. PMID:27434682

  14. Characterization of Growth and Reproduction Performance, Transgene Integration, Expression, and Transmission Patterns in Transgenic Pigs Produced by piggyBac Transposition-Mediated Gene Transfer.

    PubMed

    Zeng, Fang; Li, Zicong; Cai, Gengyuan; Gao, Wenchao; Jiang, Gelong; Liu, Dewu; Urschitz, Johann; Moisyadi, Stefan; Wu, Zhenfang

    2016-10-01

    Previously we successfully produced a group of EGFP-expressing founder transgenic pigs by a newly developed efficient and simple pig transgenesis method based on cytoplasmic injection of piggyBac plasmids. In this study, we investigated the growth and reproduction performance and characterized the transgene insertion, transmission, and expression patterns in transgenic pigs generated by piggyBac transposition. Results showed that transgene has no injurious effect on the growth and reproduction of transgenic pigs. Multiple copies of monogenic EGFP transgene were inserted at noncoding sequences of host genome, and passed from founder transgenic pigs to their transgenic offspring in segregation or linkage manner. The EGFP transgene was ubiquitously expressed in transgenic pigs, and its expression intensity was associated with transgene copy number but not related to its promoter DNA methylation level. To the best of our knowledge, this is first study that fully described the growth and reproduction performance, transgene insertion, expression, and transmission profiles in transgenic pigs produced by piggyBac system. It not only demonstrates that piggyBac transposition-mediated gene transfer is an effective and favorable approach for pig transgenesis, but also provides scientific information for understanding the transgene insertion, expression and transmission patterns in transgenic animals produced by piggyBac transposition. PMID:27565868

  15. Birth of a chimeric primate gene by capture of the transposase gene from a mobile element.

    PubMed

    Cordaux, Richard; Udit, Swalpa; Batzer, Mark A; Feschotte, Cédric

    2006-05-23

    The emergence of new genes and functions is of central importance to the evolution of species. The contribution of various types of duplications to genetic innovation has been extensively investigated. Less understood is the creation of new genes by recycling of coding material from selfish mobile genetic elements. To investigate this process, we reconstructed the evolutionary history of SETMAR, a new primate chimeric gene resulting from fusion of a SET histone methyltransferase gene to the transposase gene of a mobile element. We show that the transposase gene was recruited as part of SETMAR 40-58 million years ago, after the insertion of an Hsmar1 transposon downstream of a preexisting SET gene, followed by the de novo exonization of previously noncoding sequence and the creation of a new intron. The original structure of the fusion gene is conserved in all anthropoid lineages, but only the N-terminal half of the transposase is evolving under strong purifying selection. In vitro assays show that this region contains a DNA-binding domain that has preserved its ancestral binding specificity for a 19-bp motif located within the terminal-inverted repeats of Hsmar1 transposons and their derivatives. The presence of these transposons in the human genome constitutes a potential reservoir of approximately 1,500 perfect or nearly perfect SETMAR-binding sites. Our results not only provide insight into the conditions required for a successful gene fusion, but they also suggest a mechanism by which the circuitry underlying complex regulatory networks may be rapidly established. PMID:16672366

  16. Germline transformation of the diamondback moth, Plutella xylostella L., using the piggyBac transposable element.

    PubMed

    Martins, S; Naish, N; Walker, A S; Morrison, N I; Scaife, S; Fu, G; Dafa'alla, T; Alphey, L

    2012-08-01

    The diamondback moth, Plutella xylostella, is one of the most economically important agricultural pests. The larvae of this moth cause damage by feeding on the foliage of cruciferous vegetables such as cabbage, broccoli, cauliflower and rapeseed. Control generally comprises chemical treatment; however, the diamondback moth is renowned for rapid development of resistance to pesticides. Other methods, such as biological control, have not been able to provide adequate protection. Germline transformation of pest insects has become available in recent years as an enabling technology for new genetics-based control methods, such as the Release of Insects carrying a Dominant Lethal (RIDL(®) ). In the present study, we report the first transformation of the diamondback moth, using the piggyBac transposable element, by embryo microinjection. In generating transgenic strains using four different constructs, the function of three regulatory sequences in this moth was demonstrated in driving expression of fluorescent proteins. The transformation rates achieved, 0.48-0.68%, are relatively low compared with those described in other Lepidoptera, but not prohibitive, and are likely to increase with experience. We anticipate that germline transformation of the diamondback moth will permit the development of RIDL strains for use against this pest and facilitate the wider use of this species as a model organism for basic studies.

  17. Efficient Production of Fluorescent Transgenic Rats using the piggyBac Transposon

    PubMed Central

    Li, Tianda; Shuai, Ling; Mao, Junjie; Wang, Xuepeng; Wang, Mei; Zhang, Xinxin; Wang, Leyun; Li, Yanni; Li, Wei; Zhou, Qi

    2016-01-01

    Rats with fluorescent markers are of great value for studies that trace lineage-specific development, particularly those assessing the differentiation potential of embryonic stem cells (ESCs). The piggyBac (PB) transposon is widely used for the efficient introduction of genetic modifications into genomes, and has already been successfully used to produce transgenic mice and rats. Here, we generated transgenic rats carrying either the desRed fluorescent protein (RFP) gene or the enhanced green fluorescent protein (eGFP) gene by injecting pronuclei with PB plasmids. We showed that the transgenic rats expressed the RFP or eGFP gene in many organs and had the capability to transmit the marker gene to the next generation through germline integration. In addition, rat embryonic stem cells (ESCs) carrying an RFP reporter gene can be derived from the blastocysts of the transgenic rats. Moreover, the RFP gene can be detected in chimeras derived from RFP ESCs via blastocyst injection. This work suggests that PB-mediated transgenesis is a powerful tool to generate transgenic rats expressing fluorescent proteins with high efficiency, and this technique can be used to derive rat ESCs expressing a reporter protein. PMID:27624004

  18. Efficient Production of Fluorescent Transgenic Rats using the piggyBac Transposon.

    PubMed

    Li, Tianda; Shuai, Ling; Mao, Junjie; Wang, Xuepeng; Wang, Mei; Zhang, Xinxin; Wang, Leyun; Li, Yanni; Li, Wei; Zhou, Qi

    2016-01-01

    Rats with fluorescent markers are of great value for studies that trace lineage-specific development, particularly those assessing the differentiation potential of embryonic stem cells (ESCs). The piggyBac (PB) transposon is widely used for the efficient introduction of genetic modifications into genomes, and has already been successfully used to produce transgenic mice and rats. Here, we generated transgenic rats carrying either the desRed fluorescent protein (RFP) gene or the enhanced green fluorescent protein (eGFP) gene by injecting pronuclei with PB plasmids. We showed that the transgenic rats expressed the RFP or eGFP gene in many organs and had the capability to transmit the marker gene to the next generation through germline integration. In addition, rat embryonic stem cells (ESCs) carrying an RFP reporter gene can be derived from the blastocysts of the transgenic rats. Moreover, the RFP gene can be detected in chimeras derived from RFP ESCs via blastocyst injection. This work suggests that PB-mediated transgenesis is a powerful tool to generate transgenic rats expressing fluorescent proteins with high efficiency, and this technique can be used to derive rat ESCs expressing a reporter protein. PMID:27624004

  19. Simple piggyBac transposon-based mammalian cell expression system for inducible protein production

    PubMed Central

    Li, Zhijie; Michael, Iacovos P.; Zhou, Dongxia; Nagy, Andras; Rini, James M.

    2013-01-01

    Reported here is a piggyBac transposon-based expression system for the generation of doxycycline-inducible, stably transfected mammalian cell cultures for large-scale protein production. The system works with commonly used adherent and suspension-adapted mammalian cell lines and requires only a single transfection step. Moreover, the high uniform expression levels observed among clones allow for the use of stable bulk cell cultures, thereby eliminating time-consuming cloning steps. Under continuous doxycycline induction, protein expression levels have been shown to be stable for at least 2 mo in the absence of drug selection. The high efficiency of the system also allows for the generation of stable bulk cell cultures in 96-well format, a capability leading to the possibility of generating stable cell cultures for entire families of membrane or secreted proteins. Finally, we demonstrate the utility of the system through the large-scale production (140–750 mg scale) of an endoplasmic reticulum-resident fucosyltransferase and two potential anticancer protein therapeutic agents. PMID:23476064

  20. Transposon domestication versus mutualism in ciliate genome rearrangements.

    PubMed

    Vogt, Alexander; Goldman, Aaron David; Mochizuki, Kazufumi; Landweber, Laura F

    2013-01-01

    Ciliated protists rearrange their genomes dramatically during nuclear development via chromosome fragmentation and DNA deletion to produce a trimmer and highly reorganized somatic genome. The deleted portion of the genome includes potentially active transposons or transposon-like sequences that reside in the germline. Three independent studies recently showed that transposase proteins of the DDE/DDD superfamily are indispensible for DNA processing in three distantly related ciliates. In the spirotrich Oxytricha trifallax, high copy-number germline-limited transposons mediate their own excision from the somatic genome but also contribute to programmed genome rearrangement through a remarkable transposon mutualism with the host. By contrast, the genomes of two oligohymenophorean ciliates, Tetrahymena thermophila and Paramecium tetraurelia, encode homologous PiggyBac-like transposases as single-copy genes in both their germline and somatic genomes. These domesticated transposases are essential for deletion of thousands of different internal sequences in these species. This review contrasts the events underlying somatic genome reduction in three different ciliates and considers their evolutionary origins and the relationships among their distinct mechanisms for genome remodeling.

  1. Domestic Violence

    MedlinePlus

    ... AQ FREQUENTLY ASKED QUESTIONS FAQ083 WOMEN’S HEALTH Domestic Violence • What is domestic violence? • What are the types of abuse? • How can ... available to help abused women? What is domestic violence? Domestic violence is a pattern of threatening or ...

  2. piggyBac as a high-capacity transgenesis and gene-therapy vector in human cells and mice.

    PubMed

    Li, Rongbo; Zhuang, Yuan; Han, Min; Xu, Tian; Wu, Xiaohui

    2013-05-01

    The stable genomic integration and expression of a large transgene is a major hurdle in gene therapy. We show that the modified piggyBac (PB) transposon system can be used to introduce a 207 kb genomic DNA fragment containing the RORγ/γt locus into human cells and mice. PB-mediated transgenesis results in a single copy of a stably inherited and expressed transgene. These results indicate that PB could serve as an effective high-capacity vector for functional analysis of the mammalian genome and for gene therapy in human cells.

  3. Identification of multiple binding sites for the THAP domain of the Galileo transposase in the long terminal inverted-repeats.

    PubMed

    Marzo, Mar; Liu, Danxu; Ruiz, Alfredo; Chalmers, Ronald

    2013-08-01

    Galileo is a DNA transposon responsible for the generation of several chromosomal inversions in Drosophila. In contrast to other members of the P-element superfamily, it has unusually long terminal inverted-repeats (TIRs) that resemble those of Foldback elements. To investigate the function of the long TIRs we derived consensus and ancestral sequences for the Galileo transposase in three species of Drosophilids. Following gene synthesis, we expressed and purified their constituent THAP domains and tested their binding activity towards the respective Galileo TIRs. DNase I footprinting located the most proximal DNA binding site about 70 bp from the transposon end. Using this sequence we identified further binding sites in the tandem repeats that are found within the long TIRs. This suggests that the synaptic complex between Galileo ends may be a complicated structure containing higher-order multimers of the transposase. We also attempted to reconstitute Galileo transposition in Drosophila embryos but no events were detected. Thus, although the limited numbers of Galileo copies in each genome were sufficient to provide functional consensus sequences for the THAP domains, they do not specify a fully active transposase. Since the THAP recognition sequence is short, and will occur many times in a large genome, it seems likely that the multiple binding sites within the long, internally repetitive, TIRs of Galileo and other Foldback-like elements may provide the transposase with its binding specificity.

  4. Lens epithelium-derived growth factor/p75 interacts with the transposase-derived DDE domain of PogZ.

    PubMed

    Bartholomeeusen, Koen; Christ, Frauke; Hendrix, Jelle; Rain, Jean-Christophe; Emiliani, Stéphane; Benarous, Richard; Debyser, Zeger; Gijsbers, Rik; De Rijck, Jan

    2009-04-24

    Lens epithelium-derived growth factor/p75 (LEDGF/p75) is a prominent cellular interaction partner of human immunodeficiency virus-1 (HIV-1) integrase, tethering the preintegration complex to the host chromosome. In light of the development of LEDGF/p75-integrase interaction inhibitors, it is essential to understand the cell biology of LEDGF/p75. We identified pogZ as new cellular interaction partner of LEDGF/p75. Analogous to lentiviral integrase, pogZ, a domesticated transposase, carries a DDE domain, the major determinant for LEDGF/p75 interaction. Using different in vitro and in vivo approaches, we corroborated the interaction between the C terminus of LEDGF/p75 and the DDE domain of pogZ, revealing an overlap in the binding of pogZ and HIV-1 integrase. Competition experiments showed that integrase is efficient in displacing pogZ from LEDGF/p75. Moreover, pogZ does not seem to play a role as a restriction factor of HIV. The finding that LEDGF/p75 is capable of interacting with a DDE domain protein that is not a lentiviral integrase points to a profound role of LEDGF/p75 in DDE domain protein function.

  5. Counterselection and co-delivery of transposon and transposase functions for Sleeping Beauty-mediated transposition in cultured mammalian cells.

    PubMed

    Converse, Andrea D; Belur, Lalitha R; Gori, Jennifer L; Liu, Geyi; Amaya, Felipe; Aguilar-Cordova, Estuardo; Hackett, Perry B; McIvor, R Scott

    2004-12-01

    Sleeping Beauty (SB) is a gene-insertion system reconstructed from transposon sequences found in teleost fish and is capable of mediating the transposition of DNA sequences from transfected plasmids into the chromosomes of vertebrate cell populations. The SB system consists of a transposon, made up of a gene of interest flanked by transposon inverted repeats, and a source of transposase. Here we carried out a series of studies to further characterize SB-mediated transposition as a tool for gene transfer to chromosomes and ultimately for human gene therapy. Transfection of mouse 3T3 cells, HeLa cells, and human A549 lung carcinoma cells with a transposon containing the neomycin phosphotransferase (NEO) gene resulted in a several-fold increase in drug-resistant colony formation when co-transfected with a plasmid expressing the SB transposase. A transposon containing a methotrexate-resistant dihydrofolate reductase gene was also found to confer an increased frequency of methotrexate-resistant colony formation when co-transfected with SB transposase-encoding plasmid. A plasmid containing a herpes simplex virus thymidine kinase gene as well as a transposon containing a NEO gene was used for counterselection against random recombinants (NEO+TK+) in medium containing G418 plus ganciclovir. Effective counterselection required a recovery period of 5 days after transfection before shifting into medium containing ganciclovir to allow time for transiently expressed thymidine kinase activity to subside in cells not stably transfected. Southern analysis of clonal isolates indicated a shift from random recombination events toward transposition events when clones were isolated in medium containing ganciclovir as well as G418. We found that including both transposon and transposase functions on the same plasmid substantially increased the stable gene transfer frequency in Huh7 human hepatoma cells. The results from these experiments contribute technical and conceptual insight into

  6. TnpPred: A Web Service for the Robust Prediction of Prokaryotic Transposases

    PubMed Central

    Riadi, Gonzalo; Medina-Moenne, Cristobal; Holmes, David S.

    2012-01-01

    Transposases (Tnps) are enzymes that participate in the movement of insertion sequences (ISs) within and between genomes. Genes that encode Tnps are amongst the most abundant and widely distributed genes in nature. However, they are difficult to predict bioinformatically and given the increasing availability of prokaryotic genomes and metagenomes, it is incumbent to develop rapid, high quality automatic annotation of ISs. This need prompted us to develop a web service, termed TnpPred for Tnp discovery. It provides better sensitivity and specificity for Tnp predictions than given by currently available programs as determined by ROC analysis. TnpPred should be useful for improving genome annotation. The TnpPred web service is freely available for noncommercial use. PMID:23251097

  7. TnpPred: A Web Service for the Robust Prediction of Prokaryotic Transposases.

    PubMed

    Riadi, Gonzalo; Medina-Moenne, Cristobal; Holmes, David S

    2012-01-01

    Transposases (Tnps) are enzymes that participate in the movement of insertion sequences (ISs) within and between genomes. Genes that encode Tnps are amongst the most abundant and widely distributed genes in nature. However, they are difficult to predict bioinformatically and given the increasing availability of prokaryotic genomes and metagenomes, it is incumbent to develop rapid, high quality automatic annotation of ISs. This need prompted us to develop a web service, termed TnpPred for Tnp discovery. It provides better sensitivity and specificity for Tnp predictions than given by currently available programs as determined by ROC analysis. TnpPred should be useful for improving genome annotation. The TnpPred web service is freely available for noncommercial use. PMID:23251097

  8. Domestic Violence

    MedlinePlus

    Domestic violence is a type of abuse. It usually involves a spouse or partner, but it can also be ... child, elderly relative, or other family member. Domestic violence may include Physical violence that can lead to ...

  9. Domestic violence

    MedlinePlus

    Intimate partner violence; Spousal abuse; Elder abuse; Child abuse; Sexual abuse - domestic violence ... Domestic violence can include any of these behaviors: Physical abuse, including hitting, kicking, biting, slapping, choking, or attacking with ...

  10. The Folding of the Specific DNA Recognition Subdomain of the Sleeping Beauty Transposase Is Temperature-Dependent and Is Required for Its Binding to the Transposon DNA

    PubMed Central

    Leighton, Gage O.; Konnova, Tatiana A.; Idiyatullin, Bulat; Hurr, Sophia H.; Zuev, Yuriy F.; Nesmelova, Irina V.

    2014-01-01

    The reaction of DNA transposition begins when the transposase enzyme binds to the transposon DNA. Sleeping Beauty is a member of the mariner family of DNA transposons. Although it is an important tool in genetic applications and has been adapted for human gene therapy, its molecular mechanism remains obscure. Here, we show that only the folded conformation of the specific DNA recognition subdomain of the Sleeping Beauty transposase, the PAI subdomain, binds to the transposon DNA. Furthermore, we show that the PAI subdomain is well folded at low temperatures, but the presence of unfolded conformation gradually increases at temperatures above 15°C, suggesting that the choice of temperature may be important for the optimal transposase activity. Overall, the results provide a molecular-level insight into the DNA recognition by the Sleeping Beauty transposase. PMID:25375127

  11. Generation of Naïve Bovine Induced Pluripotent Stem Cells Using PiggyBac Transposition of Doxycycline-Inducible Transcription Factors.

    PubMed

    Kawaguchi, Takamasa; Tsukiyama, Tomoyuki; Kimura, Koji; Matsuyama, Shuichi; Minami, Naojiro; Yamada, Masayasu; Imai, Hiroshi

    2015-01-01

    Generation of pluripotent stem cells (PSCs) in large domestic animals has achieved only limited success; most of the PSCs obtained to date have been classified as primed PSCs, which possess very little capacity to produce chimeric offspring. By contrast, mouse PSCs have been classified as naïve PSCs that can contribute to most of the tissues of chimeras, including germ cells. Here, we describe the generation of two different types of bovine induced pluripotent stem cells (biPSCs) from amnion cells, achieved through introduction of piggyBac vectors containing doxycycline-inducible transcription factors (Oct3/4, Sox2, Klf4, and c-Myc). One type of biPSCs, cultured in medium supplemented with knockout serum replacement (KSR), FGF2, and bovine leukemia inhibitory factor (bLIF), had a flattened morphology like human PSCs; these were classified as primed-type. The other type biPSCs, cultured in KSR, bLIF, Mek/Erk inhibitor, GSK3 inhibitor and forskolin, had a compact morphology like mouse PSCs; these were classified as naïve-type. Cells could easily be switched between these two types of biPSCs by changing the culture conditions. Both types of biPSCs had strong alkaline phosphatase activity, expressed pluripotent markers (OCT3/4, NANOG, REX1, ESRRβ, STELLA, and SOCS3), and formed embryoid bodies that gave rise to differentiated cells from all three embryonic germ layers. However, only naïve-type biPSCs showed the hallmarks of naïve mouse PSCs, such as LIF-dependent proliferation, lack of FGF5 expression, and active XIST expression with two active X chromosomes. Furthermore, naïve-type biPSCs could contribute to the inner cell mass (ICM) of host blastocysts and most tissues within chimeric embryos. This is the first report of generation of biPSCs with several characteristics similar to those of naïve mouse PSCs and a demonstrated potential to contribute to chimeras. PMID:26287611

  12. A green fluorescent protein solubility screen in E. coli reveals domain boundaries of the GTP-binding domain in the P element transposase

    PubMed Central

    Sabogal, Alex; Rio, Donald C

    2010-01-01

    Guanosine triphosphate (GTP) binding and hydrolysis events often act as molecular switches in proteins, modulating conformational changes between active and inactive states in many signaling molecules and transport systems. The P element transposase of Drosophila melanogaster requires GTP binding to proceed along its reaction pathway, following initial site-specific DNA binding. GTP binding is unique to P elements and may represent a novel form of transpositional regulation, allowing the bound transposase to find a second site, looping the transposon DNA for strand cleavage and excision. The GTP-binding activity has been previously mapped to the central portion of the transposase protein; however, the P element transposase contains little sequence identity with known GTP-binding folds. To identify soluble, active transposase domains, a GFP solubility screen was used testing the solubility of random P element gene fragments in E. coli. The screen produced a single clone spanning known GTP-binding residues in the central portion of the transposase coding region. This clone, amino acids 275–409 in the P element transposase, was soluble, highly expressed in E.coli and active for GTP-binding activity, therefore is a candidate for future biochemical and structural studies. In addition, the chimeric screen revealed a minimal N-terminal THAP DNA-binding domain attached to an extended leucine zipper coiled-coil dimerization domain in the P element transposase, precisely delineating the DNA-binding and dimerization activities on the primary sequence. This study highlights the use of a GFP-based solubility screen on a large multidomain protein to identify highly expressed, soluble truncated domain subregions. PMID:20842711

  13. Structural basis of hAT transposon end recognition by Hermes, an octameric DNA transposase from Musca domestica.

    PubMed

    Hickman, Alison B; Ewis, Hosam E; Li, Xianghong; Knapp, Joshua A; Laver, Thomas; Doss, Anna-Louise; Tolun, Gökhan; Steven, Alasdair C; Grishaev, Alexander; Bax, Ad; Atkinson, Peter W; Craig, Nancy L; Dyda, Fred

    2014-07-17

    Hermes is a member of the hAT transposon superfamily that has active representatives, including McClintock's archetypal Ac mobile genetic element, in many eukaryotic species. The crystal structure of the Hermes transposase-DNA complex reveals that Hermes forms an octameric ring organized as a tetramer of dimers. Although isolated dimers are active in vitro for all the chemical steps of transposition, only octamers are active in vivo. The octamer can provide not only multiple specific DNA-binding domains to recognize repeated subterminal sequences within the transposon ends, which are important for activity, but also multiple nonspecific DNA binding surfaces for target capture. The unusual assembly explains the basis of bipartite DNA recognition at hAT transposon ends, provides a rationale for transposon end asymmetry, and suggests how the avidity provided by multiple sites of interaction could allow a transposase to locate its transposon ends amidst a sea of chromosomal DNA.

  14. Integration Profile and Safety of an Adenovirus Hybrid-Vector Utilizing Hyperactive Sleeping Beauty Transposase for Somatic Integration

    PubMed Central

    Zhang, Wenli; Muck-Hausl, Martin; Wang, Jichang; Sun, Chuanbo; Gebbing, Maren; Miskey, Csaba; Ivics, Zoltan; Izsvak, Zsuzsanna; Ehrhardt, Anja

    2013-01-01

    We recently developed adenovirus/transposase hybrid-vectors utilizing the previously described hyperactive Sleeping Beauty (SB) transposase HSB5 for somatic integration and we could show stabilized transgene expression in mice and a canine model for hemophilia B. However, the safety profile of these hybrid-vectors with respect to vector dose and genotoxicity remains to be investigated. Herein, we evaluated this hybrid-vector system in C57Bl/6 mice with escalating vector dose settings. We found that in all mice which received the hyperactive SB transposase, transgene expression levels were stabilized in a dose-dependent manner and that the highest vector dose was accompanied by fatalities in mice. To analyze potential genotoxic side-effects due to somatic integration into host chromosomes, we performed a genome-wide integration site analysis using linker-mediated PCR (LM-PCR) and linear amplification-mediated PCR (LAM-PCR). Analysis of genomic DNA samples obtained from HSB5 treated female and male mice revealed a total of 1327 unique transposition events. Overall the chromosomal distribution pattern was close-to-random and we observed a random integration profile with respect to integration into gene and non-gene areas. Notably, when using the LM-PCR protocol, 27 extra-chromosomal integration events were identified, most likely caused by transposon excision and subsequent transposition into the delivered adenoviral vector genome. In total, this study provides a careful evaluation of the safety profile of adenovirus/Sleeping Beauty transposase hybrid-vectors. The obtained information will be useful when designing future preclinical studies utilizing hybrid-vectors in small and large animal models. PMID:24124483

  15. Self-inflicted wounds, template-directed gap repair and a recombination hotspot. Effects of the mariner transposase.

    PubMed Central

    Lohe, A R; Timmons, C; Beerman, I; Lozovskaya, E R; Hartl, D L

    2000-01-01

    Aberrant repair products of mariner transposition occur at a frequency of approximately 1/500 per target element per generation. Among 100 such mutations in the nonautonomous element peach, most had aberrations in the 5' end of peach (40 alleles), in the 3' end of peach (11 alleles), or a deletion of peach with or without deletion of flanking genomic DNA (29 alleles). Most mariner mutations can be explained by exonuclease "nibble" and host-mediated repair of the double-stranded gap created by the transposase, in contrast to analogous mutations in the P element. In mariner, mutations in the 5' inverted repeat are smaller and more frequent than those in the 3' inverted repeat, but secondary mutations in target elements with a 5' lesion usually had 3' lesions resembling those normally found at the 5' end. We suggest that the mariner transposase distinguishes between the 5' and 3' ends of the element, and that the 5' end is relatively more protected after strand scission. We also find: (1) that homolog-dependent gap repair is a frequent accompaniment to mariner excision, estimated as 30% of all excision events; and (2) that mariner is a hotspot of recombination in Drosophila females, but only in the presence of functional transposase. PMID:10655218

  16. An atypical AAA+ ATPase assembly controls efficient transposition through DNA remodeling and transposase recruitment

    PubMed Central

    Arias-Palomo, Ernesto; Berger, James M.

    2015-01-01

    Transposons are ubiquitous genetic elements that drive genome rearrangements, evolution, and the spread of infectious disease and drug-resistance. Many transposons, such as Mu, Tn7 and IS21, require regulatory AAA+ ATPases for function. We use x-ray crystallography and cryo-electron microscopy to show that the ATPase subunit of IS21, IstB, assembles into a clamshell-shaped decamer that sandwiches DNA between two helical pentamers of ATP-associated AAA+ domains, sharply bending the duplex into a 180° U-turn. Biochemical studies corroborate key features of the structure, and further show that the IS21 transposase, IstA, recognizes the IstB•DNA complex and promotes its disassembly by stimulating ATP hydrolysis. Collectively, these studies reveal a distinct manner of higher-order assembly and client engagement by a AAA+ ATPase and suggest a mechanistic model where IstB binding and subsequent DNA bending primes a selected insertion site for efficient transposition. PMID:26276634

  17. Full-length Dysferlin Transfer by the Hyperactive Sleeping Beauty Transposase Restores Dysferlin-deficient Muscle

    PubMed Central

    Escobar, Helena; Schöwel, Verena; Spuler, Simone; Marg, Andreas; Izsvák, Zsuzsanna

    2016-01-01

    Dysferlin-deficient muscular dystrophy is a progressive disease characterized by muscle weakness and wasting for which there is no treatment. It is caused by mutations in DYSF, a large, multiexonic gene that forms a coding sequence of 6.2 kb. Sleeping Beauty (SB) transposon is a nonviral gene transfer vector, already used in clinical trials. The hyperactive SB system consists of a transposon DNA sequence and a transposase protein, SB100X, that can integrate DNA over 10 kb into the target genome. We constructed an SB transposon-based vector to deliver full-length human DYSF cDNA into dysferlin-deficient H2K A/J myoblasts. We demonstrate proper dysferlin expression as well as highly efficient engraftment (>1,100 donor-derived fibers) of the engineered myoblasts in the skeletal muscle of dysferlin- and immunodeficient B6.Cg-Dysfprmd Prkdcscid/J (Scid/BLA/J) mice. Nonviral gene delivery of full-length human dysferlin into muscle cells, along with a successful and efficient transplantation into skeletal muscle are important advances towards successful gene therapy of dysferlin-deficient muscular dystrophy. PMID:26784637

  18. Full-length Dysferlin Transfer by the Hyperactive Sleeping Beauty Transposase Restores Dysferlin-deficient Muscle.

    PubMed

    Escobar, Helena; Schöwel, Verena; Spuler, Simone; Marg, Andreas; Izsvák, Zsuzsanna

    2016-01-19

    Dysferlin-deficient muscular dystrophy is a progressive disease characterized by muscle weakness and wasting for which there is no treatment. It is caused by mutations in DYSF, a large, multiexonic gene that forms a coding sequence of 6.2 kb. Sleeping Beauty (SB) transposon is a nonviral gene transfer vector, already used in clinical trials. The hyperactive SB system consists of a transposon DNA sequence and a transposase protein, SB100X, that can integrate DNA over 10 kb into the target genome. We constructed an SB transposon-based vector to deliver full-length human DYSF cDNA into dysferlin-deficient H2K A/J myoblasts. We demonstrate proper dysferlin expression as well as highly efficient engraftment (>1,100 donor-derived fibers) of the engineered myoblasts in the skeletal muscle of dysferlin- and immunodeficient B6.Cg-Dysf(prmd) Prkdc(scid)/J (Scid/BLA/J) mice. Nonviral gene delivery of full-length human dysferlin into muscle cells, along with a successful and efficient transplantation into skeletal muscle are important advances towards successful gene therapy of dysferlin-deficient muscular dystrophy.

  19. Full-length Dysferlin Transfer by the Hyperactive Sleeping Beauty Transposase Restores Dysferlin-deficient Muscle.

    PubMed

    Escobar, Helena; Schöwel, Verena; Spuler, Simone; Marg, Andreas; Izsvák, Zsuzsanna

    2016-01-01

    Dysferlin-deficient muscular dystrophy is a progressive disease characterized by muscle weakness and wasting for which there is no treatment. It is caused by mutations in DYSF, a large, multiexonic gene that forms a coding sequence of 6.2 kb. Sleeping Beauty (SB) transposon is a nonviral gene transfer vector, already used in clinical trials. The hyperactive SB system consists of a transposon DNA sequence and a transposase protein, SB100X, that can integrate DNA over 10 kb into the target genome. We constructed an SB transposon-based vector to deliver full-length human DYSF cDNA into dysferlin-deficient H2K A/J myoblasts. We demonstrate proper dysferlin expression as well as highly efficient engraftment (>1,100 donor-derived fibers) of the engineered myoblasts in the skeletal muscle of dysferlin- and immunodeficient B6.Cg-Dysf(prmd) Prkdc(scid)/J (Scid/BLA/J) mice. Nonviral gene delivery of full-length human dysferlin into muscle cells, along with a successful and efficient transplantation into skeletal muscle are important advances towards successful gene therapy of dysferlin-deficient muscular dystrophy. PMID:26784637

  20. TrAmplification of Human Dental Follicle Cells by piggyBac Transposon - Mediated Reversible Immortalization System

    PubMed Central

    Wu, Yan; Feng, Ge; Song, Jinlin; Zhang, Yuanyuan; Yu, Yong; Huang, Lan; Zheng, Leilei; Deng, Feng

    2015-01-01

    Dental follicle cells (DFCs) are the precursor cells of periodontium. Under certain differentiation conditions, DFCs can be induced to differentiate into chondrogenic, osteogenic and adipogenic cells. However, DFCs has limited lifespan in vitro, so it’s difficult to harvest enough cells for basic research and translational application. pMPH86 is a piggyBac transposon-mediated vector which contains SV40 T-Ag cassette that can be removed by flippase recognition target (FRT) recombinase. Here we demonstrated the pMPH86 can effectively amplify human DFCs through reversible immortalization. The immortalized DFCs (iDFCs) exhibit higher proliferate activity, which can be reversed to its original level before immortalization when deimmortalized by FLP recombinase. The iDFCs and deimmortalized DFCs (dDFCs) express most DFC markers and maintain multiple differentiation potential in vitro as they can be induced by BMP9 to differentiate into chondrogenic, osteogenic and adipogenic cells evidenced by gene expression and protein marker. We also proved telomerase activity of iDFCs are significantly increased and maintained at a high level, while the telomerase activity of primary DFCs was relatively low and decreased with every passage. After SV40 T-Ag was removed to deimmortalize the cells, telomerase activity was reduced to its original level before immortalization and decreased with passages just the same as primary DFCs. These results suggest that piggyBac immortalization system could be a potential strategy to amplify primary cells, which is critical for regenerative research and further clinical application. PMID:26172849

  1. Domestic Violence

    MedlinePlus

    ... to anyone regardless of race, age, sexual orientation, religion, or gender. Domestic violence affects people of all ... ABOUT The Attorney General Budget & Performance Strategic Plans History AGENCIES BUSINESS Business Opportunities Small & Disadvantaged Business Grants ...

  2. The N-terminal zinc finger domain of Tgf2 transposase contributes to DNA binding and to transposition activity

    PubMed Central

    Jiang, Xia-Yun; Hou, Fei; Shen, Xiao-Dan; Du, Xue-Di; Xu, Hai-Li; Zou, Shu-Ming

    2016-01-01

    Active Hobo/Activator/Tam3 (hAT) transposable elements are rarely found in vertebrates. Previously, goldfish Tgf2 was found to be an autonomously active vertebrate transposon that is efficient at gene-transfer in teleost fish. However, little is known about Tgf2 functional domains required for transposition. To explore this, we first predicted in silico a zinc finger domain in the N-terminus of full length Tgf2 transposase (L-Tgf2TPase). Two truncated recombinant Tgf2 transposases with deletions in the N-terminal zinc finger domain, S1- and S2-Tgf2TPase, were expressed in bacteria from goldfish cDNAs. Both truncated Tgf2TPases lost their DNA-binding ability in vitro, specifically at the ends of Tgf2 transposon than native L-Tgf2TPase. Consequently, S1- and S2-Tgf2TPases mediated gene transfer in the zebrafish genome in vivo at a significantly (p < 0.01) lower efficiency (21%–25%), in comparison with L-Tgf2TPase (56% efficiency). Compared to L-Tgf2TPase, truncated Tgf2TPases catalyzed imprecise excisions with partial deletion of TE ends and/or plasmid backbone insertion/deletion. The gene integration into the zebrafish genome mediated by truncated Tgf2TPases was imperfect, creating incomplete 8-bp target site duplications at the insertion sites. These results indicate that the zinc finger domain in Tgf2 transposase is involved in binding to Tgf2 terminal sequences, and loss of those domains has effects on TE transposition. PMID:27251101

  3. The N-terminal zinc finger domain of Tgf2 transposase contributes to DNA binding and to transposition activity.

    PubMed

    Jiang, Xia-Yun; Hou, Fei; Shen, Xiao-Dan; Du, Xue-Di; Xu, Hai-Li; Zou, Shu-Ming

    2016-01-01

    Active Hobo/Activator/Tam3 (hAT) transposable elements are rarely found in vertebrates. Previously, goldfish Tgf2 was found to be an autonomously active vertebrate transposon that is efficient at gene-transfer in teleost fish. However, little is known about Tgf2 functional domains required for transposition. To explore this, we first predicted in silico a zinc finger domain in the N-terminus of full length Tgf2 transposase (L-Tgf2TPase). Two truncated recombinant Tgf2 transposases with deletions in the N-terminal zinc finger domain, S1- and S2-Tgf2TPase, were expressed in bacteria from goldfish cDNAs. Both truncated Tgf2TPases lost their DNA-binding ability in vitro, specifically at the ends of Tgf2 transposon than native L-Tgf2TPase. Consequently, S1- and S2-Tgf2TPases mediated gene transfer in the zebrafish genome in vivo at a significantly (p < 0.01) lower efficiency (21%-25%), in comparison with L-Tgf2TPase (56% efficiency). Compared to L-Tgf2TPase, truncated Tgf2TPases catalyzed imprecise excisions with partial deletion of TE ends and/or plasmid backbone insertion/deletion. The gene integration into the zebrafish genome mediated by truncated Tgf2TPases was imperfect, creating incomplete 8-bp target site duplications at the insertion sites. These results indicate that the zinc finger domain in Tgf2 transposase is involved in binding to Tgf2 terminal sequences, and loss of those domains has effects on TE transposition. PMID:27251101

  4. Expression of the transposase gene tnpA of Tn4652 is positively affected by integration host factor.

    PubMed

    Hõrak, R; Kivisaar, M

    1998-06-01

    Tn4652 is a derivative of the toluene degradation transposon Tn4651 that belongs to the Tn3 family of transposons (M. Tsuda and T. Iino, Mol. Gen. Genet. 210:270-276, 1987). We have sequenced the transposase gene tnpA of transposon Tn4652 and mapped its promoter to the right end of the element. The deduced amino acid sequence of tnpA revealed 96.2% identity with the putative transposase of Tn5041. Homology with other Tn3 family transposases was only moderate (about 20 to 24% identity), suggesting that Tn4652 and Tn5041 are distantly related members of the Tn3 family. Functional analysis of the tnpA promoter revealed that it is active in Pseudomonas putida but silent in Escherichia coli, indicating that some P. putida-specific factor is required for the transcription from this promoter. Additionally, tnpA promoter activity was shown to be modulated by integration host factor (IHF). The presence of an IHF-binding site upstream of the tnpA promoter enhanced the promoter activity. The positive role of IHF was also confirmed by the finding that the enhancing effect of IHF was not detected in the P. putida ihfA-deficient strain A8759. Moreover, the Tn4652 terminal sequences had a negative effect on transcription from the tnpA promoter in the ihfA-defective strain. This finding suggests that IHF not only enhances transcription from the tnpA promoter but also alleviates the negative effect of terminal sequences of Tn4652 on the promoter activity. Also, an in vitro binding assay demonstrated that both ends of Tn4652 bind IHF from a cell lysate of E. coli. PMID:9603867

  5. The wing of the enhancer-binding domain of Mu phage transposase is flexible and is essential for efficient transposition.

    PubMed

    Clubb, R T; Mizuuchi, M; Huth, J R; Omichinski, J G; Savilahti, H; Mizuuchi, K; Clore, G M; Gronenborn, A M

    1996-02-01

    A tetramer of the Mu transposase (MuA) pairs the recombination sites, cleaves the donor DNA, and joins these ends to a target DNA by strand transfer. Juxtaposition of the recombination sites is accomplished by the assembly of a stable synaptic complex of MuA protein and Mu DNA. This initial critical step is facilitated by the transient binding of the N-terminal domain of MuA to an enhancer DNA element within the Mu genome (called the internal activation sequence, IAS). Recently we solved the three-dimensional solution structure of the enhancer-binding domain of Mu phage transposase (residues 1-76, MuA76) and proposed a model for its interaction with the IAS element. Site-directed mutagenesis coupled with an in vitro transposition assay has been used to assess the validity of the model. We have identified five residues on the surface of MuA that are crucial for stable synaptic complex formation but dispensable for subsequent events in transposition. These mutations are located in the loop (wing) structure and recognition helix of the MuA76 domain of the transposase and do not seriously perturb the structure of the domain. Furthermore, in order to understand the dynamic behavior of the MuA76 domain prior to stable synaptic complex formation, we have measured heteronuclear 15N relaxation rates for the unbound MuA76 domain. In the DNA free state the backbone atoms of the helix-turn-helix motif are generally immobilized whereas the residues in the wing are highly flexible on the pico- to nanosecond time scale. Together these studies define the surface of MuA required for enhancement of transposition in vitro and suggest that a flexible loop in the MuA protein required for DNA recognition may become structurally ordered only upon DNA binding. PMID:8577730

  6. Tc1-like Transposase Thm3 of Silver Carp (Hypophthalmichthys molitrix) Can Mediate Gene Transposition in the Genome of Blunt Snout Bream (Megalobrama amblycephala)

    PubMed Central

    Guo, Xiu-Ming; Zhang, Qian-Qian; Sun, Yi-Wen; Jiang, Xia-Yun; Zou, Shu-Ming

    2015-01-01

    Tc1-like transposons consist of an inverted repeat sequence flanking a transposase gene that exhibits similarity to the mobile DNA element, Tc1, of the nematode, Caenorhabditis elegans. They are widely distributed within vertebrate genomes including teleost fish; however, few active Tc1-like transposases have been discovered. In this study, 17 Tc1-like transposon sequences were isolated from 10 freshwater fish species belonging to the families Cyprinidae, Adrianichthyidae, Cichlidae, and Salmonidae. We conducted phylogenetic analyses of these sequences using previously isolated Tc1-like transposases and report that 16 of these elements comprise a new subfamily of Tc1-like transposons. In particular, we show that one transposon, Thm3 from silver carp (Hypophthalmichthys molitrix; Cyprinidae), can encode a 335-aa transposase with apparently intact domains, containing three to five copies in its genome. We then coinjected donor plasmids harboring 367 bp of the left end and 230 bp of the right end of the nonautonomous silver carp Thm1 cis-element along with capped Thm3 transposase RNA into the embryos of blunt snout bream (Megalobrama amblycephala; one- to two-cell embryos). This experiment revealed that the average integration rate could reach 50.6% in adult fish. Within the blunt snout bream genome, the TA dinucleotide direct repeat, which is the signature of Tc1-like family of transposons, was created adjacent to both ends of Thm1 at the integration sites. Our results indicate that the silver carp Thm3 transposase can mediate gene insertion by transposition within the genome of blunt snout bream genome, and that this occurs with a TA position preference. PMID:26438298

  7. Electrochemically modified carbon and chromium surfaces for AFM imaging of double-strand DNA interaction with transposase protein.

    PubMed

    Esnault, Charles; Chénais, Benoît; Casse, Nathalie; Delorme, Nicolas; Louarn, Guy; Pilard, Jean-François

    2013-02-01

    Carbon and chromium surfaces were modified by electrochemical reduction of a diazonium salt formed in situ from the sulfanilic acid. The organic layer formed was activated by phosphorus pentachloride (PCl(5)) to form a benzene sulfonil chloride (Ar-SO(2)Cl). An electrochemical study of the blocking effect and the activity of this surface was carried out on a carbon electrode. The chromium surface study was completed by X-ray photoelectron spectroscopy and atomic force microscopy to characterize the formation of a compact monolayer (0.8 nm height and roughness 0.2-0.3 nm). The compactness and the activity of this organic monolayer allowed us to affix a length dsDNA with the aim of analyzing the formation of a complex between dsDNA and a protein. The interaction of a transposase protein with its target dsDNA was investigated. The direct imaging of the nucleoproteic complex considered herein gives new insights in the comprehension of transposase-DNA interaction in agreement with biochemical data.

  8. Sense overlapping transcripts in IS1341-type transposase genes are functional non-coding RNAs in archaea

    PubMed Central

    Gomes-Filho, José Vicente; Zaramela, Livia Soares; Italiani, Valéria Cristina da Silva; Baliga, Nitin S; Vêncio, Ricardo Z N; Koide, Tie

    2015-01-01

    The existence of sense overlapping transcripts that share regulatory and coding information in the same genomic sequence shows an additional level of prokaryotic gene expression complexity. Here we report the discovery of ncRNAs associated with IS1341-type transposase (tnpB) genes, at the 3'-end of such elements, with examples in archaea and bacteria. Focusing on the model haloarchaeon Halobacterium salinarum NRC-1, we show the existence of sense overlapping transcripts (sotRNAs) for all its IS1341-type transposases. Publicly available transcriptome compendium show condition-dependent differential regulation between sotRNAs and their cognate genes. These sotRNAs allowed us to find a UUCA tetraloop motif that is present in other archaea (ncRNA family HgcC) and in a H. salinarum intergenic ncRNA derived from a palindrome associated transposable elements (PATE). Overexpression of one sotRNA and the PATE-derived RNA harboring the tetraloop motif improved H. salinarum growth, indicating that these ncRNAs are functional. PMID:25806405

  9. PBmice: an integrated database system of piggyBac (PB) insertional mutations and their characterizations in mice

    PubMed Central

    Sun, Ling V.; Jin, Ke; Liu, Yiming; Yang, Wenwei; Xie, Xing; Ye, Lin; Wang, Li; Zhu, Lin; Ding, Sheng; Su, Yi; Zhou, Jie; Han, Min; Zhuang, Yuan; Xu, Tian; Wu, Xiaohui; Gu, Ning; Zhong, Yang

    2008-01-01

    DNA transposon piggyBac (PB) is a newly established mutagen for large-scale mutagenesis in mice. We have designed and implemented an integrated database system called PBmice (PB Mutagenesis Information CEnter) for storing, retrieving and displaying the information derived from PB insertions (INSERTs) in the mouse genome. This system is centered on INSERTs with information including their genomic locations and flanking genomic sequences, the expression levels of the hit genes, and the expression patterns of the trapped genes if a trapping vector was used. It also archives mouse phenotyping data linked to INSERTs, and allows users to conduct quick and advanced searches for genotypic and phenotypic information relevant to a particular or a set of INSERT(s). Sequence-based information can be cross-referenced with other genomic databases such as Ensembl, BLAST and GBrowse tools used in PBmice offer enhanced search and display for additional information relevant to INSERTs. The total number and genomic distribution of PB INSERTs, as well as the availability of each PB insertional LINE can also be viewed with user-friendly interfaces. PBmice is freely available at http://www.idmshanghai.cn/PBmice or http://www.scbit.org/PBmice/. PMID:17932058

  10. piggyBac transposon plus insulators overcome epigenetic silencing to provide for stable signaling pathway reporter cell lines.

    PubMed

    Mossine, Valeri V; Waters, James K; Hannink, Mark; Mawhinney, Thomas P

    2013-01-01

    Genetically modified hematopoietic progenitors represent an important testing platform for a variety of cell-based therapies, pharmaceuticals, diagnostics and other applications. Stable expression of a transfected gene of interest in the cells is often obstructed by its silencing. DNA transposons offer an attractive non-viral alternative of transgene integration into the host genome, but their broad applicability to leukocytes and other "transgene unfriendly" cells has not been fully demonstrated. Here we assess stability of piggyBac transposon-based reporter expression in murine prostate adenocarcinoma TRAMP-C2, human monocyte THP-1 and erythroleukemia K562 cell lines, along with macrophages and dendritic cells (DCs) that have differentiated from the THP-1 transfects. The most efficient and stable reporter activity was observed for combinations of the transposon inverted terminal repeats and one 5'- or two cHS4 core insulators flanking a green fluorescent protein reporter construct, with no detectable silencing over 10 months of continuous cell culture in absence of any selective pressure. In monocytic THP-1 cells, the functional activity of luciferase reporters for NF-κB, Nrf2, or HIF-1α has not decreased over time and was retained following differentiation into macrophages and DCs, as well. These results imply pB as a versatile tool for gene integration in monocytic cells in general, and as a convenient access route to DC-based signaling pathway reporters suitable for high-throughput assays, in particular. PMID:24376882

  11. The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor.

    PubMed

    Jin, Z; Maiti, S; Huls, H; Singh, H; Olivares, S; Mátés, L; Izsvák, Z; Ivics, Z; Lee, D A; Champlin, R E; Cooper, L J N

    2011-09-01

    Sleeping Beauty (SB3) transposon and transposase constitute a DNA plasmid system used for therapeutic human cell genetic engineering. Here we report a comparison of SB100X, a newly developed hyperactive SB transposase, to a previous generation SB11 transposase to achieve stable expression of a CD19-specific chimeric antigen receptor (CAR3) in primary human T cells. The electro-transfer of SB100X expressed from a DNA plasmid or as an introduced mRNA species had superior transposase activity in T cells based on the measurement of excision circles released after transposition and emergence of CAR expression on T cells selectively propagated upon CD19+ artificial antigen-presenting cells. Given that T cells modified with SB100X and SB11 integrate on average one copy of the CAR transposon in each T-cell genome, the improved transposition mediated by SB100X apparently leads to an augmented founder effect of electroporated T cells with durable integration of CAR. In aggregate, SB100X improves SB transposition in primary human T cells and can be titrated with an SB transposon plasmid to improve the generation of CD19-specific CAR+ T cells. PMID:21451576

  12. Purified mariner (Mos1) transposase catalyzes the integration of marked elements into the germ-line of the yellow fever mosquito, Aedes aegypti.

    PubMed

    Coates, C J; Jasinskiene, N; Morgan, D; Tosi, L R; Beverley, S M; James, A A

    2000-11-01

    Derivatives of the mariner transposable element, Mos1, from Drosophila mauritiana, can integrate into the germ-line of the yellow fever mosquito, Aedes aegypti. Previously, the transposase required to mobilize Mos1 was provided in trans by a helper plasmid expressing the enzyme under the control of the D. psuedoobscura heat-shock protein 82 promoter. Here we tested whether purified recombinant Mos1 transposase could increase the recovery of Ae. aegypti transformants. Mos1 transposase was injected into white-eyed, kh(w)/kh(w), Ae. aegypti embryos with a Mos1 donor plasmid containing a copy of the wild-type allele of the D. melanogaster cinnabar gene. Transformed mosquitoes were recognized by partial restoration of eye color in the G(1) animals and confirmed by Southern analyses of genomic DNA. At Mos1 transposase concentrations approaching 100 nM, the rate of germ-line transformants arising from independent insertions in G(0) animals was elevated 2-fold compared to that seen in experiments with helper plasmids. Furthermore, the recovery of total G(1) transformants was increased 7.5-fold over the frequency seen with co-injected helper plasmid. Southern blot analyses and gene amplification experiments confirmed the integration of the transposons into the mosquito genome, although not all integrations were of the expected cut-and-paste type transposition. The increased frequency of germ-line integrations obtained with purified transposase will facilitate the generation of Mos1 transgenic mosquitoes and the application of transgenic approaches to the biology of this important vector of multiple pathogens. PMID:10989286

  13. Comparative Analysis of piggyBac, CRISPR/Cas9 and TALEN Mediated BAC Transgenesis in the Zygote for the Generation of Humanized SIRPA Rats.

    PubMed

    Jung, Chris J; Ménoret, Séverine; Brusselle, Lucas; Tesson, Laurent; Usal, Claire; Chenouard, Vanessa; Remy, Séverine; Ouisse, Laure-Hélène; Poirier, Nicolas; Vanhove, Bernard; de Jong, Pieter J; Anegon, Ignacio

    2016-01-01

    BAC transgenic mammalian systems offer an important platform for recapitulating human gene expression and disease modeling. While the larger body mass, and greater genetic and physiologic similarity to humans render rats well suited for reproducing human immune diseases and evaluating therapeutic strategies, difficulties of generating BAC transgenic rats have hindered progress. Thus, an efficient method for BAC transgenesis in rats would be valuable. Immunodeficient mice carrying a human SIRPA transgene have previously been shown to support improved human cell hematopoiesis. Here, we have generated for the first time, human SIRPA BAC transgenic rats, for which the gene is faithfully expressed, functionally active, and germline transmissible. To do this, human SIRPA BAC was modified with elements to work in coordination with genome engineering technologies-piggyBac, CRISPR/Cas9 or TALEN. Our findings show that piggyBac transposition is a more efficient approach than the classical BAC transgenesis, resulting in complete BAC integration with predictable end sequences, thereby permitting precise assessment of the integration site. Neither CRISPR/Cas9 nor TALEN increased BAC transgenesis. Therefore, an efficient generation of human SIRPA transgenic rats using piggyBac opens opportunities for expansion of humanized transgenic rat models in the future to advance biomedical research and therapeutic applications. PMID:27530248

  14. Comparative Analysis of piggyBac, CRISPR/Cas9 and TALEN Mediated BAC Transgenesis in the Zygote for the Generation of Humanized SIRPA Rats

    PubMed Central

    Jung, Chris J.; Ménoret, Séverine; Brusselle, Lucas; Tesson, Laurent; Usal, Claire; Chenouard, Vanessa; Remy, Séverine; Ouisse, Laure-Hélène; Poirier, Nicolas; Vanhove, Bernard; de Jong, Pieter J.; Anegon, Ignacio

    2016-01-01

    BAC transgenic mammalian systems offer an important platform for recapitulating human gene expression and disease modeling. While the larger body mass, and greater genetic and physiologic similarity to humans render rats well suited for reproducing human immune diseases and evaluating therapeutic strategies, difficulties of generating BAC transgenic rats have hindered progress. Thus, an efficient method for BAC transgenesis in rats would be valuable. Immunodeficient mice carrying a human SIRPA transgene have previously been shown to support improved human cell hematopoiesis. Here, we have generated for the first time, human SIRPA BAC transgenic rats, for which the gene is faithfully expressed, functionally active, and germline transmissible. To do this, human SIRPA BAC was modified with elements to work in coordination with genome engineering technologies-piggyBac, CRISPR/Cas9 or TALEN. Our findings show that piggyBac transposition is a more efficient approach than the classical BAC transgenesis, resulting in complete BAC integration with predictable end sequences, thereby permitting precise assessment of the integration site. Neither CRISPR/Cas9 nor TALEN increased BAC transgenesis. Therefore, an efficient generation of human SIRPA transgenic rats using piggyBac opens opportunities for expansion of humanized transgenic rat models in the future to advance biomedical research and therapeutic applications. PMID:27530248

  15. Amino acid sequence homology between Piv, an essential protein in site-specific DNA inversion in Moraxella lacunata, and transposases of an unusual family of insertion elements.

    PubMed Central

    Lenich, A G; Glasgow, A C

    1994-01-01

    Deletion analysis of the subcloned DNA inversion region of Moraxella lacunata indicates that Piv is the only M. lacunata-encoded factor required for site-specific inversion of the tfpQ/tfpI pilin segment. The predicted amino acid sequence of Piv shows significant homology solely with the transposases/integrases of a family of insertion sequence elements, suggesting that Piv is a novel site-specific recombinase. Images PMID:8021196

  16. Fis negatively affects binding of Tn4652 transposase by out-competing IHF from the left end of Tn4652.

    PubMed

    Teras, Riho; Jakovleva, Julia; Kivisaar, Maia

    2009-04-01

    Transposition activity in bacteria is generally maintained at a low level. The activity of mobile DNA elements can be controlled by bacterially encoded global regulators. Regulation of transposition of Tn4652 in Pseudomonas putida is one such example. Activation of transposition of Tn4652 in starving bacteria requires the stationary-phase sigma factor RpoS and integration host factor (IHF). IHF plays a dual role in Tn4652 translocation by activating transcription of the transposase gene tnpA of the transposon and facilitating TnpA binding to the inverted repeats of the transposon. Our previous results have indicated that besides IHF some other P. putida-encoded global regulator(s) might bind to the ends of Tn4652 and regulate transposition activity. In this study, employing a DNase I footprint assay we have identified a binding site of P. putida Fis (factor for inversion stimulation) centred 135 bp inside the left end of Tn4652. Our results of gel mobility shift and DNase I footprint studies revealed that Fis out-competes IHF from the left end of Tn4652, thereby abolishing the binding of TnpA. Thus, the results obtained in this study indicate that the transposition of Tn4652 is regulated by the cellular amount of P. putida global regulators Fis and IHF. PMID:19332822

  17. From crop domestication to super-domestication.

    PubMed

    Vaughan, D A; Balázs, E; Heslop-Harrison, J S

    2007-11-01

    Research related to crop domestication has been transformed by technologies and discoveries in the genome sciences as well as information-related sciences that are providing new tools for bioinformatics and systems' biology. Rapid progress in archaeobotany and ethnobotany are also contributing new knowledge to understanding crop domestication. This sense of rapid progress is encapsulated in this Special Issue, which contains 18 papers by scientists in botanical, crop sciences and related disciplines on the topic of crop domestication. One paper focuses on current themes in the genetics of crop domestication across crops, whereas other papers have a crop or geographic focus. One feature of progress in the sciences related to crop domestication is the availability of well-characterized germplasm resources in the global network of genetic resources centres (genebanks). Germplasm in genebanks is providing research materials for understanding domestication as well as for plant breeding. In this review, we highlight current genetic themes related to crop domestication. Impressive progress in this field in recent years is transforming plant breeding into crop engineering to meet the human need for increased crop yield with the minimum environmental impact - we consider this to be 'super-domestication'. While the time scale of domestication of 10 000 years or less is a very short evolutionary time span, the details emerging of what has happened and what is happening provide a window to see where domestication might - and can - advance in the future.

  18. Natural Stilbenoids Isolated from Grapevine Exhibiting Inhibitory Effects against HIV-1 Integrase and Eukaryote MOS1 Transposase In Vitro Activities

    PubMed Central

    Chaignepain, Stéphane; Subra, Frederic; Munir, Soundasse; Delelis, Olivier; Lesbats, Paul; Calmels, Christina; Andreola, Marie-Line; Merillon, Jean-Michel; Auge-Gouillou, Corinne; Parissi, Vincent

    2013-01-01

    Polynucleotidyl transferases are enzymes involved in several DNA mobility mechanisms in prokaryotes and eukaryotes. Some of them such as retroviral integrases are crucial for pathogenous processes and are therefore good candidates for therapeutic approaches. To identify new therapeutic compounds and new tools for investigating the common functional features of these proteins, we addressed the inhibition properties of natural stilbenoids deriving from resveratrol on two models: the HIV-1 integrase and the eukaryote MOS-1 transposase. Two resveratrol dimers, leachianol F and G, were isolated for the first time in Vitis along with fourteen known stilbenoids: E-resveratrol, E-piceid, E-pterostilbene, E-piceatannol, (+)-E-ε-viniferin, E-ε-viniferinglucoside, E-scirpusin A, quadragularin A, ampelopsin A, pallidol, E-miyabenol C, E-vitisin B, hopeaphenol, and isohopeaphenol and were purified from stalks of Vitis vinifera (Vitaceae), and moracin M from stem bark of Milliciaexelsa (Moraceae). These compounds were tested in in vitro and in vivo assays reproducing the activity of both enzymes. Several molecules presented significant inhibition on both systems. Some of the molecules were found to be active against both proteins while others were specific for one of the two models. Comparison of the differential effects of the molecules suggested that the compounds could target specific intermediate nucleocomplexes of the reactions. Additionally E-pterostilbene was found active on the early lentiviral replication steps in lentiviruses transduced cells. Consequently, in addition to representing new original lead compounds for further modelling of new active agents against HIV-1 integrase, these molecules could be good tools for identifying such reaction intermediates in DNA mobility processes. PMID:24312275

  19. Mobile Minos elements from Drosophila hydei encode a two-exon transposase with similarity to the paired DNA-binding domain.

    PubMed Central

    Franz, G; Loukeris, T G; Dialektaki, G; Thompson, C R; Savakis, C

    1994-01-01

    Elements related to the Tc1-like Minos mobile element have been cloned from Drosophila hydei and sequenced. Southern blot and sequence analyses show that (i) the elements are actively transposing in the Drosophila hydei germ line, (ii) they are characterized by a striking degree of sequence and size homogeneity, and (iii) like Tc1, they insert at a TA dinucleotide that is probably duplicated during the process. The nucleotide sequences of two elements, Minos-2 and Minos-3, differ at only one position from each other and contain two nonoverlapping open reading frames that are separated by a putative 60-nucleotide intron. The amino-terminal part of the Minos putative transposase shows sequence similarity to the paired DNA-binding domain. Forced transcription of a modified Minos element that was introduced into the Drosophila melanogaster germ line by P element-mediated transformation resulted in the production of accurately spliced polyadenylylated RNA molecules. It is proposed that Minos-2 and/or Minos-3 may encode an active transposase containing an amino-terminal DNA-binding domain that is distantly related to the paired DNA-binding domain. Images PMID:8197129

  20. Neuronal precursor-restricted transduction via in utero CNS gene delivery of a novel bipartite HSV amplicon/transposase hybrid vector.

    PubMed

    Bowers, William J; Mastrangelo, Michael A; Howard, Darlene F; Southerland, Hilary A; Maguire-Zeiss, Kathleen A; Federoff, Howard J

    2006-03-01

    The ability to modify genetically in utero the precursors of neuronal lineage contributing to multiple postmitotic cell types in the adult central nervous system would provide a means to evaluate strategies to ameliorate conditions affecting cellular patterning, metabolism, or survival. The herpes simplex virus (HSV)-derived amplicon, a vector devoid of viral genes and with the largest known payload capacity, normally exists episomally within nuclei of transduced cells, thus precluding conveyance during mitosis. Herein, we modify the Tc1-like Sleeping Beauty (SB) transposon system to create an integrating amplicon vector platform wherein provision of transposase in trans effectively catalyzes integration of a transgenomic segment. Cotransduction with a Rous sarcoma virus promoter-driven beta-galactosidase-neomycin (betageo) fusion flanked by SB terminal repeats (HSVT-betageo) and a second expressing the SB transposase gene under HSV immediate-early 4/5 gene promoter control (HSVsb) resulted in integration and extension of expression duration. Most notably, in utero intraventricular application led to extensive transgene expression within neuronal precursors and their derivatives without attendant adverse consequences, suggesting this new platform could be used to evaluate prenatally the function of gene products in neuronal lineages and evaluate therapeutic strategies for correction of genetic abnormalities affecting the developing CNS. PMID:16412694

  1. Prevalence of mobile genetic elements and transposase genes in Vibrio alginolyticus from the southern coastal region of China and their role in horizontal gene transfer.

    PubMed

    Luo, Peng; Jiang, Haiying; Wang, Yanhong; Su, Ting; Hu, Chaoqun; Ren, Chunhua; Jiang, Xiao

    2012-12-01

    Vibrio alginolyticus has high genetic diversity, but little is known about the means by which it has been acquired. In this study, the distributions of mobile genetic elements (MGEs), including integrating conjugative elements (ICEs), superintegron-like cassettes (SICs), insertion sequences (ISs), and two types of transposase genes (valT1 and valT2), in 192 strains of V. alginolyticus were investigated. ICE, SIC, and IS elements, valT1, and valT2 were detected in 8.9%, 13.0%, 4.7%, 9.4%, and 2.6% of the strains, respectively. Blast searches and phylogenetic analysis of the acquired sequences of the ICE, SIC, IS elements and transposase genes showed that the corresponding homologues were bacterial and derived from extensive sources. The high prevalences of these MGEs in V. alginolyticus implied the extensive and frequent exchange of genes with environmental bacteria and that these elements strongly contribute to the genetic and phenotypic diversity of the bacterium. To our knowledge, this is the first report of V. alginolyticus harboring ICE and SIC elements.

  2. Clustered regulatory interspaced short palindromic repeats (CRISPR)-mediated mutagenesis and phenotype rescue by piggyBac transgenesis in a nonmodel Drosophila species.

    PubMed

    Tanaka, R; Murakami, H; Ote, M; Yamamoto, D

    2016-08-01

    How behavioural diversity emerged in evolution is an unexplored subject in biology. To tackle this problem, genes and circuits for a behaviour need to be determined in different species for phylogenetic comparisons. The recently developed clustered regulatory interspaced short palindromic repeats/CRISPR associated protein9 (CRISPR/Cas9) system made such a challenge possible by providing the means to induce mutations in a gene of interest in any organism. Aiming at elucidating diversification in genetic and neural networks for courtship behaviour, we attempted to generate a genetic tool kit in Drosophila subobscura, a nonmodel species distantly related to the genetic model Drosophila melanogaster. Here we report the generation of yellow (y) and white mutations with the aid of the CRISPR/Cas9 system, and the rescue of the y mutant phenotype by germline transformation of the newly established y mutant fly line with a y(+) -marked piggyBac vector. This successful mutagenesis and transformation in D. subobscura open up an avenue for comprehensive genetic analyses of higher functions in this and other nonmodel Drosophila species, representing a key step toward systematic comparisons of genes and circuitries underlying behaviour amongst species. PMID:27015359

  3. Combination of the clustered regularly interspaced short palindromic repeats (CRISPR)-associated 9 technique with the piggybac transposon system for mouse in utero electroporation to study cortical development.

    PubMed

    Cheng, Man; Jin, Xubin; Mu, Lili; Wang, Fangyu; Li, Wei; Zhong, Xiaoling; Liu, Xuan; Shen, Wenchen; Liu, Ying; Zhou, Yan

    2016-09-01

    In utero electroporation (IUE) is commonly used to study cortical development of cerebrum by downregulating or overexpressing genes of interest in neural progenitor cells (NPCs) of small mammals. However, exogenous plasmids are lost or diluted over time. Furthermore, gene knockdown based on short-hairpin RNAs may exert nonspecific effects that lead to aberrant neuronal migration. Genomic engineering by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system has great research and therapeutic potentials. Here we integrate the CRISPR/Cas9 components into the piggyBac (PB) transposon system (the CRISPR/Cas9-PB toolkit) for cortical IUEs. The mouse Sry-related HMG box-2 (Sox2) gene was selected as the target for its application. Most transduced cortical NPCs were depleted of SOX2 protein as early as 3 days post-IUE, whereas expressions of SOX1 and PAX6 remained intact. Furthermore, both the WT Cas9 and the D10A nickase mutant Cas9n showed comparable knockout efficiency. Transduced cortical cells were purified with fluorescence-activated cell sorting, and effective gene editing at the Sox2 loci was confirmed. Thus, application of the CRISPR/Cas9-PB toolkit in IUE is a promising strategy to study gene functions in cortical NPCs and their progeny. © 2016 Wiley Periodicals, Inc. PMID:27317429

  4. Combination of the clustered regularly interspaced short palindromic repeats (CRISPR)-associated 9 technique with the piggybac transposon system for mouse in utero electroporation to study cortical development.

    PubMed

    Cheng, Man; Jin, Xubin; Mu, Lili; Wang, Fangyu; Li, Wei; Zhong, Xiaoling; Liu, Xuan; Shen, Wenchen; Liu, Ying; Zhou, Yan

    2016-09-01

    In utero electroporation (IUE) is commonly used to study cortical development of cerebrum by downregulating or overexpressing genes of interest in neural progenitor cells (NPCs) of small mammals. However, exogenous plasmids are lost or diluted over time. Furthermore, gene knockdown based on short-hairpin RNAs may exert nonspecific effects that lead to aberrant neuronal migration. Genomic engineering by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system has great research and therapeutic potentials. Here we integrate the CRISPR/Cas9 components into the piggyBac (PB) transposon system (the CRISPR/Cas9-PB toolkit) for cortical IUEs. The mouse Sry-related HMG box-2 (Sox2) gene was selected as the target for its application. Most transduced cortical NPCs were depleted of SOX2 protein as early as 3 days post-IUE, whereas expressions of SOX1 and PAX6 remained intact. Furthermore, both the WT Cas9 and the D10A nickase mutant Cas9n showed comparable knockout efficiency. Transduced cortical cells were purified with fluorescence-activated cell sorting, and effective gene editing at the Sox2 loci was confirmed. Thus, application of the CRISPR/Cas9-PB toolkit in IUE is a promising strategy to study gene functions in cortical NPCs and their progeny. © 2016 Wiley Periodicals, Inc.

  5. Stable, Nonviral Expression of Mutated Tumor Neoantigen-specific T-cell Receptors Using the Sleeping Beauty Transposon/Transposase System.

    PubMed

    Deniger, Drew C; Pasetto, Anna; Tran, Eric; Parkhurst, Maria R; Cohen, Cyrille J; Robbins, Paul F; Cooper, Laurence Jn; Rosenberg, Steven A

    2016-06-01

    Neoantigens unique to each patient's tumor can be recognized by autologous T cells through their T-cell receptor (TCR) but the low frequency and/or terminal differentiation of mutation-specific T cells in tumors can limit their utility as adoptive T-cell therapies. Transfer of TCR genes into younger T cells from peripheral blood with a high proliferative potential could obviate this problem. We generated a rapid, cost-effective strategy to genetically engineer cancer patient T cells with TCRs using the clinical Sleeping Beauty transposon/transposase system. Patient-specific TCRs reactive against HLA-A*0201-restriced neoantigens AHNAK(S2580F) or ERBB2(H473Y) or the HLA-DQB*0601-restricted neoantigen ERBB2IP(E805G) were assembled with murine constant chains and cloned into Sleeping Beauty transposons. Patient peripheral blood lymphocytes were coelectroporated with SB11 transposase and Sleeping Beauty transposon, and transposed T cells were enriched by sorting on murine TCRβ (mTCRβ) expression. Rapid expansion of mTCRβ(+) T cells with irradiated allogeneic peripheral blood lymphocytes feeders, OKT3, interleukin-2 (IL-2), IL-15, and IL-21 resulted in a preponderance of effector (CD27(-)CD45RA(-)) and less-differentiated (CD27(+)CD45RA(+)) T cells. Transposed T cells specifically mounted a polyfunctional response against cognate mutated neoantigens and tumor cell lines. Thus, Sleeping Beauty transposition of mutation-specific TCRs can facilitate the use of personalized T-cell therapy targeting unique neoantigens.

  6. A Broad Range of Dose Optima Achieve High-level, Long-term Gene Expression After Hydrodynamic Delivery of Sleeping Beauty Transposons Using Hyperactive SB100x Transposase

    PubMed Central

    Podetz-Pedersen, Kelly M; Olson, Erik R; Somia, Nikunj V; Russell, Stephen J; McIvor, R Scott

    2016-01-01

    The Sleeping Beauty (SB) transposon system has been shown to enable long-term gene expression by integrating new sequences into host cell chromosomes. We found that the recently reported SB100x hyperactive transposase conferred a surprisingly high level of long-term expression after hydrodynamic delivery of luciferase-encoding reporter transposons in the mouse. We conducted dose-ranging studies to determine the effect of varying the amount of SB100x transposase-encoding plasmid (pCMV-SB100x) at a set dose of luciferase transposon and of varying the amount of transposon-encoding DNA at a set dose of pCMV-SB100x in hydrodynamically injected mice. Animals were immunosuppressed using cyclophosphamide in order to prevent an antiluciferase immune response. At a set dose of transposon DNA (25 µg), we observed a broad range of pCMV-SB100x doses (0.1–2.5 µg) conferring optimal levels of long-term expression (>1011 photons/second/cm2). At a fixed dose of 0.5 μg of pCMV-SB100x, maximal long-term luciferase expression (>1010 photons/second/cm2) was achieved at a transposon dose of 5–125 μg. We also found that in the linear range of transposon doses (100 ng), co-delivering the CMV-SB100x sequence on the same plasmid was less effective in achieving long-term expression than delivery on separate plasmids. These results show marked flexibility in the doses of SB transposon plus pCMV-SB100x that achieve maximal SB-mediated gene transfer efficiency and long-term gene expression after hydrodynamic DNA delivery to mouse liver. PMID:26784638

  7. A Broad Range of Dose Optima Achieve High-level, Long-term Gene Expression After Hydrodynamic Delivery of Sleeping Beauty Transposons Using Hyperactive SB100x Transposase.

    PubMed

    Podetz-Pedersen, Kelly M; Olson, Erik R; Somia, Nikunj V; Russell, Stephen J; McIvor, R Scott

    2016-01-19

    The Sleeping Beauty (SB) transposon system has been shown to enable long-term gene expression by integrating new sequences into host cell chromosomes. We found that the recently reported SB100x hyperactive transposase conferred a surprisingly high level of long-term expression after hydrodynamic delivery of luciferase-encoding reporter transposons in the mouse. We conducted dose-ranging studies to determine the effect of varying the amount of SB100x transposase-encoding plasmid (pCMV-SB100x) at a set dose of luciferase transposon and of varying the amount of transposon-encoding DNA at a set dose of pCMV-SB100x in hydrodynamically injected mice. Animals were immunosuppressed using cyclophosphamide in order to prevent an antiluciferase immune response. At a set dose of transposon DNA (25 µg), we observed a broad range of pCMV-SB100x doses (0.1-2.5 µg) conferring optimal levels of long-term expression (>10(11) photons/second/cm(2)). At a fixed dose of 0.5 μg of pCMV-SB100x, maximal long-term luciferase expression (>10(10) photons/second/cm(2)) was achieved at a transposon dose of 5-125 μg. We also found that in the linear range of transposon doses (100 ng), co-delivering the CMV-SB100x sequence on the same plasmid was less effective in achieving long-term expression than delivery on separate plasmids. These results show marked flexibility in the doses of SB transposon plus pCMV-SB100x that achieve maximal SB-mediated gene transfer efficiency and long-term gene expression after hydrodynamic DNA delivery to mouse liver.

  8. A Broad Range of Dose Optima Achieve High-level, Long-term Gene Expression After Hydrodynamic Delivery of Sleeping Beauty Transposons Using Hyperactive SB100x Transposase.

    PubMed

    Podetz-Pedersen, Kelly M; Olson, Erik R; Somia, Nikunj V; Russell, Stephen J; McIvor, R Scott

    2016-01-01

    The Sleeping Beauty (SB) transposon system has been shown to enable long-term gene expression by integrating new sequences into host cell chromosomes. We found that the recently reported SB100x hyperactive transposase conferred a surprisingly high level of long-term expression after hydrodynamic delivery of luciferase-encoding reporter transposons in the mouse. We conducted dose-ranging studies to determine the effect of varying the amount of SB100x transposase-encoding plasmid (pCMV-SB100x) at a set dose of luciferase transposon and of varying the amount of transposon-encoding DNA at a set dose of pCMV-SB100x in hydrodynamically injected mice. Animals were immunosuppressed using cyclophosphamide in order to prevent an antiluciferase immune response. At a set dose of transposon DNA (25 µg), we observed a broad range of pCMV-SB100x doses (0.1-2.5 µg) conferring optimal levels of long-term expression (>10(11) photons/second/cm(2)). At a fixed dose of 0.5 μg of pCMV-SB100x, maximal long-term luciferase expression (>10(10) photons/second/cm(2)) was achieved at a transposon dose of 5-125 μg. We also found that in the linear range of transposon doses (100 ng), co-delivering the CMV-SB100x sequence on the same plasmid was less effective in achieving long-term expression than delivery on separate plasmids. These results show marked flexibility in the doses of SB transposon plus pCMV-SB100x that achieve maximal SB-mediated gene transfer efficiency and long-term gene expression after hydrodynamic DNA delivery to mouse liver. PMID:26784638

  9. Stable, Nonviral Expression of Mutated Tumor Neoantigen-specific T-cell Receptors Using the Sleeping Beauty Transposon/Transposase System.

    PubMed

    Deniger, Drew C; Pasetto, Anna; Tran, Eric; Parkhurst, Maria R; Cohen, Cyrille J; Robbins, Paul F; Cooper, Laurence Jn; Rosenberg, Steven A

    2016-06-01

    Neoantigens unique to each patient's tumor can be recognized by autologous T cells through their T-cell receptor (TCR) but the low frequency and/or terminal differentiation of mutation-specific T cells in tumors can limit their utility as adoptive T-cell therapies. Transfer of TCR genes into younger T cells from peripheral blood with a high proliferative potential could obviate this problem. We generated a rapid, cost-effective strategy to genetically engineer cancer patient T cells with TCRs using the clinical Sleeping Beauty transposon/transposase system. Patient-specific TCRs reactive against HLA-A*0201-restriced neoantigens AHNAK(S2580F) or ERBB2(H473Y) or the HLA-DQB*0601-restricted neoantigen ERBB2IP(E805G) were assembled with murine constant chains and cloned into Sleeping Beauty transposons. Patient peripheral blood lymphocytes were coelectroporated with SB11 transposase and Sleeping Beauty transposon, and transposed T cells were enriched by sorting on murine TCRβ (mTCRβ) expression. Rapid expansion of mTCRβ(+) T cells with irradiated allogeneic peripheral blood lymphocytes feeders, OKT3, interleukin-2 (IL-2), IL-15, and IL-21 resulted in a preponderance of effector (CD27(-)CD45RA(-)) and less-differentiated (CD27(+)CD45RA(+)) T cells. Transposed T cells specifically mounted a polyfunctional response against cognate mutated neoantigens and tumor cell lines. Thus, Sleeping Beauty transposition of mutation-specific TCRs can facilitate the use of personalized T-cell therapy targeting unique neoantigens. PMID:26945006

  10. Domestic Communication Satellites

    ERIC Educational Resources Information Center

    Horowitz, Andrew

    1974-01-01

    A discussion of the Federal Communications Commission's new policy on domestic satellites in light of our 1) military and economic history; 2) corporate interests; 3) citizen surveillance; and 4) media control. (HB)

  11. Domestic violence in America.

    PubMed

    Bash, K L; Jones, F

    1994-09-01

    Domestic violence is an underrecognized problem of immense cost. It is a crime; its victims must be identified and protected. The medical and judicial communities share responsibility in addressing this issue and providing support for victims. The role of health care workers in recognizing and preventing domestic violence cannot be overestimated. Direct questioning of patients, especially about the source of any injuries and about safety at home, is the first step in uncovering abuse. Educational programs for health care providers and the general public can change society's view and tolerance of this problem. Physicians must take an active role in changing community attitudes about domestic violence and in instituting programs to reduce its incidence. Medical treatment of the injuries resulting from domestic violence is not sufficient. Abused women need the care of a team of professionals who can address psychological, emotional, and physical injuries. They must also be provided with safe housing and financial and legal assistance in order to escape the abusive relationship. Physicians and legislators must work together to effect change. Domestic violence is a public health menace. We need to break the cycle of abuse that has become an integral part of our society.

  12. Cloning of a CACTA transposon-like insertion in intron I of tomato invertase Lin5 gene and identification of transposase-like sequences of Solanaceae species.

    PubMed

    Proels, Reinhard K; Roitsch, Thomas

    2006-03-01

    Very few CACTA transposon-like sequences have been described in Solanaceae species. Sequence information has been restricted to partial transposase (TPase)-like fragments, and no target gene of CACTA-like transposon insertion has been described in tomato to date. In this manuscript, we report on a CACTA transposon-like insertion in intron I of tomato (Lycopersicon esculentum) invertase gene Lin5 and TPase-like sequences of several Solanaceae species. Consensus primers deduced from the TPase region of the tomato CACTA transposon-like element allowed the amplification of similar sequences from various Solanaceae species of different subfamilies including Solaneae (Solanum tuberosum), Cestreae (Nicotiana tabacum) and Datureae (Datura stramonium). This demonstrates the ubiquitous presence of CACTA-like elements in Solanaceae genomes. The obtained partial sequences are highly conserved, and allow further detection and detailed analysis of CACTA-like transposons throughout Solanaceae species. CACTA-like transposon sequences make possible the evaluation of their use for genome analysis, functional studies of genes and the evolutionary relationships between plant species.

  13. The Mu transposase tetramer is inactive in unassisted strand transfer: an auto-allosteric effect of Mu A promotes the reaction in the absence of Mu B.

    PubMed

    Wu, Z; Chaconas, G

    1997-03-21

    A tetramer of the Mu transposase is the structural and functional core in all three stable higher-order nucleoprotein complexes (Type 0, Type 1 and Type 2 transpososomes) generated in a defined in vitro strand transfer reaction. Although functional in donor cleavage, we report here that contrary to previous belief, the Mu A tetramer is incapable of unassisted strand transfer. The Mu B protein is required to stimulate the tetramer for intermolecular strand transfer. In the absence of Mu B protein we show that additional Mu A molecules must be added to the core tetramer to stimulate intramolecular strand transfer. Mapping experiments indicate that domain II of the assisting Mu A mediates functional interactions with the core tetramer. The recipient site for Mu A stimulated strand transfer on the A tetramer is likely in domain II and is clearly different from the domain IIIb site used by the Mu B protein. The Mu accessory end binding sites and the Mu enhancer are not required in the Mu A assisted strand transfer, suggesting that helper A molecules in solution can interact with the core tetramer to stimulate the reaction. Finally, we argue that the strand transfer activity and protein sites for target interaction reside within the core tetramer; hence the role of the stimulatory A molecules appears to be limited to that of an auto-allosteric effector.

  14. Domestication genomics: evidence from animals.

    PubMed

    Wang, Guo-Dong; Xie, Hai-Bing; Peng, Min-Sheng; Irwin, David; Zhang, Ya-Ping

    2014-02-01

    Animal domestication has far-reaching significance for human society. The sequenced genomes of domesticated animals provide critical resources for understanding the genetic basis of domestication. Various genomic analyses have shed a new light on the mechanism of artificial selection and have allowed the mapping of genes involved in important domestication traits. Here, we summarize the published genomes of domesticated animals that have been generated over the past decade, as well as their origins, from a phylogenomic point of view. This review provides a general description of the genomic features encountered under a two-stage domestication process. We also introduce recent findings for domestication traits based on results from genome-wide association studies and selective-sweep scans for artificially selected genomic regions. Particular attention is paid to issues relating to the costs of domestication and the convergent evolution of genes between domesticated animals and humans.

  15. Domestic Research at Rand.

    ERIC Educational Resources Information Center

    Rand Corp., Santa Monica, CA.

    This third descriptive inventory of the Rand Corporation's domestic non-military research covers all projects since 1972, as well as its special 1969-1975 program for New York City. For each of over 300 research projects in nine program areas, brief summaries identify the project title and sponsor, beginning and ending dates, person-years of work…

  16. Domestic Communications Satellites.

    ERIC Educational Resources Information Center

    Network Project Notebook, 1972

    1972-01-01

    The June, 1972 Federal Communications Commission's (FCC) decision allowed an "open skies" policy in regard to domestic communication satellites and raised Liberal opposition to a situation where exclusive and unchecked communications power is now in the hands of private entrepreneurs, primarily the big Defense Department oriented aerospace…

  17. Domestication and plant genomes.

    PubMed

    Tang, Haibao; Sezen, Uzay; Paterson, Andrew H

    2010-04-01

    The techniques of plant improvement have been evolving with the advancement of technology, progressing from crop domestication by Neolithic humans to scientific plant breeding, and now including DNA-based genotyping and genetic engineering. Archeological findings have shown that early human ancestors often unintentionally selected for and finally fixed a few major domestication traits over time. Recent advancement of molecular and genomic tools has enabled scientists to pinpoint changes to specific chromosomal regions and genetic loci that are responsible for dramatic morphological and other transitions that distinguish crops from their wild progenitors. Extensive studies in a multitude of additional crop species, facilitated by rapid progress in sequencing and resequencing(s) of crop genomes, will further our understanding of the genomic impact from both the unusual population history of cultivated plants and millennia of human selection.

  18. Solution structure of the Mu end DNA-binding ibeta subdomain of phage Mu transposase: modular DNA recognition by two tethered domains.

    PubMed Central

    Schumacher, S; Clubb, R T; Cai, M; Mizuuchi, K; Clore, G M; Gronenborn, A M

    1997-01-01

    The phage Mu transposase (MuA) binds to the ends of the Mu genome during the assembly of higher order nucleoprotein complexes. We investigate the structure and function of the MuA end-binding domain (Ibetagamma). The three-dimensional solution structure of the Ibeta subdomain (residues 77-174) has been determined using multidimensional NMR spectroscopy. It comprises five alpha-helices, including a helix-turn-helix (HTH) DNA-binding motif formed by helices 3 and 4, and can be subdivided into two interacting structural elements. The structure has an elongated disc-like appearance from which protrudes the recognition helix of the HTH motif. The topology of helices 2-4 is very similar to that of helices 1-3 of the previously determined solution structure of the MuA Igamma subdomain and to that of the homeodomain family of HTH DNA-binding proteins. We show that each of the two subdomains binds to one half of the 22 bp recognition sequence, Ibeta to the more conserved Mu end distal half (beta subsite) and Igamma to the Mu end proximal half (gamma subsite) of the consensus Mu end-binding site. The complete Ibetagamma domain binds the recognition sequence with a 100- to 1000-fold higher affinity than the two subdomains independently, indicating a cooperative effect. Our results show that the Mu end DNA-binding domain of MuA has a modular organization, with each module acting on a specific part of the 22 bp binding site. Based on the present binding data and the structures of the Ibeta and Igamma subdomains, a model for the interaction of the complete Ibetagamma domain with DNA is proposed. PMID:9405381

  19. Deletion of the GI-2 integrase and the wbkA flanking transposase improves the stability of Brucella melitensis Rev 1 vaccine

    PubMed Central

    2013-01-01

    Brucella melitensis Rev 1 is the best vaccine available for the prophylaxis of small ruminant brucellosis and, indirectly, for reducing human brucellosis. However, Rev 1 shows anomalously high rates of spontaneous dissociation from smooth (S) to rough (R) bacteria, the latter being inefficacious as vaccines. This S-R instability results from the loss of the O-polysaccharide. To overcome this problem, we investigated whether some recently described mechanisms promoting mutations in O-polysaccharide genes were involved in Rev 1 S-R dissociation. We found that a proportion of Rev 1 R mutants result from genome rearrangements affecting the wbo O-polysaccharide loci of genomic island GI-2 and the wbkA O-polysaccharide glycosyltransferase gene of the wbk region. Accordingly, we mutated the GI-2 int gene and the wbk IS transposase involved in those arrangements, and found that these Rev 1 mutants maintained the S phenotype and showed lower dissociation levels. Combining these two mutations resulted in a strain (Rev 2) displaying a 95% decrease in dissociation with respect to parental Rev 1 under conditions promoting dissociation. Rev 2 did not differ from Rev 1 in the characteristics used in Rev 1 typing (growth rate, colonial size, reactivity with O-polysaccharide antibodies, phage, dye and antibiotic susceptibility). Moreover, Rev 2 and Rev 1 showed similar attenuation and afforded similar protection in the mouse model of brucellosis vaccines. We conclude that mutations targeting genes and DNA sequences involved in spontaneous O-polysaccharide loss enhance the stability of a critical vaccine phenotype and complement the empirical stabilization precautions taken during S Brucella vaccine production. PMID:24176078

  20. Domestic Violence: Protecting Yourself and Your Children

    MedlinePlus

    MENU Return to Web version Domestic Violence: Protecting Yourself and Your Children Domestic Violence: Protecting Yourself and Your Children What is domestic violence? Domestic violence is abuse by a ...

  1. Genetic perspectives on crop domestication.

    PubMed

    Gross, Briana L; Olsen, Kenneth M

    2010-09-01

    The process of crop domestication has long been a topic of active research for biologists, anthropologists and others. Genetic data have proved a powerful resource for drawing inferences on questions regarding the geographical origins of crops, the numbers of independent domestication events for a given crop species, the specific molecular changes underlying domestication traits, and the nature of artificial selection during domestication and subsequent crop improvement. We would argue that these genetic inferences are fundamentally compatible with recent archaeological data that support a view of domestication as a geographically diffuse, gradual process. In this review, we summarize methodologies ranging from quantitative trait locus mapping to resequencing used in genetic analyses of crop evolution. We also highlight recent major insights regarding the timing and spatial patterning of crop domestication and the distinct genetic underpinnings of domestication, diversification and improvement traits.

  2. Animal evolution during domestication: the domesticated fox as a model

    PubMed Central

    Trut, Lyudmila; Oskina, Irina; Kharlamova, Anastasiya

    2009-01-01

    Summary We review the evolution of domestic animals, emphasizing the effect of the earliest steps of domestication on its course. Using the first domesticated species, the dog (Canis familiaris) as an illustration, we describe the evolutionary specificities of the historical domestication, such as the high level and wide range of diversity. We suggest that the process of earliest domestication via unconscious and later conscious selection of human-defined behavioral traits may accelerate phenotypic variations. The review is based on the results of the long-term experiment designed to reproduce early mammalian domestication in the silver fox (Vulpes vulpes) selected for tameability, or amenability to domestication. We describe changes in behavior, morphology and physiology that appeared in the fox during its selection for tameability and that were similar to those observed in the domestic dog. Based on the experimental fox data and survey of relevant data, we discuss the developmental, genetic and possible molecular-genetic mechanisms of these changes. We assign the causative role in evolutionary transformation of domestic animals to selection for behavior and to the neurospecific regulatory genes it affects. PMID:19260016

  3. Genetic analysis of wheat domestication and evolution under domestication.

    PubMed

    Peleg, Zvi; Fahima, Tzion; Korol, Abraham B; Abbo, Shahal; Saranga, Yehoshua

    2011-10-01

    Wheat is undoubtedly one of the world's major food sources since the dawn of Near Eastern agriculture and up to the present day. Morphological, physiological, and genetic modifications involved in domestication and subsequent evolution under domestication were investigated in a tetraploid recombinant inbred line population, derived from a cross between durum wheat and its immediate progenitor wild emmer wheat. Experimental data were used to test previous assumptions regarding a protracted domestication process. The brittle rachis (Br) spike, thought to be a primary characteristic of domestication, was mapped to chromosome 2A as a single gene, suggesting, in light of previously reported Br loci (homoeologous group 3), a complex genetic model involved in spike brittleness. Twenty-seven quantitative trait loci (QTLs) conferring threshability and yield components (kernel size and number of kernels per spike) were mapped. The large number of QTLs detected in this and other studies suggests that following domestication, wheat evolutionary processes involved many genomic changes. The Br gene did not show either genetic (co-localization with QTLs) or phenotypic association with threshability or yield components, suggesting independence of the respective loci. It is argued here that changes in spike threshability and agronomic traits (e.g. yield and its components) are the outcome of plant evolution under domestication, rather than the result of a protracted domestication process. Revealing the genomic basis of wheat domestication and evolution under domestication, and clarifying their inter-relationships, will improve our understanding of wheat biology and contribute to further crop improvement.

  4. Animal evolution during domestication: the domesticated fox as a model.

    PubMed

    Trut, Lyudmila; Oskina, Irina; Kharlamova, Anastasiya

    2009-03-01

    We review the evolution of domestic animals, emphasizing the effect of the earliest steps of domestication on its course. Using the first domesticated species, the dog (Canis familiaris), for illustration, we describe the evolutionary peculiarities during the historical domestication, such as the high level and wide range of diversity. We suggest that the process of earliest domestication via unconscious and later conscious selection of human-defined behavioral traits may accelerate phenotypic variations. The review is based on the results of a long-term experiment designed to reproduce early mammalian domestication in the silver fox (Vulpes vulpes) selected for tameability or amenability to domestication. We describe changes in behavior, morphology and physiology that appeared in the fox during its selection for tameability, which were similar to those observed in the domestic dog. Based on the data of the fox experiment and survey of relevant data, we discuss the developmental, genetic and possible molecular genetic mechanisms underlying these changes. We ascribe the causative role in evolutionary transformation of domestic animals to the selection for behavior and to the neurospecific regulatory genes it affects.

  5. Core questions in domestication research.

    PubMed

    Zeder, Melinda A

    2015-03-17

    The domestication of plants and animals is a key transition in human history, and its profound and continuing impacts are the focus of a broad range of transdisciplinary research spanning the physical, biological, and social sciences. Three central aspects of domestication that cut across and unify this diverse array of research perspectives are addressed here. Domestication is defined as a distinctive coevolutionary, mutualistic relationship between domesticator and domesticate and distinguished from related but ultimately different processes of resource management and agriculture. The relative utility of genetic, phenotypic, plastic, and contextual markers of evolving domesticatory relationships is discussed. Causal factors are considered, and two leading explanatory frameworks for initial domestication of plants and animals, one grounded in optimal foraging theory and the other in niche-construction theory, are compared.

  6. Domestic and Domesticating Education in the Late Victorian City

    ERIC Educational Resources Information Center

    Heggie, Vanessa

    2011-01-01

    This article explores the various types of domestic education, particularly cookery, available in Manchester between 1870 and 1902. The work of the two local School Boards and the Manchester School of Domestic Economy are shown as part of a complicated network of provision--a mixed economy of welfare, including enthusiastic philanthropists and…

  7. [Domestic violence: any progress?].

    PubMed

    Henrion, Roger

    2014-01-01

    Since the publication of the French national survey of violence against women in 2000, the fight against domestic violence has made steady progress. Knowledge of the phenomenon has significantly improved. A nationwide study of murders and manslaughters perpetrated by one partner of a couple against the other has been published annually since 2006. In 2012, domestic violence resulted in the deaths of 314 persons: 166 women, 31 men, 25 children, 9 collateral victims, 14 rivals, and two former spouses killed by their ex-fathers in law. In addition, 67 perpetrators committed suicide (51 men and3 women). The number of victims fluctuates from year to year but has remained fairly stable since 2006 (n=168). Legislation has improved significantly: eight new laws have been passed since 2004, all designed to protect women and to ensure that violent men are restrained and treated. New measures to inform and protect women have been implemented and others have been improved, such as the anonymous helpline (phone no 3919, "domestic violence information"). An inter-ministerial committee on the protection of women from violence and the prevention of human trafficking (MIPROF) was created on 3 January 2013. A website entitled "Stop violence against women " (Stop violences faites aux femmes) is now available. The "Imminent Danger" mobile phone system, designed to alert police if a suspected or known perpetrator breaches restraint conditions, will be extended to the entire country from January 2014. Referees charged with coordinating comprehensive long-tern care of women victims have been deployed at the county level. Information centers on the rights of women and families (CIDFF) now form a local nationwide network. Routine interviews with a midwife during the fourth month of pregnancy, focusing on the woman's emotional, economic and social conditions, have been implemented in 21 % of maternity units and should gradually be generalized. The authorities who have enforced the law have

  8. [Domestic violence: any progress?].

    PubMed

    Henrion, Roger

    2014-01-01

    Since the publication of the French national survey of violence against women in 2000, the fight against domestic violence has made steady progress. Knowledge of the phenomenon has significantly improved. A nationwide study of murders and manslaughters perpetrated by one partner of a couple against the other has been published annually since 2006. In 2012, domestic violence resulted in the deaths of 314 persons: 166 women, 31 men, 25 children, 9 collateral victims, 14 rivals, and two former spouses killed by their ex-fathers in law. In addition, 67 perpetrators committed suicide (51 men and3 women). The number of victims fluctuates from year to year but has remained fairly stable since 2006 (n=168). Legislation has improved significantly: eight new laws have been passed since 2004, all designed to protect women and to ensure that violent men are restrained and treated. New measures to inform and protect women have been implemented and others have been improved, such as the anonymous helpline (phone no 3919, "domestic violence information"). An inter-ministerial committee on the protection of women from violence and the prevention of human trafficking (MIPROF) was created on 3 January 2013. A website entitled "Stop violence against women " (Stop violences faites aux femmes) is now available. The "Imminent Danger" mobile phone system, designed to alert police if a suspected or known perpetrator breaches restraint conditions, will be extended to the entire country from January 2014. Referees charged with coordinating comprehensive long-tern care of women victims have been deployed at the county level. Information centers on the rights of women and families (CIDFF) now form a local nationwide network. Routine interviews with a midwife during the fourth month of pregnancy, focusing on the woman's emotional, economic and social conditions, have been implemented in 21 % of maternity units and should gradually be generalized. The authorities who have enforced the law have

  9. Replacement Cost of Domestic Crude

    1994-12-01

    The DEEPWATER model forecasts the replacement cost of domestic crude oil for 13 offshore regions in the lower 48 states. The replacement cost of domestic crude oil is the constant or levelized selling price that will recover the full expense of exploration, development, and productions with a reasonable return on capital.

  10. The Complexities of Domestic Violence

    ERIC Educational Resources Information Center

    Dutton, Donald G.

    2007-01-01

    Comments on the article by Robert Bornstein, "The complex relationship between dependency and domestic violence: Converging psychological factors and social forces." Although a more focused examination of the psychological factors involved in domestic violence is welcome, there are some factual errors in Bornstein's article that need attention and…

  11. Plant domestication slows pest evolution.

    PubMed

    Turcotte, Martin M; Lochab, Amaneet K; Turley, Nash E; Johnson, Marc T J

    2015-09-01

    Agricultural practices such as breeding resistant varieties and pesticide use can cause rapid evolution of pest species, but it remains unknown how plant domestication itself impacts pest contemporary evolution. Using experimental evolution on a comparative phylogenetic scale, we compared the evolutionary dynamics of a globally important economic pest - the green peach aphid (Myzus persicae) - growing on 34 plant taxa, represented by 17 crop species and their wild relatives. Domestication slowed aphid evolution by 13.5%, maintained 10.4% greater aphid genotypic diversity and 5.6% higher genotypic richness. The direction of evolution (i.e. which genotypes increased in frequency) differed among independent domestication events but was correlated with specific plant traits. Individual-based simulation models suggested that domestication affects aphid evolution directly by reducing the strength of selection and indirectly by increasing aphid density and thus weakening genetic drift. Our results suggest that phenotypic changes during domestication can alter pest evolutionary dynamics.

  12. The perpetrators of domestic violence.

    PubMed

    Romans, S E; Poore, M R; Martin, J L

    2000-11-01

    There has been little useful research in recent years into those who perpetrate domestic violence. Domestic violence is always anchored in a social context in which the aspirations of men and women are dealt with unequally. The majority of perpetrators of domestic violence are men. Perpetrators are often young, troubled, unemployed, and of low self-esteem; they have often experienced abuse (of various types) themselves. However, these factors do not justify their abusive behaviour. General practitioners and other health workers have a responsibility to broach the subject of domestic violence with both perpetrators and victims. They are in a key position to break the silence that allows it to continue. Programs for stopping domestic violence can be effective for those who are motivated to change their behaviour and see the programs through to completion.

  13. The Paramecium germline genome provides a niche for intragenic parasitic DNA: evolutionary dynamics of internal eliminated sequences.

    PubMed

    Arnaiz, Olivier; Mathy, Nathalie; Baudry, Céline; Malinsky, Sophie; Aury, Jean-Marc; Denby Wilkes, Cyril; Garnier, Olivier; Labadie, Karine; Lauderdale, Benjamin E; Le Mouël, Anne; Marmignon, Antoine; Nowacki, Mariusz; Poulain, Julie; Prajer, Malgorzata; Wincker, Patrick; Meyer, Eric; Duharcourt, Sandra; Duret, Laurent; Bétermier, Mireille; Sperling, Linda

    2012-01-01

    Insertions of parasitic DNA within coding sequences are usually deleterious and are generally counter-selected during evolution. Thanks to nuclear dimorphism, ciliates provide unique models to study the fate of such insertions. Their germline genome undergoes extensive rearrangements during development of a new somatic macronucleus from the germline micronucleus following sexual events. In Paramecium, these rearrangements include precise excision of unique-copy Internal Eliminated Sequences (IES) from the somatic DNA, requiring the activity of a domesticated piggyBac transposase, PiggyMac. We have sequenced Paramecium tetraurelia germline DNA, establishing a genome-wide catalogue of -45,000 IESs, in order to gain insight into their evolutionary origin and excision mechanism. We obtained direct evidence that PiggyMac is required for excision of all IESs. Homology with known P. tetraurelia Tc1/mariner transposons, described here, indicates that at least a fraction of IESs derive from these elements. Most IES insertions occurred before a recent whole-genome duplication that preceded diversification of the P. aurelia species complex, but IES invasion of the Paramecium genome appears to be an ongoing process. Once inserted, IESs decay rapidly by accumulation of deletions and point substitutions. Over 90% of the IESs are shorter than 150 bp and present a remarkable size distribution with a -10 bp periodicity, corresponding to the helical repeat of double-stranded DNA and suggesting DNA loop formation during assembly of a transpososome-like excision complex. IESs are equally frequent within and between coding sequences; however, excision is not 100% efficient and there is selective pressure against IES insertions, in particular within highly expressed genes. We discuss the possibility that ancient domestication of a piggyBac transposase favored subsequent propagation of transposons throughout the germline by allowing insertions in coding sequences, a fraction of the

  14. Reproduction in domestic buffalo.

    PubMed

    Perera, B M A O

    2008-07-01

    The domestic buffalo is an indispensable livestock resource to millions of smallholder farmers in developing countries, particularly in Asia. Although its reproductive biology is basically similar to that of cattle, there are important differences and unique characteristics that need to be considered in order to apply modern reproductive technologies to improve its productivity. Under most smallholder production systems, the reproductive efficiency of buffalo is compromised by factors related to climate, management, nutrition and diseases. However, when managed and fed properly, buffalo can have good fertility and provide milk, calves and draught power over a long productive life. The basic technical problems associated with artificial insemination in buffalo were largely overcome two decades ago, but the technology has not had the expected impact in some developing countries, because largely of infrastructural and logistic problems. Approaches involving the use of hormones for treating anoestrus and for synchronizing oestrus have had varying rates of success, depending on the protocols used and the incidence of underlying problems that cause infertility. Embryo technologies such as multiple ovulation embryo transfer, in vitro embryo production, cryopreservation and cloning are being intensively studied but have had far lower success rates than in cattle. Improving the productivity of buffalo requires an understanding of their potential and limitations under each farming system, development of simple intervention strategies to ameliorate deficiencies in management, nutrition and healthcare, followed by judicious application of reproductive technologies that are sustainable with the resources available to buffalo farmers.

  15. Reproduction in domestic buffalo.

    PubMed

    Perera, B M A O

    2008-07-01

    The domestic buffalo is an indispensable livestock resource to millions of smallholder farmers in developing countries, particularly in Asia. Although its reproductive biology is basically similar to that of cattle, there are important differences and unique characteristics that need to be considered in order to apply modern reproductive technologies to improve its productivity. Under most smallholder production systems, the reproductive efficiency of buffalo is compromised by factors related to climate, management, nutrition and diseases. However, when managed and fed properly, buffalo can have good fertility and provide milk, calves and draught power over a long productive life. The basic technical problems associated with artificial insemination in buffalo were largely overcome two decades ago, but the technology has not had the expected impact in some developing countries, because largely of infrastructural and logistic problems. Approaches involving the use of hormones for treating anoestrus and for synchronizing oestrus have had varying rates of success, depending on the protocols used and the incidence of underlying problems that cause infertility. Embryo technologies such as multiple ovulation embryo transfer, in vitro embryo production, cryopreservation and cloning are being intensively studied but have had far lower success rates than in cattle. Improving the productivity of buffalo requires an understanding of their potential and limitations under each farming system, development of simple intervention strategies to ameliorate deficiencies in management, nutrition and healthcare, followed by judicious application of reproductive technologies that are sustainable with the resources available to buffalo farmers. PMID:18638124

  16. Mutualism breakdown in breadfruit domestication.

    PubMed

    Xing, Xiaoke; Koch, Alexander M; Jones, A Maxwell P; Ragone, Diane; Murch, Susan; Hart, Miranda M

    2012-03-22

    During the process of plant domestication, below-ground communities are rarely considered. Some studies have attempted to understand the changes in root symbionts owing to domestication, but little is known about how it influences mycorrhizal response in domesticated crops. We hypothesized that selection for above-ground traits may also result in decreased mycorrhizal abundance in roots. Breadfruit (Artocarpus sp.) has a long domestication history, with a strong geographical movement of cultivars from west to east across the Melanesian and Polynesian islands. Our results clearly show a decrease in arbuscular mycorrhizas (AMs) along a domestication gradient from wild to recently derived cultivars. We showed that the vesicular and arbuscular colonization rate decreased significantly in more recently derived breadfruit cultivars. In addition, molecular analyses of breadfruit roots indicated that AM fungal species richness also responded along the domestication gradient. These results suggest that human-driven selection for plant cultivars can have unintended effects on below-ground mutualists, with potential impacts on the stress tolerance of crops and long-term food security.

  17. gDNA Enrichment by a Transposase-based Technology for NGS Analysis of the Whole Sequence of BRCA1, BRCA2, and 9 Genes Involved in DNA Damage Repair

    PubMed Central

    Chevrier, Sandy; Boidot, Romain

    2014-01-01

    The widespread use of Next Generation Sequencing has opened up new avenues for cancer research and diagnosis. NGS will bring huge amounts of new data on cancer, and especially cancer genetics. Current knowledge and future discoveries will make it necessary to study a huge number of genes that could be involved in a genetic predisposition to cancer. In this regard, we developed a Nextera design to study 11 complete genes involved in DNA damage repair. This protocol was developed to safely study 11 genes (ATM, BARD1, BRCA1, BRCA2, BRIP1, CHEK2, PALB2, RAD50, RAD51C, RAD80, and TP53) from promoter to 3'-UTR in 24 patients simultaneously. This protocol, based on transposase technology and gDNA enrichment, gives a great advantage in terms of time for the genetic diagnosis thanks to sample multiplexing. This protocol can be safely used with blood gDNA. PMID:25350069

  18. gDNA enrichment by a transposase-based technology for NGS analysis of the whole sequence of BRCA1, BRCA2, and 9 genes involved in DNA damage repair.

    PubMed

    Chevrier, Sandy; Boidot, Romain

    2014-10-06

    The widespread use of Next Generation Sequencing has opened up new avenues for cancer research and diagnosis. NGS will bring huge amounts of new data on cancer, and especially cancer genetics. Current knowledge and future discoveries will make it necessary to study a huge number of genes that could be involved in a genetic predisposition to cancer. In this regard, we developed a Nextera design to study 11 complete genes involved in DNA damage repair. This protocol was developed to safely study 11 genes (ATM, BARD1, BRCA1, BRCA2, BRIP1, CHEK2, PALB2, RAD50, RAD51C, RAD80, and TP53) from promoter to 3'-UTR in 24 patients simultaneously. This protocol, based on transposase technology and gDNA enrichment, gives a great advantage in terms of time for the genetic diagnosis thanks to sample multiplexing. This protocol can be safely used with blood gDNA.

  19. Child Domestic Work. Innocenti Digest 5.

    ERIC Educational Resources Information Center

    Black, Maggie, Comp.; Blagbrough, Jonathan, Comp.

    Child "domestics" or "domestic workers" are defined here as children under the age of 18 who work in other people's households, doing domestic chores, caring for children and running errands, among other tasks. This digest focuses mainly on the situation of live-in child domestics, that is, children who work full time in exchange for room, board,…

  20. Multi-perpetrator domestic violence.

    PubMed

    Salter, Michael

    2014-04-01

    A significant proportion of reports of domestic violence against women involve multiple perpetrators. Although the number of perpetrators has been consistently identified as a measure of abuse severity, only a minority of studies of domestic violence examine the role of multiple offenders. Data on multi-perpetrator domestic violence (MDV) is frequently removed from analysis in domestic violence studies, or multi-perpetrator incidents are treated as single-perpetrator incidents. However, the available research links MDV to negative mental and physical health outcomes, intimate partner homicide, homelessness among women, and severe mental illness and suicidality. This article reviews the available prevalence data on MDV and draws together research on the contexts in which MDV takes place. It highlights two groups that are particularly vulnerable to MDV: (1) girls and women partnered to members of gangs and organized crime groups and (2) girls and women in some ethnic minority communities. While discussions of honor in relation to domestic violence are often racialized in Western media, this article highlights the cross-cultural role of masculine honor in collective violence against women in the working class and impoverished communities of majority cultures as well as in migrant and ethnic minority communities. It is clear that such complex forms of violence present a range of challenges for intervention and treatment and the article emphasizes the need for specialized and coordinated modes of investigation, support, and care. PMID:24217092

  1. Multi-perpetrator domestic violence.

    PubMed

    Salter, Michael

    2014-04-01

    A significant proportion of reports of domestic violence against women involve multiple perpetrators. Although the number of perpetrators has been consistently identified as a measure of abuse severity, only a minority of studies of domestic violence examine the role of multiple offenders. Data on multi-perpetrator domestic violence (MDV) is frequently removed from analysis in domestic violence studies, or multi-perpetrator incidents are treated as single-perpetrator incidents. However, the available research links MDV to negative mental and physical health outcomes, intimate partner homicide, homelessness among women, and severe mental illness and suicidality. This article reviews the available prevalence data on MDV and draws together research on the contexts in which MDV takes place. It highlights two groups that are particularly vulnerable to MDV: (1) girls and women partnered to members of gangs and organized crime groups and (2) girls and women in some ethnic minority communities. While discussions of honor in relation to domestic violence are often racialized in Western media, this article highlights the cross-cultural role of masculine honor in collective violence against women in the working class and impoverished communities of majority cultures as well as in migrant and ethnic minority communities. It is clear that such complex forms of violence present a range of challenges for intervention and treatment and the article emphasizes the need for specialized and coordinated modes of investigation, support, and care.

  2. State Support of Domestic Production

    SciTech Connect

    Amy Wright

    2007-12-30

    This project was developed in response to a cooperative agreement offering by the U.S. Department of Energy (DOE) and the National Energy Technology Laboratory (NETL) under the State Support of Domestic Production DE-FC26-04NT15456. The Interstate Oil and Gas Compact Commission (IOGCC) performed efforts in support of State programs related to the security, reliability and growth if our nation's domestic production of oil and natural gas. The project objectives were to improve the States ability to monitor the security of oil and gas operations; to maximize the production of domestic oil and natural gas thereby minimizing the threat to national security posed by interruptions in energy imports; to assist States in developing and maintaining high standards of environmental protection; to assist in addressing issues that limit the capacity of the industry; to promote the deployment of the appropriate application of technology for regulatory efficiency; and to inform the public about emerging energy issues.

  3. Ovarian disorders in domestic animals.

    PubMed Central

    MacLachlan, N J

    1987-01-01

    The histologic appearance of the ovaries and persistence of corpora lutea vary considerably among domestic animals, particularly between spontaneous and induced ovulators. The seasonally polyestrous mare has a variety of unique characteristics in ovarian structure and general reproductive function. Among the anomalies of ovarian development is the bovine freemartin with gonads containing a mixture of male and female elements. A variety of ovarian cysts occur in domestic animals, and persistent corpora lutea with associated reproductive perturbations occur in several species. Ovarian tumors are relatively uncommon in domestic animals, with most examples described in dogs, cats, and horses. These ovarian neoplasms are generally classified as epithelial, germ cell, or sex cord-stromal tumors. PMID:3665869

  4. Cohabitation Duration and Transient Domesticity.

    PubMed

    Golub, Andrew; Reid, Megan; Strickler, Jennifer; Dunlap, Eloise

    2013-01-01

    Research finds that many impoverished urban Black adults engage in a pattern of partnering and family formation involving a succession of short cohabitations yielding children, a paradigm referred to as transient domesticity. Researchers have identified socioeconomic status, cultural adaptations, and urbanicity as explanations for aspects of this pattern. We used longitudinal data from the 2001 Survey of Income and Program Participation to analyze variation in cohabitation and marriage duration by race/ethnicity, income, and urban residence. Proportional hazards regression indicated that separation risk is greater among couples that are cohabiting, below 200% of the federal poverty line, and Black but is not greater among urban dwellers. This provides empirical demographic evidence to support the emerging theory of transient domesticity and suggests that both socioeconomic status and race explain this pattern. We discuss the implications of these findings for understanding transient domesticity and make recommendations for using the Survey of Income and Program Participation to further study this family formation paradigm.

  5. Assessment of Domestic Appliance Noise.

    NASA Astrophysics Data System (ADS)

    Brooks, Jeanette Rosamond

    Available from UMI in association with The British Library. The aims of this study were: (i) to identify the factors involved in eliciting a subjective reaction to domestic appliance noise, (ii) to identify the noise index (or indices) that correlate highly with a subjective reaction to the noise, and (iii) to investigate the contribution of domestic appliance noise to an individual's daily noise dose. Two series of experimental studies were carried out using several examples of each of five types of domestic appliances. One determined the index values of domestic appliance noise--namely L_{WA} (using ISO 3741), L_{pA} , L_{pD}, L _{p}, PNL, L_{Aeq, 30 sec}, L_{Amax} and L_{AX}; the other determined subjective reactions to domestic appliance noise (judgements of noisiness, annoyance, the acceptability of the appliance noise and appraisals of usefulness). The success or failure of the research hypotheses was assessed statistically by analysis of variance, regression analysis, log linear analysis, Hotelling test, bootstrapping, t-test and post-hoc comparisons. Ratings of annoyance, noisiness and the acceptability of the noise of the appliance were found to be interrelated and interdependent, and not influenced by appraisals of usefulness of the appliances. Noisiness ratings were the most consistent of the subjective ratings investigated, and were influenced by the duration of the exposure, and the actual appliance type under investigation. Significant correlations were obtained between noisiness ratings and all the noise indices under investigation. However, statistical analysis demonstrated that L_ {WA} correlated less successfully with noisiness ratings than all other indices. L_ {Amax},L_{Aeq,30 sec }, and L_{AX} indices were the most successful. It is therefore suggested that the labelling of domestic appliance noise consist of L_{WA} and L _{Aeq} as measured in a standardised test environment. The percentage contribution of domestic appliance noise to the total

  6. Superheroes and masterminds of plant domestication.

    PubMed

    Martínez-Ainsworth, Natalia E; Tenaillon, Maud I

    2016-01-01

    Domestication is one of the most fundamental changes in the evolution of human societies. The geographical origins of domesticated plants are inferred from archaeology, ecology and genetic data. Scenarios vary among species and include single, diffuse or multiple independent domestications. Cultivated plants present a panel of traits, the "domestication syndrome" that distinguish them from their wild relatives. It encompasses yield-, food usage-, and cultivation-related traits. Most genes underlying those traits are "masterminds" affecting the regulation of gene networks. Phenotypic convergence of domestication traits across species or within species between independently domesticated forms rarely coincides with convergence at the gene level. We review here current data/models that propose a protracted transition model for domestication and investigate the impact of mating system, life cycle and gene flow on the pace of domestication. Finally, we discuss the cost of domestication, pointing to the importance of characterizing adaptive functional variation in wild resources. PMID:27317057

  7. Superheroes and masterminds of plant domestication.

    PubMed

    Martínez-Ainsworth, Natalia E; Tenaillon, Maud I

    2016-01-01

    Domestication is one of the most fundamental changes in the evolution of human societies. The geographical origins of domesticated plants are inferred from archaeology, ecology and genetic data. Scenarios vary among species and include single, diffuse or multiple independent domestications. Cultivated plants present a panel of traits, the "domestication syndrome" that distinguish them from their wild relatives. It encompasses yield-, food usage-, and cultivation-related traits. Most genes underlying those traits are "masterminds" affecting the regulation of gene networks. Phenotypic convergence of domestication traits across species or within species between independently domesticated forms rarely coincides with convergence at the gene level. We review here current data/models that propose a protracted transition model for domestication and investigate the impact of mating system, life cycle and gene flow on the pace of domestication. Finally, we discuss the cost of domestication, pointing to the importance of characterizing adaptive functional variation in wild resources.

  8. Domestic Violence. Technical Assistance Packet.

    ERIC Educational Resources Information Center

    Join Together, Boston, MA.

    Substance abuse has long been recognized as a precipitating factor in many domestic violence incidents. The main type of substance abuse is alcohol usage. Forty-six percent of the offenders reported being dependent on or abusing alcohol, while another 28% were found to be dependent on opiates, cocaine, marijuana, or inhalants. Nearly two-fifths of…

  9. Girl domestic workers in Kenya.

    PubMed

    Mzungu, M

    1999-03-01

    This article exposes the conditions among children who are forced by their poor families to assume domestic work in households in Kenya. It is an accepted practice for parents to place daughters in households to help with housework and baby-sitting. The Sinaga Women and Child Labor Resource Center in Nairobi finds this exploitative and part of a wider practice that institutionalizes violence against women. The Center was established in 1995 to challenge the practice of child domestic labor. The Center's research reveals that child domestic workers tend to come from large, poor, and rural families or from urban slums. Wages are low or exchanged for shoes, clothes, and food. The hours of work are long. Mistreatment may include sexual molestation by male household members, beatings, verbal abuse, and mistrust. There is little recourse. Complaints from child workers or others outside the household can result in further mistreatment. Action against mistreatment is complicated by the prevailing image of activists as frustrated women with vendettas against men. The Center focuses on rehabilitation, literacy training, marketable skill development, and awareness creation. Counseling includes parents, children, and employers. Public awareness campaigns have resulted in employer referrals of youth workers for training. Other groups are joining the effort to improve conditions for child domestic workers.

  10. The Japanese Domestic Labor Debate.

    ERIC Educational Resources Information Center

    Ueno, Chizuko

    The changing role of Japanese women can be seen in the stages of a domestic labor debate which occurred at three different times in the past 30 years. The first debate began with Ayako Ishigaki's (1955) insistence that women should have a job outside the home. Wartime production helped break down traditional divisions of labor by encouraging women…

  11. Genetic analysis of safflower domestication

    PubMed Central

    2014-01-01

    Background Safflower (Carthamus tinctorius L.) is an oilseed crop in the Compositae (a.k.a. Asteraceae) that is valued for its oils rich in unsaturated fatty acids. Here, we present an analysis of the genetic architecture of safflower domestication and compare our findings to those from sunflower (Helianthus annuus L.), an independently domesticated oilseed crop within the same family. We mapped quantitative trait loci (QTL) underlying 24 domestication-related traits in progeny from a cross between safflower and its wild progenitor, Carthamus palaestinus Eig. Also, we compared QTL positions in safflower against those that have been previously identified in cultivated x wild sunflower crosses to identify instances of colocalization. Results We mapped 61 QTL, the vast majority of which (59) exhibited minor or moderate phenotypic effects. The two large-effect QTL corresponded to one each for flower color and leaf spininess. A total of 14 safflower QTL colocalized with previously reported sunflower QTL for the same traits. Of these, QTL for three traits (days to flower, achene length, and number of selfed seed) had cultivar alleles that conferred effects in the same direction in both species. Conclusions As has been observed in sunflower, and unlike many other crops, our results suggest that the genetics of safflower domestication is quite complex. Moreover, our comparative mapping results indicate that safflower and sunflower exhibit numerous instances of QTL colocalization, suggesting that parallel trait transitions during domestication may have been driven, at least in part, by parallel genotypic evolution at some of the same underlying genes. PMID:24502326

  12. A pipeline strategy for grain crop domestication

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In recent decades, in the interest of diversifying the global food system, improving human nutrition, or making agriculture more sustainable, there have been many proposals for domesticating or completing the domestication of wild plants or semi-domesticated “orphan” crops. However, very few new cro...

  13. 25 CFR 11.454 - Domestic violence.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Domestic violence. 11.454 Section 11.454 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR LAW AND ORDER COURTS OF INDIAN OFFENSES AND LAW AND ORDER CODE Criminal Offenses § 11.454 Domestic violence. (a) A person who commits domestic violence...

  14. 25 CFR 11.454 - Domestic violence.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 25 Indians 1 2012-04-01 2011-04-01 true Domestic violence. 11.454 Section 11.454 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR LAW AND ORDER COURTS OF INDIAN OFFENSES AND LAW AND ORDER CODE Criminal Offenses § 11.454 Domestic violence. (a) A person who commits domestic violence by...

  15. 25 CFR 11.454 - Domestic violence.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 25 Indians 1 2014-04-01 2014-04-01 false Domestic violence. 11.454 Section 11.454 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR LAW AND ORDER COURTS OF INDIAN OFFENSES AND LAW AND ORDER CODE Criminal Offenses § 11.454 Domestic violence. (a) A person who commits domestic violence...

  16. 25 CFR 11.454 - Domestic violence.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 25 Indians 1 2013-04-01 2013-04-01 false Domestic violence. 11.454 Section 11.454 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR LAW AND ORDER COURTS OF INDIAN OFFENSES AND LAW AND ORDER CODE Criminal Offenses § 11.454 Domestic violence. (a) A person who commits domestic violence...

  17. 25 CFR 11.454 - Domestic violence.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 25 Indians 1 2011-04-01 2011-04-01 false Domestic violence. 11.454 Section 11.454 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR LAW AND ORDER COURTS OF INDIAN OFFENSES AND LAW AND ORDER CODE Criminal Offenses § 11.454 Domestic violence. (a) A person who commits domestic violence...

  18. 31 CFR 515.320 - Domestic bank.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Domestic bank. 515.320 Section 515... § 515.320 Domestic bank. The term domestic bank shall mean any branch or office within the United States of any of the following which is not a national of a designated foreign country; any bank or...

  19. 31 CFR 535.320 - Domestic bank.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Domestic bank. 535.320 Section 535... § 535.320 Domestic bank. (a) The term domestic bank shall mean any branch or office within the United States of any of the following which is not Iran or an Iranian entity: any bank or trust...

  20. 31 CFR 500.320 - Domestic bank.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Domestic bank. 500.320 Section 500... § 500.320 Domestic bank. The term domestic bank shall mean any branch or office within the United States of any of the following which is not a national of any designated foreign country: Any bank or...

  1. Risk Factors for Domestic Violence in Curacao

    ERIC Educational Resources Information Center

    van Wijk, N. Ph. L.; de Bruijn, J. G. M.

    2012-01-01

    One out of three people (25% of men, 38% of women) in Curacao have experienced some form of domestic violence at some point in their adult lives. The most significant risk factors for domestic violence in Curacao are the female gender, a young age, low education, and experiencing domestic violence victimization in childhood. Divorce, single…

  2. 31 CFR 515.320 - Domestic bank.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 31 Money and Finance:Treasury 3 2012-07-01 2012-07-01 false Domestic bank. 515.320 Section 515.320... Domestic bank. The term domestic bank shall mean any branch or office within the United States of any of the following which is not a national of a designated foreign country; any bank or trust...

  3. 31 CFR 535.320 - Domestic bank.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 31 Money and Finance:Treasury 3 2012-07-01 2012-07-01 false Domestic bank. 535.320 Section 535.320....320 Domestic bank. (a) The term domestic bank shall mean any branch or office within the United States of any of the following which is not Iran or an Iranian entity: any bank or trust...

  4. 31 CFR 515.320 - Domestic bank.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 31 Money and Finance:Treasury 3 2014-07-01 2014-07-01 false Domestic bank. 515.320 Section 515.320... Domestic bank. The term domestic bank shall mean any branch or office within the United States of any of the following which is not a national of a designated foreign country; any bank or trust...

  5. 31 CFR 535.320 - Domestic bank.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 31 Money and Finance:Treasury 3 2013-07-01 2013-07-01 false Domestic bank. 535.320 Section 535.320....320 Domestic bank. (a) The term domestic bank shall mean any branch or office within the United States of any of the following which is not Iran or an Iranian entity: any bank or trust...

  6. 31 CFR 515.320 - Domestic bank.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 31 Money and Finance:Treasury 3 2013-07-01 2013-07-01 false Domestic bank. 515.320 Section 515.320... Domestic bank. The term domestic bank shall mean any branch or office within the United States of any of the following which is not a national of a designated foreign country; any bank or trust...

  7. 31 CFR 535.320 - Domestic bank.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 31 Money and Finance:Treasury 3 2014-07-01 2014-07-01 false Domestic bank. 535.320 Section 535.320....320 Domestic bank. (a) The term domestic bank shall mean any branch or office within the United States of any of the following which is not Iran or an Iranian entity: any bank or trust...

  8. 31 CFR 515.320 - Domestic bank.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 31 Money and Finance:Treasury 3 2011-07-01 2011-07-01 false Domestic bank. 515.320 Section 515.320... Domestic bank. The term domestic bank shall mean any branch or office within the United States of any of the following which is not a national of a designated foreign country; any bank or trust...

  9. 31 CFR 535.320 - Domestic bank.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 31 Money and Finance:Treasury 3 2011-07-01 2011-07-01 false Domestic bank. 535.320 Section 535.320....320 Domestic bank. (a) The term domestic bank shall mean any branch or office within the United States of any of the following which is not Iran or an Iranian entity: any bank or trust...

  10. Genetics and consequences of crop domestication

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phenotypic variation has been manipulated by humans during crop domestication, which occurred primarily between 3,000 and 10,000 years ago in the various centers of origin around the world. The process of domestication has profound consequences on crops, where the domesticate has moderately reduced ...

  11. 19 CFR 123.13 - Foreign repairs to domestic locomotives and other domestic railroad equipment.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... domestic railroad equipment. 123.13 Section 123.13 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION... International Traffic § 123.13 Foreign repairs to domestic locomotives and other domestic railroad equipment. A report of the first arrival in the United States of a domestic locomotive or other railroad...

  12. 19 CFR 123.13 - Foreign repairs to domestic locomotives and other domestic railroad equipment.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... domestic railroad equipment. 123.13 Section 123.13 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION... International Traffic § 123.13 Foreign repairs to domestic locomotives and other domestic railroad equipment. A report of the first arrival in the United States of a domestic locomotive or other railroad...

  13. 19 CFR 123.13 - Foreign repairs to domestic locomotives and other domestic railroad equipment.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... domestic railroad equipment. 123.13 Section 123.13 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION... International Traffic § 123.13 Foreign repairs to domestic locomotives and other domestic railroad equipment. A report of the first arrival in the United States of a domestic locomotive or other railroad...

  14. 19 CFR 123.13 - Foreign repairs to domestic locomotives and other domestic railroad equipment.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... domestic railroad equipment. 123.13 Section 123.13 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION... International Traffic § 123.13 Foreign repairs to domestic locomotives and other domestic railroad equipment. A report of the first arrival in the United States of a domestic locomotive or other railroad...

  15. Genetic testing in domestic cats.

    PubMed

    Lyons, Leslie A

    2012-12-01

    Varieties of genetic tests are currently available for the domestic cat that support veterinary health care, breed management, species identification, and forensic investigations. Approximately thirty-five genes contain over fifty mutations that cause feline health problems or alterations in the cat's appearance. Specific genes, such as sweet and drug receptors, have been knocked-out of Felidae during evolution and can be used along with mtDNA markers for species identification. Both STR and SNP panels differentiate cat race, breed, and individual identity, as well as gender-specific markers to determine sex of an individual. Cat genetic tests are common offerings for commercial laboratories, allowing both the veterinary clinician and the private owner to obtain DNA test results. This article will review the genetic tests for the domestic cat, and their various applications in different fields of science. Highlighted are genetic tests specific to the individual cat, which are a part of the cat's genome.

  16. Ferrochromium from Domestic Lateritic Chromites

    NASA Astrophysics Data System (ADS)

    Nafziger, R. H.

    1988-09-01

    As part of its effort to devise suitable technology for processing low-grade domestic materials and recycling wastes, the U.S. Bureau of Mines has evaluated the feasibility of smelting a chromite concentrate derived from residues generated by the processing of nickel and cobalt from domestic later-ites. The product sought was a high-carbon ferrochromium suitable for stainless- and alloy-steel production. The concentrate was blended with re-ductants and fluxing constituents and was smelted under submerged arc conditions in a laboratory-scale, single-phase ac electric arc furnace. The results revealed that metallurgical-grade coke provides the best quality product. High-carbon ferrochromium, which met ASTM specifications except for phosphorus and sulfur, was obtained. Agglomeration of the charge materials was not required.

  17. Artificial cloning of domestic animals.

    PubMed

    Keefer, Carol L

    2015-07-21

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research.

  18. Artificial cloning of domestic animals.

    PubMed

    Keefer, Carol L

    2015-07-21

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research. PMID:26195770

  19. Artificial cloning of domestic animals

    PubMed Central

    Keefer, Carol L.

    2015-01-01

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research. PMID:26195770

  20. Evolutionary genomics of dog domestication.

    PubMed

    Wayne, Robert K; vonHoldt, Bridgett M

    2012-02-01

    We review the underlying principles and tools used in genomic studies of domestic dogs aimed at understanding the genetic changes that have occurred during domestication. We show that there are two principle modes of evolution within dogs. One primary mode that accounts for much of the remarkable diversity of dog breeds is the fixation of discrete mutations of large effect in individual lineages that are then crossed to various breed groupings. This transfer of mutations across the dog evolutionary tree leads to the appearance of high phenotypic diversity that in actuality reflects a small number of major genes. A second mechanism causing diversification involves the selective breeding of dogs within distinct phenotypic or functional groups, which enhances specific group attributes such as heading or tracking. Such progressive selection leads to a distinct genetic structure in evolutionary trees such that functional and phenotypic groups cluster genetically. We trace the origin of the nuclear genome in dogs based on haplotype-sharing analyses between dogs and gray wolves and show that contrary to previous mtDNA analyses, the nuclear genome of dogs derives primarily from Middle Eastern or European wolves, a result more consistent with the archeological record. Sequencing analysis of the IGF1 gene, which has been the target of size selection in small breeds, further supports this conclusion. Finally, we discuss how a black coat color mutation that evolved in dogs has transformed North American gray wolf populations, providing a first example of a mutation that appeared under domestication and selectively swept through a wild relative. PMID:22270221

  1. [Major domestication traits in Asian rice].

    PubMed

    Ou, Shu-Jun; Wang, Hong-Ru; Chu, Cheng-Cai

    2012-11-01

    Rice (Oryza sativa L.) is an excellent model plant in elucidation of cereal domestication. Loss of seed shattering, weakened dormancy, and changes in plant architecture were thought to be three key events in the rice domestication and creating the high-yield, uniform-germinating, and densely-planting modern rice. Loss of shattering is considered to be the direct morphological evidence for identifying domesticated rice. Two major shattering QTLs, Sh4 and qSH1, have displayed different domestication histories. Weakened seed dormancy is essential for synchronous germination in agricultural production. Genes Sdr4, qSD7-1, and qSD12 impose a global and complementary adaptation strategies in controlling seed dormancy. The prostate growth habit of wild rice is an adaptation to disturbed habitats, while the erect growth habit of rice cultivars meet the needs of compact planting, and such a plant architecture is mainly controlled by PROG1. The outcrossing habit of wild rice promotes propagation of domestication genes among different populations, while the self-pollinating habit of cultivated rice facilitates fixation of domestication genes. Currently, the researches on rice domestication mainly focus on individual genes or multiple neutral markers, and much less attention has been paid to the evolution of network controlling domestication traits. With the progress in functional genomics research, the molecular mechanism of domestication traits is emerging. Rice domestication researches based on network will be more comprehensive and better reflect rice domestica-tion process. Here, we reviewed most progresses in molecular mechanisms of rice domestication traits, in order to provide the new insights for rice domestication and molecular breeding.

  2. Renal biomarkers in domestic species.

    PubMed

    Hokamp, Jessica A; Nabity, Mary B

    2016-03-01

    Current conventional tests of kidney damage and function in blood (serum creatinine and urea nitrogen) and urine (urine protein creatinine ratio and urine specific gravity) are widely used for diagnosis and monitoring of kidney disease. However, they all have important limitations, and additional markers of glomerular filtration rate and glomerular and tubular damage are desirable, particularly for earlier detection of renal disease when therapy is most effective. Additionally, urinary markers of kidney damage and function may help localize damage to the affected portion of the kidney. In general, the presence of high- and intermediate-molecular weight proteins in the urine are indicative of glomerular damage, while low-molecular weight proteins and enzymes in the urine suggest tubular damage due to decreased reabsorption of proteins, direct tubular damage, or both. This review aims to discuss many of these new blood and urinary biomarkers in domestic veterinary species, focusing primarily on dogs and cats, how they may be used for diagnosis of renal disease, and their limitations. Additionally, a brief discussion of serum creatinine is presented, highlighting its limitations and important considerations for its improved interpretation in domestic species based on past literature and recent studies. PMID:26918420

  3. Domestic Abuse in Behshahr, Iran

    PubMed Central

    Rahmatian, Ali Akbar; Hosseini, Seyyed Ali Asghar

    2015-01-01

    Background: The United Nations in a resolution defined abuse as any violent act that is primarily or exclusively committed against females and results in physical, sexual and psychological harm. Objectives: The aim of this research was to study the contributing factors of husband’s violence against females residing in the city of Behshahr, Iran. Materials and Methods: We distributed a specifically designed questionnaire among 380 married females aged between 15 and 65 years. According to the Morgan table, the subjects were randomly selected from a list of 301000 females. Demographic data and data on spouse abuse were then analyzed using the SPSS software, Spearman and Pearson correlation coefficients. According to Cronbach’s alpha, the reliability of the questionnaire was 0.96. Results: All of the females reported at least one form of violence within the past year, with R square 0.20, indicating that the independent variable can explain 20% of the violence against females. years of marriage, female’s education, male’s addiction and the number of children each had their share in the explanation of violence against females. Conclusions: This study revealed a high prevalence of domestic violence in the sample population. Violence existed among all ages, social categories and male occupational groups, and also involved both employed and unemployed females. The situation regarding domestic abuse is similar worldwide. PMID:26834799

  4. Genetic testing in domestic cats

    PubMed Central

    Lyons, Leslie A.

    2012-01-01

    Varieties of genetic tests are currently available for the domestic cat that support veterinary health care, breed management, species identification, and forensic investigations. Approximately thirty-five genes contain over fifty mutations that cause feline health problems or alterations in the cat’s appearance. Specific genes, such as sweet and drug receptors, have been knocked-out of Felidae during evolution and can be used along with mtDNA markers for species identification. Both STR and SNP panels differentiate cat race, breed, and individual identity, as well as gender-specific markers to determine sex of an individual. Cat genetic tests are common offerings for commercial laboratories, allowing both the veterinary clinician and the private owner to obtain DNA test results. This article will review the genetic tests for the domestic cat, and their various applications in different fields of science. Highlighted are genetic tests specific to the individual cat, which are a part of the cat’s genome. PMID:22546621

  5. Wheat domestication: lessons for the future.

    PubMed

    Charmet, Gilles

    2011-03-01

    Wheat was one of the first crops to be domesticated more than 10,000 years ago in the Middle East. Molecular genetics and archaeological data have allowed the reconstruction of plausible domestication scenarios leading to modern cultivars. For diploid einkorn and tetraploid durum wheat, a single domestication event has likely occurred in the Karacadag Mountains, Turkey. Following a cross between tetraploid durum and diploid T. tauschii, the resultant hexaploid bread wheat was domesticated and disseminated around the Caucasian region. These polyploidisation events facilitated wheat domestication and created genetic bottlenecks, which excluded potentially adaptive alleles. With the urgent need to accelerate genetic progress to confront the challenges of climate change and sustainable agriculture, wild ancestors and old landraces represent a reservoir of underexploited genetic diversity that may be utilized through modern breeding methods. Understanding domestication processes may thus help identifying new strategies.

  6. Genetics and consequences of crop domestication.

    PubMed

    Flint-Garcia, Sherry A

    2013-09-01

    Phenotypic variation has been manipulated by humans during crop domestication, which occurred primarily between 3000 and 10000 years ago in the various centers of origin around the world. The process of domestication has profound consequences on crops, where the domesticate has moderately reduced genetic diversity relative to the wild ancestor across the genome, and severely reduced diversity for genes targeted by domestication. The question that remains is whether reduction in genetic diversity has affected crop production today. A case study in maize ( Zea mays ) demonstrates the application of understanding relationships between genetic diversity and phenotypic diversity in the wild ancestor and the domesticate. As an outcrossing species, maize has tremendous genetic variation. The complementary combination of genome-wide association mapping (GWAS) approaches, large HapMap data sets, and germplasm resources is leading to important discoveries of the relationship between genetic diversity and phenotypic variation and the impact of domestication on trait variation.

  7. Toward understanding dog evolutionary and domestication history.

    PubMed

    Galibert, Francis; Quignon, Pascale; Hitte, Christophe; André, Catherine

    2011-03-01

    Dog domestication was probably started very early during the Upper paleolithic period (~35,000 BP), thus well before any other animal or plant domestication. This early process, probably unconscious, is called proto-domestication to distinguish it from the real domestication process that has been dated around 14,000 BC. Genomic DNA analyses have shown recently that domestication started in the Middle East and rapidly expanded into all human populations. Nowadays, the dog population is fragmented in several hundreds of breeds well characterized by their phenotypes that offer a unique spectrum of polymorphism. More recent studies detect genetic signatures that will be useful to highlight breed history as well as the impact of domestication at the DNA level.

  8. Chicken domestication: from archeology to genomics.

    PubMed

    Tixier-Boichard, Michèle; Bed'hom, Bertrand; Rognon, Xavier

    2011-03-01

    Current knowledge on chicken domestication is reviewed on the basis of archaeological, historical and molecular data. Several domestication centres have been identified in South and South-East Asia. Gallus gallus is the major ancestor species, but Gallus sonneratii has also contributed to the genetic make-up of the domestic chicken. Genetic diversity is now distributed among traditional populations, standardized breeds and highly selected lines. Knowing the genome sequence has accelerated the identification of causal mutations determining major morphological differences between wild Gallus and domestic breeds. Comparative genome resequencing between Gallus and domestic chickens has identified 21 selective sweeps, one involving a non-synonymous mutation in the TSHR gene, which functional consequences remain to be explored. The resequencing approach could also identify candidate genes responsible of quantitative traits loci (QTL) effects in selected lines. Genomics is opening new ways to understand major switches that took place during domestication and subsequent selection.

  9. Germline modification of domestic animals

    PubMed Central

    Tang, L.; González, R.; Dobrinski, I.

    2016-01-01

    Genetically-modified domestic animal models are of increasing significance in biomedical research and agriculture. As authentic ES cells derived from domestic animals are not yet available, the prevailing approaches for engineering genetic modifications in those animals are pronuclear microinjection and somatic cell nuclear transfer (SCNT, also known as cloning). Both pronuclear microinjection and SCNT are inefficient, costly, and time-consuming. In animals produced by pronuclear microinjection, the exogenous transgene is usually inserted randomly into the genome, which results in highly variable expression patterns and levels in different founders. Therefore, significant efforts are required to generate and screen multiple founders to obtain animals with optimal transgene expression. For SCNT, specific genetic modifications (both gain-of-function and loss-of-function) can be engineered and carefully selected in the somatic cell nucleus before nuclear transfer. SCNT has been used to generate a variety of genetically modified animals such as goats, pigs, sheep and cattle; however, animals resulting from SCNT frequently suffer from developmental abnormalities associated with incomplete nuclear reprogramming. Other strategies to generate genetically-modified animals rely on the use of the spermatozoon as a natural vector to introduce genetic material into the female gamete. This sperm mediated DNA transfer (SMGT) combined with intracytoplasmatic sperm injection (ICSI) has relatively high efficiency and allows the insertion of large DNA fragments, which, in turn, enhance proper gene expression. An approach currently being developed to complement SCNT for producing genetically modified animals is germ cell transplantation using genetically modified male germline stem cells (GSCs). This approach relies on the ability of GSCs that are genetically modified in vitro to colonize the recipient testis and produce donor derived sperm upon transplantation. As the genetic change

  10. The nature of selection during plant domestication.

    PubMed

    Purugganan, Michael D; Fuller, Dorian Q

    2009-02-12

    Plant domestication is an outstanding example of plant-animal co-evolution and is a far richer model for studying evolution than is generally appreciated. There have been numerous studies to identify genes associated with domestication, and archaeological work has provided a clear understanding of the dynamics of human cultivation practices during the Neolithic period. Together, these have provided a better understanding of the selective pressures that accompany crop domestication, and they demonstrate that a synthesis from the twin vantage points of genetics and archaeology can expand our understanding of the nature of evolutionary selection that accompanies domestication.

  11. Promoting domestic violence education for nurses.

    PubMed

    Schroeder, M; Weber, J R

    1998-01-01

    Domestic violence is one of the major health problems facing families today. Women in rural areas often are an overlooked population at high risk for this problem. Domestic violence is a concern for women, who may be patients or healthcare workers. Teaching about domestic violence is a very sensitive issue because it is often difficult for the abused to admit or confront that she is being abused. The authors developed an effective interactive workshop to teach nurses how to assess and intervene with women who have experienced domestic violence.

  12. Mothers, domestic violence, and child protection.

    PubMed

    Douglas, Heather; Walsh, Tamara

    2010-05-01

    This article explores the relationship between understandings of domestic violence and the child protection response drawing on material gathered in focus groups with workers who support mothers dealing with both domestic violence and child protection issues. The interviewees expressed concern that the dynamics of domestic violence are often misunderstood and inappropriately responded to by child protection workers. This article critically examines the interviewees' concerns and concludes that to properly protect children, it is crucial that child protection workers have a clear understanding of the dynamics of and issues related to domestic violence.

  13. Microbial prevalence in domestic humidifiers.

    PubMed Central

    Burge, H A; Solomon, W R; Boise, J R

    1980-01-01

    The prevalence of viable thermophilic bacteria and actinomycetes and mesophilic fungi was examined in 145 samples from 110 domestic humidifiers. A total of 72 and 43% of furnace and console humidifier samples, respectively, contained viable thermophilic bacteria, whereas 60 and 72% of these samples produced mesophilic fungal growth. Thermophilic actinomycetes were recovered from seven humidifier samples. Efforts to detect thermophilic actinomycete antigens in 15 humidifier fluid samples were not successful. Antifoulants added to humidifier fluid reservoirs had no apparent effect on microbial frequency. Airborne microbial recoveries did not reflect patterns of humidifier contamination with respect to either kinds or numbers of microorganisms in 20 homes in which volumetric air samples were obtained during humidifier operation. PMID:7377779

  14. Reduced domestic satellite orbit spacing

    NASA Astrophysics Data System (ADS)

    Sharp, G. L.

    The demand for services provided by communications satellites in geostationary orbit is growing, and problems arise with respect to the required increase in capacity. One approach for providing such an increase involves the employment of more satellites operating at smaller orbital spacings. The present investigation is concerned with the results of technical studies conducted by the Federal Communications Commission (FCC) to determine the feasibility of reducing orbital spacings between U.S. 'domestic fixed satellites' (domsats). Attention is given to details regarding the usable orbital arc, an adjacent satellite interference model, antenna sidelobe patterns, a single entry analysis, a 4/6 GHz aggregate analysis, results for the 4/6 GHz bands, results for the 12/14 GHz bands, data services, voice services, video reception, and high power spot beams.

  15. 76 FR 36512 - USDA Increases the Domestic Sugar Overall Allotment Quantity, Reassigns Domestic Cane Sugar...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-22

    ... Office of the Secretary USDA Increases the Domestic Sugar Overall Allotment Quantity, Reassigns Domestic Cane Sugar Allotments, and Increases the Fiscal Year 2011 Raw Sugar Tariff-Rate Quota AGENCY: Office of... in the domestic sugar Overall Allotment Quantity (OAQ); a reassignment of surplus sugar...

  16. Genetic divergence in domesticated and non-domesticated gene regions of barley chromosomes.

    PubMed

    Yan, Songxian; Sun, Dongfa; Sun, Genlou

    2015-01-01

    Little is known about the genetic divergence in the chromosomal regions with domesticated and non-domesticated genes. The objective of our study is to examine the effect of natural selection on shaping genetic diversity of chromosome region with domesticated and non-domesticated genes in barley using 110 SSR markers. Comparison of the genetic diversity loss between wild and cultivated barley for each chromosome showed that chromosome 5H had the highest divergence of 35.29%, followed by 3H, 7H, 4H, 2H, 6H. Diversity ratio was calculated as (diversity of wild type - diversity of cultivated type)/diversity of wild type×100%. It was found that diversity ratios of the domesticated regions on 5H, 1H and 7H were higher than those of non-domesticated regions. Diversity ratio of the domesticated region on 2H and 4H is similar to that of non-domesticated region. However, diversity ratio of the domesticated region on 3H is lower than that of non-domesticated region. Averaged diversity among six chromosomes in domesticated region was 33.73% difference between wild and cultivated barley, and was 27.56% difference in the non-domesticated region. The outcome of this study advances our understanding of the evolution of crop chromosomes. PMID:25812037

  17. International and Domestic Evaluation: Comparisons and Insights.

    ERIC Educational Resources Information Center

    Conner, Ross F.

    1985-01-01

    The author compares his experiences evaluating Peace Corps volunteers in Kenya and evaluating a health promotion program for adolescents in Orange County, California. Doing international evaluations can improve domestic evaluations by heightening awareness of often unrecognized cultural variations in the domestic setting. (BS)

  18. 48 CFR 1529.401 - Domestic contracts.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 48 Federal Acquisition Regulations System 6 2010-10-01 2010-10-01 true Domestic contracts. 1529.401 Section 1529.401 Federal Acquisition Regulations System ENVIRONMENTAL PROTECTION AGENCY GENERAL CONTRACTING REQUIREMENTS TAXES Contract Clauses 1529.401 Domestic contracts....

  19. 7 CFR 1260.310 - Domestic assessments.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 10 2012-01-01 2012-01-01 false Domestic assessments. 1260.310 Section 1260.310... Rules and Regulations § 1260.310 Domestic assessments. (a) A $1.00 per head assessment on cattle sold... portion of the assessments which are equivalent to the producer's proportionate share of the proceeds....

  20. Domestic Violence against Married Women in Cambodia

    ERIC Educational Resources Information Center

    Yount, Kathryn M.; Carrera, Jennifer S.

    2006-01-01

    We evaluate the effects of marital resources and early-life experiences on recent domestic violence and attitudes about wife abuse among 2,074 married Cambodian women. Household standard of living was negatively associated with physical domestic violence. Women with 8-13 fewer years of schooling than their husbands more often experienced physical…

  1. Child protective services and domestic violence.

    PubMed

    Findlater, J E; Kelly, S

    1999-01-01

    Studies estimate that domestic violence is present in at least one-third of the families involved in child protective services (CPS). Yet, until recently, CPS has not directly addressed domestic violence in its handling of child abuse and neglect cases. By the same token, domestic violence programs have historically emphasized services for battered women, with limited understanding of the child safety goals of CPS. Despite these historical differences, collaborative efforts between CPS and domestic violence service programs are emerging based on a common goal of safety from violence for all family members. Innovative strategies include the use of domestic violence specialists in a variety of child protection settings for case consultation and for support to the battered women, direct referrals of battered women from domestic violence programs to family preservation services, and cross-training of CPS workers and domestic violence service providers. A survey of state CPS administrators and domestic violence coalition directors conducted for this article revealed that although there is mutual interest in greater collaboration, such efforts remain limited. New forums, such as CPS citizen review panels and community-based CPS partnerships, hold promise for further collaboration. Critical to successful strategies are supportive agency leadership, greater trust and understanding across systems, a recognition of common goals, and a willingness to change policies and practice.

  2. Empowering Women with Domestic Violence Experience

    ERIC Educational Resources Information Center

    Anczewska, Marta; Roszczynska-Michta, Joanna; Waszkiewicz, Justyna; Charzynska, Katarzyna; Czabala, Czeslaw

    2012-01-01

    It is generally held that it has been only recently that domestic violence gained appropriate attention as a major social problem. However several approaches, drawn from different theories are applicable in explaining the origin of this negative phenomenon. It is well recognized that trauma of domestic violence has destructive impact on somatic…

  3. Culture and Domestic Violence: Transforming Knowledge Development

    ERIC Educational Resources Information Center

    Bent-Goodley, Tricia B.

    2005-01-01

    Cultural competence continues to receive limited attention in domestic violence service provision from research to the evaluation of programs. Yet with changing demographics reflecting larger numbers of people of color and increasing needs for more effective responses, it is critical that we change the way we think about domestic violence. Using a…

  4. Domestic Violence against Married Women in Edirne

    ERIC Educational Resources Information Center

    Tokuc, Burcu; Ekuklu, Galip; Avcioglu, Serap

    2010-01-01

    The aim of this study is to assess the prevalence and risk factors of domestic violence against married women in Edirne, Turkey. This is a cross-sectional study which included a representative sample of the married women living in the Provincial Center of Edirne. The total past year prevalence of some forms of physical domestic violence is 34% in…

  5. Service Learning in Domestic and International Settings

    ERIC Educational Resources Information Center

    Miller, Kari Knutson; Gonzalez, Amber M.

    2009-01-01

    This paper examines pre-service teacher outcomes associated with service learning in domestic and international settings. One group of upper-division undergraduate level pre service teachers participated in supervised experiences in domestic settings. A second group of upper-division undergraduate level pre-service teachers participated in…

  6. Domestic Violence during Pregnancy in India

    ERIC Educational Resources Information Center

    Mahapatro, Meerambika; Gupta, R. N.; Gupta, Vinay; Kundu, A. S.

    2011-01-01

    Domestic violence can result in many negative health consequences for women's health and well-being. Studies on domestic violence illustrate that abused women in various settings had increased health problems such as injury, chronic pain, gastrointestinal, and gynecological signs including sexually transmitted diseases, depression, and…

  7. Responding to Domestic Violence against Women.

    ERIC Educational Resources Information Center

    Stalans, Loretta J.; Lurigio, Arthur J.

    1995-01-01

    Gives an overview of issues related to domestic violence against women as a social problem: changing responses from the legal system and the community over the course of history, possible causes of domestic violence against women, current perspectives and trends, prevalence, seriousness, and our response as a society. (LKS)

  8. Domestic Options to Offshore Oil and Gas.

    ERIC Educational Resources Information Center

    Kash, Don E.

    1983-01-01

    The continuing controversey over offshore oil/gas has given impetus to searching for domestic energy alternatives. The need for and types of several alternative sources are discussed. Indicates that the United States needs to pursue both offshore and other domestic liquid-fuel sources if it is to avoid becoming increasingly dependent on imports.…

  9. The Effect of Divorce on Domestic Crime

    ERIC Educational Resources Information Center

    Stolzenberg, Lisa; D'Alessio, Stewart J.

    2007-01-01

    Social scientists remain unsure as to whether divorce acts to alleviate domestic violence or whether ex-spouses become the targets of the displaced violence. Using data from the National Incident-Based Reporting System and the Census, this study investigates the relationship between the divorce rate and the domestic crime rate. The study…

  10. Pathobiology of avian influenza in domestic ducks

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Domestic ducks are an important source of food and income in many parts of the world. The susceptibility of domestic ducks to avian influenza (AI) viruses varies depending on many factors, including the species and the age of the ducks, the virus strain, and management practices. Although wild wat...

  11. Behavior genetics and the domestication of animals.

    PubMed

    Jensen, Per

    2014-02-01

    Across species, a similar suite of traits tends to develop in response to domestication, including modifications in behavior. Reduced fear and increased stress tolerance were central in early domestication, and many domestication-related behaviors may have developed as traits correlated to reduced fear. Genetic mechanisms involved in domestication of behavior can be investigated by using top-down or bottom-up approaches, either starting from the behavior variation and searching for underlying genes or finding selected loci and then attempting to identify the associated phenotypes. Combinations of these approaches have proven powerful, and examples of results from such studies are presented and discussed. This includes loci associated with tameness in foxes and dogs, as well as loci correlated with reduced aggression and increased sociality in chickens. Finally, some examples are provided on epigenetic mechanisms in behavior, and it is suggested that selection of favorable epigenetic variants may have been an important mechanism in domestication.

  12. A population genetics view of animal domestication.

    PubMed

    Larson, Greger; Burger, Joachim

    2013-04-01

    The fundamental shift associated with the domestication of plants and animals allowed for a dramatic increase in human population sizes and the emergence of modern society. Despite its importance and the decades of research devoted to studying it, questions regarding the origins and processes of domestication remain. Here, we review recent theoretical advances and present a perspective that underscores the crucial role that population admixture has played in influencing the genomes of domestic animals over the past 10000 years. We then discuss novel approaches to generating and analysing genetic data, emphasising the importance of an explicit hypothesis-testing approach for the inference of the origins and subsequent evolution and demography of domestic animals. By applying next-generation sequencing technology alongside appropriate biostatistical methodologies, a substantially deeper understanding of domestication is on the horizon.

  13. Getting domestication straight: ramosa1 in maize.

    PubMed

    Dempewolf, Hannes

    2010-04-01

    Knowledge of the identities and characteristics of genes that govern the dramatic phenotypic differences between cultivated plants and their wild ancestors has greatly enhanced our understanding of the domestication process. In this issue of Molecular Ecology, Sigmon & Vollbrecht report the discovery of a new maize domestication gene, ramosa1, which encodes a putative transcription factor in the ramosa developmental pathway. Ramosa1 appears to be instrumental in determining the straightness of kernel rows on the maize cob. The key domestication alleles at ramosa1 are prevalent in landraces of maize. These results reinforce findings from previous studies of crop evolution by highlighting the importance of standing genetic variation and changes in transcriptional regulators in domestication. The evolutionary genetics of domestication also provides a framework for predicting the evolutionary response of organisms to strong human-induced selection pressures over limited time intervals.

  14. The Near Eastern origin of cat domestication.

    PubMed

    Driscoll, Carlos A; Menotti-Raymond, Marilyn; Roca, Alfred L; Hupe, Karsten; Johnson, Warren E; Geffen, Eli; Harley, Eric H; Delibes, Miguel; Pontier, Dominique; Kitchener, Andrew C; Yamaguchi, Nobuyuki; O'brien, Stephen J; Macdonald, David W

    2007-07-27

    The world's domestic cats carry patterns of sequence variation in their genome that reflect a history of domestication and breed development. A genetic assessment of 979 domestic cats and their wild progenitors-Felis silvestris silvestris (European wildcat), F. s. lybica (Near Eastern wildcat), F. s. ornata (central Asian wildcat), F. s. cafra (southern African wildcat), and F. s. bieti (Chinese desert cat)-indicated that each wild group represents a distinctive subspecies of Felis silvestris. Further analysis revealed that cats were domesticated in the Near East, probably coincident with agricultural village development in the Fertile Crescent. Domestic cats derive from at least five founders from across this region, whose descendants were transported across the world by human assistance.

  15. Behavior genetics and the domestication of animals.

    PubMed

    Jensen, Per

    2014-02-01

    Across species, a similar suite of traits tends to develop in response to domestication, including modifications in behavior. Reduced fear and increased stress tolerance were central in early domestication, and many domestication-related behaviors may have developed as traits correlated to reduced fear. Genetic mechanisms involved in domestication of behavior can be investigated by using top-down or bottom-up approaches, either starting from the behavior variation and searching for underlying genes or finding selected loci and then attempting to identify the associated phenotypes. Combinations of these approaches have proven powerful, and examples of results from such studies are presented and discussed. This includes loci associated with tameness in foxes and dogs, as well as loci correlated with reduced aggression and increased sociality in chickens. Finally, some examples are provided on epigenetic mechanisms in behavior, and it is suggested that selection of favorable epigenetic variants may have been an important mechanism in domestication. PMID:25384136

  16. Sexing sperm of domestic animals.

    PubMed

    Espinosa-Cervantes, Román; Córdova-Izquierdo, Alejandro

    2013-01-01

    The ability to preselect or predetermine the sex of offspring prior to conception is a highly desired technological tool for assisted female breeding programs specifically for milk production, and in males, for meat production and increasing livestock numbers. The current technology is based on the well-known differences in X- and Y-sperm in the amount of DNA. The technology uses modified flow cytometric instrumentation for sorting X- and Y-bearing sperm. The method can be validated on the basis of live births, laboratory reanalysis of sorted sperm for DNA content, and embryo biopsy for sex determination. Currently, the sex of animals has been predetermined with 90 % accuracy by sexing spermatozoa. In the bovine breeding industry, flow cytometric sperm sexing has not fulfilled its original promise. Sexed sperm doses are too expensive for widespread application while the fertility of sexed sperm doses is lower than unsexed ones. Essentially all bovine sexed semen is frozen and then applied through artificial insemination (AI) or in vitro fertilization. There is still a need in the animal breeding industry to develop a technique for sperm sexing that provides sufficient spermatozoa for AI doses, does not compromise sperm fertility, and is widely applicable to a range of species. In this review, we will summarize the current state-of-the-art in sex preselection in domestic animals and some wildlife species using flow cytometric sperm-sorting of X from Y sperm based on DNA differences.

  17. Sexing sperm of domestic animals.

    PubMed

    Espinosa-Cervantes, Román; Córdova-Izquierdo, Alejandro

    2013-01-01

    The ability to preselect or predetermine the sex of offspring prior to conception is a highly desired technological tool for assisted female breeding programs specifically for milk production, and in males, for meat production and increasing livestock numbers. The current technology is based on the well-known differences in X- and Y-sperm in the amount of DNA. The technology uses modified flow cytometric instrumentation for sorting X- and Y-bearing sperm. The method can be validated on the basis of live births, laboratory reanalysis of sorted sperm for DNA content, and embryo biopsy for sex determination. Currently, the sex of animals has been predetermined with 90 % accuracy by sexing spermatozoa. In the bovine breeding industry, flow cytometric sperm sexing has not fulfilled its original promise. Sexed sperm doses are too expensive for widespread application while the fertility of sexed sperm doses is lower than unsexed ones. Essentially all bovine sexed semen is frozen and then applied through artificial insemination (AI) or in vitro fertilization. There is still a need in the animal breeding industry to develop a technique for sperm sexing that provides sufficient spermatozoa for AI doses, does not compromise sperm fertility, and is widely applicable to a range of species. In this review, we will summarize the current state-of-the-art in sex preselection in domestic animals and some wildlife species using flow cytometric sperm-sorting of X from Y sperm based on DNA differences. PMID:22829354

  18. Phylogeography and domestication of Chinese swamp buffalo.

    PubMed

    Yue, Xiang-Peng; Li, Ran; Xie, Wen-Mei; Xu, Ping; Chang, Ti-Cheng; Liu, Li; Cheng, Feng; Zhang, Run-Feng; Lan, Xian-Yong; Chen, Hong; Lei, Chu-Zhao

    2013-01-01

    To further probe into whether swamp buffaloes were domesticated once or multiple times in China, this survey examined the mitochondrial DNA (mtDNA) Control Region (D-loop) diversity of 471 individuals representing 22 populations of 455 Chinese swamp buffaloes and 16 river buffaloes. Phylogenetic analysis revealed that Chinese swamp buffaloes could be divided into two distinct lineages, A and B, which were defined previously. Of the two lineages, lineage A was predominant across all populations. For predominant lineage A, Southwestern buffalo populations possess the highest genetic diversity among the three hypothesized domestication centers (Southeastern, Central, and Southwestern China), suggesting Southwestern China as the most likely location for the domestication of lineage A. However, a complex pattern of diversity is detected for the lineage B, preventing the unambiguous pinpointing of the exact place of domestication center and suggesting the presence of a long-term, strong gene flow among swamp buffalo populations caused by extensive migrations of buffaloes and frequent human movements along the Yangtze River throughout history. Our current study suggests that Southwestern China is the most likely domestication center for lineage A, and may have been a primary center of swamp buffalo domestication. More archaeological and genetic evidence is needed to show the process of domestication.

  19. The Genetic Architecture of Domestication in Animals

    PubMed Central

    Wright, Dominic

    2015-01-01

    Domestication has been essential to the progress of human civilization, and the process itself has fascinated biologists for hundreds of years. Domestication has led to a series of remarkable changes in a variety of plants and animals, in what is termed the “domestication phenotype.” In domesticated animals, this general phenotype typically consists of similar changes in tameness, behavior, size/morphology, color, brain composition, and adrenal gland size. This domestication phenotype is seen in a range of different animals. However, the genetic basis of these associated changes is still puzzling. The genes for these different traits tend to be grouped together in clusters in the genome, though it is still not clear whether these clusters represent pleiotropic effects, or are in fact linked clusters. This review focuses on what is currently known about the genetic architecture of domesticated animal species, if genes of large effect (often referred to as major genes) are prevalent in driving the domestication phenotype, and whether pleiotropy can explain the loci underpinning these diverse traits being colocated. PMID:26512200

  20. The Genetic Architecture of Domestication in Animals.

    PubMed

    Wright, Dominic

    2015-01-01

    Domestication has been essential to the progress of human civilization, and the process itself has fascinated biologists for hundreds of years. Domestication has led to a series of remarkable changes in a variety of plants and animals, in what is termed the "domestication phenotype." In domesticated animals, this general phenotype typically consists of similar changes in tameness, behavior, size/morphology, color, brain composition, and adrenal gland size. This domestication phenotype is seen in a range of different animals. However, the genetic basis of these associated changes is still puzzling. The genes for these different traits tend to be grouped together in clusters in the genome, though it is still not clear whether these clusters represent pleiotropic effects, or are in fact linked clusters. This review focuses on what is currently known about the genetic architecture of domesticated animal species, if genes of large effect (often referred to as major genes) are prevalent in driving the domestication phenotype, and whether pleiotropy can explain the loci underpinning these diverse traits being colocated.

  1. Counting Child Domestic Servants in Latin America.

    PubMed

    Levison, Deborah; Langer, Anna

    2010-03-01

    This paper summarizes trends in the use of child domestic servants in six Latin American countries using IPUMS-International census samples for 1960 to 2000. Child domestics are among the most vulnerable of child workers, and the most invisible. They may be treated kindly and allowed to attend school, or they may be secluded in their employers' home, overworked, verbally abused, beaten, and unable to leave or report their difficulties to kin. Estimates and imputations are based on labor force and relationship-to-head variables. We find that domestic service makes up a substantial fraction of girls' employment in some countries. We also analyze trends in live-in versus live-out status and school enrollment of child domestic servants. While all child workers are disadvantaged in enrollment relative to non-workers, domestics are sometimes better off than non-domestic workers. In some samples, live-ins are more likely to go to school than live-out child domestics. In others, they are substantially worse off.

  2. Counting Child Domestic Servants in Latin America

    PubMed Central

    Levison, Deborah; Langer, Anna

    2012-01-01

    This paper summarizes trends in the use of child domestic servants in six Latin American countries using IPUMS-International census samples for 1960 to 2000. Child domestics are among the most vulnerable of child workers, and the most invisible. They may be treated kindly and allowed to attend school, or they may be secluded in their employers’ home, overworked, verbally abused, beaten, and unable to leave or report their difficulties to kin. Estimates and imputations are based on labor force and relationship-to-head variables. We find that domestic service makes up a substantial fraction of girls’ employment in some countries. We also analyze trends in live-in versus live-out status and school enrollment of child domestic servants. While all child workers are disadvantaged in enrollment relative to non-workers, domestics are sometimes better off than non-domestic workers. In some samples, live-ins are more likely to go to school than live-out child domestics. In others, they are substantially worse off. PMID:22844164

  3. Phylogeography and Domestication of Chinese Swamp Buffalo

    PubMed Central

    Xie, Wen-Mei; Xu, Ping; Chang, Ti-Cheng; Liu, Li; Cheng, Feng; Zhang, Run-Feng; Lan, Xian-Yong; Chen, Hong; Lei, Chu-Zhao

    2013-01-01

    To further probe into whether swamp buffaloes were domesticated once or multiple times in China, this survey examined the mitochondrial DNA (mtDNA) Control Region (D-loop) diversity of 471 individuals representing 22 populations of 455 Chinese swamp buffaloes and 16 river buffaloes. Phylogenetic analysis revealed that Chinese swamp buffaloes could be divided into two distinct lineages, A and B, which were defined previously. Of the two lineages, lineage A was predominant across all populations. For predominant lineage A, Southwestern buffalo populations possess the highest genetic diversity among the three hypothesized domestication centers (Southeastern, Central, and Southwestern China), suggesting Southwestern China as the most likely location for the domestication of lineage A. However, a complex pattern of diversity is detected for the lineage B, preventing the unambiguous pinpointing of the exact place of domestication center and suggesting the presence of a long-term, strong gene flow among swamp buffalo populations caused by extensive migrations of buffaloes and frequent human movements along the Yangtze River throughout history. Our current study suggests that Southwestern China is the most likely domestication center for lineage A, and may have been a primary center of swamp buffalo domestication. More archaeological and genetic evidence is needed to show the process of domestication. PMID:23437167

  4. The domestic natural gas shortage in China

    NASA Astrophysics Data System (ADS)

    Guo, Ting

    This thesis analyzes the domestic shortage in the Chinese natural gas market. Both the domestic supply and demand of natural gas are growing fast in China. However, the supply cannot catch up with the demand. Under the present pricing mechanism, the Chinese natural gas market cannot get the equilibrium by itself. Expensive imports are inadequate to fill the increasing gap between the domestic demand and supply. Therefore, the shortage problem occurs. Since the energy gap can result in the arrested development of economics, the shortage problem need to be solved. This thesis gives three suggestions to solve the problem: the use of Unconventional Gas, Natural Gas Storage and Pricing Reform.

  5. Molecular mechanisms involved in convergent crop domestication.

    PubMed

    Lenser, Teresa; Theißen, Günter

    2013-12-01

    Domestication has helped to understand evolution. We argue that, vice versa, novel insights into evolutionary principles could provide deeper insights into domestication. Molecular analyses have demonstrated that convergent phenotypic evolution is often based on molecular changes in orthologous genes or pathways. Recent studies have revealed that during plant domestication the causal mutations for convergent changes in key traits are likely to be located in particular genes. These insights may contribute to defining candidate genes for genetic improvement during the domestication of new plant species. Such efforts may help to increase the range of arable crops available, thus increasing crop biodiversity and food security to help meet the predicted demands of the continually growing global population under rapidly changing environmental conditions.

  6. Plant domestication through an ecological lens.

    PubMed

    Milla, Rubén; Osborne, Colin P; Turcotte, Martin M; Violle, Cyrille

    2015-08-01

    Our understanding of domestication comes largely from archeology and genetics. Here, we advocate using current ecological theory and methodologies to provide novel insights into the causes and limitations of evolution under cultivation, as well as into the wider ecological impacts of domestication. We discuss the importance of natural selection under cultivation, that is, the forces promoting differences in Darwinian fitness between plants in crop populations and of constraints, that is, limitations of diverse nature that, given values for trait X, shorten the range of variation of trait Y, during the domestication process. Throughout this opinion paper, we highlight how ecology can yield insight into the effects of domestication on plant traits, on crop feedback over ecological processes, and on how species interactions develop in croplands.

  7. Congenital and Genetic Disease in Domestic Animals

    ERIC Educational Resources Information Center

    Mulvihill, John J.

    1972-01-01

    Reviews observations on domestic animals that have led to the identification of environmental teratogens, and have provided insight into the pathogenesis of congenital defects and genetic diseases in man." (Author/AL)

  8. Women in Domestic Work: Yesterday and Today.

    ERIC Educational Resources Information Center

    Grossman, Allyson Sherman

    1980-01-01

    Provides a historical overview of private household workers and builds a demographic profile of today's domestic workers. Discusses changes in women's employment which may upgrade the status of household workers. (SK)

  9. Helping Children Exposed to Domestic Violence

    MedlinePlus

    ... withdrawal Depression or anxiety Loss of interest in school, friends or other things they enjoyed in the past Children and adolescents exposed to domestic violence should be evaluated by a trained mental health ...

  10. Community-based domestic violence services.

    PubMed

    Saathoff, A J; Stoffel, E A

    1999-01-01

    Community-based domestic violence services have grown significantly since their emergence in the 1970s. Now more than 2,000 in number, domestic violence organizations have expanded their range of programs. In addition to crisis-oriented services, such as telephone hot lines and temporary shelter, many of these agencies provide legal, health, mental health, or vocational services or referrals, and assistance in finding housing, relocating, and planning for safety. Most recently, in response to increasing knowledge about the deleterious effects of exposure to domestic violence on children, community-based service providers have developed programs addressing children's mental health, health, educational, and safety needs. This article describes and analyzes trends in service delivery by these community-based organizations to children affected by domestic violence. It concludes that, although there has been significant growth in services, substantial segments of the target population still are not reached, and most organizations do not yet have a sufficient range of services to meet children's diverse needs. Challenges posed by inadequate funding, needs for specialized staffing, and a dearth of data on the efficacy of current intervention programs hamper domestic violence service providers' ability to meet children's needs. However, this article highlights promising new directions in service delivery. Community-based domestic violence organizations increasingly are using innovative strategies to address children's service needs. These agencies are expanding community outreach efforts and attempts to educate the public and professionals about domestic violence and children. In addition, these organizations are building important collaborative relationships with other agencies concerned with children's welfare, such as child protective services, law enforcement, schools, and health care facilities. These and related developments suggest cautious optimism that future

  11. Social preferences of domestic hens for domestic vs. red junglefowl males and females.

    PubMed

    Borowicz, V A; Graves, H B

    1986-02-01

    The striking dimorphism of male and female junglefowl may have evolved in large part due to female choice of mates. However, domestic fowl hens consistently chose domestic, not junglefowl, males as well as females in preference tests. Either genetic predisposition to favor junglefowl morphological features over those of the domestic types utilized in these tests was overridden by experience or such genetic predisposition does not exist in the breed we tested. PMID:24897347

  12. Whole mitochondrial genome sequencing of domestic horses reveals incorporation of extensive wild horse diversity during domestication

    PubMed Central

    2011-01-01

    Background DNA target enrichment by micro-array capture combined with high throughput sequencing technologies provides the possibility to obtain large amounts of sequence data (e.g. whole mitochondrial DNA genomes) from multiple individuals at relatively low costs. Previously, whole mitochondrial genome data for domestic horses (Equus caballus) were limited to only a few specimens and only short parts of the mtDNA genome (especially the hypervariable region) were investigated for larger sample sets. Results In this study we investigated whole mitochondrial genomes of 59 domestic horses from 44 breeds and a single Przewalski horse (Equus przewalski) using a recently described multiplex micro-array capture approach. We found 473 variable positions within the domestic horses, 292 of which are parsimony-informative, providing a well resolved phylogenetic tree. Our divergence time estimate suggests that the mitochondrial genomes of modern horse breeds shared a common ancestor around 93,000 years ago and no later than 38,000 years ago. A Bayesian skyline plot (BSP) reveals a significant population expansion beginning 6,000-8,000 years ago with an ongoing exponential growth until the present, similar to other domestic animal species. Our data further suggest that a large sample of wild horse diversity was incorporated into the domestic population; specifically, at least 46 of the mtDNA lineages observed in domestic horses (73%) already existed before the beginning of domestication about 5,000 years ago. Conclusions Our study provides a window into the maternal origins of extant domestic horses and confirms that modern domestic breeds present a wide sample of the mtDNA diversity found in ancestral, now extinct, wild horse populations. The data obtained allow us to detect a population expansion event coinciding with the beginning of domestication and to estimate both the minimum number of female horses incorporated into the domestic gene pool and the time depth of the

  13. The domestication of Amazonia before European conquest

    PubMed Central

    Clement, Charles R.; Denevan, William M.; Heckenberger, Michael J.; Junqueira, André Braga; Neves, Eduardo G.; Teixeira, Wenceslau G.; Woods, William I.

    2015-01-01

    During the twentieth century, Amazonia was widely regarded as relatively pristine nature, little impacted by human history. This view remains popular despite mounting evidence of substantial human influence over millennial scales across the region. Here, we review the evidence of an anthropogenic Amazonia in response to claims of sparse populations across broad portions of the region. Amazonia was a major centre of crop domestication, with at least 83 native species containing populations domesticated to some degree. Plant domestication occurs in domesticated landscapes, including highly modified Amazonian dark earths (ADEs) associated with large settled populations and that may cover greater than 0.1% of the region. Populations and food production expanded rapidly within land management systems in the mid-Holocene, and complex societies expanded in resource-rich areas creating domesticated landscapes with profound impacts on local and regional ecology. ADE food production projections support estimates of at least eight million people in 1492. By this time, highly diverse regional systems had developed across Amazonia where subsistence resources were created with plant and landscape domestication, including earthworks. This review argues that the Amazonian anthrome was no less socio-culturally diverse or populous than other tropical forested areas of the world prior to European conquest. PMID:26202998

  14. The domestication of Amazonia before European conquest.

    PubMed

    Clement, Charles R; Denevan, William M; Heckenberger, Michael J; Junqueira, André Braga; Neves, Eduardo G; Teixeira, Wenceslau G; Woods, William I

    2015-08-01

    During the twentieth century, Amazonia was widely regarded as relatively pristine nature, little impacted by human history. This view remains popular despite mounting evidence of substantial human influence over millennial scales across the region. Here, we review the evidence of an anthropogenic Amazonia in response to claims of sparse populations across broad portions of the region. Amazonia was a major centre of crop domestication, with at least 83 native species containing populations domesticated to some degree. Plant domestication occurs in domesticated landscapes, including highly modified Amazonian dark earths (ADEs) associated with large settled populations and that may cover greater than 0.1% of the region. Populations and food production expanded rapidly within land management systems in the mid-Holocene, and complex societies expanded in resource-rich areas creating domesticated landscapes with profound impacts on local and regional ecology. ADE food production projections support estimates of at least eight million people in 1492. By this time, highly diverse regional systems had developed across Amazonia where subsistence resources were created with plant and landscape domestication, including earthworks. This review argues that the Amazonian anthrome was no less socio-culturally diverse or populous than other tropical forested areas of the world prior to European conquest.

  15. The domestication of Amazonia before European conquest.

    PubMed

    Clement, Charles R; Denevan, William M; Heckenberger, Michael J; Junqueira, André Braga; Neves, Eduardo G; Teixeira, Wenceslau G; Woods, William I

    2015-08-01

    During the twentieth century, Amazonia was widely regarded as relatively pristine nature, little impacted by human history. This view remains popular despite mounting evidence of substantial human influence over millennial scales across the region. Here, we review the evidence of an anthropogenic Amazonia in response to claims of sparse populations across broad portions of the region. Amazonia was a major centre of crop domestication, with at least 83 native species containing populations domesticated to some degree. Plant domestication occurs in domesticated landscapes, including highly modified Amazonian dark earths (ADEs) associated with large settled populations and that may cover greater than 0.1% of the region. Populations and food production expanded rapidly within land management systems in the mid-Holocene, and complex societies expanded in resource-rich areas creating domesticated landscapes with profound impacts on local and regional ecology. ADE food production projections support estimates of at least eight million people in 1492. By this time, highly diverse regional systems had developed across Amazonia where subsistence resources were created with plant and landscape domestication, including earthworks. This review argues that the Amazonian anthrome was no less socio-culturally diverse or populous than other tropical forested areas of the world prior to European conquest. PMID:26202998

  16. 19 CFR 175.3 - Domestic interested party.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 2 2010-04-01 2010-04-01 false Domestic interested party. 175.3 Section 175.3... TREASURY (CONTINUED) PETITIONS BY DOMESTIC INTERESTED PARTIES Request for Classification, Appraised Value and Rate of Duty § 175.3 Domestic interested party. “Domestic interested party”, when used in...

  17. Domesticated animals and human infectious diseases of zoonotic origins: domestication time matters.

    PubMed

    Morand, Serge; McIntyre, K Marie; Baylis, Matthew

    2014-06-01

    The rate of emergence for emerging infectious diseases has increased dramatically over the last century, and research findings have implicated wildlife as an importance source of novel pathogens. However, the role played by domestic animals as amplifiers of pathogens emerging from the wild could also be significant, influencing the human infectious disease transmission cycle. The impact of domestic hosts on human disease emergence should therefore be ascertained. Here, using three independent datasets we showed positive relationships between the time since domestication of the major domesticated mammals and the total number of parasites or infectious diseases they shared with humans. We used network analysis, to better visualize the overall interactions between humans and domestic animals (and amongst animals) and estimate which hosts are potential sources of parasites/pathogens for humans (and for all other hosts) by investigating the network architecture. We used centrality, a measure of the connection amongst each host species (humans and domestic animals) in the network, through the sharing of parasites/pathogens, where a central host (i.e. high value of centrality) is the one that is infected by many parasites/pathogens that infect many other hosts in the network. We showed that domesticated hosts that were associated a long time ago with humans are also the central ones in the network and those that favor parasites/pathogens transmission not only to humans but also to all other domesticated animals. These results urge further investigation of the diversity and origin of the infectious diseases of domesticated animals in their domestication centres and the dispersal routes associated with human activities. Such work may help us to better understand how domesticated animals have bridged the epidemiological gap between humans and wildlife.

  18. Domestic violence in Iranian infertile women

    PubMed Central

    Sheikhan, Zohre; Ozgoli, Giti; Azar, Mahyar; Alavimajd, Hamid

    2014-01-01

    Background: Millions of men and women suffer from infertility worldwide. In many cultures, infertile women are at risk of social and emotional problems. Infertility may affect the public health in many countries. Domestic violence is the intentional use of physical force, power or threat against oneself, another person or another group or community which leads to injury, death, mental harm, lack of development or deprivation. This study aimed to assess the prevalence of domestic violence against infertile women who referred to the infertility centres of Tehran, Iran in 2011. Methods: This was cross- sectional descriptive study conducted on 400 infertile women who were selected through convenient sampling method. The questionnaire used in this study included two sections: a demographic section with questions about demographic characteristics of the infertile women and their husbands; and the domestic violence questionnaire with questions about physical, emotional and sexual violence. Data were analysed by SPSS16; descriptive statistics, Spearman’s test, t- test, one-way analysis of variance (ANOVA) and logistic regression were used for data analysis. Results: Four hundred women with the average age of 30.50 ± 6.16 years participated in the study; of whom, 34.7% experienced domestic violence physical violence (5.3%), emotional violence (74.3%) and sexual violence (47.3%). Domestic violence was significantly associated with unwanted marriage, number of IVFs, drug abuse, emotional status of the women, smoking and addiction or drug abuse of the spouse, mental and physical diseases of the husband (p< 0.05). Conclusion: Many of the current problems in this society, particularly in families are due to the transition of the society from a traditional model to a modern one. The majority of the infertile women experience violence in Iran. Domestic violence against infertile women is a problem that should not be ignored. Clinicians should identify abused women. Providing

  19. From wild animals to domestic pets, an evolutionary view of domestication

    PubMed Central

    Driscoll, Carlos A.; Macdonald, David W.; O'Brien, Stephen J.

    2009-01-01

    Artificial selection is the selection of advantageous natural variation for human ends and is the mechanism by which most domestic species evolved. Most domesticates have their origin in one of a few historic centers of domestication as farm animals. Two notable exceptions are cats and dogs. Wolf domestication was initiated late in the Mesolithic when humans were nomadic hunter-gatherers. Those wolves less afraid of humans scavenged nomadic hunting camps and over time developed utility, initially as guards warning of approaching animals or other nomadic bands and soon thereafter as hunters, an attribute tuned by artificial selection. The first domestic cats had limited utility and initiated their domestication among the earliest agricultural Neolithic settlements in the Near East. Wildcat domestication occurred through a self-selective process in which behavioral reproductive isolation evolved as a correlated character of assortative mating coupled to habitat choice for urban environments. Eurasian wildcats initiated domestication and their evolution to companion animals was initially a process of natural, rather than artificial, selection over time driven during their sympatry with forbear wildcats. PMID:19528637

  20. From wild animals to domestic pets, an evolutionary view of domestication.

    PubMed

    Driscoll, Carlos A; Macdonald, David W; O'Brien, Stephen J

    2009-06-16

    Artificial selection is the selection of advantageous natural variation for human ends and is the mechanism by which most domestic species evolved. Most domesticates have their origin in one of a few historic centers of domestication as farm animals. Two notable exceptions are cats and dogs. Wolf domestication was initiated late in the Mesolithic when humans were nomadic hunter-gatherers. Those wolves less afraid of humans scavenged nomadic hunting camps and over time developed utility, initially as guards warning of approaching animals or other nomadic bands and soon thereafter as hunters, an attribute tuned by artificial selection. The first domestic cats had limited utility and initiated their domestication among the earliest agricultural Neolithic settlements in the Near East. Wildcat domestication occurred through a self-selective process in which behavioral reproductive isolation evolved as a correlated character of assortative mating coupled to habitat choice for urban environments. Eurasian wildcats initiated domestication and their evolution to companion animals was initially a process of natural, rather than artificial, selection over time driven during their sympatry with forbear wildcats.

  1. 26 CFR 1.904(g)-1 - Overall domestic loss and the overall domestic loss account.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... loss account. 1.904(g)-1 Section 1.904(g)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... States § 1.904(g)-1 Overall domestic loss and the overall domestic loss account. For further guidance, see § 1.904(g)-1T....

  2. Early Holocene chicken domestication in northern China.

    PubMed

    Xiang, Hai; Gao, Jianqiang; Yu, Baoquan; Zhou, Hui; Cai, Dawei; Zhang, Youwen; Chen, Xiaoyong; Wang, Xi; Hofreiter, Michael; Zhao, Xingbo

    2014-12-01

    Chickens represent by far the most important poultry species, yet the number, locations, and timings of their domestication have remained controversial for more than a century. Here we report ancient mitochondrial DNA sequences from the earliest archaeological chicken bones from China, dating back to ∼ 10,000 B.P. The results clearly show that all investigated bones, including the oldest from the Nanzhuangtou site, are derived from the genus Gallus, rather than any other related genus, such as Phasianus. Our analyses also suggest that northern China represents one region of the earliest chicken domestication, possibly dating as early as 10,000 y B.P. Similar to the evidence from pig domestication, our results suggest that these early domesticated chickens contributed to the gene pool of modern chicken populations. Moreover, our results support the idea that multiple members of the genus Gallus, specifically Gallus gallus and Gallus sonneratii contributed to the gene pool of the modern domestic chicken. Our results provide further support for the growing evidence of an early mixed agricultural complex in northern China.

  3. Storytelling and story testing in domestication

    PubMed Central

    Gerbault, Pascale; Allaby, Robin G.; Boivin, Nicole; Rudzinski, Anna; Grimaldi, Ilaria M.; Pires, J. Chris; Climer Vigueira, Cynthia; Dobney, Keith; Gremillion, Kristen J.; Barton, Loukas; Arroyo-Kalin, Manuel; Purugganan, Michael D.; Rubio de Casas, Rafael; Bollongino, Ruth; Burger, Joachim; Fuller, Dorian Q.; Bradley, Daniel G.; Balding, David J.; Richerson, Peter J.; Gilbert, M. Thomas P.; Larson, Greger; Thomas, Mark G.

    2014-01-01

    The domestication of plants and animals marks one of the most significant transitions in human, and indeed global, history. Traditionally, study of the domestication process was the exclusive domain of archaeologists and agricultural scientists; today it is an increasingly multidisciplinary enterprise that has come to involve the skills of evolutionary biologists and geneticists. Although the application of new information sources and methodologies has dramatically transformed our ability to study and understand domestication, it has also generated increasingly large and complex datasets, the interpretation of which is not straightforward. In particular, challenges of equifinality, evolutionary variance, and emergence of unexpected or counter-intuitive patterns all face researchers attempting to infer past processes directly from patterns in data. We argue that explicit modeling approaches, drawing upon emerging methodologies in statistics and population genetics, provide a powerful means of addressing these limitations. Modeling also offers an approach to analyzing datasets that avoids conclusions steered by implicit biases, and makes possible the formal integration of different data types. Here we outline some of the modeling approaches most relevant to current problems in domestication research, and demonstrate the ways in which simulation modeling is beginning to reshape our understanding of the domestication process. PMID:24753572

  4. Storytelling and story testing in domestication.

    PubMed

    Gerbault, Pascale; Allaby, Robin G; Boivin, Nicole; Rudzinski, Anna; Grimaldi, Ilaria M; Pires, J Chris; Climer Vigueira, Cynthia; Dobney, Keith; Gremillion, Kristen J; Barton, Loukas; Arroyo-Kalin, Manuel; Purugganan, Michael D; Rubio de Casas, Rafael; Bollongino, Ruth; Burger, Joachim; Fuller, Dorian Q; Bradley, Daniel G; Balding, David J; Richerson, Peter J; Gilbert, M Thomas P; Larson, Greger; Thomas, Mark G

    2014-04-29

    The domestication of plants and animals marks one of the most significant transitions in human, and indeed global, history. Traditionally, study of the domestication process was the exclusive domain of archaeologists and agricultural scientists; today it is an increasingly multidisciplinary enterprise that has come to involve the skills of evolutionary biologists and geneticists. Although the application of new information sources and methodologies has dramatically transformed our ability to study and understand domestication, it has also generated increasingly large and complex datasets, the interpretation of which is not straightforward. In particular, challenges of equifinality, evolutionary variance, and emergence of unexpected or counter-intuitive patterns all face researchers attempting to infer past processes directly from patterns in data. We argue that explicit modeling approaches, drawing upon emerging methodologies in statistics and population genetics, provide a powerful means of addressing these limitations. Modeling also offers an approach to analyzing datasets that avoids conclusions steered by implicit biases, and makes possible the formal integration of different data types. Here we outline some of the modeling approaches most relevant to current problems in domestication research, and demonstrate the ways in which simulation modeling is beginning to reshape our understanding of the domestication process.

  5. Reshaping of the maize transcriptome by domestication.

    PubMed

    Swanson-Wagner, Ruth; Briskine, Roman; Schaefer, Robert; Hufford, Matthew B; Ross-Ibarra, Jeffrey; Myers, Chad L; Tiffin, Peter; Springer, Nathan M

    2012-07-17

    Through domestication, humans have substantially altered the morphology of Zea mays ssp. parviglumis (teosinte) into the currently recognizable maize. This system serves as a model for studying adaptation, genome evolution, and the genetics and evolution of complex traits. To examine how domestication has reshaped the transcriptome of maize seedlings, we used expression profiling of 18,242 genes for 38 diverse maize genotypes and 24 teosinte genotypes. We detected evidence for more than 600 genes having significantly different expression levels in maize compared with teosinte. Moreover, more than 1,100 genes showed significantly altered coexpression profiles, reflective of substantial rewiring of the transcriptome since domestication. The genes with altered expression show a significant enrichment for genes previously identified through population genetic analyses as likely targets of selection during maize domestication and improvement; 46 genes previously identified as putative targets of selection also exhibit altered expression levels and coexpression relationships. We also identified 45 genes with altered, primarily higher, expression in inbred relative to outcrossed teosinte. These genes are enriched for functions related to biotic stress and may reflect responses to the effects of inbreeding. This study not only documents alterations in the maize transcriptome following domestication, identifying several genes that may have contributed to the evolution of maize, but highlights the complementary information that can be gained by combining gene expression with population genetic analyses.

  6. Reproductive Isolation during Domestication[W

    PubMed Central

    Dempewolf, Hannes; Hodgins, Kathryn A.; Rummell, Sonja E.; Ellstrand, Norman C.; Rieseberg, Loren H.

    2012-01-01

    It has been hypothesized that reproductive isolation should facilitate evolution under domestication. However, a systematic comparison of reproductive barrier strength between crops and their progenitors has not been conducted to test this hypothesis. Here, we present a systematic survey of reproductive barriers between 32 economically important crop species and their progenitors to better understand the role of reproductive isolation during the domestication process. We took a conservative approach, avoiding those types of reproductive isolation that are poorly known for these taxa (e.g., differences in flowering time). We show that the majority of crops surveyed are isolated from their progenitors by one or more reproductive barriers, despite the fact that the most important reproductive barrier in natural systems, geographical isolation, was absent, at least in the initial stages of domestication for most species. Thus, barriers to reproduction between crops and wild relatives are closely associated with domestication and may facilitate it, thereby raising the question whether reproductive isolation could be viewed as a long-overlooked “domestication trait.” Some of the reproductive barriers observed (e.g., polyploidy and uniparental reproduction), however, may have been favored for reasons other than, or in addition to, their effects on gene flow. PMID:22773750

  7. Human tuberculosis predates domestication in ancient Syria.

    PubMed

    Baker, Oussama; Lee, Oona Y-C; Wu, Houdini H T; Besra, Gurdyal S; Minnikin, David E; Llewellyn, Gareth; Williams, Christopher M; Maixner, Frank; O'Sullivan, Niall; Zink, Albert; Chamel, Bérénice; Khawam, Rima; Coqueugniot, Eric; Helmer, Daniel; Le Mort, Françoise; Perrin, Pascale; Gourichon, Lionel; Dutailly, Bruno; Pálfi, György; Coqueugniot, Hélène; Dutour, Olivier

    2015-06-01

    The question of pre-neolithic tuberculosis is still open in paleopathological perspective. One of the major interests is to explore what type of infection could have existed around the early stage of animal domestication. Paleopathological lesions evoking skeletal TB were observed on five human skeletons coming from two PPNB sites in Syria, which belongs to the geographical cradle of agriculture. These sites represent respectively pre-domestication phase (Dja'de el Mughara, Northern Syria, 8800-8300 BCE cal.) and early domestication phase (Tell Aswad, Southern Syria, 8200-7600 BCE cal.). MicroCT scan analyses were performed on two specimens (one per site) and revealed microscopic changes in favor of TB infection. Detection of lipid biomarkers is positive for two specimens (one per site). Initial molecular analysis further indicates the presence of TB in one individual from Dja'de. Interestingly, no morphological evidence of TB was observed on animal remains of wild and newly domesticated species, discovered in these sites. These observations strongly suggest the presence of human tuberculosis before domestication and at its early stages.

  8. Domestic utility attitudes toward foreign uranium supply

    SciTech Connect

    Not Available

    1981-06-01

    The current embargo on the enrichment of foreign-origin uranium for use in domestic utilization facilities is scheduled to be removed in 1984. The pending removal of this embargo, complicated by a depressed worldwide market for uranium, has prompted consideration of a new or extended embargo within the US Government. As part of its on-going data collection activities, Nuclear Resources International (NRI) has surveyed 50 domestic utility/utility holding companies (representing 60 lead operator-utilities) on their foreign uranium purchase strategies and intentions. The most recent survey was conducted in early May 1981. A number of qualitative observations were made during the course of the survey. The major observations are: domestic utility views toward foreign uranium purchase are dynamic; all but three utilities had some considered foreign purchase strategy; some utilities have problems with buying foreign uranium from particular countries; an inducement is often required by some utilities to buy foreign uranium; opinions varied among utilities concerning the viability of the domestic uranium industry; and many utilities could have foreign uranium fed through their domestic uranium contracts (indirect purchases). The above observations are expanded in the final section of the report. However, it should be noted that two of the observations are particularly important and should be seriously considered in formulation of foreign uranium import restrictions. These important observations are the dynamic nature of the subject matter and the potentially large and imbalanced effect the indirect purchases could have on utility foreign uranium procurement.

  9. Human tuberculosis predates domestication in ancient Syria.

    PubMed

    Baker, Oussama; Lee, Oona Y-C; Wu, Houdini H T; Besra, Gurdyal S; Minnikin, David E; Llewellyn, Gareth; Williams, Christopher M; Maixner, Frank; O'Sullivan, Niall; Zink, Albert; Chamel, Bérénice; Khawam, Rima; Coqueugniot, Eric; Helmer, Daniel; Le Mort, Françoise; Perrin, Pascale; Gourichon, Lionel; Dutailly, Bruno; Pálfi, György; Coqueugniot, Hélène; Dutour, Olivier

    2015-06-01

    The question of pre-neolithic tuberculosis is still open in paleopathological perspective. One of the major interests is to explore what type of infection could have existed around the early stage of animal domestication. Paleopathological lesions evoking skeletal TB were observed on five human skeletons coming from two PPNB sites in Syria, which belongs to the geographical cradle of agriculture. These sites represent respectively pre-domestication phase (Dja'de el Mughara, Northern Syria, 8800-8300 BCE cal.) and early domestication phase (Tell Aswad, Southern Syria, 8200-7600 BCE cal.). MicroCT scan analyses were performed on two specimens (one per site) and revealed microscopic changes in favor of TB infection. Detection of lipid biomarkers is positive for two specimens (one per site). Initial molecular analysis further indicates the presence of TB in one individual from Dja'de. Interestingly, no morphological evidence of TB was observed on animal remains of wild and newly domesticated species, discovered in these sites. These observations strongly suggest the presence of human tuberculosis before domestication and at its early stages. PMID:25819157

  10. Early Holocene chicken domestication in northern China

    PubMed Central

    Xiang, Hai; Gao, Jianqiang; Yu, Baoquan; Zhou, Hui; Cai, Dawei; Zhang, Youwen; Chen, Xiaoyong; Wang, Xi; Hofreiter, Michael; Zhao, Xingbo

    2014-01-01

    Chickens represent by far the most important poultry species, yet the number, locations, and timings of their domestication have remained controversial for more than a century. Here we report ancient mitochondrial DNA sequences from the earliest archaeological chicken bones from China, dating back to ∼10,000 B.P. The results clearly show that all investigated bones, including the oldest from the Nanzhuangtou site, are derived from the genus Gallus, rather than any other related genus, such as Phasianus. Our analyses also suggest that northern China represents one region of the earliest chicken domestication, possibly dating as early as 10,000 y B.P. Similar to the evidence from pig domestication, our results suggest that these early domesticated chickens contributed to the gene pool of modern chicken populations. Moreover, our results support the idea that multiple members of the genus Gallus, specifically Gallus gallus and Gallus sonneratii contributed to the gene pool of the modern domestic chicken. Our results provide further support for the growing evidence of an early mixed agricultural complex in northern China. PMID:25422439

  11. Domestic violence against married women in Edirne.

    PubMed

    Tokuç, Burcu; Ekuklu, Galip; Avcioglu, Serap

    2010-05-01

    The aim of this study is to assess the prevalence and risk factors of domestic violence against married women in Edirne, Turkey. This is a cross-sectional study which included a representative sample of the married women living in the Provincial Center of Edirne. The total past year prevalence of some forms of physical domestic violence is 34% in the last 12 months. 93% of women reported that they have been experiencing different forms of verbal and psychological domestic violence. The important risk factors for physical domestic violence were being Roma woman (OR = 2.97, 95% CI: 1.44-6.12), living with more than four people in the household (OR = 2.29, 95% CI: 1.21-4.36), being unemployed (OR = 2.15, 95% CI: 1.06-4.37), and got married only with her families' decision (OR = 4.60, 95% CI: 1.42-14.80). Our findings conclude that patriarchal and traditional values, women's lack of financial autonomy, and low socioeconomic status are the risk factors for physical domestic violence.

  12. The dead end of domestic violence: spotlight on children's narratives during forensic investigations following domestic homicide.

    PubMed

    Katz, Carmit

    2014-12-01

    The current study provides an in-depth exploration of the narratives of children who witnessed their father killing their mother. This exploration was conducted using a thematic analysis of the children's forensic interviews based on seven investigative interviews that were conducted with children following the domestic homicide. Investigative interviews were selected for study only for substantiated cases and only if the children disclosed the domestic homicide. All of the investigative interviews were conducted within 24h of the domestic homicide. Thematic analysis revealed the following four key categories: the domestic homicide as the dead end of domestic violence, what I did when daddy killed mommy, that one time that daddy killed mommy, and mommy will feel better and will go back home. The discussion examines the multiple layers of this phenomenon as revealed in the children's narratives and its consequences for professionals within the legal and clinical contexts.

  13. Programmed Rearrangement in Ciliates: Paramecium.

    PubMed

    Betermier, Mireille; Duharcourt, Sandra

    2014-12-01

    Programmed genome rearrangements in the ciliate Paramecium provide a nice illustration of the impact of transposons on genome evolution and plasticity. During the sexual cycle, development of the somatic macronucleus involves elimination of ∼30% of the germline genome, including repeated DNA (e.g., transposons) and ∼45,000 single-copy internal eliminated sequences (IES). IES excision is a precise cut-and-close process, in which double-stranded DNA cleavage at IES ends depends on PiggyMac, a domesticated piggyBac transposase. Genome-wide analysis has revealed that at least a fraction of IESs originate from Tc/mariner transposons unrelated to piggyBac. Moreover, genomic sequences with no transposon origin, such as gene promoters, can be excised reproducibly as IESs, indicating that genome rearrangements contribute to the control of gene expression. How the system has evolved to allow elimination of DNA sequences with no recognizable conserved motif has been the subject of extensive research during the past two decades. Increasing evidence has accumulated for the participation of noncoding RNAs in epigenetic control of elimination for a subset of IESs, and in trans-generational inheritance of alternative rearrangement patterns. This chapter summarizes our current knowledge of the structure of the germline and somatic genomes for the model species Paramecium tetraurelia, and describes the DNA cleavage and repair factors that constitute the IES excision machinery. We present an overview of the role of specialized RNA interference machineries and their associated noncoding RNAs in the control of DNA elimination. Finally, we discuss how RNA-dependent modification and/or remodeling of chromatin may guide PiggyMac to its cognate cleavage sites. PMID:26104450

  14. Domestic violence: impact on psychiatric medicine.

    PubMed

    Owens, P L

    1995-10-01

    The 1984 Attorney General's Task Force Report on Domestic Violence attested: Anyone who lives in a violent home experiences an essential loss. The one place on earth where they should feel safe and secure has become a place of danger...the shadow of domestic violence has fallen across their lives and they are forever changed. One report states that the victims of the crime of domestic violence include "not only the people who die from injuries, but the family members who daily endure the psychological, emotional and spiritual abuse and pass on the emotional scars and violent behavior to one generation after another." If we are to stop this cycle of violence, we as physicians must step forward and assume our personal and professional responsibilities. PMID:7474954

  15. Domestication of Mahseer (Tor soro) in Indonesia.

    PubMed

    Gustiano, R; Kontara, E K; Wahyuningsih, H; Subagja, J; Asih, S; Saputr, A

    2013-01-01

    Mahseer is an economically important fish in the world with a distribution from the Himalayan Mountains up to Southeast Asia. One species of Masheer is Tor soro in the Indonesian Archipelago. For a long time, T. soro has been collected from the wild and now is becoming rare. In addition, the degradation of its habitat is another problem. Hence, it is urgent that this species is saved from extinction. Domestication of T. soro from North Sumatra has been conducted since 1996. This paper reports the results of domestication of T. soro starting from collecting live specimens, genetic and morphometric characterizations, evaluation to breeding, and growing out. In 2010, domestication was successful to produce a second generation of young broodstocks and now the breeding technology is being shared with several provinces where T. soro exist.

  16. The domestication and evolutionary ecology of apples.

    PubMed

    Cornille, Amandine; Giraud, Tatiana; Smulders, Marinus J M; Roldán-Ruiz, Isabel; Gladieux, Pierre

    2014-02-01

    The cultivated apple is a major fruit crop in temperate zones. Its wild relatives, distributed across temperate Eurasia and growing in diverse habitats, represent potentially useful sources of diversity for apple breeding. We review here the most recent findings on the genetics and ecology of apple domestication and its impact on wild apples. Genetic analyses have revealed a Central Asian origin for cultivated apple, together with an unexpectedly large secondary contribution from the European crabapple. Wild apple species display strong population structures and high levels of introgression from domesticated apple, and this may threaten their genetic integrity. Recent research has revealed a major role of hybridization in the domestication of the cultivated apple and has highlighted the value of apple as an ideal model for unraveling adaptive diversification processes in perennial fruit crops. We discuss the implications of this knowledge for apple breeding and for the conservation of wild apples.

  17. piggyBac-based insertional mutagenesis and enhancer detection as a tool for functional insect genomics.

    PubMed Central

    Horn, Carsten; Offen, Nils; Nystedt, Sverker; Häcker, Udo; Wimmer, Ernst A

    2003-01-01

    Transposon mutagenesis provides a fundamental tool for functional genomics. Here we present a non-species-specific, combined enhancer detection and binary expression system based on the transposable element piggyBac: For the different components of this insertional mutagenesis system, we used widely applicable transposons and distinguishable broad-range transformation markers, which should enable this system to be operational in nonmodel arthropods. In a pilot screen in Drosophila melanogaster, piggyBac mutator elements on the X chromosome were mobilized in males by a Hermes-based jumpstarter element providing piggyBac transposase activity under control of the alpha1-tubulin promoter. As primary reporters in the piggyBac mutator elements, we employed the heterologous transactivators GAL4delta or tTA. To identify larval and adult enhancer detectors, strains carrying UASp-EYFP or TRE-EYFP as secondary reporter elements were used. Tissue-specific enhancer activities were readily observed in the GAL4delta/UASp-based systems, but only rarely in the tTA/TRE system. Novel autosomal insertions were recovered with an average jumping rate of 80%. Of these novel insertions, 3.8% showed homozygous lethality, which was reversible by piggyBac excision. Insertions were found in both coding and noncoding regions of characterized genes and also in noncharacterized and non-P-targeted CG-number genes. This indicates that piggyBac will greatly facilitate the intended saturation mutagenesis in Drosophila. PMID:12618403

  18. [Nursing discourse on domestic violence in Taiwan].

    PubMed

    Chen, Yu-Hsiu; Huang, Joh-Jong

    2009-06-01

    The purpose of this article was to perform a critical appraisal of nursing discourses on domestic violence published in journals between 1999 and 2007 in Taiwan. All searched materials were retrieved from official websites using key words "domestic violence" or "marital violence". Ten related articles were obtained in all. The most significant finding was related to autonomic nursing intervention, which can effectively meet the needs of abused women. Such an approach differs from traditional order-based nursing activities in the patriarchal practicum of medical teams and offers a promising way to reform medical team hierarchies.

  19. The Gender Wage Gap and Domestic Violence.

    PubMed

    Aizer, Anna

    2010-09-01

    Three quarters of all violence against women is perpetrated by domestic partners. This study exploits exogenous changes in the demand for labor in female-dominated industries to estimate the impact of the male-female wage gap on domestic violence. Decreases in the wage gap reduce violence against women, consistent with a household bargaining model. These findings shed new light on the health production process as well as observed income gradients in health and suggest that in addition to addressing concerns of equity and efficiency, pay parity can also improve the health of American women via reductions in violence. PMID:25110354

  20. Domestic violence among Iraqi refugees in Syria.

    PubMed

    Tappis, Hannah; Biermann, Elizabeth; Glass, Nancy; Tileva, Margarita; Doocy, Shannon

    2012-01-01

    A domestic violence questionnaire was administered to 701 adult females in a sample of 813 Iraqi households in Syria; unmarried women and women whose husbands were away were excluded, yielding a final sample of 486. Lifetime physical, verbal, or emotional abuse was reported by 30%, and approximately 20% experienced abuse within the past year. Non-Damascus residence, children <18 years in the household, no financial challenges upon arrival, and borrowing money in Syria were associated with increased risk of domestic violence within the past year. Support services are inadequate and should be expanded; and longer-term prevention measures also should be implemented.

  1. Multidisciplinary perspectives on banana (Musa spp.) domestication.

    PubMed

    Perrier, Xavier; De Langhe, Edmond; Donohue, Mark; Lentfer, Carol; Vrydaghs, Luc; Bakry, Frédéric; Carreel, Françoise; Hippolyte, Isabelle; Horry, Jean-Pierre; Jenny, Christophe; Lebot, Vincent; Risterucci, Ange-Marie; Tomekpe, Kodjo; Doutrelepont, Hugues; Ball, Terry; Manwaring, Jason; de Maret, Pierre; Denham, Tim

    2011-07-12

    Original multidisciplinary research hereby clarifies the complex geodomestication pathways that generated the vast range of banana cultivars (cvs). Genetic analyses identify the wild ancestors of modern-day cvs and elucidate several key stages of domestication for different cv groups. Archaeology and linguistics shed light on the historical roles of people in the movement and cultivation of bananas from New Guinea to West Africa during the Holocene. The historical reconstruction of domestication processes is essential for breeding programs seeking to diversify and improve banana cvs for the future.

  2. The Gender Wage Gap and Domestic Violence

    PubMed Central

    Aizer, Anna

    2014-01-01

    Three quarters of all violence against women is perpetrated by domestic partners. This study exploits exogenous changes in the demand for labor in female-dominated industries to estimate the impact of the male-female wage gap on domestic violence. Decreases in the wage gap reduce violence against women, consistent with a household bargaining model. These findings shed new light on the health production process as well as observed income gradients in health and suggest that in addition to addressing concerns of equity and efficiency, pay parity can also improve the health of American women via reductions in violence. PMID:25110354

  3. Cat scratch disease from a domestic dog.

    PubMed

    Chen, Tun-Chieh; Lin, Wei-Ru; Lu, Po-Liang; Lin, Chun-Yu; Chen, Yen-Hsu

    2007-02-01

    Cat scratch disease (CSD), caused by Bartonella henselae, is a zoonosis and characterized by self-limited lymphadenopathy. It is transmitted commonly by scratch or bite from cats or kitten. We report an unusual case of CSD caused by a domestic dog scratch that we believe is the first report in Taiwan. A 23-year-old healthy woman developed cervical lymphadenopathy, mild fever, headache, and malaise 3 days after dog scratch. Her symptoms improved after azithromycin treatment. Serology proved B. henselae infection. The owners of a domestic dog might be at risk of "cat" scratch disease.

  4. Multidisciplinary perspectives on banana (Musa spp.) domestication

    PubMed Central

    Perrier, Xavier; De Langhe, Edmond; Donohue, Mark; Lentfer, Carol; Vrydaghs, Luc; Bakry, Frédéric; Carreel, Françoise; Hippolyte, Isabelle; Horry, Jean-Pierre; Jenny, Christophe; Lebot, Vincent; Risterucci, Ange-Marie; Tomekpe, Kodjo; Doutrelepont, Hugues; Ball, Terry; Manwaring, Jason; de Maret, Pierre; Denham, Tim

    2011-01-01

    Original multidisciplinary research hereby clarifies the complex geodomestication pathways that generated the vast range of banana cultivars (cvs). Genetic analyses identify the wild ancestors of modern-day cvs and elucidate several key stages of domestication for different cv groups. Archaeology and linguistics shed light on the historical roles of people in the movement and cultivation of bananas from New Guinea to West Africa during the Holocene. The historical reconstruction of domestication processes is essential for breeding programs seeking to diversify and improve banana cvs for the future. PMID:21730145

  5. A comparative view of the evolution of grasses under domestication.

    PubMed

    Glémin, Sylvain; Bataillon, Thomas

    2009-01-01

    Crop grasses were among the first plants to be domesticated c. 12,000 yr ago, and they still represent the main staple crops for humans. During domestication, as did many other crops, grasses went through dramatic genetic and phenotypic changes. The recent massive increase in genomic data has provided new tools to investigate the genetic basis and consequences of domestication. Beyond the genetics of domestication, many aspects of grass biology, including their phylogeny and developmental biology, are also increasingly well studied, offering a unique opportunity to analyse the domestication process in a comparative way. Taking such a comparative point of view, we review the history of domesticated grasses and how domestication affected their phenotypic and genomic diversity. Considering recent theoretical developments and the accumulation of genetic data, we revisit more specifically the role of mating systems in the domestication process. We close by suggesting future directions for the study of domestication in grasses.

  6. When domestic goes capital: Juror decision making in capital murder trials involving domestic homicide.

    PubMed

    Richards, Tara N; Smith, M Dwayne; Fogel, Sondra J; Bjerregaard, Beth

    2015-08-01

    Prior research suggests that homicide cases involving familial offenders and victims are subject to a "domestic discount" that reduces sentencing severity. However, the operation of a domestic discount in regard to death penalty sentencing has been rarely examined. The current research uses a near-population of jury decisions in capital murder trials conducted in North Carolina from 1991 to 2009 (n = 800), and a series of logistic regression analyses to determine whether there is (a) a direct effect between offender-victim relationship (e.g., domestic, friend/acquaintance, and stranger) and jury decision making, and/or (b) whether domestic offender-victim relationship (as well as other offender-victim relationships) moderates the effect of legal and extralegal case characteristics on jury assessment of the death penalty. Our findings revealed no empirical support for a "domestic discount" whereby juries are less likely to impose death sentences in cases involving domestic homicides. However, substantial differences in predictors of death sentencing were found across offender-victim dyads; most notably, domestic homicide cases demonstrated the most legalistic model of jury decisions to impose death sentences.

  7. Sunflower domestication alleles support single domestication center in eastern North America

    PubMed Central

    Blackman, Benjamin K.; Scascitelli, Moira; Kane, Nolan C.; Luton, Harry H.; Rasmussen, David A.; Bye, Robert A.; Lentz, David L.; Rieseberg, Loren H.

    2011-01-01

    Phylogenetic analyses of genes with demonstrated involvement in evolutionary transitions can be an important means of resolving conflicting hypotheses about evolutionary history or process. In sunflower, two genes have previously been shown to have experienced selective sweeps during its early domestication. In the present study, we identified a third candidate early domestication gene and conducted haplotype analyses of all three genes to address a recent, controversial hypothesis about the origin of cultivated sunflower. Although the scientific consensus had long been that sunflower was domesticated once in eastern North America, the discovery of pre-Columbian sunflower remains at archaeological sites in Mexico led to the proposal of a second domestication center in southern Mexico. Previous molecular studies with neutral markers were consistent with the former hypothesis. However, only two indigenous Mexican cultivars were included in these studies, and their provenance and genetic purity have been questioned. Therefore, we sequenced regions of the three candidate domestication genes containing SNPs diagnostic for domestication from large, newly collected samples of Mexican sunflower landraces and Mexican wild populations from a broad geographic range. The new germplasm also was genotyped for 12 microsatellite loci. Our evidence from multiple evolutionarily important loci and from neutral markers supports a single domestication event for extant cultivated sunflower in eastern North America. PMID:21844335

  8. Sunflower domestication alleles support single domestication center in eastern North America.

    PubMed

    Blackman, Benjamin K; Scascitelli, Moira; Kane, Nolan C; Luton, Harry H; Rasmussen, David A; Bye, Robert A; Lentz, David L; Rieseberg, Loren H

    2011-08-23

    Phylogenetic analyses of genes with demonstrated involvement in evolutionary transitions can be an important means of resolving conflicting hypotheses about evolutionary history or process. In sunflower, two genes have previously been shown to have experienced selective sweeps during its early domestication. In the present study, we identified a third candidate early domestication gene and conducted haplotype analyses of all three genes to address a recent, controversial hypothesis about the origin of cultivated sunflower. Although the scientific consensus had long been that sunflower was domesticated once in eastern North America, the discovery of pre-Columbian sunflower remains at archaeological sites in Mexico led to the proposal of a second domestication center in southern Mexico. Previous molecular studies with neutral markers were consistent with the former hypothesis. However, only two indigenous Mexican cultivars were included in these studies, and their provenance and genetic purity have been questioned. Therefore, we sequenced regions of the three candidate domestication genes containing SNPs diagnostic for domestication from large, newly collected samples of Mexican sunflower landraces and Mexican wild populations from a broad geographic range. The new germplasm also was genotyped for 12 microsatellite loci. Our evidence from multiple evolutionarily important loci and from neutral markers supports a single domestication event for extant cultivated sunflower in eastern North America.

  9. When domestic goes capital: Juror decision making in capital murder trials involving domestic homicide.

    PubMed

    Richards, Tara N; Smith, M Dwayne; Fogel, Sondra J; Bjerregaard, Beth

    2015-08-01

    Prior research suggests that homicide cases involving familial offenders and victims are subject to a "domestic discount" that reduces sentencing severity. However, the operation of a domestic discount in regard to death penalty sentencing has been rarely examined. The current research uses a near-population of jury decisions in capital murder trials conducted in North Carolina from 1991 to 2009 (n = 800), and a series of logistic regression analyses to determine whether there is (a) a direct effect between offender-victim relationship (e.g., domestic, friend/acquaintance, and stranger) and jury decision making, and/or (b) whether domestic offender-victim relationship (as well as other offender-victim relationships) moderates the effect of legal and extralegal case characteristics on jury assessment of the death penalty. Our findings revealed no empirical support for a "domestic discount" whereby juries are less likely to impose death sentences in cases involving domestic homicides. However, substantial differences in predictors of death sentencing were found across offender-victim dyads; most notably, domestic homicide cases demonstrated the most legalistic model of jury decisions to impose death sentences. PMID:25844513

  10. Evidence of long-term gene flow and selection during domestication from analyses of Eurasian wild and domestic pig genomes.

    PubMed

    Frantz, Laurent A F; Schraiber, Joshua G; Madsen, Ole; Megens, Hendrik-Jan; Cagan, Alex; Bosse, Mirte; Paudel, Yogesh; Crooijmans, Richard P M A; Larson, Greger; Groenen, Martien A M

    2015-10-01

    Traditionally, the process of domestication is assumed to be initiated by humans, involve few individuals and rely on reproductive isolation between wild and domestic forms. We analyzed pig domestication using over 100 genome sequences and tested whether pig domestication followed a traditional linear model or a more complex, reticulate model. We found that the assumptions of traditional models, such as reproductive isolation and strong domestication bottlenecks, are incompatible with the genetic data. In addition, our results show that, despite gene flow, the genomes of domestic pigs have strong signatures of selection at loci that affect behavior and morphology. We argue that recurrent selection for domestic traits likely counteracted the homogenizing effect of gene flow from wild boars and created 'islands of domestication' in the genome. Our results have major ramifications for the understanding of animal domestication and suggest that future studies should employ models that do not assume reproductive isolation.

  11. Police Attitudes toward Domestic Violence Offenders

    ERIC Educational Resources Information Center

    Logan, T. K.; Shannon, Lisa; Walker, Robert

    2006-01-01

    Police attitudes are important in facilitating a sense of safety and comfort in women seeking justice-system support for protection from partner violence. This study examined police attitudes toward sanctions and treatment for domestic violence offenders compared with other violent and nonviolent offenders. In addition, police attitudes toward…

  12. 7 CFR 983.12 - Domestic shipments.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Domestic shipments. 983.12 Section 983.12 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE PISTACHIOS GROWN IN...

  13. Attitudes toward Domestic Violence: A Cultural Perspective

    ERIC Educational Resources Information Center

    Wallach, Helene S.; Weingram, Ziv; Avitan, Orli

    2010-01-01

    This study examines the effect of acculturation on the attitudes held by Ethiopian Jews in Israel toward domestic violence (DV). The study findings revealed the following: Ethiopians who immigrated to Israel (n = 31) held more lenient attitudes toward DV than Israeli born Jews (n = 62), which supported the hypothesis that culture influences…

  14. Interstitial Spaces: Visual Culture, Domesticity, and Metaphor

    ERIC Educational Resources Information Center

    Sanders, James H., III; Buenger, Allison

    2010-01-01

    Allison Buenger graduated from the Ohio State University with a bachelor of fine arts degree in ceramics in December 2009. Her works morph domestic objects into vessels of meaning, sculptures displayed in larger installations of textiles and found objects. Originally from Northeast Ohio, Allison resides in Columbus, Ohio, and works at Open Door…

  15. Children's Actions when Experiencing Domestic Violence

    ERIC Educational Resources Information Center

    Overlien, Carolina; Hyden, Margareta

    2009-01-01

    The aim of this article is, by analysing children's discourses, to investigate their actions or absence of actions during a domestic violence episode. The empirical data are recorded group therapy sessions and individual interviews with children who have grown up experiencing their fathers' violence against their mothers. The analysis shows that…

  16. Domestic violence as a development issue.

    PubMed

    Jacobson, R

    1993-06-01

    Northern development agencies must provide practical support to the efforts of southern women to deal with domestic violence. In some ways, domestic violence wears the same face in the north and in the south: it is not an "abnormal" occurrence, it is not confined to any particular socioeconomic class, and it is a complex occurrence in which other women are sometimes complicities. Northern analyses, however, often refer to "battered wives," whereas in other parts of the world assaults may be made by brothers, uncles, or male in-laws. Development initiatives may also affect those power relations that trigger domestic violence. In Sri Lanka, a credit scheme allowed women who processed cashew nuts for export to expand their efforts and achieve remarkable increases in income and in living conditions. This increase, however, was taking place in the context of male unemployment (the men would not engage in the "women's work" of nut processing). Men in the villages where credit was available, therefore, consumed more alcohol. Thus, interventions are rarely gender-neutral. This must be recognized by those who prepare material on domestic violence.

  17. Displaying Geographically-Based Domestic Statistics

    NASA Technical Reports Server (NTRS)

    Quann, J.; Dalton, J.; Banks, M.; Helfer, D.; Szczur, M.; Winkert, G.; Billingsley, J.; Borgstede, R.; Chen, J.; Chen, L.; Fuh, J.; Cyprych, E.

    1982-01-01

    Decision Information Display System (DIDS) is rapid-response information-retrieval and color-graphics display system. DIDS transforms tables of geographically-based domestic statistics (such as population or unemployment by county, energy usage by county, or air-quality figures) into high-resolution, color-coded maps on television display screen.

  18. Quantity Discrimination in Domestic Rats, Rattus norvegicus.

    PubMed

    Cox, Laura; Montrose, V Tamara

    2016-01-01

    Quantity discrimination is a basic form of numerical competence where an animal distinguishes which of two amounts is greater in size. Whilst quantity discrimination in rats has been investigated via training paradigms, rats' natural quantity discrimination abilities without explicit training for a desired response have not been explored. This study investigated domestic rats' ability to perform quantity discrimination. Domestic rats ( n = 12) were examined for their ability to distinguish the larger amount under nine quantity comparisons. One-sample t -tests identified a significant preference for the larger quantity in comparisons of 1 vs. 2, 2 vs. 3, 3 vs. 5, 3 vs. 8, 4 vs. 6, and 4 vs. 8. No preference between quantities was found for comparisons of 3 vs. 4, 4 vs. 5 and 5 vs. 6. Overall, this study drew two key conclusions. Firstly, that domestic rats are capable of performing quantity discrimination without extensive training. Secondly, as subjects adhered to Weber's law, it was concluded that the approximate number system underpins domestic rats' ability to perform spontaneous quantity discrimination. PMID:27527223

  19. Abiotrophy in domestic animals: a review.

    PubMed Central

    de Lahunta, A

    1990-01-01

    This review of abiotrophies in domestic animals has been organized by the predominate anatomical location of the lesion. Secondary considerations include the major signs of the clinical disorder and special neuropathological features. Those abiotrophies that have an established genetic basis are identified but the review includes degenerative disorders in which the etiology is not yet established. PMID:2407332

  20. Domestic Violence Encountered among Kurdish Women

    ERIC Educational Resources Information Center

    Ali, Sirwan Kamil

    2015-01-01

    Background and objective; There is growing recognition that violence against women has a large public health impact, in addition to being a gross violation of women's human rights. The study's aims were: To show the types of domestic abuse encountered by Kurdish women, and study the relationship between them. Methods; The study conducted in the…

  1. Domestic violence. Risk factors and outcomes.

    PubMed Central

    Berrios, D. C.; Grady, D.

    1991-01-01

    Domestic violence is a pervasive and frequently unrecognized cause of injury among women. We reviewed data from standardized interviews with 218 women who presented to an emergency department with injuries due to domestic violence. Victims ranged in age from 16 to 66 years and constituted a wide range of socioeconomic and ethnic backgrounds. Domestic violence often resulted in severe injury; 28% of the women interviewed required admission to hospital for injuries, and 13% required major surgical treatment. The typical presentation was injuries to the face, skull, eyes, extremities, and upper torso. A third of the cases involved a weapon, such as a knife, club, or gun. In all, 10% of the victims were pregnant at the time of abuse, and 10% reported that their children had also been abused by the batterer. Most victims (86%) had suffered at least one previous incident of abuse, and about 40% had previously required medical care for abuse. Victim recognition and referral to appropriate agencies could be improved if primary care physicians were more aware of the prevalence, severity, frequency of occurrence, and typical presentation of domestic violence. PMID:1926841

  2. The Discourses of British Domestic Sitcom.

    ERIC Educational Resources Information Center

    Bee, Jim

    This analysis of the British domestic situation comedy (sitcom) as a genre begins by noting that it is considered basically taxonomical, using the categories of character, narrative, and theme to develop understanding of its discursive strategies. Considered to be preliminary and tentative, the results of this analysis suggest that: (1) the sitcom…

  3. Domestic Violence, Personal Control, and Gender.

    ERIC Educational Resources Information Center

    Umberson, Debra; Anderson, Kristin; Glick, Jennifer; Shapiro, Adam

    1998-01-01

    Explores how domestic violence is related to personal control. Finds that individuals who have initiated violence against a partner do not differ from individuals who have nonviolent relationships in feelings of personal control. Experiencing violence at the hands of a partner has more significant adverse effects on sense of personal control for…

  4. Psychopathology in Women Arrested for Domestic Violence

    ERIC Educational Resources Information Center

    Stuart, Gregory L.; Moore, Todd M.; Gordon, Kristina Coop; Ramsey, Susan E.; Kahler, Christopher W.

    2006-01-01

    This study examined the prevalence of psychopathology among women arrested for violence and whether the experience of intimate partner violence (IPV) was associated with Axis I psychopathology. Women who were arrested for domestic violence perpetration and court referred to violence intervention programs (N=103) completed measures of IPV…

  5. 7 CFR 983.12 - Domestic shipments.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 8 2012-01-01 2012-01-01 false Domestic shipments. 983.12 Section 983.12 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE PISTACHIOS GROWN IN...

  6. 7 CFR 983.12 - Domestic shipments.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 8 2011-01-01 2011-01-01 false Domestic shipments. 983.12 Section 983.12 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE PISTACHIOS GROWN IN...

  7. 7 CFR 983.12 - Domestic shipments.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 8 2013-01-01 2013-01-01 false Domestic shipments. 983.12 Section 983.12 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (MARKETING AGREEMENTS AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE PISTACHIOS GROWN IN...

  8. 7 CFR 983.12 - Domestic shipments.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 8 2014-01-01 2014-01-01 false Domestic shipments. 983.12 Section 983.12 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (MARKETING AGREEMENTS AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE PISTACHIOS GROWN IN...

  9. The TOEFL and Domestic Students: Conclusively Inappropriate

    ERIC Educational Resources Information Center

    Johnson, Dixon C.

    1977-01-01

    This experiment tested college students whose first language is English with the TOEFL examination. The major conclusion was that TOEFL scores do not relate to academic aptitude or performance of domestic students and to evaluate English competency of native speakers with this test is inappropriate. (CHK)

  10. Two maternal origins of Chinese domestic goose.

    PubMed

    Li, H F; Zhu, W Q; Chen, K W; H, Y; Xu, W J; Song, W

    2011-12-01

    China is particularly rich in goose genetic resources. Systematic study of the genetic diversity and origin of Chinese domestic geese will provide an important scientific basis for the conservation and utilization of these resources and for human history. The 521-bp control region (D-loop) of mitochondrial DNA from 26 goose breeds and 6 Landaise geese were sequenced. The results showed that the average haplotype diversity and nucleotide diversity of Chinese domestic geese were 0.1384 and 0.00029, respectively. Shared haplotype analysis and systematic evolution analysis revealed that Chinese domestic geese had 2 maternal origins. The Yili goose breed originated from the Greylag goose (Anser anser), and the other 25 domestic goose breeds originated from the swan goose (Anser cygnoides). An interesting finding was that 1 Linxian white goose and 1 Wanxi white goose shared the same H4 haplotype with the Rhine goose and the Landaise goose, which originated from the Greylag goose (A. anser). Further research on this finding is planned.

  11. Lifelong Learning To Reduce Domestic Violence.

    ERIC Educational Resources Information Center

    Miller, Clinton E.; Mullins, Barbara K.

    2002-01-01

    Domestic violence is a legal, social, and public health issue. Increased public awareness has improved laws and training. Educational programs for both offenders and public officials (police, social workers, and health care providers) are addressing the issue. (Contains 19 references.) (SK)

  12. Pervasive domestication of defective prophages by bacteria

    PubMed Central

    Bobay, Louis-Marie; Touchon, Marie; Rocha, Eduardo P. C.

    2014-01-01

    Integrated phages (prophages) are major contributors to the diversity of bacterial gene repertoires. Domestication of their components is thought to have endowed bacteria with molecular systems involved in secretion, defense, warfare, and gene transfer. However, the rates and mechanisms of domestication remain unknown. We used comparative genomics to study the evolution of prophages within the bacterial genome. We identified over 300 vertically inherited prophages within enterobacterial genomes. Some of these elements are very old and might predate the split between Escherichia coli and Salmonella enterica. The size distribution of prophage elements is bimodal, suggestive of rapid prophage inactivation followed by much slower genetic degradation. Accordingly, we observed a pervasive pattern of systematic counterselection of nonsynonymous mutations in prophage genes. Importantly, such patterns of purifying selection are observed not only on accessory regions but also in core phage genes, such as those encoding structural and lysis components. This suggests that bacterial hosts select for phage-associated functions. Several of these conserved prophages have gene repertoires compatible with described functions of adaptive prophage-derived elements such as bacteriocins, killer particles, gene transfer agents, or satellite prophages. We suggest that bacteria frequently domesticate their prophages. Most such domesticated elements end up deleted from the bacterial genome because they are replaced by analogous functions carried by new prophages. This puts the bacterial genome in a state of continuous flux of acquisition and loss of phage-derived adaptive genes. PMID:25092302

  13. Quantity Discrimination in Domestic Rats, Rattus norvegicus

    PubMed Central

    Cox, Laura; Montrose, V. Tamara

    2016-01-01

    Simple Summary Quantity discrimination involves distinguishing which of two quantities is greater. This discrimination between larger and smaller quantities has only been demonstrated in rats post extensive training. We tested whether domestic rats could perform quantity discrimination without explicit training. We found that rats could distinguish the greater amount in comparisons of 1 vs. 2, 2 vs. 3, 3 vs. 5, 3 vs. 8, 4 vs. 6, and 4 vs. 8. Rats could not distinguish between 3 vs. 4, 4 vs. 5 and 5 vs. 6. We also found that as the ratio between quantities became finer the choice of the larger quantity decreased. We conclude that rats can perform quantity discrimination without extensive training and that their quantity discrimination ability is influenced by the ratio between quantities. Abstract Quantity discrimination is a basic form of numerical competence where an animal distinguishes which of two amounts is greater in size. Whilst quantity discrimination in rats has been investigated via training paradigms, rats’ natural quantity discrimination abilities without explicit training for a desired response have not been explored. This study investigated domestic rats’ ability to perform quantity discrimination. Domestic rats (n = 12) were examined for their ability to distinguish the larger amount under nine quantity comparisons. One-sample t-tests identified a significant preference for the larger quantity in comparisons of 1 vs. 2, 2 vs. 3, 3 vs. 5, 3 vs. 8, 4 vs. 6, and 4 vs. 8. No preference between quantities was found for comparisons of 3 vs. 4, 4 vs. 5 and 5 vs. 6. Overall, this study drew two key conclusions. Firstly, that domestic rats are capable of performing quantity discrimination without extensive training. Secondly, as subjects adhered to Weber’s law, it was concluded that the approximate number system underpins domestic rats’ ability to perform spontaneous quantity discrimination. PMID:27527223

  14. 49 CFR 526.3 - Transfer of vehicle from non-domestic to domestic fleet.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... UNDER THE AUTOMOBILE FUEL EFFICIENCY ACT OF 1980 § 526.3 Transfer of vehicle from non-domestic to... type and displacement, transmission class, car line designation, and fuel economy; (2) The projected...

  15. 49 CFR 526.3 - Transfer of vehicle from non-domestic to domestic fleet.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... UNDER THE AUTOMOBILE FUEL EFFICIENCY ACT OF 1980 § 526.3 Transfer of vehicle from non-domestic to... type and displacement, transmission class, car line designation, and fuel economy; (2) The projected...

  16. 49 CFR 526.3 - Transfer of vehicle from non-domestic to domestic fleet.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... UNDER THE AUTOMOBILE FUEL EFFICIENCY ACT OF 1980 § 526.3 Transfer of vehicle from non-domestic to... type and displacement, transmission class, car line designation, and fuel economy; (2) The projected...

  17. International and domestic regulator issues facing the Canadian MSAT system

    NASA Technical Reports Server (NTRS)

    Bahman, Azarbar; Langlois, Jacques R.; Frank, Christopher J.

    1988-01-01

    International and domestic regulatory issues which affect the implementation of a mobile satellite system (MSAT) over North America are addressed. WARC-MOB-87, MSAT frequency co-ordination, frequency sharing and key Canadian domestic issues are discussed.

  18. The goat domestication process inferred from large-scale mitochondrial DNA analysis of wild and domestic individuals

    PubMed Central

    Naderi, Saeid; Rezaei, Hamid-Reza; Pompanon, François; Blum, Michael G. B.; Negrini, Riccardo; Naghash, Hamid-Reza; Balkız, Özge; Mashkour, Marjan; Gaggiotti, Oscar E.; Ajmone-Marsan, Paolo; Kence, Aykut; Vigne, Jean-Denis; Taberlet, Pierre

    2008-01-01

    The emergence of farming during the Neolithic transition, including the domestication of livestock, was a critical point in the evolution of human kind. The goat (Capra hircus) was one of the first domesticated ungulates. In this study, we compared the genetic diversity of domestic goats to that of the modern representatives of their wild ancestor, the bezoar, by analyzing 473 samples collected over the whole distribution range of the latter species. This partly confirms and significantly clarifies the goat domestication scenario already proposed by archaeological evidence. All of the mitochondrial DNA haplogroups found in current domestic goats have also been found in the bezoar. The geographic distribution of these haplogroups in the wild ancestor allowed the localization of the main domestication centers. We found no haplotype that could have been domesticated in the eastern half of the Iranian Plateau, nor further to the east. A signature of population expansion in bezoars of the C haplogroup suggests an early domestication center on the Central Iranian Plateau (Yazd and Kerman Provinces) and in the Southern Zagros (Fars Province), possibly corresponding to the management of wild flocks. However, the contribution of this center to the current domestic goat population is rather low (1.4%). We also found a second domestication center covering a large area in Eastern Anatolia, and possibly in Northern and Central Zagros. This last domestication center is the likely origin of almost all domestic goats today. This finding is consistent with archaeological data identifying Eastern Anatolia as an important domestication center. PMID:19004765

  19. The goat domestication process inferred from large-scale mitochondrial DNA analysis of wild and domestic individuals.

    PubMed

    Naderi, Saeid; Rezaei, Hamid-Reza; Pompanon, François; Blum, Michael G B; Negrini, Riccardo; Naghash, Hamid-Reza; Balkiz, Ozge; Mashkour, Marjan; Gaggiotti, Oscar E; Ajmone-Marsan, Paolo; Kence, Aykut; Vigne, Jean-Denis; Taberlet, Pierre

    2008-11-18

    The emergence of farming during the Neolithic transition, including the domestication of livestock, was a critical point in the evolution of human kind. The goat (Capra hircus) was one of the first domesticated ungulates. In this study, we compared the genetic diversity of domestic goats to that of the modern representatives of their wild ancestor, the bezoar, by analyzing 473 samples collected over the whole distribution range of the latter species. This partly confirms and significantly clarifies the goat domestication scenario already proposed by archaeological evidence. All of the mitochondrial DNA haplogroups found in current domestic goats have also been found in the bezoar. The geographic distribution of these haplogroups in the wild ancestor allowed the localization of the main domestication centers. We found no haplotype that could have been domesticated in the eastern half of the Iranian Plateau, nor further to the east. A signature of population expansion in bezoars of the C haplogroup suggests an early domestication center on the Central Iranian Plateau (Yazd and Kerman Provinces) and in the Southern Zagros (Fars Province), possibly corresponding to the management of wild flocks. However, the contribution of this center to the current domestic goat population is rather low (1.4%). We also found a second domestication center covering a large area in Eastern Anatolia, and possibly in Northern and Central Zagros. This last domestication center is the likely origin of almost all domestic goats today. This finding is consistent with archaeological data identifying Eastern Anatolia as an important domestication center.

  20. 32 CFR 635.29 - Domestic violence and protection orders.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 4 2014-07-01 2013-07-01 true Domestic violence and protection orders. 635.29... ENFORCEMENT AND CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.29 Domestic violence... permitted by law and regulation. AR 608-18 contains additional information about domestic violence...

  1. 32 CFR 635.29 - Domestic violence and protection orders.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 4 2011-07-01 2011-07-01 false Domestic violence and protection orders. 635.29... ENFORCEMENT AND CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.29 Domestic violence... permitted by law and regulation. AR 608-18 contains additional information about domestic violence...

  2. 32 CFR 635.29 - Domestic violence and protection orders.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 4 2013-07-01 2013-07-01 false Domestic violence and protection orders. 635.29... ENFORCEMENT AND CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.29 Domestic violence... permitted by law and regulation. AR 608-18 contains additional information about domestic violence...

  3. 32 CFR 635.29 - Domestic violence and protection orders.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 4 2010-07-01 2010-07-01 true Domestic violence and protection orders. 635.29... ENFORCEMENT AND CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.29 Domestic violence... permitted by law and regulation. AR 608-18 contains additional information about domestic violence...

  4. 32 CFR 635.29 - Domestic violence and protection orders.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 4 2012-07-01 2011-07-01 true Domestic violence and protection orders. 635.29... ENFORCEMENT AND CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.29 Domestic violence... permitted by law and regulation. AR 608-18 contains additional information about domestic violence...

  5. 45 CFR 1160.5 - Eligibility for domestic exhibitions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... AND ARTIFACTS INDEMNITY ACT § 1160.5 Eligibility for domestic exhibitions. An indemnity agreement for...-owned objects; (B) Exhibitions outside of the United States of domestic-owned objects; or (C) Exhibitions in the United States of both foreign- and domestic-owned objects, with the foreign-owned...

  6. 29 CFR 552.3 - Domestic service employment.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Domestic service employment. 552.3 Section 552.3 Labor Regulations Relating to Labor (Continued) WAGE AND HOUR DIVISION, DEPARTMENT OF LABOR REGULATIONS APPLICATION OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE General Regulations § 552.3 Domestic...

  7. 29 CFR 552.101 - Domestic service employment.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE Interpretations § 552.101 Domestic service... regulations issued under the Social Security Act (20 CFR 404.1057) and from “the generally accepted meaning... 29 Labor 3 2012-07-01 2012-07-01 false Domestic service employment. 552.101 Section 552.101...

  8. 29 CFR 552.3 - Domestic service employment.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Domestic service employment. 552.3 Section 552.3 Labor Regulations Relating to Labor (Continued) WAGE AND HOUR DIVISION, DEPARTMENT OF LABOR REGULATIONS APPLICATION OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE General Regulations § 552.3 Domestic...

  9. 29 CFR 552.3 - Domestic service employment.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Domestic service employment. 552.3 Section 552.3 Labor Regulations Relating to Labor (Continued) WAGE AND HOUR DIVISION, DEPARTMENT OF LABOR REGULATIONS APPLICATION OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE General Regulations § 552.3 Domestic...

  10. 29 CFR 552.101 - Domestic service employment.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE Interpretations § 552.101 Domestic service... regulations issued under the Social Security Act (20 CFR 404.1057) and from “the generally accepted meaning... 29 Labor 3 2011-07-01 2011-07-01 false Domestic service employment. 552.101 Section 552.101...

  11. 29 CFR 552.3 - Domestic service employment.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Domestic service employment. 552.3 Section 552.3 Labor Regulations Relating to Labor (Continued) WAGE AND HOUR DIVISION, DEPARTMENT OF LABOR REGULATIONS APPLICATION OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE General Regulations § 552.3 Domestic...

  12. 29 CFR 552.3 - Domestic service employment.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Domestic service employment. 552.3 Section 552.3 Labor Regulations Relating to Labor (Continued) WAGE AND HOUR DIVISION, DEPARTMENT OF LABOR REGULATIONS APPLICATION OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE General Regulations § 552.3 Domestic...

  13. 29 CFR 552.101 - Domestic service employment.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE Interpretations § 552.101 Domestic service... regulations issued under the Social Security Act (20 CFR 404.1057) and from “the generally accepted meaning... 29 Labor 3 2010-07-01 2010-07-01 false Domestic service employment. 552.101 Section 552.101...

  14. 29 CFR 552.101 - Domestic service employment.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE Interpretations § 552.101 Domestic service... regulations issued under the Social Security Act (20 CFR 404.1057) and from “the generally accepted meaning... 29 Labor 3 2013-07-01 2013-07-01 false Domestic service employment. 552.101 Section 552.101...

  15. Outsourcing of Domestic Tasks and Time-Saving Effects

    ERIC Educational Resources Information Center

    van der Lippe, Tanja; Tijdens, Kea; de Ruijter, Esther

    2004-01-01

    The increased participation of women in paid labor has changed the organization of domestic work. This article deals with a strategy to cope with remaining domestic duties; to what extent are domestic tasks outsourced, what are the main determinants, and does it indeed save time spent on housework? Five outsourcing options are investigated:…

  16. 48 CFR 252.206-7000 - Domestic source restriction.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Domestic source... of Provisions And Clauses 252.206-7000 Domestic source restriction. As prescribed at 206.302-3-70, use the following provision: Domestic Source Restriction (DEC 1991) This solicitation is restricted...

  17. The complex domestication history of the common bean.

    PubMed

    Gaut, Brandon S

    2014-07-01

    A new study reports the genome of common bean (Phaseolus vulgaris) and genome-wide resequencing data from both wild and domesticated accessions. These data confirm that common bean was domesticated at least twice, in Mesoamerica and South America, and also provide a framework to identify genes that contributed to the phenotypic changes associated with domestication.

  18. 75 FR 62303 - National Domestic Violence Awareness Month, 2010

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-08

    ... Documents#0;#0; ] Proclamation 8575 of October 1, 2010 National Domestic Violence Awareness Month, 2010 By... Violence Against Women Act (VAWA), we have broken the silence surrounding domestic violence to reach... are critical achievements, domestic violence remains a devastating public health crisis when one...

  19. 20 CFR 404.1038 - Domestic employees under age 18.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 20 Employees' Benefits 2 2010-04-01 2010-04-01 false Domestic employees under age 18. 404.1038 Section 404.1038 Employees' Benefits SOCIAL SECURITY ADMINISTRATION FEDERAL OLD-AGE, SURVIVORS AND... from Employment § 404.1038 Domestic employees under age 18. Domestic services you perform in a...

  20. 20 CFR 404.1038 - Domestic employees under age 18.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 20 Employees' Benefits 2 2014-04-01 2014-04-01 false Domestic employees under age 18. 404.1038 Section 404.1038 Employees' Benefits SOCIAL SECURITY ADMINISTRATION FEDERAL OLD-AGE, SURVIVORS AND... from Employment § 404.1038 Domestic employees under age 18. Domestic services you perform in a...

  1. 20 CFR 404.1038 - Domestic employees under age 18.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 20 Employees' Benefits 2 2012-04-01 2012-04-01 false Domestic employees under age 18. 404.1038 Section 404.1038 Employees' Benefits SOCIAL SECURITY ADMINISTRATION FEDERAL OLD-AGE, SURVIVORS AND... from Employment § 404.1038 Domestic employees under age 18. Domestic services you perform in a...

  2. 20 CFR 404.1038 - Domestic employees under age 18.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 20 Employees' Benefits 2 2011-04-01 2011-04-01 false Domestic employees under age 18. 404.1038 Section 404.1038 Employees' Benefits SOCIAL SECURITY ADMINISTRATION FEDERAL OLD-AGE, SURVIVORS AND... from Employment § 404.1038 Domestic employees under age 18. Domestic services you perform in a...

  3. 20 CFR 404.1038 - Domestic employees under age 18.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 20 Employees' Benefits 2 2013-04-01 2013-04-01 false Domestic employees under age 18. 404.1038 Section 404.1038 Employees' Benefits SOCIAL SECURITY ADMINISTRATION FEDERAL OLD-AGE, SURVIVORS AND... from Employment § 404.1038 Domestic employees under age 18. Domestic services you perform in a...

  4. Precinct Domestic Violence Teams: Whose Goals Should Determine Program Activities?

    ERIC Educational Resources Information Center

    Weisz, Arlene N.; Black, Beverly M.; Nahan, Neva

    2005-01-01

    This article describes an urban community's attempts to increase domestic violence survivors' participation in the criminal justice system by combining social work advocacy, specialized police officers, and prosecutors into precinct domestic violence teams. An analysis of the outcomes of 1,057 domestic violence reports found that the presence of…

  5. Longitudinal Effects of Domestic Violence on Employment and Welfare Outcomes

    ERIC Educational Resources Information Center

    Lindhorst, Taryn; Oxford, Monica; Gillmore, Mary Rogers

    2007-01-01

    This study uses longitudinal data spanning 13 years from a study of 234 adolescent mothers to evaluate the effects of cumulative domestic violence on employment and welfare use before and after welfare reform. Domestic violence increased the odds of unemployment after welfare reform, but not before; domestic violence had no effect on welfare use…

  6. Perceptions of Domestic Violence: A Dialogue with African American Women

    ERIC Educational Resources Information Center

    Bent-Goodley, Tricia B.

    2004-01-01

    Although empirical research has accumulated over the past 20 years regarding African Americans and domestic violence, many questions remain about African American perceptions of domestic violence. This article explores African American women's perceptions about domestic violence through three focus groups held at a New York social services agency.…

  7. Domestic Violence among the Black Poor: Intersectionality and Social Justice

    ERIC Educational Resources Information Center

    Conwill, William Louis

    2010-01-01

    There are striking gender, race, and class variations in rates of domestic violence. Some leading family theorists called for an intersectional analysis of how gender, race and class systems interact to improve domestic violence theory. This article improves domestic violence theory by: 1) using the discourse, or language, of intersectionality; 2)…

  8. Losing out on Both Counts: Disabled Women and Domestic Violence

    ERIC Educational Resources Information Center

    Thiara, Ravi K.; Hague, Gill; Mullender, Audrey

    2011-01-01

    The links between disability and domestic violence have been under-examined to date, leading to the marginalisation of disabled women affected by domestic violence in theory, politics, and practice. This paper draws on the findings from the first national study in the United Kingdom of the needs of disabled women experiencing domestic violence and…

  9. Domestic Violence against People with Disabilities: Prevalence and Trend Analyses

    ERIC Educational Resources Information Center

    Lin, Jin-Ding; Lin, Lan-Ping; Lin, Pei-Ying; Wu, Jia-Lin; Li, Chien-De; Kuo, Fang-Yu

    2010-01-01

    The present study analyzed national data from "Domestic Violence Report System" derived primarily from the Council of Domestic Violence and Sexual Assaults Prevention, Ministry of the Interior, Taiwan, to describe the reported prevalence of domestic violence in people with disabilities and to examine the time-effect on the prevalence from years…

  10. 34 CFR 7.3 - Determination as to domestic rights.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 34 Education 1 2013-07-01 2013-07-01 false Determination as to domestic rights. 7.3 Section 7.3 Education Office of the Secretary, Department of Education EMPLOYEE INVENTIONS § 7.3 Determination as to domestic rights. The determination of the ownership of the domestic right, title, and interest in and to...

  11. 34 CFR 7.3 - Determination as to domestic rights.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 34 Education 1 2014-07-01 2014-07-01 false Determination as to domestic rights. 7.3 Section 7.3 Education Office of the Secretary, Department of Education EMPLOYEE INVENTIONS § 7.3 Determination as to domestic rights. The determination of the ownership of the domestic right, title, and interest in and to...

  12. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 1 2011-04-01 2011-04-01 false Aircraft carrying domestic (stopover) passengers... Passengers § 122.88 Aircraft carrying domestic (stopover) passengers. Airlines that commingle domestic... continuing on another aircraft to a second U.S. destination) with international passengers who are...

  13. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 1 2012-04-01 2012-04-01 false Aircraft carrying domestic (stopover) passengers... Passengers § 122.88 Aircraft carrying domestic (stopover) passengers. Airlines that commingle domestic... continuing on another aircraft to a second U.S. destination) with international passengers who are...

  14. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 1 2014-04-01 2014-04-01 false Aircraft carrying domestic (stopover) passengers... Passengers § 122.88 Aircraft carrying domestic (stopover) passengers. Airlines that commingle domestic... continuing on another aircraft to a second U.S. destination) with international passengers who are...

  15. The Role of Teachers in Helping Children of Domestic Violence.

    ERIC Educational Resources Information Center

    Kearney, Margaret

    1999-01-01

    Discusses domestic violence as a social issue and the roles of school, social worker, and teacher in combating domestic violence. Focuses on teachers' roles in identifying domestic violence and includes questions to assist in identification. Suggests ways to respond to a child's disclosure of violence, and classroom strategies to establish a…

  16. Energy use of icemaking in domestic refrigerators

    SciTech Connect

    Meier, A.; Martinez, M.S.

    1996-02-01

    This study was designed to develop and test a procedure to measure the electrical consumption of ice making in domestic refrigerators. The Department of Energy (DOE) test procedure was modified to include the energy used for icemaking in conventional refrigerators and those equipped with automatic icemakers. The procedure assumed that 500 grams of ice would be produced daily. Using the new test procedure and the existing DOE test (as a benchmark), four refrigerators equipped with automatic icemakers were tested for ice-making energy use. With the revised test, gross electricity consumption increased about 10% (100 kWh/yr) due to automatic icemaking but about 5% (55 kWh/yr) could be attributed to the special features of the automatic icemaker. The test also confirmed the feasibility of establishing procedures for measuring energy use of specific loads and other activities related to domestic refrigerators. Field testing and subsequent retesting revealed a 14% increase in energy use.

  17. Advanced high capacity domestic satellite communications system

    NASA Astrophysics Data System (ADS)

    Iso, Akio; Kohiyama, Kenji; Odate, Hitoshi; Ishida, Noriaki

    This paper describes a concept of multibeam high capacity transmission possible with a 30/20 GHz and 50/40 GHz domestic satellite communication system. The relationship between satellite antenna pointing accuracy and multi-beam antenna interference, as well as the relationship between satellite antenna pointing accuracy and multi-satellite interference are looked at. The ultra high capacity domestic satellite communication system will have multi-beam antennas with a 76.0 dB at both 20 GHz and 40 GHz. These antennas will provide 4950 beams that approximately correspond to the number of end office of the Japanese telephone network, and have a pointing accuracy of 0.005 degrees. This system will be equipped with 9900 30/20 GHz and 50/40 GHz transponder channels with bit rates of 800 Mbps. Its capacity will be 119 Tbps through use of 15 large communication satellite platforms.

  18. China's future domestic satellite communications systems

    NASA Astrophysics Data System (ADS)

    Liang, G.; Yaokun, Y.

    1985-01-01

    Some ideas about China's future domestic satellite communications system are discussed. The following objectives are considered in the design of the new domestic satellite communications system: (1) the satellite communications system should link the entire country and form a unified national network with the existing ground communications system, (2) based on the geographic situation, the satellite communications system should be primarily small and scattered in the west, and large and concentrated in the east, (3) the Telephone business should be the primary business of the satellite communications system. It should also be capable of transmitting television, telegram, data, facsimile, and broadcasting, (4) the communications system should be primarily designed as a single jump system. It should not exceed two jumps, and (5) the parameters of the communications system should be in agreement with CCIR and CCITT recommendations.

  19. Death by attack from a domestic buffalo.

    PubMed

    Bakkannavar, Shankar M; Monteiro, Francis N P; Bhagavath, Prashantha; Pradeep Kumar, G

    2010-02-01

    Attacks on humans by domestic animals causing fatal injuries are not uncommon in rural areas of India. But injuries due to buffalo gore are rarely observed in villages and are different from other casualties like stab injuries, road fatalities, etc. As the victims of buffalo attack are usually recovered from the fields or forest, the investigating officer could be mislead as to the nature of infliction of fatal injuries to a possible homicide. The injuries caused by the horns of buffaloes are of various shapes, sizes and directions. They are violent and goring in nature. The wound sustained may be contusions, lacerations, criss-cross wounds, penetration of body cavities, and sometimes fractures. In the absence of any eye witness, it becomes very difficult to believe the unsuspecting domestic water buffalo as attacker. This case is reported for its rarity, for the awareness of the possible injuries in such unnatural deaths, and factors predisposing to a buffalo attack.

  20. The magnetic compass of domestic chickens.

    PubMed

    Denzau, Susanne; Nießner, Christine; Rogers, Lesley J; Wiltschko, Wolfgang

    2013-11-01

    In a recent paper, we showed that domestic chickens can be trained to search for a social stimulus in specific magnetic directions. Chickens can hardly fly and have only small home ranges, hence their having a functional magnetic compass may seem rather surprising. Yet considering the natural habitat of their ancestors and their lifestyle until recently, the advantages of a magnetic compass become evident. PMID:24753787

  1. Domesticated Animal Biobanking: Land of Opportunity.

    PubMed

    Groeneveld, Linn F; Gregusson, Sigbjørn; Guldbrandtsen, Bernt; Hiemstra, Sipke J; Hveem, Kristian; Kantanen, Juha; Lohi, Hannes; Stroemstedt, Lina; Berg, Peer

    2016-07-01

    In the past decade, biobanking has fuelled great scientific advances in the human medical sector. Well-established domesticated animal biobanks and integrated networks likewise harbour immense potential for great scientific advances with broad societal impacts, which are currently not being fully realised. Political and scientific leaders as well as journals and ethics committees should help to ensure that we are well equipped to meet future demands in livestock production, animal models, and veterinary care of companion animals. PMID:27467395

  2. Origin and domestication of papaya Yh chromosome.

    PubMed

    VanBuren, Robert; Zeng, Fanchang; Chen, Cuixia; Zhang, Jisen; Wai, Ching Man; Han, Jennifer; Aryal, Rishi; Gschwend, Andrea R; Wang, Jianping; Na, Jong-Kuk; Huang, Lixian; Zhang, Lingmao; Miao, Wenjing; Gou, Jiqing; Arro, Jie; Guyot, Romain; Moore, Richard C; Wang, Ming-Li; Zee, Francis; Charlesworth, Deborah; Moore, Paul H; Yu, Qingyi; Ming, Ray

    2015-04-01

    Sex in papaya is controlled by a pair of nascent sex chromosomes. Females are XX, and two slightly different Y chromosomes distinguish males (XY) and hermaphrodites (XY(h)). The hermaphrodite-specific region of the Y(h) chromosome (HSY) and its X chromosome counterpart were sequenced and analyzed previously. We now report the sequence of the entire male-specific region of the Y (MSY). We used a BAC-by-BAC approach to sequence the MSY and resequence the Y regions of 24 wild males and the Y(h) regions of 12 cultivated hermaphrodites. The MSY and HSY regions have highly similar gene content and structure, and only 0.4% sequence divergence. The MSY sequences from wild males include three distinct haplotypes, associated with the populations' geographic locations, but gene flow is detected for other genomic regions. The Y(h) sequence is highly similar to one Y haplotype (MSY3) found only in wild dioecious populations from the north Pacific region of Costa Rica. The low MSY3-Y(h) divergence supports the hypothesis that hermaphrodite papaya is a product of human domestication. We estimate that Y(h) arose only ∼ 4000 yr ago, well after crop plant domestication in Mesoamerica >6200 yr ago but coinciding with the rise of the Maya civilization. The Y(h) chromosome has lower nucleotide diversity than the Y, or the genome regions that are not fully sex-linked, consistent with a domestication bottleneck. The identification of the ancestral MSY3 haplotype will expedite investigation of the mutation leading to the domestication of the hermaphrodite Y(h) chromosome. In turn, this mutation should identify the gene that was affected by the carpel-suppressing mutation that was involved in the evolution of males.

  3. Origin and domestication of papaya Yh chromosome.

    PubMed

    VanBuren, Robert; Zeng, Fanchang; Chen, Cuixia; Zhang, Jisen; Wai, Ching Man; Han, Jennifer; Aryal, Rishi; Gschwend, Andrea R; Wang, Jianping; Na, Jong-Kuk; Huang, Lixian; Zhang, Lingmao; Miao, Wenjing; Gou, Jiqing; Arro, Jie; Guyot, Romain; Moore, Richard C; Wang, Ming-Li; Zee, Francis; Charlesworth, Deborah; Moore, Paul H; Yu, Qingyi; Ming, Ray

    2015-04-01

    Sex in papaya is controlled by a pair of nascent sex chromosomes. Females are XX, and two slightly different Y chromosomes distinguish males (XY) and hermaphrodites (XY(h)). The hermaphrodite-specific region of the Y(h) chromosome (HSY) and its X chromosome counterpart were sequenced and analyzed previously. We now report the sequence of the entire male-specific region of the Y (MSY). We used a BAC-by-BAC approach to sequence the MSY and resequence the Y regions of 24 wild males and the Y(h) regions of 12 cultivated hermaphrodites. The MSY and HSY regions have highly similar gene content and structure, and only 0.4% sequence divergence. The MSY sequences from wild males include three distinct haplotypes, associated with the populations' geographic locations, but gene flow is detected for other genomic regions. The Y(h) sequence is highly similar to one Y haplotype (MSY3) found only in wild dioecious populations from the north Pacific region of Costa Rica. The low MSY3-Y(h) divergence supports the hypothesis that hermaphrodite papaya is a product of human domestication. We estimate that Y(h) arose only ∼ 4000 yr ago, well after crop plant domestication in Mesoamerica >6200 yr ago but coinciding with the rise of the Maya civilization. The Y(h) chromosome has lower nucleotide diversity than the Y, or the genome regions that are not fully sex-linked, consistent with a domestication bottleneck. The identification of the ancestral MSY3 haplotype will expedite investigation of the mutation leading to the domestication of the hermaphrodite Y(h) chromosome. In turn, this mutation should identify the gene that was affected by the carpel-suppressing mutation that was involved in the evolution of males. PMID:25762551

  4. Lousicidal effect of deltamethrin in domestic pigeons.

    PubMed

    Sivajothi, S; Reddy, B Sudhakara

    2016-09-01

    Present study reports the effect of deltamethrin against louse infestation in domestic pigeons. In this study, twelve pigeons infested with Columbicola columbae louse were sprayed with deltamethrin (12.5 %) and another twelve pigeons were kept as control group. The lice intensity was reduced 89 %, 98 % and 100 % on 1st, 7th and 14th day of post treatment in treatment group. Out of twelve pigeons, four pigeons showed sneezing and temporary ocular inflammations after spraying.

  5. Domesticated Animal Biobanking: Land of Opportunity

    PubMed Central

    Groeneveld, Linn F.; Hveem, Kristian; Kantanen, Juha; Lohi, Hannes; Stroemstedt, Lina; Berg, Peer

    2016-01-01

    In the past decade, biobanking has fuelled great scientific advances in the human medical sector. Well-established domesticated animal biobanks and integrated networks likewise harbour immense potential for great scientific advances with broad societal impacts, which are currently not being fully realised. Political and scientific leaders as well as journals and ethics committees should help to ensure that we are well equipped to meet future demands in livestock production, animal models, and veterinary care of companion animals. PMID:27467395

  6. Bilateral song production in domestic canaries.

    PubMed

    Suthers, Roderick A; Vallet, Eric; Tanvez, Aurélie; Kreutzer, Michel

    2004-09-01

    We studied the mechanism of song production in the outbred common or domestic canary (Serinus canaria). The contribution that each side of the syrinx makes to song was investigated by observing the effect of unilaterally occluding the left or right primary bronchus, followed by section of the ipsilateral branch of the tracheosyringeal nerve. In other birds with a bilaterally intact vocal system we monitored airflow through each side of the syrinx, together with subsyringeal pressure, during spontaneous song. Song production by domestic canaries is not strongly lateralized as it is in the conspecific song-bred waterslager strain. Some syllables are produced entirely on the left or right side of the syrinx, whereas others contain sequential contributions from each side. Low fundamental frequencies are produced with the left syrinx and high frequencies by the right syrinx, increasing the frequency range of domestic canary song compared to that of the waterslager strain. Midrange frequencies can be generated by either side. Syllables at repetition rates below about 25 s(-1) were accompanied by minibreaths, which were usually bilateral. Unilateral minibreaths were typically on the left side. At higher syllable repetition rates, minibreaths were replaced by a respiratory pattern of pulsatile expiration. Our data show that strong unilateral dominance in song production, present in the waterslager strain, is not a trait of the species as a whole and that the pattern of song lateralization can be altered by selective breeding for particular song characteristics.

  7. Domestic animals as models for biomedical research

    PubMed Central

    Andersson, Leif

    2016-01-01

    Domestic animals are unique models for biomedical research due to their long history (thousands of years) of strong phenotypic selection. This process has enriched for novel mutations that have contributed to phenotype evolution in domestic animals. The characterization of such mutations provides insights in gene function and biological mechanisms. This review summarizes genetic dissection of about 50 genetic variants affecting pigmentation, behaviour, metabolic regulation, and the pattern of locomotion. The variants are controlled by mutations in about 30 different genes, and for 10 of these our group was the first to report an association between the gene and a phenotype. Almost half of the reported mutations occur in non-coding sequences, suggesting that this is the most common type of polymorphism underlying phenotypic variation since this is a biased list where the proportion of coding mutations are inflated as they are easier to find. The review documents that structural changes (duplications, deletions, and inversions) have contributed significantly to the evolution of phenotypic diversity in domestic animals. Finally, we describe five examples of evolution of alleles, which means that alleles have evolved by the accumulation of several consecutive mutations affecting the function of the same gene. PMID:26479863

  8. Domestication changes innate constraints for birdsong learning.

    PubMed

    Kagawa, Hiroko; Suzuki, Kenta; Takahasi, Miki; Okanoya, Kazuo

    2014-07-01

    Birdsongs are acquired by imitating the sounds produced by conspecifics. Within a species, songs diverge by cultural transmission, but the range of species-specific features is restricted by innate constraints. Bengalese finches (Lonchura striata var. domestica) are a domesticated strain of the wild White-rumped munia (Lonchura striata). The songs of the domesticated strain have more tonal sounds and more variable sequences than those of the wild strain. We compared the features of songs that were produced by normal birds, isolation-reared birds, and cross-fostered birds in both White-rumped munias and Bengalese finches to identify differences in the genetic and environmental factors of their songs. Factor analyses were conducted based on 17 song measurements. We found that isolated songs differed from normal and cross-fostered songs, especially in unstable prosodic features. In addition, there were significant differences in sound property of mean frequency between the two strains regardless of the rearing conditions. Thus, innate constraints that partially determine birdsong phenotypes may be altered through domestication.

  9. US domestic and international regulatory issues

    NASA Technical Reports Server (NTRS)

    Levin, Lon C.; Nash, Dennis C.

    1993-01-01

    The U.S. domestic and international regulatory and policy milestones since 1982, when NASA filed its petition with the Federal Communications Commission (FCC) to establish the U.S. domestic Mobile Satellite Service (MSS), are described. In 1985, the FCC proposed to establish MSS services and allocate spectrum for such service. In 1986, the FCC allocated L-band spectrum for MSS. In 1987, at the Mobile World Administrative Radio Conference (MOB WARC-87), despite U.S., Canadian, and Mexican efforts, the WARC did not adopt a multi-service, generic MSS allocation. In 1989, the FCC licensed the first MSS system. After two decisions by the U.S. Court of Appeals, the FCC's licensing actions remain intact. The FCC also has permitted Comsat to provide international aeronautical and land MSS via the Inmarsat system. Inmarsat, however, may not serve the domestic U.S. market. In 1991, the FCC accepted applications for MSS systems, most of which were non-geostationary proposals, for operation in the Radiodetermination Satellite Service (RDSS) bands, and the VHF and UHF bands. In 1992, the FCC proposed rules for non-geostationary MSS systems and applied a negotiated rulemaking procedure to each. Also in 1992, the U.S. position for flexibility in existing MSS bands and for additional worldwide MSS allocations was adopted in large part at the 1992 World Administrative Radio Conference (WARC-92).

  10. Domestication changes innate constraints for birdsong learning.

    PubMed

    Kagawa, Hiroko; Suzuki, Kenta; Takahasi, Miki; Okanoya, Kazuo

    2014-07-01

    Birdsongs are acquired by imitating the sounds produced by conspecifics. Within a species, songs diverge by cultural transmission, but the range of species-specific features is restricted by innate constraints. Bengalese finches (Lonchura striata var. domestica) are a domesticated strain of the wild White-rumped munia (Lonchura striata). The songs of the domesticated strain have more tonal sounds and more variable sequences than those of the wild strain. We compared the features of songs that were produced by normal birds, isolation-reared birds, and cross-fostered birds in both White-rumped munias and Bengalese finches to identify differences in the genetic and environmental factors of their songs. Factor analyses were conducted based on 17 song measurements. We found that isolated songs differed from normal and cross-fostered songs, especially in unstable prosodic features. In addition, there were significant differences in sound property of mean frequency between the two strains regardless of the rearing conditions. Thus, innate constraints that partially determine birdsong phenotypes may be altered through domestication. PMID:24793499

  11. Economic valuation of domestic water uses.

    PubMed

    Justes, Antón; Barberán, Ramón; Farizo, Begoña A

    2014-02-15

    There are many reasons of concern about the quality of water for domestic uses. The strategic goals of water tariffs must include savings, efficient management and equity in order to optimise availability of the best quality water. The main domestic uses of water are food preparation, personal hygiene and household cleaning; not all of them need the same quality, and for some of them there are even potential substitutes. In order to contribute to this debate, we need to know how these different uses are valued by direct users. This article focuses on eliciting values for different domestic uses, using discrete choice experiments. As far as we know, this is the first time that a differentiated valuation per use has been defined. The paper introduces three innovations in the design of the experiment. The design takes into account decisive variables in household water consumption, such as family size, together with the consumption level, and the options offered in the choice task are combinations of different prices for each of the uses. A latent class model with both common and class-specific random parameters is applied to better account for the influence of heterogeneity in the respondents' choices. The results show that the most valued use depends on the consumption level class.

  12. 26 CFR 1.904(g)-1T - Overall domestic loss and the overall domestic loss account (temporary).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Without the United States § 1.904(g)-1T Overall domestic loss and the overall domestic loss account..., net capital losses, U.S. source losses, and separate limitation losses, and the recapture of separate... any capital gains or losses, the amount of the taxpayer's domestic loss shall be determined by...

  13. Genomics and the Contrasting Dynamics of Annual and Perennial Domestication.

    PubMed

    Gaut, Brandon S; Díez, Concepción M; Morrell, Peter L

    2015-12-01

    Plant domestication modifies a wild species genetically for human use. Among thousands of domesticated plants, a major distinction is the difference between annual and perennial life cycles. The domestication of perennials is expected to follow different processes than annuals, with distinct genetic outcomes. Here we examine domestication from a population genetics perspective, with a focus on three issues: genetic bottlenecks during domestication, introgression as a source of local adaptation, and genetic load. These three issues have been studied nominally in major annual crops but even less extensively in perennials. Here we highlight lessons from annual plants, motivations to study these issues in perennial plants, and new approaches that may lead to further progress.

  14. Spouse abuse and other domestic violence.

    PubMed

    Dickstein, L J

    1988-12-01

    Concern about the different forms of domestic violence continues to escalate. Beginning with identification of child abuse in the 1960s; spouse abuse, primarily of women, in the 1970s; and, most recently, identification of the rising incidence of elder abuse and neglect, the medical community, state, local, and federal governmental agencies and the public continue to promote joint programs to identify, guide to treatment, and simultaneously develop prevention and early intervention programs. Emphasis initially on the use of legal systems to stop, the abuse must almost be mandatory, because numerous studies show that treatment is most successful when abusers are forced to admit to themselves and others that they have, in fact, committed crimes. For women victims, safe refuge, self-help, and advocacy-support groups were found to be effective, whereas children first need the same protection and a great deal of empathy and explanation. Psychiatrists' roles lie in the important area of early diagnosis and treatment, as most domestic abuse victims do not readily admit to this violence, primarily out of shame, guilt, and fear. Numerous studies demonstrate that following a protocol with every patient, in every setting and under every circumstance, psychiatrists must ask about domestic violence when they least suspect it and when other diagnoses are obvious. The multiple etiologies include general sociocultural pressures, such as poverty and crowding, stereotypic sex role socialization, alcohol and drug abuse, history of head injury, and personal childhood abuse. Psychiatric treatment modalities must occur within a framework of acknowledging that domestic violence victims suffer from post-traumatic stress disorder. Psychiatrists can serve as leaders in coordinating multi-pronged treatment options for the victims: advocacy groups; alcohol and drug detoxification; and individual, couple, and family therapy. Psychiatrists can also serve as consultants, leaders, and educators

  15. The impact of domestication on resistance to two generalist herbivores across 29 independent domestication events.

    PubMed

    Turcotte, Martin M; Turley, Nash E; Johnson, Marc T J

    2014-11-01

    The domestication of crops is among the most important innovations in human history. Here, we test the hypothesis that cultivation and artificial selection for increased productivity of crops reduced plant defenses against herbivores. We compared the performance of two economically important generalist herbivores - the leaf-chewing beet armyworm (Spodoptera exigua) and the phloem-feeding green peach aphid (Myzus persicae) - across 29 crop species and their closely related wild relatives. We also measured putative morphological and chemical defensive traits and correlated them with herbivore performance. We show that, on average, domestication significantly reduced resistance to S. exigua, but not M. persicae, and that most independent domestication events did not cause differences in resistance to either herbivore. In addition, we found that multiple plant traits predicted resistance to S. exigua and M. persicae, and that domestication frequently altered the strength and direction of correlations between these traits and herbivore performance. Our results show that domestication can alter plant defenses, but does not cause strong allocation tradeoffs as predicted by plant defense theory. These results have important implications for understanding the evolutionary ecology of species interactions and for the search for potential resistance traits to be targeted in crop breeding.

  16. Comparative analysis of the domestic cat genome reveals genetic signatures underlying feline biology and domestication.

    PubMed

    Montague, Michael J; Li, Gang; Gandolfi, Barbara; Khan, Razib; Aken, Bronwen L; Searle, Steven M J; Minx, Patrick; Hillier, LaDeana W; Koboldt, Daniel C; Davis, Brian W; Driscoll, Carlos A; Barr, Christina S; Blackistone, Kevin; Quilez, Javier; Lorente-Galdos, Belen; Marques-Bonet, Tomas; Alkan, Can; Thomas, Gregg W C; Hahn, Matthew W; Menotti-Raymond, Marilyn; O'Brien, Stephen J; Wilson, Richard K; Lyons, Leslie A; Murphy, William J; Warren, Wesley C

    2014-12-01

    Little is known about the genetic changes that distinguish domestic cat populations from their wild progenitors. Here we describe a high-quality domestic cat reference genome assembly and comparative inferences made with other cat breeds, wildcats, and other mammals. Based upon these comparisons, we identified positively selected genes enriched for genes involved in lipid metabolism that underpin adaptations to a hypercarnivorous diet. We also found positive selection signals within genes underlying sensory processes, especially those affecting vision and hearing in the carnivore lineage. We observed an evolutionary tradeoff between functional olfactory and vomeronasal receptor gene repertoires in the cat and dog genomes, with an expansion of the feline chemosensory system for detecting pheromones at the expense of odorant detection. Genomic regions harboring signatures of natural selection that distinguish domestic cats from their wild congeners are enriched in neural crest-related genes associated with behavior and reward in mouse models, as predicted by the domestication syndrome hypothesis. Our description of a previously unidentified allele for the gloving pigmentation pattern found in the Birman breed supports the hypothesis that cat breeds experienced strong selection on specific mutations drawn from random bred populations. Collectively, these findings provide insight into how the process of domestication altered the ancestral wildcat genome and build a resource for future disease mapping and phylogenomic studies across all members of the Felidae. PMID:25385592

  17. Comparative analysis of the domestic cat genome reveals genetic signatures underlying feline biology and domestication.

    PubMed

    Montague, Michael J; Li, Gang; Gandolfi, Barbara; Khan, Razib; Aken, Bronwen L; Searle, Steven M J; Minx, Patrick; Hillier, LaDeana W; Koboldt, Daniel C; Davis, Brian W; Driscoll, Carlos A; Barr, Christina S; Blackistone, Kevin; Quilez, Javier; Lorente-Galdos, Belen; Marques-Bonet, Tomas; Alkan, Can; Thomas, Gregg W C; Hahn, Matthew W; Menotti-Raymond, Marilyn; O'Brien, Stephen J; Wilson, Richard K; Lyons, Leslie A; Murphy, William J; Warren, Wesley C

    2014-12-01

    Little is known about the genetic changes that distinguish domestic cat populations from their wild progenitors. Here we describe a high-quality domestic cat reference genome assembly and comparative inferences made with other cat breeds, wildcats, and other mammals. Based upon these comparisons, we identified positively selected genes enriched for genes involved in lipid metabolism that underpin adaptations to a hypercarnivorous diet. We also found positive selection signals within genes underlying sensory processes, especially those affecting vision and hearing in the carnivore lineage. We observed an evolutionary tradeoff between functional olfactory and vomeronasal receptor gene repertoires in the cat and dog genomes, with an expansion of the feline chemosensory system for detecting pheromones at the expense of odorant detection. Genomic regions harboring signatures of natural selection that distinguish domestic cats from their wild congeners are enriched in neural crest-related genes associated with behavior and reward in mouse models, as predicted by the domestication syndrome hypothesis. Our description of a previously unidentified allele for the gloving pigmentation pattern found in the Birman breed supports the hypothesis that cat breeds experienced strong selection on specific mutations drawn from random bred populations. Collectively, these findings provide insight into how the process of domestication altered the ancestral wildcat genome and build a resource for future disease mapping and phylogenomic studies across all members of the Felidae.

  18. The impact of domestication on resistance to two generalist herbivores across 29 independent domestication events.

    PubMed

    Turcotte, Martin M; Turley, Nash E; Johnson, Marc T J

    2014-11-01

    The domestication of crops is among the most important innovations in human history. Here, we test the hypothesis that cultivation and artificial selection for increased productivity of crops reduced plant defenses against herbivores. We compared the performance of two economically important generalist herbivores - the leaf-chewing beet armyworm (Spodoptera exigua) and the phloem-feeding green peach aphid (Myzus persicae) - across 29 crop species and their closely related wild relatives. We also measured putative morphological and chemical defensive traits and correlated them with herbivore performance. We show that, on average, domestication significantly reduced resistance to S. exigua, but not M. persicae, and that most independent domestication events did not cause differences in resistance to either herbivore. In addition, we found that multiple plant traits predicted resistance to S. exigua and M. persicae, and that domestication frequently altered the strength and direction of correlations between these traits and herbivore performance. Our results show that domestication can alter plant defenses, but does not cause strong allocation tradeoffs as predicted by plant defense theory. These results have important implications for understanding the evolutionary ecology of species interactions and for the search for potential resistance traits to be targeted in crop breeding. PMID:25039644

  19. Of Domestic and Wild Guinea Pigs: Studies in Sociophysiology, Domestication, and Social Evolution

    NASA Astrophysics Data System (ADS)

    Sachser, Norbert

    Among mammals a majority of each individual's daily expectations, motivations, and behaviors are directed to encounters with conspecifics. Therefore the knowledge of the genesis, control, and consequences of social interactions is crucial for understanding their social life. We present here our research on the sociophysiology, domestication, and social evolution of wild (Cavia aperea and Galea musteloides) and domestic (Cavia aperea f. porcellus) guinea pigs, which summarizes general rules for many group-living mammals. It is shown that social interactions have consequences not only for the individuals' reproductive success but also for their degrees of stress and welfare. The way in which individuals interact is controlled not only by the present environment but also by the previous social experiences which they have gathered during their behavioral development. Furthermore, the study of ontogeny does not begin at birth, because prenatal social factors acting on pregnant females can also affect the way in which the offspring will interact when adult. In addition, to understand the genesis of interactions between domesticated animals implies knowledge of the behavioral and physiological changes which occurred during the process of domestication. Finally, understanding the social interactions among individuals of the wild ancestor of the domesticated form requires knowledge of how their behavior patterns were brought about by natural selection during the process of social evolution.

  20. Comparative analysis of the domestic cat genome reveals genetic signatures underlying feline biology and domestication

    PubMed Central

    Li, Gang; Gandolfi, Barbara; Khan, Razib; Aken, Bronwen L.; Searle, Steven M. J.; Minx, Patrick; Hillier, LaDeana W.; Koboldt, Daniel C.; Davis, Brian W.; Driscoll, Carlos A.; Barr, Christina S.; Blackistone, Kevin; Quilez, Javier; Lorente-Galdos, Belen; Marques-Bonet, Tomas; Alkan, Can; Thomas, Gregg W. C.; Hahn, Matthew W.; Menotti-Raymond, Marilyn; O’Brien, Stephen J.; Wilson, Richard K.; Lyons, Leslie A.; Murphy, William J.; Warren, Wesley C.

    2014-01-01

    Little is known about the genetic changes that distinguish domestic cat populations from their wild progenitors. Here we describe a high-quality domestic cat reference genome assembly and comparative inferences made with other cat breeds, wildcats, and other mammals. Based upon these comparisons, we identified positively selected genes enriched for genes involved in lipid metabolism that underpin adaptations to a hypercarnivorous diet. We also found positive selection signals within genes underlying sensory processes, especially those affecting vision and hearing in the carnivore lineage. We observed an evolutionary tradeoff between functional olfactory and vomeronasal receptor gene repertoires in the cat and dog genomes, with an expansion of the feline chemosensory system for detecting pheromones at the expense of odorant detection. Genomic regions harboring signatures of natural selection that distinguish domestic cats from their wild congeners are enriched in neural crest-related genes associated with behavior and reward in mouse models, as predicted by the domestication syndrome hypothesis. Our description of a previously unidentified allele for the gloving pigmentation pattern found in the Birman breed supports the hypothesis that cat breeds experienced strong selection on specific mutations drawn from random bred populations. Collectively, these findings provide insight into how the process of domestication altered the ancestral wildcat genome and build a resource for future disease mapping and phylogenomic studies across all members of the Felidae. PMID:25385592

  1. Genetic analyses reveal independent domestication origins of Eurasian reindeer.

    PubMed

    Røed, Knut H; Flagstad, Oystein; Nieminen, Mauri; Holand, Oystein; Dwyer, Mark J; Røv, Nils; Vilà, Carles

    2008-08-22

    Although there is little doubt that the domestication of mammals was instrumental for the modernization of human societies, even basic features of the path towards domestication remain largely unresolved for many species. Reindeer are considered to be in the early phase of domestication with wild and domestic herds still coexisting widely across Eurasia. This provides a unique model system for understanding how the early domestication process may have taken place. We analysed mitochondrial sequences and nuclear microsatellites in domestic and wild herds throughout Eurasia to address the origin of reindeer herding and domestication history. Our data demonstrate independent origins of domestic reindeer in Russia and Fennoscandia. This implies that the Saami people of Fennoscandia domesticated their own reindeer independently of the indigenous cultures in western Russia. We also found that augmentation of local reindeer herds by crossing with wild animals has been common. However, some wild reindeer populations have not contributed to the domestic gene pool, suggesting variation in domestication potential among populations. These differences may explain why geographically isolated indigenous groups have been able to make the technological shift from mobile hunting to large-scale reindeer pastoralism independently.

  2. Domestic outsourcing and multitasking: How much do they really contribute?

    PubMed

    Sullivan, Oriel; Gershuny, Jonathan

    2013-09-01

    The bulk of responsibility for domestic work and childcare in heterosexual couples falls on women. But the means they find to cope with this load, and how these means relate to the factors underpinning the division of labor are not often studied. Two much-cited ways of reducing overall work time are purchasing domestic assistance (outsourcing) and the multitasking of domestic/caring tasks. Using UK 2000/2001 time-use data (N=4196 couples), we find domestic outsourcing is related to having dependent children and to partners' resources, but has little impact on the total domestic/caring workload of either partner. Nor can outsourcing account for the reduction in women's unpaid labor with increasing economic resources. Wives spend more time multitasking than husbands, but their proportion of multitasked domestic time is similar, and is not affected by resources or dependent children. Domestic multitasking seems to be more related to opportunity (time at home) than to time pressure.

  3. Eastern North America as an independent center of plant domestication.

    PubMed

    Smith, Bruce D

    2006-08-15

    The status of eastern North America as an independent center of plant domestication has recently been called into question by a number of genetic and archaeological studies, which suggest that the region may not have witnessed the independent domestication of local crop plants, but rather may have been on the receiving end of domesticated crop plants introduced from Mexico. Here, I provide a synthesis of the currently available archaeological and genetic evidence from both eastern North America and Mexico regarding the spatial and temporal context of initial domestication of the four plant species identified as potential eastern North American domesticates: marshelder (Iva annua), chenopod (Chenopodium berlandieri), squash (Cucurbita pepo), and sunflower (Helianthus annuus). Genetic and archaeological evidence provides strong support for the independent domestication of all four of these plant species in the eastern United States and reconfirms the region as one of the world's independent centers of domestication.

  4. Programmed Genome Rearrangements in Tetrahymena.

    PubMed

    Yao, Meng-Chao; Chao, Ju-Lan; Cheng, Chao-Yin

    2014-12-01

    Ciliates are champions in programmed genome rearrangements. They carry out extensive restructuring during differentiation to drastically alter the complexity, relative copy number, and arrangement of sequences in the somatic genome. This chapter focuses on the model ciliate Tetrahymena, perhaps the simplest and best-understood ciliate studied. It summarizes past studies on various genome rearrangement processes and describes in detail the remarkable progress made in the past decade on the understanding of DNA deletion and other processes. The process occurs at thousands of specific sites to remove defined DNA segments that comprise roughly one-third of the genome including all transposons. Interestingly, this DNA rearranging process is a special form of RNA interference. It involves the production of double-stranded RNA and small RNA that guides the formation of heterochromatin. A domesticated piggyBac transposase is believed to cut off the marked chromatin, and the retained sequences are joined together through nonhomologous end-joining processes. Many of the proteins and DNA players involved have been analyzed and are described. This link provides possible explanations for the evolution, mechanism, and functional roles of the process. The article also discusses the interactions between parental and progeny somatic nuclei that affect the selection of sequences for deletion, and how the specific deletion boundaries are determined after heterochromatin marking.

  5. Domestic satellite services for rural areas

    NASA Astrophysics Data System (ADS)

    Briskman, R. D.

    1984-03-01

    It is pointed out that rural areas can be served by a domestic satellite communications system in an efficient and economical manner. To accomplish such efficiency and economy, the engineering parameters of the satellite communications system must be analyzed and selected with a view toward achieving the desired performance at minimum total cost. The equipment for an entire rural satellite communication system serving 1200 communities can be acquired for approximately $200 million (1983 dollars). An identical system, however, could also be implemented at much lower capital costs by leasing space segment capacity from existing satellite systems (Briskman and Savage, 1983).

  6. [Domestic violence: a bibliographic and bibliometric review].

    PubMed

    Rodríguez Franco, Luís; López-Cepero, Javier; Rodríguez Díaz, Francisco Javier

    2009-05-01

    Violence among relatives and emotionally linked people has recently made a huge social impact. Professionals have suggested diverse concepts to explain the issue, but they have not yet reached an agreement about these concepts. The present work focuses on the scientific yield associated with the keywords "Domestic Violence", perhaps the most commonly used to refer to violence in romantic couples. A series of related publications is analyzed and data about publication years, countries, languages, sample genders, age groups, most prolific journals and authors, and victim-abuser relationships is provided, along with a reflection upon the correctness of the usage of the keyword.

  7. Lousicidal effect of deltamethrin in domestic pigeons.

    PubMed

    Sivajothi, S; Reddy, B Sudhakara

    2016-09-01

    Present study reports the effect of deltamethrin against louse infestation in domestic pigeons. In this study, twelve pigeons infested with Columbicola columbae louse were sprayed with deltamethrin (12.5 %) and another twelve pigeons were kept as control group. The lice intensity was reduced 89 %, 98 % and 100 % on 1st, 7th and 14th day of post treatment in treatment group. Out of twelve pigeons, four pigeons showed sneezing and temporary ocular inflammations after spraying. PMID:27605793

  8. Stalking as a variant of domestic violence.

    PubMed

    Kurt, J L

    1995-01-01

    Much of what is presently known about stalking in a domestic context has been depicted by the popular press, typically following a tragic outcome, and suggests that it is a problem of increasing dimensions. However, scientific literature on this subject is quite limited. This article provides an overview of scientific data related to stalking and associated psychiatric syndromes, including erotomania. It reviews the current antistalking legislation and the National Institute of Justice Model Anti-Stalking Code. Four case studies of stalkers with psychotic disorders versus personality disorders are presented, and the differential diagnoses are discussed. The implications of diagnostic classification, with respect to criminal responsibility, are also discussed.

  9. Locus-specific view of flax domestication history.

    PubMed

    Fu, Yong-Bi; Diederichsen, Axel; Allaby, Robin G

    2012-01-01

    Crop domestication has been inferred genetically from neutral markers and increasingly from specific domestication-associated loci. However, some crops are utilized for multiple purposes that may or may not be reflected in a single domestication-associated locus. One such example is cultivated flax (Linum usitatissimum L.), the earliest oil and fiber crop, for which domestication history remains poorly understood. Oil composition of cultivated flax and pale flax (L. bienne Mill.) indicates that the sad2 locus is a candidate domestication locus associated with increased unsaturated fatty acid production in cultivated flax. A phylogenetic analysis of the sad2 locus in 43 pale and 70 cultivated flax accessions established a complex domestication history for flax that has not been observed previously. The analysis supports an early, independent domestication of a primitive flax lineage, in which the loss of seed dispersal through capsular indehiscence was not established, but increased oil content was likely occurred. A subsequent flax domestication process occurred that probably involved multiple domestications and includes lineages that contain oil, fiber, and winter varieties. In agreement with previous studies, oil rather than fiber varieties occupy basal phylogenetic positions. The data support multiple paths of flax domestication for oil-associated traits before selection of the other domestication-associated traits of seed dispersal loss and fiber production. The sad2 locus is less revealing about the origin of winter tolerance. In this case, a single domestication-associated locus is informative about the history of domesticated forms with the associated trait while partially informative on forms less associated with the trait.

  10. 49 CFR 526.3 - Transfer of vehicle from non-domestic to domestic fleet.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... value added), provide the following information: (1) A description of the model type, including engine...; (4) The percentage of the cost to the manufacturer attributable to value added in the United States... take effect, a description of the nature of the action, and the percentage change in domestic...

  11. 49 CFR 526.3 - Transfer of vehicle from non-domestic to domestic fleet.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... value added), provide the following information: (1) A description of the model type, including engine...; (4) The percentage of the cost to the manufacturer attributable to value added in the United States... take effect, a description of the nature of the action, and the percentage change in domestic...

  12. Investigating the origins of horse domestication.

    PubMed

    Levine, M A

    1999-04-01

    Before the development of firearms, the horse was crucial to warfare and, before the invention of the steam engine, it was the fastest and most reliable form of land transport. It is crucial to the life of nomadic pastoralists on the Eurasian steppe and played a major role in the evolution of human society during the Bronze Age and Iron Age. Understanding the human past requires knowledge of the origins and development of horse husbandry. The problem of being able to identify the early stages of horse domestication is one that many researchers have grappled with for the most part unsuccessfully. Until recently the most important criteria used had been that of increased relative abundance. That is, around 3500 BC, in some parts of Eurasia, there was an apparent increase in the proportions of horse bones and teeth found in archaeological deposits by comparison with preceding periods. However, other evidence suggests that the observed increase during the Copper Age could be explained as well, or even better, by increased hunting rather than by domestication. PMID:11314236

  13. Investigating the origins of horse domestication.

    PubMed

    Levine, M A

    1999-04-01

    Before the development of firearms, the horse was crucial to warfare and, before the invention of the steam engine, it was the fastest and most reliable form of land transport. It is crucial to the life of nomadic pastoralists on the Eurasian steppe and played a major role in the evolution of human society during the Bronze Age and Iron Age. Understanding the human past requires knowledge of the origins and development of horse husbandry. The problem of being able to identify the early stages of horse domestication is one that many researchers have grappled with for the most part unsuccessfully. Until recently the most important criteria used had been that of increased relative abundance. That is, around 3500 BC, in some parts of Eurasia, there was an apparent increase in the proportions of horse bones and teeth found in archaeological deposits by comparison with preceding periods. However, other evidence suggests that the observed increase during the Copper Age could be explained as well, or even better, by increased hunting rather than by domestication.

  14. Fragrance chemicals in domestic and occupational products.

    PubMed

    Rastogi, S C; Heydorn, S; Johansen, J D; Basketter, D A

    2001-10-01

    Epidemiological studies have described an increasing prevalence of fragrance allergy and indicated an association with hand eczema. 59 domestic and occupational products intended for hand exposure were subjected to gas chromatography-mass spectrometric (GC-MS) analyses to test the hypothesis that fragrance chemicals known to have the potential to cause contact allergy but not included in fragrance mix (FM) may be common ingredients in these products. A quantitative analysis of 19 selected fragrances was performed by GC-MS. Further analysis of GC-MS data revealed the presence of 43 other fragrance chemicals/groups of fragrance chemicals in the products investigated. Among the 19 target substances the most commonly detected were limonene in 78%, linalool in 61% and citronellol in 47% of the products investigated. The FM ingredients were present in these products with the following frequencies: oak moss (evernic acid methylester) 2%, cinnamic alcohol 2%, cinnamic aldehyde (cinnamal) 3%, isoeugenol 5%, alpha-amylcinnamic aldehyde (amyl cinnamal) 8%, hydroxycitronellal 12%, eugenol 27%, and geraniol 41%. Thus, the chemical analyses of domestic and occupational products indicates that investigation of potential contact allergy related to these products types should consider fragrance allergens additional to those in the FM, since these may occur with high frequency.

  15. Phylogeography and origin of Chinese domestic chicken.

    PubMed

    Wu, Y P; Huo, J H; Xie, J F; Liu, L X; Wei, Q P; Xie, M G; Kang, Z F; Ji, H Y; Ma, Y H

    2014-04-01

    The loss of local chicken breeds as result of replacement with cosmopolitan breeds indicates the need for conservation measures to protect the future of local genetic stocks. The aim of this study is to describe the patterns of polymorphism of the hypervariable control region of mitochondrial DNA (HVR1) in domestic chicken in China's Jiangxi province to investigate genetic diversity, genetic structure and phylo-dynamics. To this end, we sequenced the mtDNA HVR1 in 231 chickens including 22 individuals which belonged to previously published sequences. A neighbor-joining tree revealed that these samples clustered into five lineages (Lineages A, B, C, E and G). The highest haplotype diversity and nucleotide diversity were both found in Anyi tile-liked gray breed. We estimated that the most recent common ancestor of the local chicken existed approximately 16 million years ago. The mismatch distribution analysis showed two major peaks at positions 4 and 9, while the neutrality test (Tajima's D = -2.19, p < 0.05) and Fu's F-statistics (-8.59, p < 0.05) revealed a significant departure from the neutrality assumption. These results support the idea that domestication of chickens facilitated population increases. Results of a global AMOVA indicated that there was no obvious geographic structure among the local chicken breeds analyzed in this study. The data obtained in this study will assist future conservation management of local breeds and also reveals intriguing implications for the history of human population movements and commerce. PMID:23617370

  16. Cogasification of coal and other domestic fuels

    SciTech Connect

    Green, A.; Mullin, J.; Zanardi, M.; Peres, S.

    1996-12-31

    Almost all new additions to electrical generation in the USA are natural gas combined cycle systems (NGCC) systems. This trend reflects the development of high efficiency gas turbines (GT), low capital, operation and maintenance of NGCC systems and optimism as to natural gas resources. With utility deregulation these developments will seriously restrict long term use of coal and other solid fuels unless a los cost integrated gasifier (IG) fed by low cost feedstocks can be coupled with a CC system. This study mainly considers on-site cogasification of coal with other domestic fuels in an indirectly heated gasifier as a long term strategy for lowering the effective costs of IGGT systems. The authors also consider cocombustion of coal with other low cost domestic fuels as a near term strategy for minimizing fuel costs for competitiveness under utility deregulation. These fuel blending approaches both make use of common fast copyrolysis processes. They examine fast copyrolysis from a molecular point of view searching for advantageous feedstock blends. The authors conclude that blending coal with complementary coals, biomass, MSW or natural gas would be useful in near term cocombustion systems and long term integrated cogasification combined cycle or cogeneration systems.

  17. Phylogeography and origin of Chinese domestic chicken.

    PubMed

    Wu, Y P; Huo, J H; Xie, J F; Liu, L X; Wei, Q P; Xie, M G; Kang, Z F; Ji, H Y; Ma, Y H

    2014-04-01

    The loss of local chicken breeds as result of replacement with cosmopolitan breeds indicates the need for conservation measures to protect the future of local genetic stocks. The aim of this study is to describe the patterns of polymorphism of the hypervariable control region of mitochondrial DNA (HVR1) in domestic chicken in China's Jiangxi province to investigate genetic diversity, genetic structure and phylo-dynamics. To this end, we sequenced the mtDNA HVR1 in 231 chickens including 22 individuals which belonged to previously published sequences. A neighbor-joining tree revealed that these samples clustered into five lineages (Lineages A, B, C, E and G). The highest haplotype diversity and nucleotide diversity were both found in Anyi tile-liked gray breed. We estimated that the most recent common ancestor of the local chicken existed approximately 16 million years ago. The mismatch distribution analysis showed two major peaks at positions 4 and 9, while the neutrality test (Tajima's D = -2.19, p < 0.05) and Fu's F-statistics (-8.59, p < 0.05) revealed a significant departure from the neutrality assumption. These results support the idea that domestication of chickens facilitated population increases. Results of a global AMOVA indicated that there was no obvious geographic structure among the local chicken breeds analyzed in this study. The data obtained in this study will assist future conservation management of local breeds and also reveals intriguing implications for the history of human population movements and commerce.

  18. Serum Biochemical Phenotypes in the Domestic Dog.

    PubMed

    Chang, Yu-Mei; Hadox, Erin; Szladovits, Balazs; Garden, Oliver A

    2016-01-01

    The serum or plasma biochemical profile is essential in the diagnosis and monitoring of systemic disease in veterinary medicine, but current reference intervals typically take no account of breed-specific differences. Breed-specific hematological phenotypes have been documented in the domestic dog, but little has been published on serum biochemical phenotypes in this species. Serum biochemical profiles of dogs in which all measurements fell within the existing reference intervals were retrieved from a large veterinary database. Serum biochemical profiles from 3045 dogs were retrieved, of which 1495 had an accompanying normal glucose concentration. Sixty pure breeds plus a mixed breed control group were represented by at least 10 individuals. All analytes, except for sodium, chloride and glucose, showed variation with age. Total protein, globulin, potassium, chloride, creatinine, cholesterol, total bilirubin, ALT, CK, amylase, and lipase varied between sexes. Neutering status significantly impacted all analytes except albumin, sodium, calcium, urea, and glucose. Principal component analysis of serum biochemical data revealed 36 pure breeds with distinctive phenotypes. Furthermore, comparative analysis identified 23 breeds with significant differences from the mixed breed group in all biochemical analytes except urea and glucose. Eighteen breeds were identified by both principal component and comparative analysis. Tentative reference intervals were generated for breeds with a distinctive phenotype identified by comparative analysis and represented by at least 120 individuals. This is the first large-scale analysis of breed-specific serum biochemical phenotypes in the domestic dog and highlights potential genetic components of biochemical traits in this species. PMID:26919479

  19. Serum Biochemical Phenotypes in the Domestic Dog

    PubMed Central

    Chang, Yu-Mei; Hadox, Erin; Szladovits, Balazs; Garden, Oliver A.

    2016-01-01

    The serum or plasma biochemical profile is essential in the diagnosis and monitoring of systemic disease in veterinary medicine, but current reference intervals typically take no account of breed-specific differences. Breed-specific hematological phenotypes have been documented in the domestic dog, but little has been published on serum biochemical phenotypes in this species. Serum biochemical profiles of dogs in which all measurements fell within the existing reference intervals were retrieved from a large veterinary database. Serum biochemical profiles from 3045 dogs were retrieved, of which 1495 had an accompanying normal glucose concentration. Sixty pure breeds plus a mixed breed control group were represented by at least 10 individuals. All analytes, except for sodium, chloride and glucose, showed variation with age. Total protein, globulin, potassium, chloride, creatinine, cholesterol, total bilirubin, ALT, CK, amylase, and lipase varied between sexes. Neutering status significantly impacted all analytes except albumin, sodium, calcium, urea, and glucose. Principal component analysis of serum biochemical data revealed 36 pure breeds with distinctive phenotypes. Furthermore, comparative analysis identified 23 breeds with significant differences from the mixed breed group in all biochemical analytes except urea and glucose. Eighteen breeds were identified by both principal component and comparative analysis. Tentative reference intervals were generated for breeds with a distinctive phenotype identified by comparative analysis and represented by at least 120 individuals. This is the first large-scale analysis of breed-specific serum biochemical phenotypes in the domestic dog and highlights potential genetic components of biochemical traits in this species. PMID:26919479

  20. Quality Assessment of Domesticated Animal Genome Assemblies

    PubMed Central

    Seemann, Stefan E.; Anthon, Christian; Palasca, Oana; Gorodkin, Jan

    2015-01-01

    The era of high-throughput sequencing has made it relatively simple to sequence genomes and transcriptomes of individuals from many species. In order to analyze the resulting sequencing data, high-quality reference genome assemblies are required. However, this is still a major challenge, and many domesticated animal genomes still need to be sequenced deeper in order to produce high-quality assemblies. In the meanwhile, ironically, the extent to which RNAseq and other next-generation data is produced frequently far exceeds that of the genomic sequence. Furthermore, basic comparative analysis is often affected by the lack of genomic sequence. Herein, we quantify the quality of the genome assemblies of 20 domesticated animals and related species by assessing a range of measurable parameters, and we show that there is a positive correlation between the fraction of mappable reads from RNAseq data and genome assembly quality. We rank the genomes by their assembly quality and discuss the implications for genotype analyses. PMID:27279738

  1. Comparative endocrinology of domestic and nondomestic felids.

    PubMed

    Brown, Janine L

    2006-07-01

    The ability to track gonadal and adrenal activity via hormones is key to optimizing health and reproduction. Through decades of study, a great deal has been learned about the biology of female domestic cats, including endocrine function. More recently, comparative endocrine studies have greatly expanded our knowledge base of nondomestic felids as well. The latter has been possible largely through the development of noninvasive fecal steroid metabolite analysis techniques, which currently is the method of choice for monitoring endocrine function in wildlife species, including felids. It now is well-recognized that a range in endocrine patterns exists among Felidae, with many traits and mechanisms being uncommon, if not unique. There is a high degree of variability in the type of ovulation (spontaneous versus induced) expressed across the taxon. Even within species, some individuals exhibit ovulation that is only induced, whereas others ovulate spontaneously as well. Steroid metabolism also differs in that metabolites are excreted almost exclusively in feces, with very little steroid found in urine. Across species there are marked differences in seasonal and social influences on reproduction, adrenal responses to husbandry practices, and ovarian responses to assisted reproductive procedures. This means that developing strategies to improve health and reproduction of felids must be done on a species by species basis. This paper summarizes current knowledge on the reproductive endocrinology of female domestic and nondomestic cats, and describes how the rapidly growing endocrine database is aiding ex situ management efforts.

  2. Transcriptome Analysis in Domesticated Species: Challenges and Strategies

    PubMed Central

    Hekman, Jessica P.; Johnson, Jennifer L.; Kukekova, Anna V.

    2015-01-01

    Domesticated species occupy a special place in the human world due to their economic and cultural value. In the era of genomic research, domesticated species provide unique advantages for investigation of diseases and complex phenotypes. RNA sequencing, or RNA-seq, has recently emerged as a new approach for studying transcriptional activity of the whole genome, changing the focus from individual genes to gene networks. RNA-seq analysis in domesticated species may complement genome-wide association studies of complex traits with economic importance or direct relevance to biomedical research. However, RNA-seq studies are more challenging in domesticated species than in model organisms. These challenges are at least in part associated with the lack of quality genome assemblies for some domesticated species and the absence of genome assemblies for others. In this review, we discuss strategies for analyzing RNA-seq data, focusing particularly on questions and examples relevant to domesticated species. PMID:26917953

  3. Mitochondrial DNA cleavage patterns distinguish independent origin of Chinese domestic geese and Western domestic geese.

    PubMed

    Shi, X-W; Wang, J-W; Zeng, F-T; Qiu, X-P

    2006-06-01

    It has generally been assumed, based on morphology, that Chinese domestic goose breeds were derived from the swan goose (Anser cygnoides) and that European and American breeds were derived from the graylag goose (Anser anser). To test the validity of this assumption, we investigated the mtDNA cleavage patterns of 16 Chinese breeds and 2 European breeds as well as hybrids produced between a Chinese breed and a European breed. After 224 mtDNAs, isolated from the Chinese and European breeds, were digested by 19 restriction endonucleases, variations of the cleavage patterns were observed for four enzymes (EcoRV, HaeII, HincII, and KpnI). All Chinese breeds and their maternal hybrids except the Yili breed showed an identical haplotype, named haplotype I or the Chinese haplotype; the European breeds and the Yili breed showed another haplotype, named haplotype II or the western haplotype. None of the haplotype found in the Chinese type was detectable in the western type and vice versa. The two haplotypes were found to differ from each other at 8.0% of the sites surveyed and with a 0.72% sequence divergence. Using 2% substitution per million years calibrated from the genera Anser and Branta, the two domestic geese haplotypes were estimated to have diverged approximately 360,000 years ago, well outside the 3000-6000 years in domestic history. Our findings provide the first molecular genetic evidence to support the dual origin assumption of domestic geese in the world. Meanwhile, the four mtDNA restriction fragment length polymorphisms can be used as maternal genetic markers to distinguish the two types of domestic geese.

  4. The Complex History of the Domestication of Rice

    PubMed Central

    Sweeney, Megan; McCouch, Susan

    2007-01-01

    Background Rice has been found in archaeological sites dating to 8000 bc, although the date of rice domestication is a matter of continuing debate. Two species of domesticated rice, Oryza sativa (Asian) and Oryza glaberrima (African) are grown globally. Numerous traits separate wild and domesticated rices including changes in: pericarp colour, dormancy, shattering, panicle architecture, tiller number, mating type and number and size of seeds. Scope Genetic studies using diverse methodologies have uncovered a deep population structure within domesticated rice. Two main groups, the indica and japonica subspecies, have been identified with several subpopulations existing within each group. The antiquity of the divide has been estimated at more than 100 000 years ago. This date far precedes domestication, supporting independent domestications of indica and japonica from pre-differentiated pools of the wild ancestor. Crosses between subspecies display sterility and segregate for domestication traits, indicating that different populations are fixed for different networks of alleles conditioning these traits. Numerous domestication QTLs have been identified in crosses between the subspecies and in crosses between wild and domesticated accessions of rice. Many of the QTLs cluster in the same genomic regions, suggesting that a single gene with pleiotropic effects or that closely linked clusters of genes underlie these QTL. Recently, several domestication loci have been cloned from rice, including the gene controlling pericarp colour and two loci for shattering. The distribution and evolutionary history of these genes gives insight into the domestication process and the relationship between the subspecies. Conclusions The evolutionary history of rice is complex, but recent work has shed light on the genetics of the transition from wild (O. rufipogon and O. nivara) to domesticated (O. sativa) rice. The types of genes involved and the geographic and genetic distribution of

  5. Women's lived experiences of domestic violence during pregnancy (2).

    PubMed

    Baird, Kathleen

    2015-10-01

    This paper is a follow up paper to a study which explored women's experiences of domestic violence before, during and after pregnancy. Findings from this study suggested that women would like midwives to be able to recognise the signs of domestic violence and to be able to offer them an appropriate response and support. Midwives are well placed to recognise the signs of domestic violence and provide appropriate support. This paper addresses some of the challenges and dilemmas for midwives when identifying and supporting women who have experienced domestic violence and provides some key messages for midwifery practice. PMID:26638654

  6. Coat color variation at the beginning of horse domestication.

    PubMed

    Ludwig, Arne; Pruvost, Melanie; Reissmann, Monika; Benecke, Norbert; Brockmann, Gudrun A; Castaños, Pedro; Cieslak, Michael; Lippold, Sebastian; Llorente, Laura; Malaspinas, Anna-Sapfo; Slatkin, Montgomery; Hofreiter, Michael

    2009-04-24

    The transformation of wild animals into domestic ones available for human nutrition was a key prerequisite for modern human societies. However, no other domestic species has had such a substantial impact on the warfare, transportation, and communication capabilities of human societies as the horse. Here, we show that the analysis of ancient DNA targeting nuclear genes responsible for coat coloration allows us to shed light on the timing and place of horse domestication. We conclude that it is unlikely that horse domestication substantially predates the occurrence of coat color variation, which was found to begin around the third millennium before the common era. PMID:19390039

  7. Domestic group, status of women, and fertility.

    PubMed

    Patel, T

    1982-01-01

    An attempt is made in this paper to explain the fertility behavior of Patels; a major peasant caste in village Mogra in Rajasthan, India. The Patels make up 162 of the 404 households in the village and nearly 50% of the village's population of 2610. The Patels claim to have migrated from Gujarat a few centuries ago. This paper is based on a study of 140 randomly selected Patel households comprising 168 couples by using the method of intensive fieldwork over a period of about 6 months. Every household in Mogra, with the possible exception of 1 household of a Bania trader, is involved in agriculture in 1 capacity or another. The Patels are the major landowning caste, and all but 2 of the households in the sample of 140 own land. The Patels marry early, and the practice of child marriage remains common today. Being a patrilineal people, Patels consider sons as heirs to family property and status. Of the 140 households in the sample, 65 are complex in composition while 75 are simple. All the simple households are consequences of the partition of complex households. The consummation of marriage marks an immediate loss of independence to the woman. The Patels of Mogra have sound economic reasons for having a large number of children. They prefer to use family labor to the maximum possible extent. When labor is the major input, the land can absorb whatever labor is available. A great demand exists for labor in the domestic sphere as well. Compared to the returns, the cost of bringing up children is negligible. The expenditure on their clothing and education is minimal. Economics alone do not explain high fertility. Sons are essential to continue the line and for old age security. The marriage of daughters brings in bride price and religious merit. 1 son may find the burden of carrying out the kinship obligations inherited from his parents too high. When there are several sons, they can share the burden among themselves. Religious values and the obligations of kinship and

  8. Cardiovascular histamine receptors in the domestic chicken.

    PubMed

    Chand, N; Eyre, P

    1975-08-01

    The effects of mepyramine (H1-antagonist) and burimamide (H2-antagonist) were studied on histamine, 2-methylhistamine (a selective H1-agonist), 4-methylhistamine (a selective H2-agonist) and acetylcholine-induced changes in systemic arterial and central venous pressure and respiration in anaesthetized chickens. The result of this study suggested a predominance of H1 and some H2 histamine receptors in the cardiovascular system of domestic fowl where both are mediating systemic hypotension. There also appears to be predominance of H1 receptors mediating venous hypertension and respiratory apnoea to large doses of histamine and 2-methylhistamine. In addition, a possible involvement of H2-receptors in the cardiovascular system of chicken is suggested by the finding that burimamide always blocked mepyramine potentiated secondary pressor response to histamine and its analogues.

  9. An advanced domestic satellite communications system

    NASA Technical Reports Server (NTRS)

    1980-01-01

    An updated traffic projection for U.S. domestic satellite communications service covering a period of 15 years; mid-1980 to mid-1995 was prepared. This model takes into account expected technology advances and reductions in transmission costs, legislative and regulatory changes permitting increased competition, and rising energy costs which will encourage more extensive substitution of telecommunications for travel. The historical development and current status of satellite systems are discussed as well as the characteristics of follow-on systems. Orbital arc utilization, spacecraft configuration for single shuttle launch, Earth station configuration, and system costs are examined. Areas which require technology development include multiple beam frequency reuse antennas, on-board switching, intersatellite links, and ka-band operation. Packing and deployment schemes for enclosing the satellite within the shuttle orbiter bay must also be devised.

  10. Domestic violence and employment: a qualitative study.

    PubMed

    Swanberg, Jennifer E; Logan, T K

    2005-01-01

    This exploratory study sought to gather detailed information about how domestic violence affects women's employment, specifically to identify the types of job interference tactics used by abusers and their consequences on women's job performance; identify and understand the context associated with disclosure about victimization to employers and coworkers; and identify the supports offered to employees after disclosure. Qualitative analyses, guided by grounded theory, revealed that perpetrators exhibited job interference behaviors before, during, and after work. Abuser tactics reduced women's job performance as measured by absenteeism, tardiness, job leavings, and terminations. Among women who disclosed victimization to employers, informal and formal job supports were offered. Workplace supports led to short-term job retention, but fear and safety issues mitigated employers' attempts to retain workers.

  11. Pluripotent Stem Cells from Domesticated Mammals.

    PubMed

    Ezashi, Toshihiko; Yuan, Ye; Roberts, R Michael

    2016-01-01

    This review deals with the latest advances in the study of embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) from domesticated species, with a focus on pigs, cattle, sheep, goats, horses, cats, and dogs. Whereas the derivation of fully pluripotent ESC from these species has proved slow, reprogramming of somatic cells to iPSC has been more straightforward. However, most of these iPSC depend on the continued expression of the introduced transgenes, a major drawback to their utility. The persistent failure in generating ESC and the dependency of iPSC on ectopic genes probably stem from an inability to maintain the stability of the endogenous gene networks necessary to maintain pluripotency. Based on work in humans and rodents, achievement of full pluripotency will likely require fine adjustments in the growth factors and signaling inhibitors provided to the cells. Finally, we discuss the future utility of these cells for biomedical and agricultural purposes. PMID:26566158

  12. Emotional Intelligence of Women Who Experience Domestic Violence.

    PubMed

    Tsirigotis, Konstantinos; Łuczak, Joanna

    2016-03-01

    Violence in family constitutes serious social and psychological problem with harmful consequences leading, among others, to changes in emotional functioning of victim and, secondarily, also perpetrator. The aim of this study was to examine emotional intelligence of women experiencing domestic violence. INTE, i.e. Polish version of "Assessing Emotional Scale" by Schutte, was used to study two groups of women. Study (criterion) group included 40 women aged 23-47 years (mean age 35.28) using assistance of Crisis Intervention Centre due to experienced domestic violence. Reference (control) group was well-matched in terms of socio-demographic characteristics and consisted of 140 women not experiencing domestic violence. Study women experiencing domestic violence have significantly lower scores on all INTE indicators (general score, Factor I and Factor II). Women not experiencing domestic violence achieved significantly higher scores on Factor I than on Factor II. In this group all INTE components (general score, Factor I, Factor II) are positively correlated, whereas in group of women experiencing domestic violence there is no significant correlation between Factor I and Factor II and coefficients are lower. Emotional intelligence of study women experiencing domestic violence is lower than emotional intelligence of women not experiencing domestic violence. Their abilities and skills making up emotional intelligence are also less developed. The internal structure of emotional intelligence of study women experiencing domestic violence differs from emotional intelligence of women not experiencing domestic violence. It seems advisable to consider emotional intelligence in the process of providing women experiencing domestic violence with psychosocial help.

  13. Emotional Intelligence of Women Who Experience Domestic Violence.

    PubMed

    Tsirigotis, Konstantinos; Łuczak, Joanna

    2016-03-01

    Violence in family constitutes serious social and psychological problem with harmful consequences leading, among others, to changes in emotional functioning of victim and, secondarily, also perpetrator. The aim of this study was to examine emotional intelligence of women experiencing domestic violence. INTE, i.e. Polish version of "Assessing Emotional Scale" by Schutte, was used to study two groups of women. Study (criterion) group included 40 women aged 23-47 years (mean age 35.28) using assistance of Crisis Intervention Centre due to experienced domestic violence. Reference (control) group was well-matched in terms of socio-demographic characteristics and consisted of 140 women not experiencing domestic violence. Study women experiencing domestic violence have significantly lower scores on all INTE indicators (general score, Factor I and Factor II). Women not experiencing domestic violence achieved significantly higher scores on Factor I than on Factor II. In this group all INTE components (general score, Factor I, Factor II) are positively correlated, whereas in group of women experiencing domestic violence there is no significant correlation between Factor I and Factor II and coefficients are lower. Emotional intelligence of study women experiencing domestic violence is lower than emotional intelligence of women not experiencing domestic violence. Their abilities and skills making up emotional intelligence are also less developed. The internal structure of emotional intelligence of study women experiencing domestic violence differs from emotional intelligence of women not experiencing domestic violence. It seems advisable to consider emotional intelligence in the process of providing women experiencing domestic violence with psychosocial help. PMID:25982081

  14. Chewing rates among domestic dog breeds

    PubMed Central

    Gerstner, Geoffrey E.; Cooper, Meghan; Helvie, Peter

    2010-01-01

    The mammalian masticatory rhythm is produced by a brainstem timing network. The rhythm is relatively fixed within individual animals but scales allometrically with body mass (Mb) across species. It has been hypothesized that sensory feedback and feed-forward adjust the rhythm to match the jaw's natural resonance frequency, with allometric scaling being an observable consequence. However, studies performed with adult animals show that the rhythm is not affected by jaw mass manipulations, indicating that either developmental or evolutionary mechanisms are required for allometry to become manifest. The present study was performed to tease out the relative effects of development versus natural selection on chewing rate allometry. Thirty-one dog breeds and 31 mass-matched non-domestic mammalian species with a range in Mb from ∼2 kg to 50 kg were studied. Results demonstrated that the chewing rhythm did not scale with Mb among dog breeds (R=0.299, P>0.10) or with jaw length (Lj) (R=0.328, P>0.05). However, there was a significant relationship between the chewing rhythm and Mb among the non-domestic mammals (R=0.634, P<0.001). These results indicate that scaling is not necessary in the adult animal. We conclude that the central timing network and related sensorimotor systems may be necessary for rhythm generation but they do not explain the 1/3rd to 1/4th allometric scaling observed among adult mammals. The rhythm of the timing network is either adjusted to the physical parameters of the jaw system during early development only, is genetically determined independently of the jaw system or is uniquely hard-wired among dogs and laboratory rodents. PMID:20543125

  15. Blood steroid concentrations in domestic Mongolian horses.

    PubMed

    Haffner, John C; Fecteau, Kellie A; Eiler, Hugo; Tserendorj, Tsek; Hoffman, Rhonda M; Oliver, Jack W

    2010-07-01

    Traditionally, analysis of blood cortisol alone has been used to evaluate adrenal function. Currently, multisteroid analyses are considered more informative than analysis of a single hormone to assess adrenal function. The objective of the present research was to create a database for steroid reference values for domestic Mongolian horses. Seven adrenal steroid levels were determined in the blood of 18 colts, 34 stallions, 25 geldings, 17 fillies, and 29 mares. Results were as follows (lowest and highest group median, in nanograms per milliliter): progesterone: <0.030 (fillies), 4.30 (mares), and 0.070 (all horses); 17-OH-progesterone: 0.070 (colts), 0.520 (mares), and 0.110 (all horses); androstenedione: 0.101 (colts), 0.256 (stallions), and 0.181 (all horses); testosterone: <0.040 (mares, stallions, and fillies), 0.040 (geldings and colts), and <0.40 (all horses); estradiol: 0.066 (stallions), 0.093 (fillies), and 0.085 (all horses); cortisol: 23.040 (colts), 70.210 (geldings), and 50.770 (all horses); and aldosterone: 0.018 (colts), 0.297 (geldings), and 0.191 (all horses). Overall medians indicate that cortisol (98.70%) is the predominant steroid, followed by aldosterone (0.37%), androstenedione (0.35%), 17-OH-progesterone (0.21%), estradiol (0.17%), progesterone (0.14%), and testosterone (0.06%). This information provides adrenal and gonadal steroid reference concentrations to assist in physiological characterization and diagnosis of endocrine disorders in domestic Mongolian horses.

  16. 45 CFR 1370.4 - State domestic violence coalition grants.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 45 Public Welfare 4 2010-10-01 2010-10-01 false State domestic violence coalition grants. 1370.4... DEVELOPMENT SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS § 1370.4 State domestic violence coalition grants....

  17. 45 CFR 1370.4 - State domestic violence coalition grants.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 45 Public Welfare 4 2011-10-01 2011-10-01 false State domestic violence coalition grants. 1370.4... DEVELOPMENT SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS § 1370.4 State domestic violence coalition grants....

  18. 45 CFR 1370.4 - State domestic violence coalition grants.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 45 Public Welfare 4 2013-10-01 2013-10-01 false State domestic violence coalition grants. 1370.4... DEVELOPMENT SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS § 1370.4 State domestic violence coalition grants....

  19. 45 CFR 1370.4 - State domestic violence coalition grants.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 45 Public Welfare 4 2014-10-01 2014-10-01 false State domestic violence coalition grants. 1370.4... DEVELOPMENT SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS § 1370.4 State domestic violence coalition grants....

  20. 45 CFR 1370.4 - State domestic violence coalition grants.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 45 Public Welfare 4 2012-10-01 2012-10-01 false State domestic violence coalition grants. 1370.4... DEVELOPMENT SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAMS § 1370.4 State domestic violence coalition grants....

  1. 50 CFR 28.42 - Impounding of domestic animals.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 9 2014-10-01 2014-10-01 false Impounding of domestic animals. 28.42... VIOLATIONS OF PARTS 25, 26, AND 27 Impoundment Procedures § 28.42 Impounding of domestic animals. (a) Any animal trespassing on the lands of any national wildlife refuge may be impounded and disposed of...

  2. 25 CFR 91.11 - Domestic animals in village reserves.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 25 Indians 1 2012-04-01 2011-04-01 true Domestic animals in village reserves. 91.11 Section 91.11... VILLAGES, OSAGE RESERVATION, OKLAHOMA § 91.11 Domestic animals in village reserves. (a) No livestock shall... owner of the animal, if known, by certified mail or by posting in the village square. The notice...

  3. 50 CFR 28.42 - Impounding of domestic animals.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 9 2013-10-01 2013-10-01 false Impounding of domestic animals. 28.42... VIOLATIONS OF PARTS 25, 26, AND 27 Impoundment Procedures § 28.42 Impounding of domestic animals. (a) Any animal trespassing on the lands of any national wildlife refuge may be impounded and disposed of...

  4. 25 CFR 91.11 - Domestic animals in village reserves.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 25 Indians 1 2014-04-01 2014-04-01 false Domestic animals in village reserves. 91.11 Section 91.11... VILLAGES, OSAGE RESERVATION, OKLAHOMA § 91.11 Domestic animals in village reserves. (a) No livestock shall... owner of the animal, if known, by certified mail or by posting in the village square. The notice...

  5. 50 CFR 28.42 - Impounding of domestic animals.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 9 2012-10-01 2012-10-01 false Impounding of domestic animals. 28.42... VIOLATIONS OF PARTS 25, 26, AND 27 Impoundment Procedures § 28.42 Impounding of domestic animals. (a) Any animal trespassing on the lands of any national wildlife refuge may be impounded and disposed of...

  6. 25 CFR 91.11 - Domestic animals in village reserves.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 25 Indians 1 2011-04-01 2011-04-01 false Domestic animals in village reserves. 91.11 Section 91.11... VILLAGES, OSAGE RESERVATION, OKLAHOMA § 91.11 Domestic animals in village reserves. (a) No livestock shall... owner of the animal, if known, by certified mail or by posting in the village square. The notice...

  7. 50 CFR 28.42 - Impounding of domestic animals.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 8 2011-10-01 2011-10-01 false Impounding of domestic animals. 28.42... VIOLATIONS OF PARTS 25, 26, AND 27 Impoundment Procedures § 28.42 Impounding of domestic animals. (a) Any animal trespassing on the lands of any national wildlife refuge may be impounded and disposed of...

  8. The domestic dog: man's best friend in the genomic era

    PubMed Central

    2011-01-01

    The domestic dog genome - shaped by domestication, adaptation to human-dominated environments and artificial selection - encodes tremendous phenotypic diversity. Recent developments have improved our understanding of the genetics underlying this diversity, unleashing the dog as an important model organism for complex-trait analysis. PMID:21338479

  9. Hope and Healing for Children Affected by Domestic Violence

    ERIC Educational Resources Information Center

    Polites, Andrea; Kuchar, Karen; Bigelow, Shauna

    2010-01-01

    Domestic violence is a devastating social problem that leaves an enduring, negative impact on all family members, especially the victims and their children. The costs to children and to society as a whole are enormous. Children who have witnessed domestic violence or have been threatened or abused by a parent are at great risk for emotional and…

  10. Domestic Violence and Sexual Assault Service Goal Priorities

    ERIC Educational Resources Information Center

    Macy, Rebecca J.; Johns, Natalie; Rizo, Cynthia F.; Martin, Sandra L.; Giattina, Mary

    2011-01-01

    We investigated agency directors' perspectives about how service goals should be prioritized for domestic violence and sexual assault service subtypes, including crisis, legal advocacy, medical advocacy, counseling, support group, and shelter services. A sample of 97 (94% response rate) North Carolina domestic violence and/or sexual assault agency…

  11. 29 CFR 552.101 - Domestic service employment.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... OF THE FAIR LABOR STANDARDS ACT TO DOMESTIC SERVICE Interpretations § 552.101 Domestic service... Act (20 CFR 404.1057) and from “the generally accepted meaning” of the term. Accordingly, the term... regulations issued under the Social Security Act (20 CFR 404.1057) and from “the generally accepted...

  12. What Would They Do? Latino Church Leaders and Domestic Violence

    ERIC Educational Resources Information Center

    Behnke, Andrew O.; Ames, Natalie; Hancock, Tina U.

    2012-01-01

    Understanding what Latino church leaders believe about domestic violence, and what they do when they confront it, is a key step in developing programs to help them engage in domestic violence prevention and intervention activities in their congregations. This article presents the findings from an exploratory study of 28 Latino church leaders. The…

  13. 25 CFR 91.11 - Domestic animals in village reserves.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 25 Indians 1 2013-04-01 2013-04-01 false Domestic animals in village reserves. 91.11 Section 91.11 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR TRIBAL GOVERNMENT GOVERNMENT OF INDIAN VILLAGES, OSAGE RESERVATION, OKLAHOMA § 91.11 Domestic animals in village reserves. (a) No livestock...

  14. Genetic structure and domestication history of the grape

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The grape is one of the earliest domesticated fruit crops and, since antiquity, it has been widely cultivated and prized for its fruit and wine. Here, we characterize genome-wide patterns of genetic variation in over 1,000 samples of the domesticated grape, Vitis vinifera subsp. vinifera, and its wi...

  15. 12 CFR 614.4232 - Loans to domestic lessors.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 6 2010-01-01 2010-01-01 false Loans to domestic lessors. 614.4232 Section 614.4232 Banks and Banking FARM CREDIT ADMINISTRATION FARM CREDIT SYSTEM LOAN POLICIES AND OPERATIONS Loan Terms and Conditions § 614.4232 Loans to domestic lessors. Loans and financial assistance extended...

  16. Understanding How Domestic Violence Affects Behavior in High School Students

    ERIC Educational Resources Information Center

    Frank, Malika

    2011-01-01

    This paper will provide the reader with an understanding of how domestic violence affects the behavior of high school students. The presentation is designed to provide the reader with a working definition of domestic violence, the rate of occurrence and its effects on high school students. Additionally the paper will summarize the negative effects…

  17. 50 CFR 28.42 - Impounding of domestic animals.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 6 2010-10-01 2010-10-01 false Impounding of domestic animals. 28.42... VIOLATIONS OF PARTS 25, 26, AND 27 Impoundment Procedures § 28.42 Impounding of domestic animals. (a) Any animal trespassing on the lands of any national wildlife refuge may be impounded and disposed of...

  18. 25 CFR 91.11 - Domestic animals in village reserves.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Domestic animals in village reserves. 91.11 Section 91.11... VILLAGES, OSAGE RESERVATION, OKLAHOMA § 91.11 Domestic animals in village reserves. (a) No livestock shall... owner of the animal, if known, by certified mail or by posting in the village square. The notice...

  19. Chicken domestication: an updated perspective based on mitochondrial genomes.

    PubMed

    Miao, Y-W; Peng, M-S; Wu, G-S; Ouyang, Y-N; Yang, Z-Y; Yu, N; Liang, J-P; Pianchou, G; Beja-Pereira, A; Mitra, B; Palanichamy, M G; Baig, M; Chaudhuri, T K; Shen, Y-Y; Kong, Q-P; Murphy, R W; Yao, Y-G; Zhang, Y-P

    2013-03-01

    Domestic chickens (Gallus gallus domesticus) fulfill various roles ranging from food and entertainment to religion and ornamentation. To survey its genetic diversity and trace the history of domestication, we investigated a total of 4938 mitochondrial DNA (mtDNA) fragments including 2843 previously published and 2095 de novo units from 2044 domestic chickens and 51 red junglefowl (Gallus gallus). To obtain the highest possible level of molecular resolution, 50 representative samples were further selected for total mtDNA genome sequencing. A fine-gained mtDNA phylogeny was investigated by defining haplogroups A-I and W-Z. Common haplogroups A-G were shared by domestic chickens and red junglefowl. Rare haplogroups H-I and W-Z were specific to domestic chickens and red junglefowl, respectively. We re-evaluated the global mtDNA profiles of chickens. The geographic distribution for each of major haplogroups was examined. Our results revealed new complexities of history in chicken domestication because in the phylogeny lineages from the red junglefowl were mingled with those of the domestic chickens. Several local domestication events in South Asia, Southwest China and Southeast Asia were identified. The assessment of chicken mtDNA data also facilitated our understanding about the Austronesian settlement in the Pacific. PMID:23211792

  20. Domestic Refrigeration, Freezer, and Window Air Conditioner Service. Teacher Edition.

    ERIC Educational Resources Information Center

    Clemons, Mark

    This curriculum guide contains six units of instruction for a course in domestic refrigerator, freezer, and window air conditioner service. The units cover the following topics: (1) service fundamentals; (2) mechanical components and functions; (3) electrical components and control devices; (4) refrigerator and freezer service; (5) domestic ice…

  1. Impact of a Rural Domestic Violence Prevention Campaign.

    ERIC Educational Resources Information Center

    Gadomski, Anne M.; Tripp, Maria; Wolff, Debra A.; Lewis, Carol; Jenkins, Paul

    2001-01-01

    A 7-month public health information campaign used radio advertising, mass media articles, mailings, and posters to address attitudes and behavioral intentions toward domestic violence in a rural county. The campaign raised public awareness, particularly among men; increased stated intentions to intervene in a neighbor's domestic violence; and…

  2. An Integrated Hypothesis on the Domestication of Bactris gasipaes

    PubMed Central

    Dufour, Dominique; van Zonneveld, Maarten; Escobar Salamanca, Andrés Felipe; Giraldo Toro, Andrés; Rivera, Andrés; Salazar Duque, Hector; Suárez Baron, Harold; Gallego, Gerardo; Scheldeman, Xavier; Gonzalez Mejia, Alonso

    2015-01-01

    Peach palm (Bactris gasipaes Kunth) has had a central place in the livelihoods of people in the Americas since pre-Columbian times, notably for its edible fruits and multi-purpose wood. The botanical taxon includes both domesticated and wild varieties. Domesticated var gasipaes is believed to derive from one or more of the three wild types of var. chichagui identified today, although the exact dynamics and location of the domestication are still uncertain. Drawing on a combination of molecular and phenotypic diversity data, modeling of past climate suitability and existing literature, we present an integrated hypothesis about peach palm’s domestication. We support a single initial domestication event in south western Amazonia, giving rise to var. chichagui type 3, the putative incipient domesticate. We argue that subsequent dispersal by humans across western Amazonia, and possibly into Central America allowed for secondary domestication events through hybridization with resident wild populations, and differential human selection pressures, resulting in the diversity of present-day landraces. The high phenotypic diversity in the Ecuadorian and northern Peruvian Amazon suggest that human selection of different traits was particularly intense there. While acknowledging the need for further data collection, we believe that our results contribute new insights and tools to understand domestication and dispersal patterns of this important native staple, as well as to plan for its conservation. PMID:26658881

  3. A Comparison of International and Domestic Tertiary Students in Australia

    ERIC Educational Resources Information Center

    Khawaja, Nigar G.; Dempsey, Jenny

    2008-01-01

    In this study international and domestic students were compared on variables such as accommodation and financial satisfaction, social support, mismatched expectations, academic stress, dysfunctional coping, and psychological distress. International and domestic students (N = 86 for each group), enrolled at a large Australian university based in a…

  4. Genetic structure and domestication history of the grape

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The grape is one of the earliest domesticated fruit crops and, since antiquity, it has been widely cultivated and prized for its fruit and wine. We characterized genome-wide patterns of genetic variation in over 1000 samples of the domesticated grape, Vitis vinifera, and its wild relative, Vitis sy...

  5. Current perspectives and the future of domestication studies.

    PubMed

    Larson, Greger; Piperno, Dolores R; Allaby, Robin G; Purugganan, Michael D; Andersson, Leif; Arroyo-Kalin, Manuel; Barton, Loukas; Climer Vigueira, Cynthia; Denham, Tim; Dobney, Keith; Doust, Andrew N; Gepts, Paul; Gilbert, M Thomas P; Gremillion, Kristen J; Lucas, Leilani; Lukens, Lewis; Marshall, Fiona B; Olsen, Kenneth M; Pires, J Chris; Richerson, Peter J; Rubio de Casas, Rafael; Sanjur, Oris I; Thomas, Mark G; Fuller, Dorian Q

    2014-04-29

    It is difficult to overstate the cultural and biological impacts that the domestication of plants and animals has had on our species. Fundamental questions regarding where, when, and how many times domestication took place have been of primary interest within a wide range of academic disciplines. Within the last two decades, the advent of new archaeological and genetic techniques has revolutionized our understanding of the pattern and process of domestication and agricultural origins that led to our modern way of life. In the spring of 2011, 25 scholars with a central interest in domestication representing the fields of genetics, archaeobotany, zooarchaeology, geoarchaeology, and archaeology met at the National Evolutionary Synthesis Center to discuss recent domestication research progress and identify challenges for the future. In this introduction to the resulting Special Feature, we present the state of the art in the field by discussing what is known about the spatial and temporal patterns of domestication, and controversies surrounding the speed, intentionality, and evolutionary aspects of the domestication process. We then highlight three key challenges for future research. We conclude by arguing that although recent progress has been impressive, the next decade will yield even more substantial insights not only into how domestication took place, but also when and where it did, and where and why it did not.

  6. European domestic horses originated in two holocene refugia.

    PubMed

    Warmuth, Vera; Eriksson, Anders; Bower, Mim A; Cañon, Javier; Cothran, Gus; Distl, Ottmar; Glowatzki-Mullis, Marie-Louise; Hunt, Harriet; Luís, Cristina; do Mar Oom, Maria; Yupanqui, Isabel Tupac; Ząbek, Tomasz; Manica, Andrea

    2011-03-30

    The role of European wild horses in horse domestication is poorly understood. While the fossil record for wild horses in Europe prior to horse domestication is scarce, there have been suggestions that wild populations from various European regions might have contributed to the gene pool of domestic horses. To distinguish between regions where domestic populations are mainly descended from local wild stock and those where horses were largely imported, we investigated patterns of genetic diversity in 24 European horse breeds typed at 12 microsatellite loci. The distribution of high levels of genetic diversity in Europe coincides with the distribution of predominantly open landscapes prior to domestication, as suggested by simulation-based vegetation reconstructions, with breeds from Iberia and the Caspian Sea region having significantly higher genetic diversity than breeds from central Europe and the UK, which were largely forested at the time the first domestic horses appear there. Our results suggest that not only the Eastern steppes, but also the Iberian Peninsula provided refugia for wild horses in the Holocene, and that the genetic contribution of these wild populations to local domestic stock may have been considerable. In contrast, the consistently low levels of diversity in central Europe and the UK suggest that domestic horses in these regions largely derive from horses that were imported from the Eastern refugium, the Iberian refugium, or both.

  7. 43 CFR 6.7 - Domestic patent protection.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 1 2012-10-01 2011-10-01 true Domestic patent protection. 6.7 Section 6.7 Public Lands: Interior Office of the Secretary of the Interior PATENT REGULATIONS Inventions by Employees § 6.7 Domestic patent protection. (a) The Solicitor, upon determining that an invention coming...

  8. 43 CFR 6.7 - Domestic patent protection.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 43 Public Lands: Interior 1 2010-10-01 2010-10-01 false Domestic patent protection. 6.7 Section 6.7 Public Lands: Interior Office of the Secretary of the Interior PATENT REGULATIONS Inventions by Employees § 6.7 Domestic patent protection. (a) The Solicitor, upon determining that an invention...

  9. 43 CFR 6.7 - Domestic patent protection.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 43 Public Lands: Interior 1 2014-10-01 2014-10-01 false Domestic patent protection. 6.7 Section 6.7 Public Lands: Interior Office of the Secretary of the Interior PATENT REGULATIONS Inventions by Employees § 6.7 Domestic patent protection. (a) The Solicitor, upon determining that an invention...

  10. 43 CFR 6.7 - Domestic patent protection.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 43 Public Lands: Interior 1 2013-10-01 2013-10-01 false Domestic patent protection. 6.7 Section 6.7 Public Lands: Interior Office of the Secretary of the Interior PATENT REGULATIONS Inventions by Employees § 6.7 Domestic patent protection. (a) The Solicitor, upon determining that an invention...

  11. 43 CFR 6.7 - Domestic patent protection.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 1 2011-10-01 2011-10-01 false Domestic patent protection. 6.7 Section 6.7 Public Lands: Interior Office of the Secretary of the Interior PATENT REGULATIONS Inventions by Employees § 6.7 Domestic patent protection. (a) The Solicitor, upon determining that an invention...

  12. Do Workplace Flexibility Policies Influence Time Spent in Domestic Labor?

    ERIC Educational Resources Information Center

    Noonan, Mary C.; Estes, Sarah Beth; Glass, Jennifer L.

    2007-01-01

    Using data from a U.S. midwestern sample of mothers and fathers, the authors examine whether using workplace flexibility policies alters time spent in housework and child care. They hypothesize that an individual's policy use will lead to more time in domestic labor and that his or her spouse's policy use will lead to less time in domestic labor.…

  13. An Integrated Hypothesis on the Domestication of Bactris gasipaes.

    PubMed

    Galluzzi, Gea; Dufour, Dominique; Thomas, Evert; van Zonneveld, Maarten; Escobar Salamanca, Andrés Felipe; Giraldo Toro, Andrés; Rivera, Andrés; Salazar Duque, Hector; Suárez Baron, Harold; Gallego, Gerardo; Scheldeman, Xavier; Gonzalez Mejia, Alonso

    2015-01-01

    Peach palm (Bactris gasipaes Kunth) has had a central place in the livelihoods of people in the Americas since pre-Columbian times, notably for its edible fruits and multi-purpose wood. The botanical taxon includes both domesticated and wild varieties. Domesticated var gasipaes is believed to derive from one or more of the three wild types of var. chichagui identified today, although the exact dynamics and location of the domestication are still uncertain. Drawing on a combination of molecular and phenotypic diversity data, modeling of past climate suitability and existing literature, we present an integrated hypothesis about peach palm's domestication. We support a single initial domestication event in south western Amazonia, giving rise to var. chichagui type 3, the putative incipient domesticate. We argue that subsequent dispersal by humans across western Amazonia, and possibly into Central America allowed for secondary domestication events through hybridization with resident wild populations, and differential human selection pressures, resulting in the diversity of present-day landraces. The high phenotypic diversity in the Ecuadorian and northern Peruvian Amazon suggest that human selection of different traits was particularly intense there. While acknowledging the need for further data collection, we believe that our results contribute new insights and tools to understand domestication and dispersal patterns of this important native staple, as well as to plan for its conservation. PMID:26658881

  14. 7 CFR 1427.103 - Upland cotton Domestic User Agreement.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 10 2014-01-01 2014-01-01 false Upland cotton Domestic User Agreement. 1427.103... CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS COTTON Economic Adjustment Assistance to Users of Upland Cotton § 1427.103 Upland cotton Domestic User Agreement. (a) Payments...

  15. 7 CFR 1427.103 - Upland cotton Domestic User Agreement.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false Upland cotton Domestic User Agreement. 1427.103... CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS COTTON Economic Adjustment Assistance to Users of Upland Cotton § 1427.103 Upland cotton Domestic User Agreement. (a) Payments...

  16. 7 CFR 1427.103 - Upland cotton Domestic User Agreement.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 10 2011-01-01 2011-01-01 false Upland cotton Domestic User Agreement. 1427.103... CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS COTTON Economic Adjustment Assistance to Users of Upland Cotton § 1427.103 Upland cotton Domestic User Agreement. (a) Payments...

  17. 7 CFR 1427.103 - Upland cotton Domestic User Agreement.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 10 2013-01-01 2013-01-01 false Upland cotton Domestic User Agreement. 1427.103... CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS COTTON Economic Adjustment Assistance to Users of Upland Cotton § 1427.103 Upland cotton Domestic User Agreement. (a) Payments...

  18. 7 CFR 1427.103 - Upland cotton Domestic User Agreement.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 10 2012-01-01 2012-01-01 false Upland cotton Domestic User Agreement. 1427.103... CORPORATION, DEPARTMENT OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS COTTON Economic Adjustment Assistance to Users of Upland Cotton § 1427.103 Upland cotton Domestic User Agreement. (a) Payments...

  19. 32 CFR 231.5 - Procedures-domestic banks.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 2 2014-07-01 2014-07-01 false Procedures-domestic banks. 231.5 Section 231.5...) MISCELLANEOUS PROCEDURES GOVERNING BANKS, CREDIT UNIONS AND OTHER FINANCIAL INSTITUTIONS ON DOD INSTALLATIONS Guidelines § 231.5 Procedures—domestic banks. (a) General policy. Given their role in promoting morale...

  20. 32 CFR 231.5 - Procedures-domestic banks.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 2 2012-07-01 2012-07-01 false Procedures-domestic banks. 231.5 Section 231.5...) MISCELLANEOUS PROCEDURES GOVERNING BANKS, CREDIT UNIONS AND OTHER FINANCIAL INSTITUTIONS ON DOD INSTALLATIONS Guidelines § 231.5 Procedures—domestic banks. (a) General policy. Given their role in promoting morale...

  1. 32 CFR 231.5 - Procedures-domestic banks.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 2 2013-07-01 2013-07-01 false Procedures-domestic banks. 231.5 Section 231.5...) MISCELLANEOUS PROCEDURES GOVERNING BANKS, CREDIT UNIONS AND OTHER FINANCIAL INSTITUTIONS ON DOD INSTALLATIONS Guidelines § 231.5 Procedures—domestic banks. (a) General policy. Given their role in promoting morale...

  2. Assembling Webs of Support: Child Domestic Workers in India

    ERIC Educational Resources Information Center

    Wasiuzzaman, Shaziah; Wells, Karen

    2010-01-01

    This paper uses ethnographic and qualitative interview data with Muslim child domestic workers, their families and employers to investigate the social ties between young workers and their employers. Our analysis shows that working-class families use children's domestic work with middle-class families as part of a web of resources to protect them…

  3. 77 FR 60611 - National Domestic Violence Awareness Month, 2012

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-04

    .... (Presidential Sig.) [FR Doc. 2012-24682 Filed 10-3-12; 8:45 am] Billing code 3295-F3 ... Documents#0;#0; ] Proclamation 8877 of October 1, 2012 National Domestic Violence Awareness Month, 2012 By the President of the United States of America A Proclamation For far too long, domestic violence...

  4. 76 FR 62291 - National Domestic Violence Awareness Month, 2011

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-07

    .... (Presidential Sig.) [FR Doc. 2011-26147 Filed 10-6-11; 8:45 am] Billing code 3295-F2-P ... Documents#0;#0; ] Proclamation 8727 of October 3, 2011 National Domestic Violence Awareness Month, 2011 By the President of the United States of America A Proclamation During Domestic Violence Awareness...

  5. An Integrated Hypothesis on the Domestication of Bactris gasipaes.

    PubMed

    Galluzzi, Gea; Dufour, Dominique; Thomas, Evert; van Zonneveld, Maarten; Escobar Salamanca, Andrés Felipe; Giraldo Toro, Andrés; Rivera, Andrés; Salazar Duque, Hector; Suárez Baron, Harold; Gallego, Gerardo; Scheldeman, Xavier; Gonzalez Mejia, Alonso

    2015-01-01

    Peach palm (Bactris gasipaes Kunth) has had a central place in the livelihoods of people in the Americas since pre-Columbian times, notably for its edible fruits and multi-purpose wood. The botanical taxon includes both domesticated and wild varieties. Domesticated var gasipaes is believed to derive from one or more of the three wild types of var. chichagui identified today, although the exact dynamics and location of the domestication are still uncertain. Drawing on a combination of molecular and phenotypic diversity data, modeling of past climate suitability and existing literature, we present an integrated hypothesis about peach palm's domestication. We support a single initial domestication event in south western Amazonia, giving rise to var. chichagui type 3, the putative incipient domesticate. We argue that subsequent dispersal by humans across western Amazonia, and possibly into Central America allowed for secondary domestication events through hybridization with resident wild populations, and differential human selection pressures, resulting in the diversity of present-day landraces. The high phenotypic diversity in the Ecuadorian and northern Peruvian Amazon suggest that human selection of different traits was particularly intense there. While acknowledging the need for further data collection, we believe that our results contribute new insights and tools to understand domestication and dispersal patterns of this important native staple, as well as to plan for its conservation.

  6. Risk Factors and Interventions for Domestic Violence among Asian Americans.

    ERIC Educational Resources Information Center

    Kim, Irene

    This paper discusses domestic violence, examining its epidemiology for the general population and for two Asian American groups. It reviews data from 10 empirical studies on domestic violence among Asian American women. Qualitative studies stress the impact of family ties, family honor, and shame; religious values; fear of the legal system; and…

  7. Archaeological data reveal slow rates of evolution during plant domestication.

    PubMed

    Purugganan, Michael D; Fuller, Dorian Q

    2011-01-01

    Domestication is an evolutionary process of species divergence in which morphological and physiological changes result from the cultivation/tending of plant or animal species by a mutualistic partner, most prominently humans. Darwin used domestication as an analogy to evolution by natural selection although there is strong debate on whether this process of species evolution by human association is an appropriate model for evolutionary study. There is a presumption that selection under domestication is strong and most models assume rapid evolution of cultivated species. Using archaeological data for 11 species from 60 archaeological sites, we measure rates of evolution in two plant domestication traits--nonshattering and grain/seed size increase. Contrary to previous assumptions, we find the rates of phenotypic evolution during domestication are slow, and significantly lower or comparable to those observed among wild species subjected to natural selection. Our study indicates that the magnitudes of the rates of evolution during the domestication process, including the strength of selection, may be similar to those measured for wild species. This suggests that domestication may be driven by unconscious selection pressures similar to that observed for natural selection, and the study of the domestication process may indeed prove to be a valid model for the study of evolutionary change.

  8. 7 CFR 1427.102 - Eligible domestic users.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false Eligible domestic users. 1427.102 Section 1427.102... OF AGRICULTURE LOANS, PURCHASES, AND OTHER OPERATIONS COTTON Economic Adjustment Assistance to Users of Upland Cotton § 1427.102 Eligible domestic users. (a) For purposes of this subpart, a...

  9. Domestic Violence and the Impact on Young Children

    ERIC Educational Resources Information Center

    Zinke, Michelle; Zinke, Linda

    2008-01-01

    Domestic violence can be described as a pattern of intentional behaviors that includes a variety of tactics, such as physical and sexual violence, stalking, threats/intimidation, isolation, psychological attacks, and spiritual and economic abuse. Domestic violence can happen to anyone. It does not discriminate on the basis of economic status,…

  10. Sex Disparities in Arrest Outcomes for Domestic Violence

    ERIC Educational Resources Information Center

    Hamilton, Melissa; Worthen, Meredith G. F.

    2011-01-01

    Domestic violence arrests have been historically focused on protecting women and children from abusive men. Arrest patterns continue to reflect this bias with more men arrested for domestic violence compared to women. Such potential gender variations in arrest patterns pave the way to the investigation of disparities by sex of the offender in…

  11. Local initiatives: USAID / Peru supports national legislation on domestic violence.

    PubMed

    1997-01-01

    This brief article describes the activities of USAID-funded groups that promote gender equality and the protection of women and children against domestic violence in Peru. The Peruvian legislature passed Public Law 26260 in 1993, which addresses the issue of domestic violence. The Movimiento Mujers Peruanas Manuela Ramos received support from USAID to develop a registration, protection, redress, and referral system for women who suffer domestic violence. A campaign was conducted that used leaflets and radio and television to educate women about the new law and their rights under it. USAID also supported the Ombudsman's Office for Women. This office instituted a national study on domestic violence that will provide a database for monitoring reductions in domestic violence expected from the new law and the campaign. ReproSalud has a reproductive health project that USAID is funding. This group as well as 18 other groups function under an umbrella group, the Manuela Ramos. The target is poor women living in disadvantaged rural and urban areas, who are at greater risk of domestic violence. Manuela Ramos conducted participatory, qualitative research that helps women rank their reproductive health needs. All 18 organizations found domestic violence to be a reproductive health problem. One organization identified it as the most serious problem, and one that contributes to complications during delivery. Manuela Ramos works to make men more aware of the negative impact that domestic violence has on women.

  12. Current perspectives and the future of domestication studies

    PubMed Central

    Larson, Greger; Piperno, Dolores R.; Allaby, Robin G.; Purugganan, Michael D.; Andersson, Leif; Arroyo-Kalin, Manuel; Barton, Loukas; Climer Vigueira, Cynthia; Denham, Tim; Dobney, Keith; Doust, Andrew N.; Gepts, Paul; Gilbert, M. Thomas P.; Gremillion, Kristen J.; Lucas, Leilani; Lukens, Lewis; Marshall, Fiona B.; Olsen, Kenneth M.; Pires, J. Chris; Richerson, Peter J.; Rubio de Casas, Rafael; Sanjur, Oris I.; Thomas, Mark G.; Fuller, Dorian Q.

    2014-01-01

    It is difficult to overstate the cultural and biological impacts that the domestication of plants and animals has had on our species. Fundamental questions regarding where, when, and how many times domestication took place have been of primary interest within a wide range of academic disciplines. Within the last two decades, the advent of new archaeological and genetic techniques has revolutionized our understanding of the pattern and process of domestication and agricultural origins that led to our modern way of life. In the spring of 2011, 25 scholars with a central interest in domestication representing the fields of genetics, archaeobotany, zooarchaeology, geoarchaeology, and archaeology met at the National Evolutionary Synthesis Center to discuss recent domestication research progress and identify challenges for the future. In this introduction to the resulting Special Feature, we present the state of the art in the field by discussing what is known about the spatial and temporal patterns of domestication, and controversies surrounding the speed, intentionality, and evolutionary aspects of the domestication process. We then highlight three key challenges for future research. We conclude by arguing that although recent progress has been impressive, the next decade will yield even more substantial insights not only into how domestication took place, but also when and where it did, and where and why it did not. PMID:24757054

  13. The Use of Adolescents as Domestic Servants in Ibadan, Nigeria

    ERIC Educational Resources Information Center

    Okafor, Emeka Emmanuel

    2009-01-01

    The use of adolescents as domestic servants has become prevalent in most urban centers in Nigeria. This study focused on adolescents who work as domestic househelps in urban centers with special reference to Ibadan, Nigeria. The main objective of the study is to examine their mode of recruitment, the nature of their work as well the impact of such…

  14. Domestic Violence Reforms: Empty Promises or Fulfilled Expectations?

    ERIC Educational Resources Information Center

    Davis, Robert C.; Smith, Barbara

    1995-01-01

    Considers major reforms in the criminal justice system in response to domestic violence incidents: more frequent arrest, reduced control of victims in domestic court cases, more widespread use of restraining orders, and court-mandated treatment for batterers. Concludes that these do not work well across a variety of situations to reduce future…

  15. Domestic Transracial Adoption: In the Words of African American Adoptees

    ERIC Educational Resources Information Center

    Smith, Ellen M.

    2013-01-01

    Domestic transracial adoption has sparked more controversy than any other type of adoption. For the purposes of this study domestic transracial adoption is defined as European American parents adopting African American children. Many of the studies completed are contradictory and fail to control for variables that affect the outcome such as age of…

  16. Judging Risk: Key Determinants in British Domestic Violence Cases

    ERIC Educational Resources Information Center

    Robinson, Amanda L.; Howarth, Emma

    2012-01-01

    Data from the largest study to date of the working practices of British victim support workers (known as Independent Domestic Violence Advisors or IDVAs) are used to provide insight into how "risk judgments" are made in cases of domestic violence. Using data from more than 2,000 victims, this study found a convergence between actuarial data and…

  17. Modeling Prosecutors' Charging Decisions in Domestic Violence Cases

    ERIC Educational Resources Information Center

    Worrall, John L.; Ross, Jay W.; McCord, Eric S.

    2006-01-01

    Relatively little research explaining prosecutors' charging decisions in criminal cases is available. Even less has focused on charging decisions in domestic violence cases. Past studies have also relied on restrictive definitions of domestic violence, notably cases with male offenders and female victims, and they have not considered prosecutors'…

  18. 22 CFR 96.16 - Public domestic authorities.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Public domestic authorities. 96.16 Section 96.16 Foreign Relations DEPARTMENT OF STATE LEGAL AND RELATED SERVICES ACCREDITATION OF AGENCIES AND... Requirements for the Provision of Adoption Services § 96.16 Public domestic authorities. Public...

  19. Genomic DNA transposition induced by human PGBD5

    PubMed Central

    Henssen, Anton G; Henaff, Elizabeth; Jiang, Eileen; Eisenberg, Amy R; Carson, Julianne R; Villasante, Camila M; Ray, Mondira; Still, Eric; Burns, Melissa; Gandara, Jorge; Feschotte, Cedric; Mason, Christopher E; Kentsis, Alex

    2015-01-01

    Transposons are mobile genetic elements that are found in nearly all organisms, including humans. Mobilization of DNA transposons by transposase enzymes can cause genomic rearrangements, but our knowledge of human genes derived from transposases is limited. In this study, we find that the protein encoded by human PGBD5, the most evolutionarily conserved transposable element-derived gene in vertebrates, can induce stereotypical cut-and-paste DNA transposition in human cells. Genomic integration activity of PGBD5 requires distinct aspartic acid residues in its transposase domain, and specific DNA sequences containing inverted terminal repeats with similarity to piggyBac transposons. DNA transposition catalyzed by PGBD5 in human cells occurs genome-wide, with precise transposon excision and preference for insertion at TTAA sites. The apparent conservation of DNA transposition activity by PGBD5 suggests that genomic remodeling contributes to its biological function. DOI: http://dx.doi.org/10.7554/eLife.10565.001 PMID:26406119

  20. Feralisation targets different genomic loci to domestication in the chicken

    PubMed Central

    Johnsson, M.; Gering, E.; Willis, P.; Lopez, S.; Van Dorp, L.; Hellenthal, G.; Henriksen, R.; Friberg, U.; Wright, D.

    2016-01-01

    Feralisation occurs when a domestic population recolonizes the wild, escaping its previous restricted environment, and has been considered as the reverse of domestication. We have previously shown that Kauai Island's feral chickens are a highly variable and admixed population. Here we map selective sweeps in feral Kauai chickens using whole-genome sequencing. The detected sweeps were mostly unique to feralisation and distinct to those selected for during domestication. To ascribe potential phenotypic functions to these genes we utilize a laboratory-controlled equivalent to the Kauai population—an advanced intercross between Red Junglefowl and domestic layer birds that has been used previously for both QTL and expression QTL studies. Certain sweep genes exhibit significant correlations with comb mass, maternal brooding behaviour and fecundity. Our analyses indicate that adaptations to feral and domestic environments involve different genomic regions and feral chickens show some evidence of adaptation at genes associated with sexual selection and reproduction. PMID:27686863