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1

77 FR 24200 - American Drum & Pallet, Memphis, Shelby County, TN; Notice of Settlement  

Federal Register 2010, 2011, 2012, 2013

...CERCLA-04-2012-3770, FRL-9663-8] American Drum & Pallet, Memphis, Shelby County, TN; Notice...costs concerning the American Drum and Pallet Superfund Site located in Memphis, Shelby...2012-3770 or Site name American Drum & Pallet Superfund Site by one of the...

2012-04-23

2

Extensional collapse along the Sevier Desert reflection, northern Sevier Desert basin, western United States  

NASA Astrophysics Data System (ADS)

Newly released and previously published seismic reflection data from the northern Sevier Desert basin provide a complete seismic transect between the tilted western margin of the basin and the eastern breakaway zone. When tied to well and surface age data, the transect delineates a continuum of extensional fault and basin fill geometries that developed between late Oligocene and Pleistocene time across the basin. A minimum of 18 km of top-to-the-west normal displacement is estimated across the Sevier Desert from only the most conspicuous growth geometries and offsets across listric normal faults that sole downward into the Sevier Desert reflection (SDR). The SDR clearly marks a normal fault zone beneath the entire basin, where stratal truncations are imaged for 50% of the 39 km length of the reflection east of the Cricket Mountains block. Restoration of extensional displacement along this entire 39 km fault length is necessary to reconstruct the pre-Oligocene configuration and erosion level of Sevier thrust sheets across the Sevier Desert area. The SDR normal fault zone underlies the former topographic crest of the Sevier orogenic belt, where it accommodated extensional collapse after cessation of regional contractile tectonism.

Coogan, James C.; Decelles, Peter G.

1996-10-01

3

Anatomy of the middle ordovician sevier shale basin, eastern Tennessee  

NASA Astrophysics Data System (ADS)

The Sevier Shale basin in eastern Tennessee comprises one of the thickest clastic sequences (nearly 2500 m) of Middle Ordovician age in North America. The lower one-half of the sequence is composed of Lenoir, Whitesburg, Blockhouse and Sevier Formations, in ascending order. The sequence ranges in age from Whiterockian to lower Wilderness in North American stages. The Middle Ordovician sequence exhibits tidal flat (Mosheim Member of Lenoir Fm.), subtidal (main body of Lenoir Fm.), slope (Whitesburg Fm.), anoxic basin (Blockhouse Fm), turbidite and contourite (Sevier Fm.) facies. The Sevier basin evolved in five stages: First, a widespread marine transgression initiated carbonate-shelf deposition in the study area. Second, a major tectonic downwarping event caused the stable shelf to break and subside rapidly at a rate of 60-65 cm 1000 yrs -1, and areas of shelf facies became areas of slope and basin facies. Third, global transgressions maintained the deep anoxic conditions for nearly 10 Ma. Fourth, turbidites began to fill the basin from a westward-prograding submarine fan system. Fifth, contour currents reworked the turbidites and progressively ventilated the Sevier basin. The basin-filling process terminated with shallow-water/subaerial clastics at the end of Middle Ordovician.

Shanmugam, Ganapathy; Walker, Kenneth R.

1983-04-01

4

Sevier Bridge Reservoir, Juab and Sanpete Counties, Utah.  

National Technical Information Service (NTIS)

Annual total phosphorus and total nitrogen loadings to the lake were estimated and subdivided according to either point or non-point source origin. An assessment of the lake's trophic condition and limiting nutrient is also provided. All data collected by...

1977-01-01

5

OBVERSE VIEW OF TN24911, LOOKING BACK FROM THE NORTHEAST ROOM ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

OBVERSE VIEW OF TN-249-11, LOOKING BACK FROM THE NORTHEAST ROOM TO THE STAIR HALL. THE DOOR TO THE NORTH (DINING) ROOM LIES AS LEFT - Hilderbrand House, 4571 Airways Boulevard, Memphis, Shelby County, TN

6

10. Observation tower atop Clingman's Dome looking SE. Great ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

10. Observation tower atop Clingman's Dome looking SE. - Great Smoky Mountains National Park Roads & Bridges, Clingmans Dome Road, Between Newfound Gap Road & Clingmans Dome, Gatlinburg, Sevier County, TN

7

Transposon Tn5.  

PubMed

Tn5 was one of the first transposons to be identified ( 10 ). As a result of Tn5's early discovery and its simple macromolecular requirements for transposition, the Tn5 system has been a very productive tool for studying the molecular mechanism of DNA transposition. These studies are of broad value because they offer insights into DNA transposition in general, because DNA transposition is a useful model with which to understand other types of protein-DNA interactions such as retroviral DNA integration and the DNA cleavage events involved in immunoglobulin gene formation, and because Tn5-derived tools are useful adjuncts in genetic experimentation. PMID:18680433

Reznikoff, William S

2008-01-01

8

Mesozoic evolution, hinterland of the Sevier orogenic belt  

NASA Astrophysics Data System (ADS)

West of the Mesozoic foreland Sevier orogenic belt in northern Utah and southern Idaho, a regional detachment of inferred mid-Jurassic age occurs in Upper Mississippian shale and can be traced westward to the margin of the coeval magmatic arc, represented by spaced calc-alkalic intrusions. The detachment may be the oldest and structurally highest thrust of the Idaho-Wyoming thrust belt. If so, it bridges the apparent spatial and temporal gap between magmatism and thrusting, thus suggesting a direct genetic link. Thin-skinned deformation is interpreted here to be driven from the heated magmatic arc toward the colder craton by lateral tectonic compression created by lateral volume increase due to intrusion, elevated crustal geotherms, and possible thickening in the arc. The metamorphic complexes occur at the crustal interface between these two thermal-tectonic regimes and may themselves be allochthonous.

Allmendinger, Richard W.; Jordan, Teresa E.

1981-07-01

9

Is the Sevier Desert reflection of west-central Utah a normal fault?  

Microsoft Academic Search

A prominent west-dipping reflection that can be traced in seismic-reflection profiles over an area of 7000 km2 beneath the Sevier Desert basin of west-central Utah is generally referred to as the Sevier Desert detachment and is widely regarded as one of the best examples of an upper-crustal low-angle normal fault. The absence of evidence for fault-related deformation in drill cuttings

Mark H. Anders; Nicholas Christie-Blick

1994-01-01

10

Role of crustal thickening and extensional collapse in the tectonic evolution of the Sevier-Laramide orogeny, western United States  

Microsoft Academic Search

The effects of crustal thickening and extensional collapse of continental crust, combined with variations in plate-convergence forces, provide a coherent explanation for evolution of the Cretaceous-early Tertiary Sevier-Laramide orogeny. Crustal thickening of the Sevier-Laramide hinterland region was triggered by the combination of compressive plate-convergence forces and subduction-induced conductive heating of the crust. Eastward progradation of the locus of Sevier-Laramide deformation

Richard F. Livaccari

1991-01-01

11

Dune field reactivation from blowouts: Sevier Desert, UT, USA  

NASA Astrophysics Data System (ADS)

Dune field reactivation (a shift from vegetated to unvegetated state) has important economic, social, and environmental implications. In some settings reactivation is desired to preserve environmental values, but in arid regions reactivation is typically a form of land degradation. Little is known about reactivation due to a lack of published records, making modeling and prediction difficult. Here we detail dune reactivations from blowout expansion in the Sevier Desert, Utah, USA. We use historical aerial photographs and satellite imagery to track the transition from stable, vegetated dunes to actively migrating sediment in 3 locations. We outline a reactivation sequence: (i) disturbance breaches vegetation and exposes sediment, then (ii) creates a blowout with a deposition apron that (iii) advances downwind with a slipface or as a sand sheet. Most deposition aprons are not colonized by vegetation and are actively migrating. To explore causes we examine local sand flux, climate data, and stream flow. Based on available data the best explanation we can provide is that some combination of anthropogenic disturbance and climate may be responsible for the reactivations. Together, these examples provide a rare glimpse of dune field reactivation from blowouts, revealing the timescales, behaviour, and morphodynamics of devegetating dune fields.

Barchyn, Thomas E.; Hugenholtz, Chris H.

2013-12-01

12

Time of initial thrusting in the Sevier orogenic belt, Idaho-Wyoming and Utah  

NASA Astrophysics Data System (ADS)

Reexamination of the distribution of fossils found in the earliest preserved synorogenic conglomerates within the Sevier thrust belt suggests that initial thrust movement may be no older than Aptian age. This interpretation is corroborated by subsidence analyses of sedimentary sequences lying within and east of the Idaho-Wyoming and Utah thrust belts. A major episode of middle Cretaceous (Aptian Cenomanian) subsidence is interpreted as recording the initiation of thrust loading deformation in the adjacent Sevier orogenic belt. An earlier subsidence event took place during Middle Jurassic time, more than 30 m.y. prior to the Cretaceous event, and may be the result of tectonic events to the west that are unrelated to thrust deformation in the Idaho-Wyoming and Utah thrust belts. We find no verifiable evidence to support previous interpretations that Sevier belt deformation and uplift began in Late Jurassic time.

Heller, P. L.; Bowdler, S. S.; Chambers, H. P.; Coogan, J. C.; Hagen, E. S.; Shuster, M. W.; Winslow, N. S.; Lawton, T. F.

1986-05-01

13

Anatexis and metamorphism in tectonically thickened continental crust exemplified by the Sevier hinterland, western North America  

NASA Astrophysics Data System (ADS)

This paper presents a thermal and petrologic model of anatexis and metamorphism in regions of crustal thickening exemplified by the Sevier hinterland in western North America, and uses the model to examine the geological and physical processes leading to crustally derived magmatism. The results of numerical experiments show that anatexis was an inevitable end-product of Barrovian metamorphism in the thickened crust of the late Mesozoic Sevier orogenic belt and that the advection of heat across the lithosphere, in the form of mantle-derived mafic magmas, was not required for melting of metasedimentary rocks. It is suggested that, in the Sevier belt, as in other intracontinental orogenic belts, anatexis occurred in the midcrust and not at the base of the crust.

Patino Douce, Alberto E.; Humphreys, Eugene D.; Johnston, A. Dana

1990-03-01

14

The Tn5 transposon.  

PubMed

The bacterial transposon Tn5 encodes two proteins, the transposase and a related protein, the transposition inhibitor, whose relative abundance determines, in part, the frequency of Tn5 transposition. The synthesis of these proteins is programmed by a complex set of genetic regulatory elements. The host DNA methylation function, dam, inhibits transposase promoter recognition and indirectly enhances the transposition inhibitor promoter. The inhibitor lacks the N-terminal 55 amino acids of the transposase, suggesting that this sequence plays a key role in the transposition process. An intact N-terminal sequence is required for the transposase's recognition of the 19-bp end DNA sequences. This is the first critical step in the transposition process. Transposase-end DNA interaction is itself regulated by an intricate series of reactions involving several host proteins: DnaA, Dam, and Fis. The transposase is a unique protein in that it acts primarily in cis and inhibits its own activity in trans. Models to explain these properties are described. Finally circumstantial evidence suggests that transposition occurs preferentially from newly replicated DNA that has yet to be partitioned to progeny cells. This timing of transposition is likely to have a selective advantage for the host and the transposable element. PMID:7504907

Reznikoff, W S

1993-01-01

15

Is the Sevier Desert reflection of west-central Utah a normal fault?  

NASA Astrophysics Data System (ADS)

A prominent west-dipping reflection that can be traced in seismic-reflection profiles over an area of 7000 km2 beneath the Sevier Desert basin of west-central Utah is generally referred to as the Sevier Desert detachment and is widely regarded as one of the best examples of an upper-crustal low-angle normal fault. The absence of evidence for fault-related deformation in drill cuttings and core from two industry boreboles that intersect this feature casts doubt on the fault interpretation. The existing interpretation is based mainly on the observation that high-angle normal faulting is restricted largely to Tertiary sedimentary and volcanic rocks above the reflection. An alternative explanation is that the high-angle faults are related to the withdrawal of early deposited lacustrine salt, which even today is as much as 2 km thick. Reevaluation of the seismic data suggests that the Sevier Desert reflection consists of two spatially and genetically distinct segments: a shallow segment here interpreted as an unconformity between Paleozoic and Tertiary strata and a fortuitously aligned deeper segment that is traceable to mid- and lower-crustal levels and that appears to represent a thrust fault related to the Cretaceous Sevier orogeny.

Anders, Mark H.; Christie-Blick, Nicholas

1994-09-01

16

Mechanics of thrust faulting in crystalline basement, Sevier erogenic belt, Utah  

Microsoft Academic Search

Archean and Proterozoic basement was involved in foreland thrust faulting during the Sevier orogeny in northern Utah. Mesozoic structures in the Farmington Canyon Complex provide evidence for the mechanics of crystalline thrust plate transport and emplacement. Structures formed during the last stage of transport of the Farmington Canyon Complex over folded but mostly unmetamorphosed Paleozoic strata occur in the upper

Ronald L. Bruhn; Susan L. Beck

1981-01-01

17

Relations between hinterland and foreland shortening: Sevier orogeny, central North American Cordillera  

Microsoft Academic Search

The tectonic relations between foreland and hinterland deformation in noncollisional orogens are critical to understanding the overall development of orogens. The classic central Cordilleran foreland fold-and-thrust belt in the United States (Late Jurassic to early Tertiary Sevier belt) and the more internal zones to the west (central Nevada thrust belt) provide data critical to understanding the development of internal and

Wanda J. Taylor; John M. Bartley; Mark W. Martin; John W. Geissman; J. Douglas Walker; Phillip A. Armstrong; Joan E. Fryxell

2000-01-01

18

Alternatives for physically modifying John Sevier detention dam to allow fish passage  

SciTech Connect

Studies conducted in the vicinity of John Sevier Steam-Electric Plant (JSF) indicated some modification of the fish assemblage from that expected. By blocking movements of fish between Cherokee Reservoir and the upper Holston River, John Sevier detention dam has affected the fisheries in both systems. Providing passage for river-spawning fish at John Sevier detention dam might improve fish communities and fisheries in Cherokee Reservoir as well as upstream habitats. This would include enhanced reproductive success of river-spawning species found in Cherokee Reservoir (e.g., white bass and possibly striped bass and paddlefish) and repopulation of John Sevier Reservoir and the upper Holston River by several species presently found only downstream of the detention dam. TVA has identified and studied several alternatives that alone or in combination might improve the fisheries. Cost estimates were developed for three alternatives. These three alternatives with cost estimates are discussed briefly along with two other alternatives for which cost estimates have not been made. Merits of the three alternatives which have at least some possibility to improve migratory fish stocks are discussed in detail. 5 references.

Not Available

1984-09-01

19

Tn polyagglutination preceding acute leukemia.  

PubMed

Tn polyagglutination (persistent mixed-field polyagglutination) was detected in the blood of a 66-yr-old male laborer at the time of a splenectomy for life-threatening thrombocytopenia. Confirmation that the polyagglutination was caused by Tn activation was established by the use of lectins, by failure of the patient's red cells to react with sera from other patients with Tn polyagglutination, by weak aggregation with polybrene, by low red cell sialic acid levels, and by the persistence of polyagglutination over several years of testing. Two years after the discovery of the Tn polyagglutination, the patient developed acute myelomonocytic leukemia. Vigorous chemotherapy regimens resulted in clinical remission of the leukemia and the Tn polyagglutination. This report describes the first known case of Tn polyagglutination preceding the development of acute myelogenous leukemia. PMID:286607

Ness, P M; Garratty, G; Morel, P A; Perkins, H A

1979-07-01

20

75 FR 20774 - Establishment of Class E Airspace; Mountain City, TN  

Federal Register 2010, 2011, 2012, 2013

...date of a direct final rule published in the Federal Register December 7, 2009 that establishes Class E airspace at Johnson County Airport, Mountain City, TN. DATES: Effective Date: 0901 UTC, April 21, 2010. FOR FURTHER INFORMATION...

2010-04-21

21

Syntectonic infiltration by meteoric waters along the Sevier thrust front, southwest Montana  

Microsoft Academic Search

Structural, petrographic, and isotopic data for calcite veins and carbonate host-rocks from the Sevier thrust front of SW Montana record syntectonic infiltration by H2O-rich fluids with meteoric oxygen isotope compositions. Multiple generations of calcite veins record protracted fluid flow associated with regional Cretaceous contraction and subsequent Eocene extension. Vein mineralization occurred during single and multiple mineralization events, at times under

A. C. RYGEL; D. J. ANASTASIO; G. E. BEBOUT

2006-01-01

22

Relations between hinterland and foreland shortening: Sevier orogeny, central North American Cordillera  

NASA Astrophysics Data System (ADS)

The tectonic relations between foreland and hinterland deformation in noncollisional orogens are critical to understanding the overall development of orogens. The classic central Cordilleran foreland fold-and-thrust belt in the United States (Late Jurassic to early Tertiary Sevier belt) and the more internal zones to the west (central Nevada thrust belt) provide data critical to understanding the development of internal and external parts of orogens. The Garden Valley thrust system, part of the central Nevada thrust belt, crops out in south-central Nevada within a region generally considered to be the hinterland of the Jurassic to Eocene Sevier thrust belt. The thrust system consists of at least four principal thrust plates composed of strata as young as Pennsylvanian in age that are unconformably overlain by rocks as old as Oligocene, suggesting that contraction occurred between those times. New U/Pb dates on intrusions that postdate contraction, combined with new paleomagnetic data showing significant tilting of one area prior to intrusion, suggest that regionally these thrusts were active before ˜85-100 Ma. The thrust faults are characterized by long, relatively steeply dipping ramps and associated folds that are broad and open to close, upright and overturned. Although now fragmented by Cenozoic crustal extension, individual thrusts can be correlated from range to range for tens to hundreds of kilometers along strike. We correlate the structurally lowest thrust of the Garden Valley thrust system, the Golden Gate-Mount Irish thrust, southward with the Gass Peak thrust of southern Nevada. This correlation carries the following regional implications. At least some of the slip across Jurassic to mid-Cretaceous foreland thrusts in southern Nevada continues northward along the central Nevada thrust belt rather than northeastward into Utah. This continuation is consistent with age relations, which indicate that thrusts in the type Sevier belt in central Utah are synchronous with or younger than the youngest thrusts in southern Nevada. This in turn implies that geometrically similar Sevier belt thrusts in Utah must die out southward before they reach Nevada, that slip along the southern Nevada thrusts is partitioned between central Nevada and Utah thrusts, or that the Utah thrusts persist into southeastern Nevada but are located east of the longitude of the central Nevada thrust belt. As a result of overall cratonward migration of thrusting, the central Nevada thrust belt probably formed the Cordilleran foreland fold-thrust belt early in the shortening event but later lay in the hinterland of the Sevier fold-thrust belt of Idaho-Wyoming-Utah.

Taylor, Wanda J.; Bartley, John M.; Martin, Mark W.; Geissman, John W.; Walker, J. Douglas; Armstrong, Phillip A.; Fryxell, Joan E.

2000-12-01

23

The Mormon Peak and Tule Springs Detachments of Southern Nevada and Their Role in Interpreting the Subsurface Structure of the Sevier Desert Basin  

Microsoft Academic Search

The eastern Sevier front is commonly overprinted by west-dipping normal faults that have been active through much of Cenozoic time. Although examples of disparate age and geometry abound, the Sevier Desert detachment stands out as being: (1) a very large, low-angle structure, (2) probably still active, and (3) at drillable depth, and hence is a prime target for scientific drilling.

B. Wernicke; G. J. Axen

2007-01-01

24

Magnitude, Timing, and Geometry of Extension in the Southern Sevier Desert Basin From Piercing Points, Seismic-Stratigraphic Reconstruction, and Deep well Data  

Microsoft Academic Search

Palinspastic reconstruction of Mesozoic thrust sheets provides the main constraint for an estimated 47 km of Cenozoic extensional displacement along the Sevier Desert detachment (SDD) in the central Sevier Desert Basin. Hanging wall and footwall piercing points indicate that the SDD accommodated a minimum of 35 km of extensional displacement in the narrower southern part of the basin. The piercing

J. C. Coogan; P. G. Decelles

2007-01-01

25

8. Underside of bridge over Happy Hollow Road showing concrete ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. Underside of bridge over Happy Hollow Road showing concrete piers looking SSW. - Great Smoky Mountains National Park Roads & Bridges, Foothills Parkway, From Chilowee to Walland & from Cosby to I-40, Gatlinburg, Sevier County, TN

26

7. Bridge over Happy Hollow Road on unopened section of ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

7. Bridge over Happy Hollow Road on unopened section of Foothills Parkway looking ENE. - Great Smoky Mountains National Park Roads & Bridges, Foothills Parkway, From Chilowee to Walland & from Cosby to I-40, Gatlinburg, Sevier County, TN

27

6. Crenulated wall along Clingman's Dome Road looking S. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

6. Crenulated wall along Clingman's Dome Road looking S. - Great Smoky Mountains National Park Roads & Bridges, Clingmans Dome Road, Between Newfound Gap Road & Clingmans Dome, Gatlinburg, Sevier County, TN

28

8. Detail of Clingman's Dome parking area looking NW. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. Detail of Clingman's Dome parking area looking NW. - Great Smoky Mountains National Park Roads & Bridges, Clingmans Dome Road, Between Newfound Gap Road & Clingmans Dome, Gatlinburg, Sevier County, TN

29

9. Clingman's dome trail head and comfort station looking NNE. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

9. Clingman's dome trail head and comfort station looking NNE. - Great Smoky Mountains National Park Roads & Bridges, Clingmans Dome Road, Between Newfound Gap Road & Clingmans Dome, Gatlinburg, Sevier County, TN

30

5. Abandoned mule trail tunnel. 1 mile from intersection with ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

5. Abandoned mule trail tunnel. 1 mile from intersection with Newfound Gap Road looking SSE. - Great Smoky Mountains National Park Roads & Bridges, Clingmans Dome Road, Between Newfound Gap Road & Clingmans Dome, Gatlinburg, Sevier County, TN

31

8. Double arch culvert on Laurel Creek Road looking WSW. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. Double arch culvert on Laurel Creek Road looking WSW. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

32

1. View of Laurel Creek Road, revetment wall and cliff ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. View of Laurel Creek Road, revetment wall and cliff looking S. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

33

22. Cades Cove Valley from Hyatt Lane looking E. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

22. Cades Cove Valley from Hyatt Lane looking E. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

34

13. Cades Cove Road, with barbed wire fence and mountain ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

13. Cades Cove Road, with barbed wire fence and mountain view. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

35

5. Laurel Creek Road, revetment wall and creek. Great ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

5. Laurel Creek Road, revetment wall and creek. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

36

28. Ford near the end of Cades Cove Loop Road ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

28. Ford near the end of Cades Cove Loop Road looking NNW. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

37

27. Abandoned treelined road in Cades Cove looking N. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

27. Abandoned tree-lined road in Cades Cove looking N. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

38

9. Double arch culvert on Laurel Creek Road looking ENE. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

9. Double arch culvert on Laurel Creek Road looking ENE. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

39

25. Dan Lawson place in Cades Cove looking S. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

25. Dan Lawson place in Cades Cove looking S. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

40

10. Concrete bridge on road to Tremont looking S. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

10. Concrete bridge on road to Tremont looking S. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

41

16. First Baptist Church in Cades Cove looking SSW. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

16. First Baptist Church in Cades Cove looking SSW. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

42

7. Elevation of single arch stone bridge on Laurel Creek ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

7. Elevation of single arch stone bridge on Laurel Creek Road looking N. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

43

4. Ford near the northern end of road looking NNE. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

4. Ford near the northern end of road looking NNE. - Great Smoky Mountains National Park Roads & Bridges, Rich Mountain Road, Between Cades Cove & park boundary at Rich Mountain Gap, Gatlinburg, Sevier County, TN

44

23. Henry Whitehead place in Cades Cove looking ESE. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

23. Henry Whitehead place in Cades Cove looking ESE. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

45

12. Cades Cove Road, view toward entrance with ruts in ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

12. Cades Cove Road, view toward entrance with ruts in road. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

46

21. View down Hyatt Lane in Cades Cove looking NNE. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

21. View down Hyatt Lane in Cades Cove looking NNE. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

47

18. View of fields, mountains and mist in Cades Cove ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

18. View of fields, mountains and mist in Cades Cove looking SW. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

48

19. Cantilevered barn in Cades Cove looking SSW. Great ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

19. Cantilevered barn in Cades Cove looking SSW. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

49

24. View of barn, fields and Cades Cove loop road ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

24. View of barn, fields and Cades Cove loop road looking NNE. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

50

1. View of Cades Cove Valley from first overlook on ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. View of Cades Cove Valley from first overlook on Rich Mountain Road looking S. - Great Smoky Mountains National Park Roads & Bridges, Rich Mountain Road, Between Cades Cove & park boundary at Rich Mountain Gap, Gatlinburg, Sevier County, TN

51

2. View of west tunnel portal and stream on Laurel ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

2. View of west tunnel portal and stream on Laurel Creek Road looking ENE. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

52

6. 1930's revetment wall next to 1886 constructed revetment wall ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

6. 1930's revetment wall next to 1886 constructed revetment wall on Laurel Creek Road looking SSW. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

53

17. Methodist Church in Cades Cove looking NW. Great ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

17. Methodist Church in Cades Cove looking NW. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

54

26. View of Cades Cove Loop Road with fields and ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

26. View of Cades Cove Loop Road with fields and mountains looking NE. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

55

2. Switchback and General Road view looking S. Great ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

2. Switchback and General Road view looking S. - Great Smoky Mountains National Park Roads & Bridges, Rich Mountain Road, Between Cades Cove & park boundary at Rich Mountain Gap, Gatlinburg, Sevier County, TN

56

15. Cades Cove Road, mountain view with horses. Great ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

15. Cades Cove Road, mountain view with horses. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

57

14. Cades Cove Road, mountain view with single tree. ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

14. Cades Cove Road, mountain view with single tree. - Great Smoky Mountains National Park Roads & Bridges, Cades Cove Road & Laurel Creek Road, From Townsend Wye to Cades Cove, Gatlinburg, Sevier County, TN

58

Immunochemical studies on Tn erythrocyte glycoprotein.  

PubMed

Glycoproteins were extracted from membranes of erythrocytes that displayed Tn polyagglutination and were compared chemically and immunologically with glycoproteins of group O, MN cells. Tn glycoprotein had lower than normal NANA : protein and sugar : protein ratios, as revealed by direct analysis and polyacrylamide gel electrophoresis, and displayed slower immunoelectrophoretic mobility than glycoproteins of group O, MN cells. Agglutination of Tn cells by Salvia sclarea lectin was inhibited by Tn glycoprotein but not by O, MN glycoprotein. Tn and MN glycoproteins were equally potent inhibitors of influenza virus HA. Our findings indicate than Tn-specific determinants are part of the glycophorin molecule. PMID:6171313

Lee, L T; Frank, S; de Jongh, D S; Howe, C

1981-12-01

59

Transposon Tn916 Mutagenesis in Bacillus Anthracis.  

National Technical Information Service (NTIS)

Mutagenesis of Bacillus anthracis by the streptococcal tetracycline resistance transposon Tn916 is described. Tn916 was transferred from Streptococcus faecalis DS16C1 to B. anthracis VNR-1 by conjugation in a standard filter mating procedure. Tetracycline...

B. E. Ivins S. L. Welkos G. B. Knudson D. J. LeBlanc

1988-01-01

60

Sevier Laramide deformation of the continental interior from calcite twinning analysis, west-central North America  

NASA Astrophysics Data System (ADS)

Paleozoic-Mesozoic carbonates that cover cratonic western North America contain a regional layer-parallel shortening (LPS) fabric that is preserved by mechanically twinned calcite. Shortening directions are generally parallel to the Sevier thrust-transport direction (E-W) in carbonates of the Idaho-Wyoming portion of the thrust belt and within carbonates as far as 2000 km into the plate interior. The inferred calcite twinning differential stress magnitudes generally decrease across the thrust belt, and decrease exponentially away from the orogenic front into the craton. Synorogenic calcite cements and veins preserve a distinct twinning deformation history: in the thrust belt, twinning strains commonly record local, out-of-transport piggyback strain events with high differential stresses (<150 MPa), whereas in Laramide uplifts and adjacent basins as far east as the Black Hills, twinned vein calcite preserves a sub-horizontal, N-S-shortening strain, with differential stress magnitudes that decrease to the east. Deformation of the plate interior during the Sevier orogeny was dominated by E-W contraction at the plate margin, which changed into dominantly oblique contraction (˜N-S shortening) along western North America during the younger, basement-involved Laramide event.

Craddock, John P.; van der Pluijm, Ben A.

1999-05-01

61

COPY NUMBER CONTROL OF TN~ TRANSPOSITION  

Microsoft Academic Search

Transposition of Tn5 in Escherichia coli strains containing one or multiple copies of the transposable element was investigated. It was found that the overall frequency of transposition within a cell remained constant regardless of the number of copies of Tn5 present in that cell. Experiments measuring the transposition frequency of differentially marked Tn5s confirmed that the fre- quency of transposition

REID C. JOHNSON; WILLIAM S. REZNIKOFF

1984-01-01

62

Isotopic evidence for preservation of Cordilleran lithospheric mantle during the Sevier-Laramide orogeny, western United States  

Microsoft Academic Search

It has been inferred that Sevier-Laramide flat subduction imparted a large basal shear traction force to the overriding North American plate, resulting in Laramide Rocky Mountain foreland deformation and significant thinning of Cordilleran lithosphere. Additional inferred consequences include regional refrigeration of Cordilleran crust and suppression of synorogenic extensional collapse. Nd isotopic data from Cenozoic mafic volcanic rocks indicate that normal-thickness

Richard F. Livaccari; Frank V. Perry

1993-01-01

63

Generation of Tn5 insertions in streptococcal conjugative transposon Tn916.  

PubMed Central

A method has been developed for the introduction of Tn5 into Escherichia coli plasmid chimeras containing Streptococcus faecalis DNA. Tn5 could be introduced via a lambda::Tn5 delivery vehicle. The system proved to be particularly efficient and facilitated insertions at numerous sites on DNA containing the 16-kilobase conjugative transposon Tn916. It was possible to introduce some of the resulting Tn916::Tn5 derivatives back into S. faecalis by using a recently developed protoplast transformation procedure. A presumed zygotic induction resulted in insertion of the Tn916 derivatives at multiple sites in the S. faecalis chromosome. Images

Yamamoto, M; Jones, J M; Senghas, E; Gawron-Burke, C; Clewell, D B

1987-01-01

64

Generation of Tn5 insertions in streptococcal conjugative transposon Tn916.  

PubMed

A method has been developed for the introduction of Tn5 into Escherichia coli plasmid chimeras containing Streptococcus faecalis DNA. Tn5 could be introduced via a lambda::Tn5 delivery vehicle. The system proved to be particularly efficient and facilitated insertions at numerous sites on DNA containing the 16-kilobase conjugative transposon Tn916. It was possible to introduce some of the resulting Tn916::Tn5 derivatives back into S. faecalis by using a recently developed protoplast transformation procedure. A presumed zygotic induction resulted in insertion of the Tn916 derivatives at multiple sites in the S. faecalis chromosome. PMID:3038014

Yamamoto, M; Jones, J M; Senghas, E; Gawron-Burke, C; Clewell, D B

1987-05-01

65

Characterization of Tn5801.Sag, a Variant of Staphylococcus aureus Tn916 Family Transposon Tn5801 That Is Widespread in Clinical Isolates of Streptococcus agalactiae  

PubMed Central

Tn5801, originally detected in Staphylococcus aureus Mu50, is a Tn916 family element in which a unique int gene (int5801) replaces the int and xis genes in Tn916 (int916 and xis916). Among 62 tet(M)-positive tetracycline-resistant Streptococcus agalactiae isolates, 43 harbored Tn916, whereas 19 harbored a Tn5801-like element (Tn5801.Sag, ?20.6 kb). Tn5801.Sag was characterized (PCR mapping, partial sequencing, and chromosomal integration) and compared to other Tn5801-like elements. Similar to Tn5801 from S. aureus Mu50, tested in parallel, Tn5801.Sag was unable to undergo circularization and conjugal transfer.

Mingoia, Marina; Morici, Eleonora; Tili, Emily; Giovanetti, Eleonora; Montanari, Maria Pia

2013-01-01

66

Ground-water conditions and geologic reconnaissance of the Upper Sevier River basin, Utah  

USGS Publications Warehouse

The upper Sevier River basin is in south-central Utah and includes an area of about 2,400 .square miles of high plateaus and valleys. It comprises the entire Sevier River drainage basin above Kingston, including the East Fork Sevier River and its tributaries. The basin was investigated to determine general ground-water conditions, the interrelation of ground water and surface water, the effects of increasing the pumping of ground water, and the amount of ground water in storage. The basin includes four main valleys--Panguitch Valley, Circle Valley, East Fork Valley, and Grass Valley--which are drained by the Sevier River, the East Fork Sevier River, and Otter Creek. The plateaus surrounding the valleys consist of sedimentary and igneous rocks that range in age from Triassic to Quaternary. The valley fill, which is predominantly alluvial gravel, sand, silt, and clay, has a maximum thickness of more than 800 feet. The four main valleys constitute separate ground-water basins. East Fork Valley basin is divided into Emery Valley, Johns Valley, and Antimony subbasins, and Grass Valley basin is divided into Koosharem and Angle subbasins. Ground water occurs under both artesian and water-table conditions in all the basins and subbasins except Johns Valley, Emery Valley, and Angle subbasins, where water is only under water-table conditions. The. water is under artesian pressure in beds of gravel and sand confined by overlying beds of silt and clay in the downstream parts of Panguitch Valley basin, Circle Valley basin, and Antimony subbasin, and in most of Koosharem subbasin. Along the sides and upstream ends of these basins, water is usually under water-table conditions. About 1 million acre-feet of ground water that is readily available to wells is stored in the gravel and sand of the upper 200 feet of saturated valley fill. About 570,000 acre-feet is stored in Panguitch Valley basin, about 210,000 in Circle Valley basin, about 6,000 in Emery Valley subbasin, about 90,000 in Johns Valley subbasin, about 36,000 in Antimony subbasin, about 90,000 in Koosharem subbasin, and about 60,000 in Angle subbasin. Additional water, although it is not readily available to wells, is stored in beds of silt and clay. Some ground water also is available in the bedrock underlying and surrounding the basins, although the bedrock formations generally are poor aquifers. The principal source of recharge to the valley fill in the upper Sevier River basin is infiltration from streams, canals, and irrigated fields. Some ground water also miles into the valley till from the bedrock surrounding the basins. The basin contains about 300 wells, most of which are less than 4 inches in diameter, are less than 250 feet deep, and are used for domestic purposes and stock watering. More than half the wells are flowing wells in Koosharem subbasin. Approximately 82,000 acre-feet of ground water was discharged in 1962 from the valley till. Springs discharged about 33,000 acre-feet, wells about 3,000, and drains about 3,000; and evapotranspiration from phreatophyte areas about 43,000 acre-feet. Springs in bedrock discharged an additional 75,000 acre-feet. Most of the water discharged by springs, wells, and drains was used for irrigation. The ground water in the basin generally is of good chemical quality. The water is excellent for irrigation and stock but is not as desirable for most domestic and industrial uses because of its hardness. The dissolved-solids content of the ground water generally increases slightly from the upstream end of the individual ground-water basins to. the downstream end owing mostly to repeated use of the water for irrigation. Surface water and ground water in the upper Sevier River basin are interconnected, and the base flows of streams are affected by changes in groundwater levels. Increased pumping of ground water would result in (1) an increase in the recharge to the aquifers from surface-water sources or (2) a decrease

Carpenter, Carl H.; Robinson, Gerald B.; Bjorklund, Louis Jay

1967-01-01

67

Piggyback basin in the Sevier orogenic belt, Utah: Implications for development of the thrust wedge  

NASA Astrophysics Data System (ADS)

Timing data on a frontal thrust system indicate that the Sevier thrust wedge of central Utah advanced rapidly to its ultimate structural front in ˜30 m.y., between early Albian and Campanian time. It then persisted by out-of-sequence thrusting for an additional 25 m.y., until the early Eocene. The key to the history of the frontal thrust is a piggyback basin that formed on the hanging wall. Unconformity-bounded sedimentary units within the basin thin both westward and eastward onto structures generated by thrust faulting. Up to 860 m of upper Campanian to lower Eocene strata accumulated between these structures, which consist of a ramp anticline in the west and a thrust duplex in the east. The newly recognized interaction of coeval deposition and thrust deformation permits revised limits on the inception and longevity of the frontal thrust system.

Lawton, Timothy F.; Trexler, James H., Jr.

1991-08-01

68

Transposon Tn5 mutagenesis of Brucella abortus.  

PubMed

We demonstrate that transposon-mediated mutagenesis can be used to construct mutations in the pathogen Brucella abortus. We have used both a plasmid and a bacteriophage to introduce either Tn5 or Tn5 lac into the Brucella chromosome. B. abortus is naturally sensitive to kanamycin. We have selected 22 independent kanamycin-resistant colonies in strain US-19 and 19 colonies in strain 2308. We have demonstrated by Southern hybridization that Tn5 was inserted into the chromosome. PMID:2822580

Smith, L D; Heffron, F

1987-11-01

69

Presence of the Tn antigen on hematopoietic progenitors from patients with the Tn syndrome.  

PubMed Central

The Tn syndrome is an acquired clonal disorder characterized by the exposure of a normally hidden determinant, the Tn antigen, on the surface of human erythrocytes, platelets, granulocytes, and lymphocytes. Two distinct populations, Tn positive (Tn+) and Tn negative (Tn-), of mature hemopoietic cells are present in Tn patients. To determine whether the Tn antigen is already expressed on erythroid, myeloid, and pluripotent progenitors, light-density mononuclear blood cells from two patients with this syndrome were separated by fluorescent-activated cell sorting and by affinity chromatography into Tn+ and Tn- fractions, using their binding properties to Helix pomatia agglutinin (HPA). Burst-forming-unit erythroid (BFU-E), colony-forming-unit granulocyte/macrophage (CFU-GM), cells were assayed in plasma clot cultures. After 12-14 d of culture, colonies were studied by a double fluorescent labeling procedure. First, a fluorescein-conjugated HPA permitted evaluation of the presence or absence of the Tn antigen at the surface of the cells composing each colony, and second, the binding of a murine monoclonal antibody against either glycophorin A (LICR-LON-R10) or against a myeloid antigen (80H5), revealed by an indirect fluorescent procedure, was used to establish the erythroid or myeloid origin of each cell. The Tn+ fraction obtained by cell sorting gave rise to nearly 100% Tn+ colonies composed exclusively of cells bearing this antigen. The reverse was observed for the Tn- cell fraction. These results demonstrate that in the Tn syndrome, BFU-E, CFU-GM, and CFU-GEMM of the Tn+ clone express the Tn antigen at this early stage of differentiation. Images

Vainchenker, W; Vinci, G; Testa, U; Henri, A; Tabilio, A; Fache, M P; Rochant, H; Cartron, J P

1985-01-01

70

Thrust timing, growth of structural culminations, and synorogenic sedimentation in the type Sevier orogenic belt, western United States  

NASA Astrophysics Data System (ADS)

Most of the regional shortening in the type area of the Sevier orogenic belt in central Utah was accommodated by displacement on the Canyon Range (Neocomian-Aptian), Pavant (Aptian-Albian), Paxton (Cenomanian-Campanian), and Gunnison (late Campanian Paleocene) thrust systems. Inception of each thrust system generated synorogenic sediment associated with frontal thrust-tip anticlines or triangle zones and older thrust sheets that were elevated above major ramps farther toward the hinterland. The Sevier culmination, a large antiformal duplex cored by crystalline basement rocks, developed during Paxton and Gunnison thrusting west of and structurally beneath the Canyon Range and Pavant thrusts. Growth of the Sevier culmination was coeval with reactivation of the Canyon Range and Pavant thrust systems and produced a second culmination in Proterozoic Lower Cambrian rocks in the Canyon Range. These structural highs provided much of the sediment to the adjacent foreland basin from late Cenomanian to late Paleocene time, and may have helped to maintain critical taper in the thrust belt.

Decelles, Peter G.; Lawton, Timothy F.; Mitra, Gautam

1995-08-01

71

Influence of Late Cretaceous magmatism on the Sevier orogenic wedge, western Montana  

NASA Astrophysics Data System (ADS)

Late Cretaceous large-volume pluton emplacement and accompanying volcanism within the evolving western Montana thrust wedge may have played important roles in determining the geometric and kinematic development of the thrust wedge, thereby influencing patterns of sediment dispersal and subsidence in the adjacent foreland basin. Intrusion of the Upper Cretaceous Boulder batholith and coeval eruption of the Elkhorn Mountains Volcanics were focused mainly at the trailing margin of the Lombard-Eldorado allochthon. The resultant thick (16 17 km) igneous culmination evolved over a relatively short interval (80 70 Ma), thickening the orogenic wedge to the point of supercritical taper and facilitating continued motion along the Lombard-Eldorado thrust system, thrust imbrication at the wedge toe, and forelandward translation of the Helena salient. Delivery of eroded volcaniclastic detritus from the thickened thrust wedge and accelerated basin subsidence due to thrust loading resulted in accumulation of a thick (4 5 km) sequence of Campanian-Maastrichtian volcaniclastic strata in the foreland basin (Livingston Group). The structural and sedimentological effects of this structural-magmatic culmination are similar to those of basement-cored culminations elsewhere in the Sevier orogen and to the Neogene central Andean orogenic wedge.

Lageson, David R.; Schmitt, James G.; Horton, Brian K.; Kalakay, Thomas J.; Burton, Bradford R.

2001-08-01

72

Mechanics of thrust faulting in crystalline basement, Sevier erogenic belt, Utah  

NASA Astrophysics Data System (ADS)

Archean and Proterozoic basement was involved in foreland thrust faulting during the Sevier orogeny in northern Utah. Mesozoic structures in the Farmington Canyon Complex provide evidence for the mechanics of crystalline thrust plate transport and emplacement. Structures formed during the last stage of transport of the Farmington Canyon Complex over folded but mostly unmetamorphosed Paleozoic strata occur in the upper plate of the Ogden thrust fault, which was emplaced by cataclastic flow. Intraplate strain was accommodated by brittle faulting, and new greenschist facies minerals formed during hydro-thermal alteration. The Paleozoic strata directly below the thrust fault were ductilely deformed. The upper thrust plate was buckled into an anticline-syncline fold pair as it moved eastward along the Ogden thrust fault. The fold axes were oriented subparallel to the eastward direction of tectonic transport. Shortening at a high angle to the fault plane and eastward extension occurred in the cataclastic gneiss of the upper plate and the underlying sedimentary tectonites. These strains developed as the result of progressive simple shear during overthrusting. Strain due to buckling of the upper plate was superimposed on the simple shear strains in the cataclastic gneiss and caused lateral variations in the three-dimensional strain field within the thrust plate. Constrictional strain developed in the anticlinal hinge area of the thrust plate, whereas flattening strain occurred in the synclinal hinge zone.

Bruhn, Ronald L.; Beck, Susan L.

1981-05-01

73

Diversity of putative Tn 5253-like elements in Streptococcus pneumoniae  

Microsoft Academic Search

Pneumococcal resistance to tetracycline, chloramphenicol, erythromycin and clindamycin is often attributed to carriage of conjugative transposons of the Tn916 family. The less well studied conjugative transposon Tn5253 is a composite transposon consisting of a Tn916-like element inserted within the unrelated Tn5252 element, which has also been associated with chloramphenicol and tetracycline resistance. Here, carriage of the Tn5252 integrase (int5252), Tn5252-encoded

S. K. Henderson-Begg; A. P. Roberts; L. M. C. Hall

2009-01-01

74

Identification and analysis of teleost slow muscle troponin T (sTnT) and intronless TnT genes  

Microsoft Academic Search

In the present study cDNA clones representing two slow skeletal muscle troponin T genes (sTnT1sb and sTnT2sb) in the sea bream (Sparus auratus), an important aquaculture species, were isolated and characterised. A third, intronless, TnT gene (iTnTsb), which is an apparent orthologue of a previously described zebrafish TnT, was also isolated. In adult sea bream sTnT expression was restricted to

M. A. Campinho; D. M. Power; G. E. Sweeney

2005-01-01

75

The association of Tn and leukemia.  

PubMed

Tn polyagglutination or persistent mixed field polyagglutination is a rare acquired erythrocytic state which has been associated with haematologic abnormalities including four reported cases of acute leukaemia. Recent information has proven that the abnormality is clonal and the haematologic findings suggest a mutation of a somatic stem cell. The red cell membrane defect results from a deficiency of B-3-D galactosyltransferase. Tn polyagglutination has been described in patients presenting with acute leukaemia and has preceded acute leukaemia. The Tn syndrome has similarities to other forms of preleukaemic syndromes including paroxysmal nocturnal haemoglobinuria. Patients with Tn should be carefully monitored, although there is little justification for any specific therapy at the present time. PMID:6592126

Ness, P M

1983-01-01

76

Tn5tac1, a derivative of transposon Tn5 that generates conditional mutations.  

PubMed

Conditional lethal mutations are valuable for analyzing essential genes. We describe here a derivative of the bacterial transposon Tn5 called Tn5tac1 and its use in an innovative strategy for making mutations with conditional phenotypes. The 4.6-kilobase Tn5tac1 element contains a strong, regulatable, outward-facing promoter (Ptac) near one end and is polar on the expression of distal genes when the inducer of Ptac [isopropyl beta-D-thiogalactoside (IPTG)] is absent. Our results show that two unusual conditional mutant phenotypes can result from Tn5tac1 insertion in Escherichia coli: one is corrected by IPTG while the other is induced by IPTG. The broad host range of Tn5 and the conditional nature of these mutant phenotypes makes Tn5tac1 well suited for identifying essential genes in diverse bacterial species. PMID:2842773

Chow, W Y; Berg, D E

1988-09-01

77

Three-dimensional kinematic history at an oblique ramp, Leamington zone, Sevier belt, Utah  

NASA Astrophysics Data System (ADS)

Differences in kinematics in adjoining salients in fold-thrust belts are typically accommodated along bounding transverse zones. The Leamington zone is an oblique ramp that accommodates slip between structures in the Provo salient and the central Utah segment of the Sevier fold-thrust belt. The Leamington zone consists of the Leamington Canyon thrust, associated second-order asymmetric folds and an out-of-syncline reverse fault. The Leamington Canyon thrust is a rotated, SE-dipping thrust fault with top-to-southeast motion; it represents an oblique ramp of the folded Canyon Range thrust. Fold tightening of the syncline SE of the folded Leamington Canyon thrust trace during later emplacement of underlying structures, caused out-of-syncline reverse faulting in the fold-core. Analysis of the finite strain, minor fault populations, and conjugate fracture sets were used in interpreting the 3-D kinematic history along the Leamington zone. Restored maximum stretching directions of finite strain ellipsoids trend eastward overall. Inferred motion planes from populations of slickenlines on minor faults are consistent with overall southeastward motion. Acute bisectors of conjugate fracture sets may reflect more southward directions later in the deformation history. These superimposed deformations, with local relative chronologies of plastic and brittle structures using cross-cutting relationships, indicate that the Leamington zone area shows overall clockwise changes in kinematic directions from E to ESE to SSE during successive pulses of deformation. The temporal changes in kinematic directions along the Leamington zone most likely reflect variations in kinematics over the oblique ramp as the overall easterly displaced 3-D fold-thrust belt wedge interacted with the pre-existing oblique ramp structure, with small magnitudes of superimposed local vertical-axis rotations during later folding and fold-tightening of the Leamington Canyon thrust.

Kwon, Sanghoon; Mitra, Gautam

2006-03-01

78

Structural Discordance Between Neogene Detachments and Frontal Sevier Thrusts, Central Mormon Mountains, Southern Nevada  

NASA Astrophysics Data System (ADS)

Detailed geologic mapping in the Mormon Mountains of southern Nevada provides significant insight into processes of extensional tectonics developed within older compressional orogens. A newly discovered, WSW-directed low-angle normal fault, the Mormon Peak detachment, juxtaposes the highest levels of the frontal most part of the east-vergent, Mesozoic Sevier thrust belt with autochthonous crystalline basement. Palinspastic analysis suggests that the detachment initially dipped 20-25° to the west and cut discordantly across thrust faults. Nearly complete lateral removal of the hanging wall from the area has exposed a 5 km thick longitudinal cross-section through the thrust belt in the footwall, while highly attenuated remnants of the hanging wall (nowhere more than a few hundred meters thick) structurally veneer the range. The present arched configuration of the detachment resulted in part from progressive "domino-style" rotation of a few degrees while it was active, but is largely due to rotation on younger, structurally lower, basement-penetrating normal faults that initiated at high-angle. The geometry and kinematics of normal faulting in the Mormon Mountains suggest that pre-existing thrust planes are not required for the initiation of low-angle normal faults, and even where closely overlapped by extensional tectonism, need not function as a primary control of detachment geometry. Caution must thus be exercised in interpreting low-angle normal faults of uncertain tectonic heritage such as those seen in the COCORP west-central Utah and BIRP's MOIST deep-reflection profiles. Although thrust fault reactivation has reasonably been shown to be the origin of a very few low-angle normal faults, our results indicate that it may not be as fundamental a component of orogenic architecture as it is now widely perceived to be. We conclude that while in many instances thrust fault reactivation may be both a plausible and attractive hypothesis, it may never be assumed.

Wernicke, Brian; Walker, J. Douglas; Beaufait, Mark S.

1985-02-01

79

77 FR 61653 - Notice of Intent To Rule on Request To Release Airport Property at the Smyrna-Rutherford County...  

Federal Register 2010, 2011, 2012, 2013

...Notice of Intent To Rule on Request To Release Airport Property at the Smyrna-Rutherford County Airport, Smyrna, TN AGENCY: Federal Aviation Administration...of land at the Smyrna-Rutherford County Airport, Smyrna, Tennessee. This property,...

2012-10-10

80

Tn-polyagglutinability associated with acute myelomonocytic leukemia.  

PubMed

A case of Tn polyagglutinability in a patient with acute myelomonocytic leukemia is described. This represents the third report in which Tn polyagglutination has been associated with a leukemic state. Serologic recognition of Tn polyagglutination and probable causes are discussed. Known cases of Tn-polyagglutinability are reviewed. PMID:293127

Baldwin, M L; Barrasso, C; Ridolfi, R L

1979-12-01

81

Detection of the Tn antigen in leukaemia using monoclonal anti-Tn antibody and immunohistochemistry.  

PubMed

Exposure of the normally cryptic Tn antigen on haemopoietic cells leading to erythrocyte polyagglutination has been reported in a few cases of malignant or premalignant haemopoietic disorders and has been attributed to a selective deficiency of the enzyme 3-beta-D-galactosyl-transferase. A male patient presented with acute myelomonocytic leukaemia with no evidence of Tn expression but, 16 months later, in the terminal stage of the disease, the majority of the erythrocytes were found to be polyagglutinable. Tn expression was confirmed by the use of lectins and by agglutination with a Tn-specific monoclonal antibody, FBT3. Retrospective studies of stored blood and bone marrow smears were performed by immunocytochemistry using FBT3. Tn positive cells were first detected in the marrow 8 months prior to death. They increased progressively in number and, in the terminal illness, over 90% of erythroid precursors in the marrow and erythrocytes in peripheral blood and 40% of granulocyte precursors of the marrow and 10% of granulocytes in the blood were Tn positive. These observations suggest that Tn expression was present in a subclone of cells which became dominant during the course of the disease and that there may be a relationship between Tn expression and leukaemic progression. PMID:3479188

Roxby, D J; Morley, A A; Burpee, M

1987-10-01

82

Tn5-rpsL: a new derivative of transposon Tn5 useful in plasmid curing.  

PubMed

The rpsL gene of Escherichia coli was inserted into the BamHI site of transposon Tn5. This transposon was called Tn5-rpsL. Tn5-rpsL may be useful in microbiological studies when one wants to cure various bacterial genera of certain plasmid(s). A streptomycin-resistant (SmR) derivative of the host bacterial strain is first isolated. The plasmid(s) later to be cured are then labelled with Tn5-rpsL, which makes the cells Sm-sensitive. These cells can regain their resistance to Sm if they lose the Tn5-rpsL-tagged plasmid. Thus, plasmid-free bacteria are easily selected among SmR survivors. The frequency of occurrence of the plasmid-less variants of plasmid-containing wild-type Salmonella typhimurium measured by this method is given as an example. PMID:1850707

Stojiljkovi?, I; Trgovcevi?, Z; Salaj-Smic, E

1991-03-01

83

[A further case of Tn-polyagglutination].  

PubMed

Another case of Tn-polyagglutination is described where for 6 years those responses have been observed which are typical of the acquired erythrocyte changes, viz. mixed-field polyagglutination by normal adult sera of all blood groups, no response with anti-TAh, agglutination of a certain part of erythrocytes by anti-TnSs, anti-ADb, anti-AHP, agglomeration of the other part by protamin sulfate and A-like specificity. Papain treatment eliminates polyagglutination by human sera of adults. Sera of new-borns did not agglutinate Tn-erythrocytes. Mrs. B. B. 38 years old belongs to blood group 0 and shows latent signs of haemolysis as well as permanently lowered leukocyte and thrombocyte numbers. The difficulties in blood group serology in connection with polyagglutination are referred too. PMID:89994

Lahmann, N

1979-01-01

84

Thrust belt curvature: Structural and paleomagnetic analyses in the Catalunyan Pyrenees and Sevier orogen  

NASA Astrophysics Data System (ADS)

The question of how curved geologic structures form, from arcuate faults to curvilinear orogenic belts, is one that transcends scale as well as rock properties. Many scale-independent lines of evidence suggest that material within fold-thrust belts is transported in three dimensions. Paleomagnetic analysis is the primary method for determining distribution and magnitude of vertical-axis rotations in arcuate regions. Thus paleomagnetic information is essential to deciphering the three-dimensional kinematic evolution of a curved orogen. In addition, most studies of foreland fold-and-thrust belts focus on geometries and timing relationships, but do not identify the deformations internal to the thrust-bounded rock packages. The Southern Pyrenean and Sevier orogenic systems both provide an outstanding opportunity to investigate thrust-belt curvature by integrating paleomagnetic and structural studies. This dissertation addresses the interplay between three-dimensional motions and the structures they cause. In the Pyrenees, the Oliana anticline is a foreland structure related to the South Central Salient, a major curve in the thrust belt. Paleomagnetic data from the Oliana anticline document counter clockwise rotations about a vertical axis. The average rotation is R +/- Delta R = -19.6° +/- 10.5°. Based on the stratigraphic horizons that record the rotation, as well as the structural evolution of the Oliana anticline, the age of rotation is ?35 Ma. This rotation is attributed to continued motion along the Serres Marginals thrust, causing rotation along the blind thrusts that underlie the Oliana anticline. Mode I fracture data from this region highlights the progressive development of joints in growing structures. Measurements were taken from four regions around the anticline, and from four sequential synorogenic conglomerates. A consistent NW-SW orientation for the development of joints was determined by performing sequential restorations of the Oliana anticline and incrementally analyzing joint orientations. Reanalysis of previously published data from the Wyoming salient of the western United States demonstrates the relationship between irregular fault traces and the three-dimensional motions that caused them to form. Interaction between rotation, strain and net translation is a global phenomenon and suggests that geologic shortening estimates can be improved by incorporating the effects of tectonic rotations.

Sussman, Aviva Joy

2002-04-01

85

Genetic and molecular studies of a composite chromosomal element (Tn3705) containing a Tn916-modified structure (Tn3704) in Streptococcus anginosus F22.  

PubMed

The plasmid-free Streptococcus anginosus F22 contained a conjugative element, Tn3705, encoding resistance to erythromycin (Emr) and tetracycline-minocycline (Tcr-Mnr). We mapped a chromosomal region (> 52 kb) of F22, corresponding to the internal part of Tn3705. Molecular analysis of Tn3705 revealed it to be a composite structure: it included in its central part a transposon designated Tn3704 (20.3 kb +/- 0.5 kb), which had a modified structure in comparison with that of Tn916 and on which the Emr Tcr-Mn4 markers were localized. Tn3705 inserted from F22 into the chromosome of various streptococcal transconjugants as well as that of Enterococcus faecalis transconjugants without changing its structure. In contrast, from the chromosome of an E. faecalis::Tn3705 transconjugant only Tn3704 inserted, at various sites, into another E. faecalis chromosome. Sugar fermentations occurred after the insertion of Tn3704 into the chromosome of an asaccharolytic E. faecalis strain. Transposition of only Tn3704 from the chromosome of E. faecalis::Tn3705 onto pIP964, an E. faecalis hemolysin plasmid, yielded two different pIP964 derivatives. The size of the entire element Tn3705 was estimated to be about 70.0 kb by pulsed-field electrophoresis. PMID:8171124

Clermont, D; Horaud, T

1994-01-01

86

Control of Neogene Extension in Idaho and Utah by Thrust Ramps in the Sevier Orogenic Belt: Evidence From Gravity Modeling in Cache and Cottonwood Valleys  

Microsoft Academic Search

The subsurface structure of Cache and Cottonwood Valleys is largely due to two episodes of deformation. The Cretaceous to Eocene Sevier orogeny caused compressional folding and faulting across the region presently occupied by the basins. Following this, Neogene extension occurred along the Bannock detachment fault and other normal faults in Cache and Cottonwood Valleys. See-saw slippage of the west-dipping East

C. L. Groze; J. M. Kruger; M. L. Eversaul

2006-01-01

87

Tn5 transposase loops DNA in the absence of Tn5 transposon end sequences.  

PubMed

Transposases mediate transposition first by binding specific DNA end sequences that define a transposable element and then by organizing protein and DNA into a highly structured and stable nucleoprotein 'synaptic' complex. Synaptic complex assembly is a central checkpoint in many transposition mechanisms. The Tn5 synaptic complex contains two Tn5 transposase subunits and two Tn5 transposon end sequences, exhibits extensive protein-end sequence DNA contacts and is the node of a DNA loop. Using single-molecule and bulk biochemical approaches, we found that Tn5 transposase assembles a stable nucleoprotein complex in the absence of Tn5 transposon end sequences. Surprisingly, this end sequence-independent complex has structural similarities to the synaptic complex. This complex is the node of a DNA loop; transposase dimerization and DNA specificity mutants affect its assembly; and it likely has the same number of proteins and DNA molecules as the synaptic complex. Furthermore, our results indicate that Tn5 transposase preferentially binds and loops a subset of non-Tn5 end sequences. Assembly of end sequence-independent nucleoprotein complexes likely plays a role in the in vivo downregulation of transposition and the cis-transposition bias of many bacterial transposases. PMID:17074070

Adams, Christian D; Schnurr, Bernhard; Skoko, Dunja; Marko, John F; Reznikoff, William S

2006-12-01

88

Origin of the Uinta recess, Sevier fold thrust belt, Utah: influence of basin architecture on fold thrust belt geometry  

NASA Astrophysics Data System (ADS)

Structural trends in the Sevier fold-thrust belt define a pronounced concave-to-the-foreland map-view curve, the Uinta recess, in north-central Utah. This recess separates two convex-to-the-foreland curves, the Provo salient on the south and the Wyoming salient on the north. The two limbs of the recess comprise transverse zones (fault zones at a high-angle to the regional trend of the orogen) that border the flanks of the east-west-trending Uinta/Cottonwood arch. Our structural analysis indicates that the transverse zones formed during the Sevier orogeny, and that they differ markedly from each other in structural style. The Charleston transverse zone (CTZ), on the south side of the arch, initiated as a complex sinistral strike-slip fault system that defines the abrupt northern boundary of the Provo salient. The Mount Raymond transverse zone (MRTZ), on the north side of the arch, represents the region in which the southeast-verging southern limb of the gently curving Wyoming salient was tilted northwards during the Laramide phase of uplift of the Uinta/Cottonwood arch. In effect, the MRTZ represents an oblique cross section through a thrust belt. The contrasting architecture of these transverse zones demonstrates how pre-deformation basin geometry influences the geometry of a fold-thrust belt. Analysis of isopach maps indicates that, at the time the Sevier fold-thrust belt formed, the area just north of the present site of the Uinta/Cottonwood arch was a basement high, with a gently dipping north flank, and a steeply dipping south flank. Thus, predeformational sediment thickened abruptly to the south of the high and thickened gradually to the north of the high. As illustrated by sandbox models, the distance that a fold-thrust belt propagates into the foreland depends on the thickness of the sedimentary layer being deformed, so the shape of the salient mimics the longitudinal cross-sectional shape of the sedimentary basin. Where basins taper gradually along strike, the thrust belt curves gently, but where basins taper abruptly along strike, the thrust belt curves so tightly that it disarticulates and becomes bounded laterally by a strike-slip accommodation zone. The geometry of the Uinta recess provides a field example of this concept. Differential movement of Sevier thrusts led to formation of gradually curving thrusts on the north side of the high, because of the gradual slope of the high's north flank, but led to the along-strike disarticulation of thrusts on the south side of the high, because of the steep slope of the high's south flank. In effect, therefore, thrust belt map-view geometry provides insight into predeformational basin geometry.

Paulsen, Timothy; Marshak, Stephen

1999-11-01

89

77 FR 13195 - Amendment of Class E Airspace; Springfield, TN  

Federal Register 2010, 2011, 2012, 2013

...11-ASO-26] Amendment of Class E Airspace; Springfield, TN AGENCY: Federal Aviation Administration...action amends Class E Airspace in the Springfield, TN area. Aydelotte Airport has been...Rules (IFR) operations within the Springfield, TN airspace area. This action...

2012-03-06

90

78 FR 52424 - Amendment of Class E Airspace; Dayton, TN, Establishment of Class E Airspace; Cleveland, TN, and...  

Federal Register 2010, 2011, 2012, 2013

...establishes Class E Airspace at Cleveland Regional Jetport, Cleveland, TN, to accommodate area navigation (RNAV) global positioning system (GPS) SIAPs at the airport. Information regarding Bradley Memorial Hospital is added to the Cleveland, TN,...

2013-08-23

91

Bacterial transposon Tn5: evolutionary inferences.  

PubMed

Transposable elements induce spontaneous mutations, promote genome rearrangements, regulate gene expression, and participate in the horizontal spread of genes encoding traits such as antibiotic resistance among bacterial genera too distantly related to undergo homologous recombination. Here we review the bacterial transposon Tn5 and focus on those aspects of its functional organization and transposition which provide insights into how it and other elements may have arisen, proliferated, and evolved. PMID:6100989

Berg, D E; Berg, C M; Sasakawa, C

1984-09-01

92

The Mormon Peak and Tule Springs Detachments of Southern Nevada and Their Role in Interpreting the Subsurface Structure of the Sevier Desert Basin  

NASA Astrophysics Data System (ADS)

The eastern Sevier front is commonly overprinted by west-dipping normal faults that have been active through much of Cenozoic time. Although examples of disparate age and geometry abound, the Sevier Desert detachment stands out as being: (1) a very large, low-angle structure, (2) probably still active, and (3) at drillable depth, and hence is a prime target for scientific drilling. Among the closest surface-exposed analogs of the detachment are the Miocene Mormon Peak and Tule Springs detachments in southern Nevada, about 250 km SSW of the Sevier Desert. In both areas, the detachments developed within a few km of the basal Sevier thrust. The few traces of the southern Nevada detachments that were identified by reconnaissance mapping in the 1950s were first interpreted as underlying "rootless" gravity-slide masses. Detailed stratigraphic and structural mapping in the 1980s revealed the full extent of these detachments and their close relationship to the basal Sevier thrust. Construction and retrodeformation of cross sections through the two detachments demonstrated that neither fault could represent the base of a surficial slide mass. The key field relationship is the structural contiguity of the hanging walls of the detachments with large mountain range blocks in their down-dip directions. "Rootless" gravity-driven masses of equivalent or larger scale are widely documented, and in contrast to the southern Nevada detachments show clear evidence along their down-dip portions of either riding over the earth's surface (e.g. Heart Mountain, WY; Shadow Valley, CA) or having been internally shortened (e.g. Bearpaw Mountains, MT; Louisiana shelf). A rootless slide origin for the Mormon Peak detachment has been revived based on meso- and microstructural analysis of the fault surface (refs. 1, 2, and 3), but the hypothesis still fails the retrodeformation test at macroscopic scale. The retrodefomed sections indicate that the footwalls of the southern Nevada detachments expose a paleodepth range from near-surface conditions to more than 7 km, which will be useful for comparison with microstructures that are encountered along the Sevier Desert detachment where it is sampled at depth. 1: Carpenter et al., 1989, Geology 17, p. 774; 2: Anders et al., 2006, J. Geol 114, p. 645; 3: Walker et al., 2007, Geology 35, p. 259.

Wernicke, B.; Axen, G. J.

2007-12-01

93

Streptococcus pneumoniae Transposon Tn1545/Tn6003 Changes to Tn6002 Due to Spontaneous Excision in Circular Form of the erm(B)- and aphA3-Containing Macrolide-Aminoglycoside-Streptothricin (MAS) Element  

PubMed Central

The macrolide-aminoglycoside-streptothricin (MAS) element, an ?4.2-kb insertion containing erm(B) and aphA3 resistance determinants, distinguishes Streptococcus pneumoniae transposon Tn1545/Tn6003 from Tn6002. Here, it is shown to be an unstable genetic element that, although it lacks recombinase genes, can exploit long, erm(B)-containing direct repeats acting as att sites for spontaneous excision that may result in loss. Consequent to excision, which is RecA independent, Tn1545/Tn6003 changes to Tn6002. In pneumococcal populations harboring Tn1545/Tn6003, the latter appears to coexist with Tn6002.

Palmieri, Claudio; Mingoia, Marina; Massidda, Orietta; Giovanetti, Eleonora

2012-01-01

94

Streptococcus pneumoniae transposon Tn1545/Tn6003 changes to Tn6002 due to spontaneous excision in circular form of the erm(B)- and aphA3-containing macrolide-aminoglycoside-streptothricin (MAS) element.  

PubMed

The macrolide-aminoglycoside-streptothricin (MAS) element, an ?4.2-kb insertion containing erm(B) and aphA3 resistance determinants, distinguishes Streptococcus pneumoniae transposon Tn1545/Tn6003 from Tn6002. Here, it is shown to be an unstable genetic element that, although it lacks recombinase genes, can exploit long, erm(B)-containing direct repeats acting as att sites for spontaneous excision that may result in loss. Consequent to excision, which is RecA independent, Tn1545/Tn6003 changes to Tn6002. In pneumococcal populations harboring Tn1545/Tn6003, the latter appears to coexist with Tn6002. PMID:22890760

Palmieri, Claudio; Mingoia, Marina; Massidda, Orietta; Giovanetti, Eleonora; Varaldo, Pietro E

2012-11-01

95

First intercomparison of Tn-Rn discriminative measurements in China.  

PubMed

The intercomparison measurements with Tn-Rn discriminative monitor were conducted at the Radon Laboratory of Nanhua University. The intercomparison results indicates that the Tn-Rn discrimination of most monitors (including the solid-state nuclear track detector of participant A and participant B, continuous monitors of RAD7 and RTM2100) keeps well in a wide range of the ratio of Tn to Rn. There are great differences of Tn measurement results among RAD7, RTM2100 and reference instrument. The discussion on the reason of these differences reveals the necessity of improvement on monitor calibration, Tn source make and the necessity of more international intercomparisons. PMID:17526483

Qiu, S; Xiao, D; Liu, L

2008-01-01

96

High frequency transposition of the Tn5 derivative Tn5493 in Streptomyces lividans.  

PubMed

The mini-Tn5 derivative transposon Tn5493 transposes at a frequency as high as 3% in the Gram-positive bacterium Streptomyces lividans. The use of a thiostrepton-resistance gene and a temperature-sensitive delivery system carrying the transposase gene allows an easy selection for stable transposition events. Insertions into the S. lividans genome seem to be fairly random, as shown by Southern blot and pulsed-field gel electrophoresis (PFGE). Transpositions were easily mapped by PFGE using the restriction sites of rare-cutting enzymes present in the transposon. Because of its characteristics, Tn5493 is a promising tool for genetic and molecular analysis and manipulation of the antibiotic-producing streptomycetes. PMID:9266676

Volff, J N; Altenbuchner, J

1997-07-18

97

Low-temperature thermochronology of the Laramide Ranges and eastward translation of shortening in the Sevier Belt, Wyoming, Utah and Montana  

Microsoft Academic Search

This dissertation contains two studies that use very different techniques to investigate the Mesozoic and Cenozoic tectonics of the western USA. The first study investigates shortening in the Sevier thrust belt of northeast Utah and southwest Wyoming using cross sections and seismic reflection data. The second study investigates the low-temperature thermochronology of the Laramide Ranges using apatite (U-Th)\\/He dating. We

Sara Lynn Peyton

2009-01-01

98

Origin of the Uinta recess, Sevier fold–thrust belt, Utah: influence of basin architecture on fold–thrust belt geometry  

Microsoft Academic Search

Structural trends in the Sevier fold–thrust belt define a pronounced concave-to-the-foreland map-view curve, the Uinta recess, in north-central Utah. This recess separates two convex-to-the-foreland curves, the Provo salient on the south and the Wyoming salient on the north. The two limbs of the recess comprise transverse zones (fault zones at a high-angle to the regional trend of the orogen) that

Timothy Paulsen; Stephen Marshak

1999-01-01

99

Layer-parallel shortening across the Sevier fold-thrust belt and Laramide foreland of Wyoming: spatial and temporal evolution of a complex geodynamic system  

NASA Astrophysics Data System (ADS)

Varying patterns of layer-parallel shortening (LPS) and vertical-axis rotations from the thin-skin Sevier fold-thrust belt to the thick-skin Laramide foreland of Wyoming are quantified from integrated structural, anisotropy of magnetic susceptibility (AMS), and paleomagnetic analyses. Within the Sevier belt, widespread early LPS was accommodated by spaced cleavage, fracture sets, minor folds, and minor faults. LPS directions are subperpendicular to structural trends of systematically curved thrust sheets of the Wyoming salient, reflecting a combination of primary dispersion and secondary rotation during thrusting. Within the Laramide foreland, limited LPS was accommodated mostly by minor faults with conjugate wedge and strike-slip geometries. LPS directions in gentler fold limbs vary from perpendicular to acute with structural trends of variably oriented, anastomosing basement-cored arches. Steep forelimbs display more complex relations, including younger fault sets that developed during evolving stress states and localized vertical-axis rotations. Although internal strain is limited, weak AMS lineations defined by kinked and rotated phyllosilicates are widely developed and consistently oriented perpendicular to measured LPS directions. Palinspastically restored LPS directions, corrected for paleomagnetically determined vertical-axis rotations, vary on average from W-E in the Sevier belt to WSW-ENE in the Laramide foreland. In detail, LPS directions display deflections related to primary sedimentary wedge geometry and basement fabrics. LPS in the Sevier belt is interpreted to partly reflect stress transmitted from the hinterland through the growing orogenic wedge and topographic stress along the front of the wedge. LPS in the Laramide foreland is interpreted to partly reflect basal traction during flat-slab subduction beneath thick cratonic lithosphere, with spatial-temporal variations in stress trajectories related to basement heterogeneities and evolving fault systems. Evidence for significant late N-S Laramide shortening, as proposed by multi-stage shortening models, is not observed.

Weil, Arlo Brandon; Yonkee, W. Adolph

2012-12-01

100

Licensing the TN-40 at Prairie Island  

SciTech Connect

Northern States Power Company (SNP) is currently in the process of licensing an independent spent-fuel storage installation (ISFSI) at its Prairie Island nuclear generating plant. The proposed facility would utilize the TN-40 dry-metal storage casks, each holding 40 Prairie Island spent-fuel assemblies. The ISFSI consists of two concrete pads with a combined capacity of 48 casks for a total capacity of 1,920 fuel assemblies. In addition to a 10CRF72 license from the US Nuclear Regulatory Commission (NRC), a certificate of need from the Minnesota Public Utilities Commission is required by state law.

Kapitz, J.; McCarten, L. (Northern States Power Co., Welch, MN (United States))

1991-11-01

101

The Drinfeld double gl(n) ? tn  

NASA Astrophysics Data System (ADS)

We present a self-dual Drinfeld double structure underlying the An series of simple Lie algebras. Such double is constructed through a central extension tn of gl(n), and is obtained by pairing two disjoint solvable subalgebras coming from positive and negative roots. The Cartan-Weyl basis of gl(n) is shown to be completely determined by the compatibility conditions in the double. A natural Lie bialgebra structure on gl(n) is obtained that offers a new perspective for the construction of its quantum deformations.

Ballesteros, A.; Celeghini, E.; del Olmo, M. A.

2006-07-01

102

Transposons Tn1696 and Tn21 and their integrons In4 and In2 have independent origins.  

PubMed

The first 13.6 kb of the mercury and multidrug resistance transposon Tn1696, which includes the class 1 integron In4, has been sequenced. In4 is 8.33 kb long and contains the 5'-conserved segment (5'-CS) and 2.24 kb of the 3'-conserved segment (3'-CS) flanking four integrated cassettes. The 3'-CS region is followed by one full copy and an adjacent partial copy of the insertion sequence IS6100 flanked, in inverse orientation, by two short segments (123 and 152 bp) from the outer right-hand end of class 1 integrons. This structure is representative of a distinct group of class 1 integrons that differs from In2, found in Tn21, and other related class 1 integrons. In4 does not include transposition genes but is bounded by characteristic 25-bp inverted repeats and flanked by a direct duplication of 5 bp of the target sequence, indicating that it was inserted by a transpositional mechanism. In4 lies between the resII and resI sites of a backbone mercury resistance transposon which is >99.5% identical to Tn5036. Although Tn21 and Tn1696 are both classified as members of the Tn21 subfamily of the Tn3 transposon family, the backbone mercury resistance transposons are only 79 to 96% identical. Tn21 also contains a region of about 0.7 kb not found in Tn1696. The integrons In2 and In4 carrying the antibiotic resistance genes have been inserted at different locations into distinct ancestral mercury resistance transposons. Thus, Tn21 and Tn1696 have independent histories and origins. Other transposons (Tn1403 and Tn1412) that include a class 1 integron also have independent origins. In all except Tn21, the integron is located within the res region of the backbone transposon. PMID:11257044

Partridge, S R; Brown, H J; Stokes, H W; Hall, R M

2001-04-01

103

Syntectonic Fluid-Rock Interactions Involving Surficial Waters in the Sevier Thrust Belt, Tendoy Mountains, Southwest Montana  

NASA Astrophysics Data System (ADS)

Calcite veins and Mississippian carbonates from the Sevier thrust front record syntectonic meteoric fluid infiltration and hydrocarbon migration. The Tendoy and Four Eyes Canyon thrust sheets were emplaced onto the western margin of the Late Cretaceous Western Interior Seaway \\{WIS\\}. Low salinity \\{Tice = -0.6° C to +3.6° C\\} and low temperature \\{110° C +/- 10\\} fluids interacted with hanging-wall carbonates at a depth of 5km. Most veins have single or multiple generations of varying apertures, composed predominately of large euhedral crystals with some finer grained layers and protolith inclusions. Orientation analysis of mutually cross-cutting, high-angle vein sets suggest development concurrent with Four Eyes Canyon thrusting but prior to Tendoy thrusting. These vein sets are generally cut by later synfolding bed-parallel shear veins. Reactivation of both the bed-parallel and bed-perpendicular vein sets \\{strike parallel and strike perpendicular\\} in the Four Eyes Canyon thrust sheet occurred subsequent to Sevier compression, creating wide, coarse crystalline veins that often transect Sevier structures. Oxygen and Carbon isotope analyses of veins allow for reconstruction of fluid-rock interactions during thrust sheet emplacement and later reactivation. All veins and variably deformed host-rocks were microsampled and analyzed for ? 18OV-SMOW and ? 13CV-PDB. Small Tendoy veins \\{1mm-1cm wide\\} have calcite ? 18O values of +8.9 to +28.8‰ and calculated fluid \\{as H2O\\} of -8.3 to +11.6‰ \\{100° C\\}, -7.3 to +12.6‰ \\{110° C\\}, and -6.3 to +13.6‰ \\{120° C\\}. Four Eyes Canyon veins \\{1cm-3m wide\\} have calcite ? 18O values of +5.9 to +17.0‰ and calculated fluid of -11.3 to -0.2‰ \\{100° C\\}, -10.3 to +0.8‰ \\{110° C\\}, and -9.3 to +1.8‰ \\{120° C\\}. While there is significant variation in ? 18O there is relatively little systematic variation seen in ? 13C. Protolith carbonate has ? 18O values of +22.2‰ +/- 3.2; and some multi-layered veins are more depleted in ? 18O in earlier-formed generations. For three sites in the Lost River Range \\{LRR\\}, Idaho, the calculated minimum fluid ? 18O is -7.5‰ \\{+150 to +250° C\\} \\{Bebout et al., 2001; GRL\\}. Although the uncertainty of the regional temperature is large, when assuming a temperature of 110° C +/- 10 the Tendoy has a minimum calculated ? 18O H2O value of -8.3 to -6.3‰ and the Four Eyes Canyon has a minimum calculated ? 18O H2O value of -11.3 to -9.3‰ . These fluid O-isotope compositions are similar to the minimum H2O ? 18O calculated for the LRR sites - all pointing to infiltration of the thrust sheets by meteoric waters, possibly relatively nearshore meteoric waters with isotopic compositions strongly influenced by the nearby WIS. Surficial fluids possibly infiltrated into the thrust sheets by topographic recharge and migrated updip towards the foreland, mixing to varying degrees with more deeply roused fluids. Smaller veins and longer travel times and distances favored more extensive fluid-rock interaction and thus more rock-controlled fluid compositions. Microfractures in veins healed by hydrocarbons indicate that hydrocarbons migrated with freshwater fluids. Calcite veins record a dynamic history of fluid pathways and fluid flow as permeability evolved during thrust emplacement.

Johnson, A. C.; Anastasio, D. J.; Bebout, G. E.

2002-05-01

104

Laserprobe 40Ar/39Ar Dating Of Strain Fringes: An Example From The Interior Of The Sevier Orogen  

NASA Astrophysics Data System (ADS)

Determining the absolute ages of deformation fabrics in metamorphic rocks is critical to understanding the nature of orogenesis in deeply exhumed ancient mountain belts. In situ isotopic analysis of fibrous strain fringes is particularly advantageous because the relationships between the analyzed mineral, deformation fabric, and isotopic age can be established, and mineral growth can be linked to the progressive strain history. Furthermore, when isotopic closure temperatures (Tc) are higher than crystallization temperatures, isotopic ages reflect deformation ages. We have applied in situ 40Ar/39Ar UV and CO2 laser techniques to dating phlogopite in quartz-phlogopite-calcite and muscovite in quartz-muscovite fibrous strain fringes in greenschist-facies metamorphic rocks from the Grouse Creek Mountains (Utah) and Black Pine Mountains (Idaho). In both cases, syntectonic mica growth occurred below the minerals Tc. Phlogopite 40Ar/39Ar ages for individual strain fringes in the Grouse Creek Mountains range from 92 Ma to 110 Ma, with the most reliable ages ranging from 101 Ma to 110 Ma (mean age, 105.0 plus or minus 5.8 Ma). Muscovite 40Ar/39Ar ages for individual strain fringes in the Black Pine Mountains range from 97 Ma to 112 Ma (mean age, 104.7 plus or minus 5.8 Ma). Strain fringes are associated with a sub-horizontal foliation and a generally N-trending elongation lineation exhibiting components of top-to-the-north simple shear and coaxial strain accommodating N-S extension and sub-vertical shortening. The new dates confirm prior correlation of fabrics between these ranges and yield a mid-Cretaceous age for the earliest ductile fabric preserved in the Raft River-Albion-Grouse Creek metamorphic core complex, in the interior (hinterland) of the retroarc Sevier orogenic belt. Mid-crustal northward flow at 105 (plus or minus 6) Ma within the interior of the Sevier orogen, coeval with east-directed shortening in the foreland and with plate convergence, records orogen-parallel synconvergent extension. Gravitational relaxation of structural culminations resulting from focused crustal shortening may be the driving mechanism for orogen-parallel flow.

Wells, M. L.; Spell, T. L.; Hoisch, T. D.; Arriola, T.; Zanetti, K. A.

2008-12-01

105

Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128).  

PubMed Central

Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6. Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure. The glycopeptides thus obtained were treated with sialidase and beta-galactosidase. The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity. The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd. 67%), 81% (calcd. 86%), and 50% (calcd. 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue. These results indicate that clusters I and II react with the antibody to the same extent. The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin.

Nakada, H; Inoue, M; Numata, Y; Tanaka, N; Funakoshi, I; Fukui, S; Mellors, A; Yamashina, I

1993-01-01

106

Transposon mutagenesis of Haemophilus paragallinarum with Tn916.  

PubMed

The Gram-positive transposon Tn916 was introduced into two strains of Haemophilus paragallinarum by electroporation of the suicide plasmid pAM120 (pGL101::Tn916). Tetracycline resistant mutants of H. paragallinarum strains Tw-1 and 221 were recovered at frequencies of 2.1 x 10(-6) and 4.5 x 10(-7) per viable cell electroporated, respectively. Tn916 generates stable single insertions within different sites of the H. paragallinarum chromosome. One Tw-1::Tn916 mutant had the Tn916 insertion in the cryptic plasmid p250. This study indicates the potential use of Tn916 as a genetic tool for insertional mutagenesis of H. paragallinarum. PMID:9054124

Gonzales, A E; Glisson, J R; Jackwood, M W

1996-02-01

107

Transposon Mutagenesis of Azospirillum brasilense and Azospirillum lipoferum: Physical Analysis of Tn5 and Tn5-Mob Insertion Mutants.  

PubMed

Tn5-induced insertion mutants were generated in Azospirillum brasilense Sp7 and A. lipoferum SpBr17 by mating with Escherichia coli strains carrying suicide plasmid vectors. The sources of Tn5 were the suicide plasmids pGS9 and pSUP2021. Kanamycin-resistant Azospirillum colonies appeared from crosses with E. coli at maximum frequencies of 10 per recipient cell. Transposon Tn5 also conferred streptomycin resistance on Azospirillum colonies as was observed earlier for Rhizobium sp. Eight Tn5-induced Km SmA. brasilense Sp7 mutants with reduced nitrogen-fixing capacity were isolated. The potential use of Tn5-Mob for labeling and mobilization of Azospirillum-indigenous plasmids was demonstrated by isolating Tn5-Mob insertions in the megaplasmids of A. brasilense Sp7. PMID:16347289

Vanstockem, M; Michiels, K; Vanderleyden, J; Van Gool, A P

1987-02-01

108

A novel transposon, Tn6009, composed of a Tn916 element linked with a Staphylococcus aureus mer operon  

Microsoft Academic Search

Objectives: The aim of this study was to characterize a novel conjugative transposon Tn6009 com- posed of a Tn916 linked to a Staphylococcus aureus mer operon in representative Gram-positive and Gram-negative bacteria isolated in Nigeria and Portugal. Methods: Eighty-three Gram-positive and 34 Gram-negative bacteria were screened for the presence of the Tn6009 using DNA-DNA hybridization, PCR, hybridization of PCR products,

Olusegun O. Soge; Nicola K. Beck; Teresa M. White; David B. No; Marilyn C. Roberts

109

Characterization of ermB Gene Transposition by Tn1545 and Tn917 in Macrolide-Resistant Streptococcus pneumoniae Isolates  

PubMed Central

In Streptococcus pneumoniae, the ermB gene is carried by transposons, such as Tn917 and Tn1545. This study investigated the relationship between macrolide resistance and the presence of the ermB gene on Tn917 or Tn1545 in 84 Japanese pneumococcal isolates. Macrolide-resistant strains were classified into two groups as follows. Group 1 (19 strains) showed a tendency to high resistance to erythromycin (MIC at which 50% of isolates are inhibited, 4 mg/liter; MIC at which 90% of isolates are inhibited [MIC90], 128 mg/liter) but susceptibility to rokitamycin (MIC90, 1 mg/liter), with the ermB gene located on Tn1545. Group 2 (65 strains) showed a tendency to high resistance to both antibiotics (MIC90s for both erythromycin and rokitamycin, >128 mg/liter), with the ermB gene located on Tn917. There were no strains with constitutive macrolide resistance in either group. All of the strains in group 2 had a deletion in the promoter region of ermB and an insertion of the TAAA motif in the leader peptide. The results of pulsed-field gel electrophoresis and serogrouping showed that Tn1545 spread clonally while Tn917 spread both horizontally and clonally. In conclusion, in Japanese macrolide-resistant S. pneumoniae isolates, the ermB gene is carried and spread primarily by Tn917.

Okitsu, Naohiro; Kaieda, Satoru; Yano, Hisakazu; Nakano, Ryuichi; Hosaka, Yoshio; Okamoto, Ryouichi; Kobayashi, Toshimitsu; Inoue, Matsuhisa

2005-01-01

110

48 CFR 1201.301-72 - TAC or TN numbering.  

Code of Federal Regulations, 2010 CFR

...Regulations System 5 2010-10-01 2010-10-01 false TAC or TN numbering. 1201.301-72 Section 1201.301-72...REGULATIONS SYSTEM Agency Acquisition Regulations 1201.301-72 TAC or TN numbering. TACs and TNs will be numbered...

2010-10-01

111

48 CFR 1201.301-72 - TAC or TN numbering.  

Code of Federal Regulations, 2010 CFR

...Regulations System 5 2009-10-01 2009-10-01 false TAC or TN numbering. 1201.301-72 Section 1201.301-72...REGULATIONS SYSTEM Agency Acquisition Regulations 1201.301-72 TAC or TN numbering. TACs and TNs will be numbered...

2009-10-01

112

Characterization of Tn916S, a Tn916-Like Element Containing the Tetracycline Resistance Determinant tet(S)  

PubMed Central

We have characterized a transferable tetracycline resistance (Tcr) element from a Streptococcus intermedius isolate. The gene responsible for this resistance was identified by PCR and Southern hybridization as tet(S). Furthermore, the genetic support for this determinant was shown to be a conjugative transposon closely related to Tn916. This element has been designated Tn916S.

Lancaster, Holli; Roberts, Adam P.; Bedi, Raman; Wilson, Michael; Mullany, Peter

2004-01-01

113

Tectonic control of nested sequence architecture in the Sego Sandstone, Neslen Formation and Upper Castlegate Sandstone (Upper Cretaceous), Sevier Foreland Basin, Utah, USA  

NASA Astrophysics Data System (ADS)

The Sego Sandstone, Neslen Formation and Upper Castlegate Sandstone are laterally equivalent formations of the Upper Cretaceous Mesaverde Group, which crop out along the Book Cliffs of eastern Utah and represent sediment shed into a foreland basin from the Sevier orogenic belt to the west. The studied interval contains three regional unconformities which bound stratigraphic sequences around 100 m thick and of one-to-three million year duration, here termed high-order Sequences I-III. The high-order sequence boundaries display increasing depth of erosion northwestward toward the Sevier orogenic belt and mark changes in sediment dispersal patterns and provenance, none of which can be explained by eustatic sea-level falls. Their formation is attributed to erosion and isostatic rebound of proximal parts of the foreland basin following thrust events in the adjacent Sevier orogenic belt. In the southeastern part of the study area, high-order Sequence II (˜Sego Sandstone) contains several (>4) nested stratigraphic sequences around 20 m thick and of hundred thousand year duration, here termed low-order sequences. The low-order sequence boundaries are disconformities rather than angular unconformities and are not marked by changes in sediment dispersal patterns or provenance. Each of these low-order sequences consists of: (a) LST of incised valleys (up to 14 m thick) filled with estuarine sandstone, and (b) TST of marine shale and sandstone. No HSTs are preserved. The origin of these sequences (eustatic/tectonic) cannot be determined. When traced to the northwest, the low-order sequence boundaries become conformable and unrecognizable, and no low-order stratigraphic sequences corresponding to those in the Sego Sandstone can be recognized within the alluvial Upper Castlegate Sandstone. The observed nested sequence architecture of the Upper Mesaverde Group is considered to be controlled by variation in subsidence rate related to the emplacement and erosion of thrust sheets in the adjacent Sevier orogenic belt. During emplacement of thrust sheets, subsidence rates in proximal areas of the basin were sufficiently high to offset any low-order base-level falls, resulting in the deposition of a conformable fluvial succession. In distal parts of the basin where long-term subsidence rates were lower, low-order base-level falls produced relative falls in sea level and generated spatially restricted low-order sequences.

Willis, A.

2000-11-01

114

Expression of the Tn antigen in myelodysplasia, lymphoma, and leukemia.  

PubMed

Expression of the normally cryptic blood group antigen Tn has occasionally been reported in hematologic disease, but the true frequency of this change is not known. A mouse monoclonal antibody (FBT3) and immunohistochemistry were used to examine expression of the Tn antigen. Expression was not detected in 35 normal bone marrow aspirates examined, but it was detected in 5 of 725 abnormal bone marrow aspirates, including 2 (3.6%) of 55 cases of de novo acute nonlymphocytic leukemia and 2 cases that terminated in acute nonlymphocytic leukemia. In two patients, one with acute myeloblastic leukemia and the other in blast transformation of chronic myeloid leukemia, the Tn antigen was expressed on 2 percent of blast cells. In one case of non-Hodgkin's lymphoma, 4 percent of normal myeloid cells expressed the antigen. In the other two cases, one of acute myelomonocytic leukemia and the other of myelodysplasia, only 2 to 8 percent of myeloid and erythroid cells initially were Tn positive. Subsequent serial immunohistochemical studies of bone marrow aspirates and peripheral blood in these two cases showed increasing numbers of Tn-positive erythroid and myeloid cells 8 to 12 months before polyagglutination was detected serologically. Tn-positive cells increased to > 90 percent in the terminal phase in both cases of both diseases. The results suggest that Tn expression in these two patients may have conferred a growth advantage to the cells and could be related to disease progression. PMID:1471247

Roxby, D J; Pfeiffer, M B; Morley, A A; Kirkland, M A

1992-01-01

115

Precise excision of bacterial transposon Tn5 in yeast.  

PubMed

We have demonstrated that precise excision of bacterial transposon Tn5 can occur in the yeast, Saccharomyces cerevisiae. Tn5 insertions in the yeast gene LYS2 were generated by transposon mutagenesis made in Escherichia coli by means of a lambda::Tn5 vector. Nine insertions of Tn5 into the structural part of the yeast LYS2 gene situated in a shuttle episomal plasmid were selected. All the plasmids with a Tn5 insertion were used to transform yeast strains carrying a deletion of the entire LYS2 gene or a deletion of the part of LYS2 overlapping the point of insertion. All insertions inactivated the LYS2 gene and were able to revert with low (about 10(-8) frequencies to lysine prototrophy. Restriction analysis of revertant plasmids revealed them to be indistinguishable from the original plasmid without Tn5 insertion. DNA sequencing of the regions containing the points of insertions, made for two revertants, proved that Tn5 excision was completely precise. PMID:2847007

Gordenin, D A; Trofimova, M V; Shaburova, O N; Pavlov, Y I; Chernoff, Y O; Chekuolene, Y V; Proscyavichus, Y Y; Sasnauskas, K V; Janulaitis, A A

1988-08-01

116

76 FR 58726 - Proposed Amendment of Class E Airspace; Springfield, TN  

Federal Register 2010, 2011, 2012, 2013

...Proposed Amendment of Class E Airspace; Springfield, TN AGENCY: Federal Aviation Administration...proposes to amend Class E Airspace in the Springfield, TN area. Aydelotte Airport has been...rules (IFR) operations within the Springfield, TN airspace area. DATES: 0901...

2011-09-22

117

Summary of reported agriculture and irrigation water use in southwestern Arkansas counties, 1991  

USGS Publications Warehouse

This report summarizes the 1991 water-use reporting through the Conservation District Offices in the following southwestern Arkansas counties: Bradley, Calhoun, Clark, Cleveland, Columbia, Dallas, Garland, Grant, Hempstead, Hot Spring, Howard, Little River, Montgomery, Nevada, Ouachita, Pike, Polk, Saline, Sevier, and Union. The number of withdrawal registrations for southwestern Arkansas counties was 132 (31 groundwater and 101 surface water). Water withdrawals reported during the registration process total 0.84 Mgal/d (none from groundwater and 0.84 Mgal/d surface water) for agriculture and 14.22 Mgal/d (1.64 Mgal/d groundwater and 12.58 Mgal/d surface water) for irrigation. The registration reports for 1991 indicate that this water was applied to 8,455 acres of land to irrigate rice, corn, sorghum, soybeans, cotton, cash grains, vegetables, sod, berries, fruit trees, timber, shrubs, and nuts as well as for the agricultural use of animal aquaculture.

Holland, T. W.; Manning, C. A.; Stafford, K. L.

1993-01-01

118

Molecular Determinants of Cardiac Myocyte Performance as Conferred by Isoform-Specific TnI Residues.  

PubMed

Troponin I (TnI) is the molecular switch of the sarcomere. Cardiac myocytes express two isoforms of TnI during development. The fetal heart expresses the slow skeletal TnI (ssTnI) isoform and shortly after birth ssTnI is completely and irreversibly replaced by the adult cardiac TnI (cTnI) isoform. These two isoforms have important functional differences; broadly, ssTnI is a positive inotrope, especially under acidic/hypoxic conditions, whereas cTnI facilitates faster relaxation performance. Evolutionary directed changes in cTnI sequence suggest cTnI evolved to favor relaxation performance in the mammalian heart. To investigate the mechanism, we focused on several notable TnI isoform and trans-species-specific residues located in TnI's helix 4 using structure/function and molecular dynamics analyses. Gene transduction of adult cardiac myocytes by cTnIs with specific helix 4 ssTnI substitutions, Q157R/A164H/E166V/H173N (QAEH), and A164H/H173N (AH), were investigated. cTnI QAEH is similar in these four residues to ssTnI and nonmammalian chordate cTnIs, whereas cTnI AH is similar to fish cTnI in these four residues. In comparison to mammalian cTnI, cTnI QAEH and cTnI AH showed increased contractility and slowed relaxation, which functionally mimicked ssTnI expressing myocytes. cTnI QAEH molecular dynamics simulations demonstrated altered intermolecular interactions between TnI helix 4 and cTnC helix A, specifically revealing a new, to our knowledge, electrostatic interaction between R171of cTnI and E15 of cTnC, which structurally phenocopied the ssTnI conformation. Free energy perturbation calculation of cTnC Ca(2+) binding for these conformations showed relative increased calcium binding for cTnI QAEH compared to cTnI. Taken together, to our knowledge, these new findings provide evidence that the evolutionary-directed coordinated acquisition of residues Q157, A164, E166, H173 facilitate enhanced relaxation performance in mammalian adult cardiac myocytes. PMID:24853739

Thompson, Brian R; Houang, Evelyne M; Sham, Yuk Y; Metzger, Joseph M

2014-05-20

119

Tn502 and Tn512 Are res Site Hunters That Provide Evidence of Resolvase-Independent Transposition to Random Sites ?  

PubMed Central

In this study, we report on the transposition behavior of the mercury(II) resistance transposons Tn502 and Tn512, which are members of the Tn5053 family. These transposons exhibit targeted and oriented insertion in the par region of plasmid RP1, since par-encoded components, namely, the ParA resolvase and its cognate res region, are essential for such transposition. Tn502 and, under some circumstances, Tn512 can transpose when par is absent, providing evidence for an alternative, par-independent pathway of transposition. We show that the alternative pathway proceeds by a two-step replicative process involving random target selection and orientation of insertion, leading to the formation of cointegrates as the predominant product of the first stage of transposition. Cointegrates remain unresolved because the transposon-encoded (TniR) recombination system is relatively inefficient, as is the host-encoded (RecA) system. In the presence of the res-ParA recombination system, TniR-mediated (and RecA-mediated) cointegrate resolution is highly efficient, enabling resolution both of cointegrates involving functional transposons (Tn502 and Tn512) and of defective elements (In0 and In2). These findings implicate the target-encoded accessory functions in the second stage of transposition as well as in the first. We also show that the par-independent pathway enables the formation of deletions in the target molecule.

Petrovski, Steve; Stanisich, Vilma A.

2010-01-01

120

A novel transposon, Tn6009, composed of a Tn916 element linked with a Staphylococcus aureus mer operon  

PubMed Central

Objectives The aim of this study was to characterize a novel conjugative transposon Tn6009 composed of a Tn916 linked to a Staphylococcus aureus mer operon in representative Gram-positive and Gram-negative bacteria isolated in Nigeria and Portugal. Methods Eighty-three Gram-positive and 34 Gram-negative bacteria were screened for the presence of the Tn6009 using DNA–DNA hybridization, PCR, hybridization of PCR products, sequencing and mating experiments by established procedures. Results Forty-three oral and 23 urine Gram-negative and Gram-positive isolates carried the Tn6009. Sequencing was performed to verify the direct linkage between the mer resistance genes and the tet(M) gene. A Nigerian Klebsiella pneumoniae, isolated from a urinary tract infection patient, and one commensal isolate from each of the other Tn6009-positive genera, Serratia liquefaciens, Pseudomonas sp., Enterococcus sp. and Streptococcus sp. isolated from the oral and urine samples of healthy Portuguese children, were able to act as donors and conjugally transfer the Tn6009 to the Enterococcus faecalis JH2-2 recipient, resulting in tetracycline- and mercury-resistant E. faecalis transconjugants. Conclusions This study reports a novel non-composite conjugative transposon Tn6009 containing a Tn916 element linked to an S. aureus mer operon carrying genes coding for inorganic mercury resistance (merA), an organic mercury resistance (merB), a regulatory protein (merR) and a mercury transporter (merT). This transposon was identified in 66 isolates from two Gram-positive and three Gram-negative genera and is the first transposon in the Tn916 family to carry the Gram-positive mer genes directly linked to the tet(M) gene.

Soge, Olusegun O.; Beck, Nicola K.; White, Teresa M.; No, David B.; Roberts, Marilyn C.

2008-01-01

121

Identification of Tn4451 and Tn4452, chloramphenicol resistance transposons from Clostridium perfringens.  

PubMed Central

The recombinant plasmids pJIR45 and pJIR97 contain the chloramphenicol resistance determinants derived from the Clostridium perfringens R plasmids pIP401 and pJIR27, respectively. Escherichia coli cultures which harbored these recombinant plasmids rapidly became chloramphenicol sensitive when grown in the absence of chloramphenicol. The loss of resistance was associated with the loss of 6.2-kilobase (kb) segments from both plasmids. Detailed restriction analysis of E. coli- and C. perfringens-derived deletion plasmids indicated that deletion of these segments was essentially precise. Transposition of the 6.2-kb segments was demonstrated by cloning the determinants into a temperature-sensitive plasmid, curing the recombinant plasmids, and selecting chloramphenicol-resistant, plasmid-free clones. Southern hybridization analysis of chromosomal DNA isolated from these recA E. coli clones indicated that the 6.2-kb segments had transposed to different sites on the chromosome. Heteroduplex analysis and restriction mapping indicated that the transposons, Tn4451 (pIP401) and Tn4452 (pJIR27), were closely related and did not contain large inverted or directly repeated sequences. These transposons represent the first transposable elements from the clostridia to be identified and characterized. Images

Abraham, L J; Rood, J I

1987-01-01

122

Expression of a Tn-like epitope by carcinoma cells.  

PubMed Central

A monoclonal antibody, FBT3, was raised against Tn positive erythrocytes and, using immunohistochemistry, fresh and fixed tissues from patients with cancer were studied to detect any expression of a Tn-like epitope. Expression was found in neoplastic cells, usually both in cytoplasm and on cell membranes, from 104 of 147 cases of carcinoma and 1 of 13 cases of lymphoma, but rarely in adjacent, morphologically normal cells. Tn expression was seen in some normal glandular cells but, unlike cancer cells, it was distributed as fine granules in a supranuclear position. Detection of a Tn-like epitope is of theoretical interest and may be of direct diagnostic value. Images Figure 1 Figure 2 Figure 3

Roxby, D. J.; Skinner, J. M.; Morley, A. A.; Weeks, S.; Burpee, M.

1987-01-01

123

Polymer-modified TN and supertwisted nematic (STN) devices  

Microsoft Academic Search

Polymer modified materials have been shown to be useful in controlling the operational voltages and speed in TN devices; and the operational voltages and the suppression of the `stripe' deformation in STN cells.

Philip J. Bos; Jianlin Li; Deng K. Yang; William Doane

1995-01-01

124

76 FR 14855 - Television Broadcasting Services; Nashville, TN  

Federal Register 2010, 2011, 2012, 2013

...Docket No. 11-29, RM-11622; DA 11-335] Television Broadcasting Services; Nashville, TN AGENCY...and 1.420. List of Subjects in 47 CFR Part 73 Television, Television broadcasting. Federal Communications...

2011-03-18

125

Tn5tac1 insertion polarity in Escherichia coli.  

PubMed

The Tn5-derived transposon Tn5tac1 carries the strong tac promoter (Ptac) facing outward at one of its ends. Expression of Ptac is under the control of the lacIq gene, also contained within the transposon. By inserting Tn5tac1 upstream from a promoterless galK gene we determined the basal level of transcription from both ends of the transposon in the absence of IPTG to be about 4% relative to the lactose promoter (Plac). As a result, derivatives of strain N100 containing these plasmids produce red colonies in MacConkey-galactose plates. Deletion of the BamHI fragment including Ptac causes galactokinase levels to drop to less than 1% of Plac, enough to render white colonies in MacConkey-galactose plates. Thus, Tn5tac1 can be used for genetic analysis under conditions in which it shows no polarity (+ IPTG), low polarity (- IPTG), or strong polarity (delta Ptac). PMID:1661429

Llosa, M; Jubete, Y; de la Cruz, F

1991-11-01

126

In vivo Tn 5 -based transposon mutagenesis of Streptomycetes  

Microsoft Academic Search

This paper reports the in vivo expression of the synthetic transposase gene tnp(a) from a hyperactive Tn5 tnp gene mutant in Streptomyces coelicolor. Using the synthetic tnp(a) gene adapted for Streptomyces codon usage, we showed random insertion of the transposon into the Streptomycetes genome. The insertion frequency for the\\u000a hyperactive Tn5 derivative is 98% of transformed S. coelicolor cells. The

Lutz Petzke; Andriy Luzhetskyy

2009-01-01

127

Transposon Tn21, flagship of the floating genome.  

PubMed

The transposon Tn21 and a group of closely related transposons (the Tn21 family) are involved in the global dissemination of antibiotic resistance determinants in gram-negative facultative bacteria. The molecular basis for their involvement is carriage by the Tn21 family of a mobile DNA element (the integron) encoding a site-specific system for the acquisition of multiple antibiotic resistance genes. The paradigm example, Tn21, also carries genes for its own transposition and a mercury resistance (mer) operon. We have compiled the entire 19,671-bp sequence of Tn21 and assessed the possible origins and functions of the genes it contains. Our assessment adds molecular detail to previous models of the evolution of Tn21 and is consistent with the insertion of the integron In2 into an ancestral Tn501-like mer transposon. Codon usage analysis indicates distinct host origins for the ancestral mer operon, the integron, and the gene cassette and two insertion sequences which lie within the integron. The sole gene of unknown function in the integron, orf5, resembles a puromycin-modifying enzyme from an antibiotic producing bacterium. A possible seventh gene in the mer operon (merE), perhaps with a role in Hg(II) transport, lies in the junction between the integron and the mer operon. Analysis of the region interrupted by insertion of the integron suggests that the putative transposition regulator, tnpM, is the C-terminal vestige of a tyrosine kinase sensor present in the ancestral mer transposon. The extensive dissemination of the Tn21 family may have resulted from the fortuitous association of a genetic element for accumulating multiple antibiotic resistances (the integron) with one conferring resistance to a toxic metal at a time when clinical, agricultural, and industrial practices were rapidly increasing the exposure to both types of selective agents. The compendium offered here will provide a reference point for ongoing observations of related elements in multiply resistant strains emerging worldwide. PMID:10477306

Liebert, C A; Hall, R M; Summers, A O

1999-09-01

128

[Transposition of the composite synthetic transposon TnV (Tn5-Rep(pSC101)) is accompanied by the formation of the mini-plasmid pTnV, containing defective Is50-elements].  

PubMed

Using thermoelimination (at 42 degrees C) of the thermoinducible coliphage P1tsCmr omega::TnV (TnV is a Tn5 derivative which contains the replication origin (Rep) of plasmid pSC101), more than 110 KmrCms Escherichia coli K12 clones were selected. It was supposed that the KmrCms phenotype could result only from insertion of TnV (Kmr) into E. coli chromosome and the loss of phage (Cms). It was found that the majority of KmrCms clones (35-90%) contained miniplasmids. Their molecular sizes did not exceed the TnV size (6.1 kb). The formation of miniplasmids called pTnV was observed both in RecA+ cells (C600) and in RecA- (HB101), more often in the latters. Interestingly, that miniplasmids of only several molecular sizes were detected: from 6.1 kb (pTnV60) to 4.35 kb (pTnV43). A restriction analysis showed that DNA of the majority of pTnV plasmids had varying deletions (0.3-1.3 kb) of mainly IS50L element which together with IS50R flank TnV. Very low transposition frequencies (approx. 10(-8) Kmr transconjugants per transferred R388) of all pTnV types (including pTnV60 plasmids containing probably microdeletions of the joining "outside" IS50's ends) suggest that pTnV plasmids are not intermediates in TnV transposition. Possibly the circularized TnV derivatives (pTnV's) are side products of the transposition resulting from the abortive attempts of an excised and autonomous transposon molecule to insert into itself. In the present paper the possible mechanisms of the origin of limited pTnV type numbers are also discussed. PMID:1963206

Borovok, I A

1990-01-01

129

Chromosomal Complementation Using Tn7 Transposon Vectors in Enterobacteriaceae  

PubMed Central

Genetic complementation in many bacteria is commonly achieved by reintroducing functional copies of the mutated or deleted genes on a recombinant plasmid. Chromosomal integration systems using the Tn7 transposon have the advantage of providing a stable single-copy integration that does not require selective pressure. Previous Tn7 systems have been developed, although none have been shown to work effectively in a variety of enterobacteria. We have developed several mini-Tn7 and transposase vectors to provide a more versatile system. Transposition of Tn7 at the chromosomal attTn7 site was achieved by a classical conjugation approach, wherein the donor strain harbored the mini-Tn7 vector and the recipient strain possessed the transposase vector. This approach was efficient for five different pathogenic enterobacterial species. Thus, this system provides a useful tool for single-copy complementation at an episomal site for research in bacterial genetics and microbial pathogenesis. Furthermore, these vectors could also be used for the introduction of foreign genes for use in biotechnology applications, vaccine development, or gene expression and gene fusion constructs.

Crepin, Sebastien; Harel, Josee

2012-01-01

130

Transposon mutagenesis in halophilic eubacteria: conjugal transfer and insertion of transposon Tn5 and Tn1732 in Halomonas elongata.  

PubMed

Molecular genetic studies of halophilic eubacteria have been limited by the lack of a suitable method for mutagenesis. To overcome this, we established a transposon mutagenesis procedure for the ectoine-producing, halophilic bacterium Halomonas elongata. We used suicide plasmids pSUP101 and pSUP102-Gm to introduce the transposons Tn5 and Tn1732 respectively into H. elongata via Escherichia coli SM10 mediated conjugation. Our finding that H. elongata is sensitive to aminoglycoside antibiotics at low salinity enabled us to apply transposons that mediate kanamycin resistance. The insertions of transposon Tn1732 occurred at different sites in the chromosome of H. elongata, as proved by Southern hybridization analysis. Phenotypic analysis revealed that different auxotrophic and salt sensitive mutants were generated by mutagenesis with transposon Tn1732. To our knowledge this is the first report of a successful application of a transposon for direct generalized mutagenesis in a halophilic eubacterium. PMID:7781978

Kunte, H J; Galinski, E A

1995-05-15

131

Tn1403, a Multiple-Antibiotic Resistance Transposon Made Up of Three Distinct Transposons?  

PubMed Central

Transposon Tn1403 from a clinical Pseudomonas strain is composed of three transposons, including Tn5393c. A related transposon Tn1404* from a plant-associated Pseudomonas strain lacks Tn5393 but includes a transposon carrying the tet(C) tetracycline resistance determinant. These compound transposons illustrate the role of preexisting transposons in generating clusters of antibiotic resistance genes.

Stokes, H. W.; Elbourne, Liam D. H.; Hall, Ruth M.

2007-01-01

132

Structural and microstructural evolution of the Rattlesnake Mountain Anticline (Wyoming, USA): New insights into the Sevier and Laramide orogenic stress build-up in the Bighorn Basin  

NASA Astrophysics Data System (ADS)

The Rocky Mountains in western US provide among the best examples of thick-skinned tectonics: following a period of thin-skinned tectonics related to the Sevier orogeny, the compressional reactivation of basement faults gave birth to the so-called Laramide uplifts/arches. The Bighorn basin, located in Wyoming, is therefore a key place to study the transition from thin- to thick-skinned tectonics in orogenic forelands, especially in terms of microstructural and stress/strain evolution. Our study focuses on a classic Laramide structure: the Rattlesnake Mountain Anticline (RMA, Wyoming, USA), a basement-cored anticline located in the western part of the Bighorn basin. Stress and strain evolution analysis in folded sedimentary layers and underlying faulted basement rocks were performed on the basis of combined analyses of fractures, fault-slip data and calcite twinning paleopiezometry. Most of the fractures are related to three main tectonic events: the Sevier thin-skinned contraction, the Laramide thick-skinned contraction, and the Basin and Range extension. Serial balanced cross-sections of RMA and displacement profiles suggest that all thrust faults were coeval, evidencing strain distribution in the basement during faulting. The comparison of RMA with another structure located in the eastern edge of the Bighorn basin, i.e. the Sheep Mountain Anticline (SMA), allows to propose a conceptual model for the geometric and kinematic evolution of Laramide-related basement-cored anticlines. Finally, the stress evolution is reconstructed at both the fold scale and the basin scale. We show that the evolution of stress trends and magnitudes was quite similar in both structures (RMA and SMA) during Laramide times (thick-skinned tectonics), in spite of different stress regimes. During Sevier (thin-skinned tectonics) and post-Laramide times, stress trends and fracture patterns were different in these two structures. These results suggest that the distance to the orogenic front influenced the fracture patterns but not the foreland stress magnitudes, which were likely controlled by the structural style.

Beaudoin, Nicolas; Leprêtre, Rémi; Bellahsen, Nicolas; Lacombe, Olivier; Amrouch, Khalid; Callot, Jean-Paul; Emmanuel, Laurent; Daniel, Jean-Marc

2012-11-01

133

Isolation of Bacteroides fragilis Mutants with In Vivo Growth Defects by Using Tn4400?, a Modified Tn4400 Transposition System, and a New Screening Method  

PubMed Central

A modified version of the Bacteroides fragilis transposon Tn4400, designated Tn4400?, enabling rapid isolation and analysis of B. fragilis mutants has been constructed. To identify potential virulence factors, Tn4400?-generated mutants were screened by a new method; this resulted in the isolation of 21 mutant strains with impaired growth characteristics on tissue culture monolayers but normal growth in rich medium anaerobically.

Tang, Yixin P.; Malamy, Michael H.

2000-01-01

134

In vivo Tn5-based transposon mutagenesis of Streptomycetes.  

PubMed

This paper reports the in vivo expression of the synthetic transposase gene tnp(a) from a hyperactive Tn5 tnp gene mutant in Streptomyces coelicolor. Using the synthetic tnp(a) gene adapted for Streptomyces codon usage, we showed random insertion of the transposon into the Streptomycetes genome. The insertion frequency for the hyperactive Tn5 derivative is 98% of transformed S. coelicolor cells. The random transposition has been confirmed by the recovery of ~1.1% of auxotrophs. The Tn5 insertions are stably inherited in the absence of apramycin selection. The transposon contains an apramycin resistance selection marker and an R6Kgamma origin of replication for transposon rescue. We identified the transposon insertion loci by random sequencing of 14 rescue plasmids. The majority of insertions (12 of 14) were mapped to putative open-reading frames on the S. coelicolor chromosome. These included two new regulatory genes affecting S. coelicolor growth and actinorhodin biosynthesis. PMID:19479250

Petzke, Lutz; Luzhetskyy, Andriy

2009-07-01

135

Tn10-derived transposons active in Bacillus subtilis.  

PubMed

Small derivatives of the Escherichia coli transposon Tn10, comprising IS10 ends and a chloramphenicol resistance gene, were introduced in Bacillus subtilis on a thermosensitive plasmid, pE194. In the presence of the Tn10 transposase gene fused to signals functional in B. subtilis, these derivatives transposed with a frequency of 10(-6) per element per generation. They had no highly preferred insertion site or region, as judged by restriction analysis of the chromosomal DNA, and generated auxotrophic and sporulation-deficient mutants with a frequency of about 1%. These results suggest that Tn10 derivatives might be a useful genetic tool in B. subtilis and possibly other gram-positive microorganisms. PMID:2174858

Petit, M A; Bruand, C; Jannière, L; Ehrlich, S D

1990-12-01

136

Magnetoresistivity below TN in CeMo 6S 8  

NASA Astrophysics Data System (ADS)

The T?1.15 K resistivity of two CeMo 6S 8 Chevrel single crystals is reported. It shows the occurrence of (presumably) antiferromagnetic (AF) order at TN?2.1 and 2.55 K, respectively, in agreement with previous results. The magnetoresistance, measured on sample 1 shows a drop below TN for a field Bc?1.25 T at 1.6 K. This behavior suggests the crossing from an AF state to a polarized state. It is discussed in connection with other results reported for this compound.

Lapierre, F.; Haen, P.; Le Berre, F.; Peña, O.; Horyn, R.

1999-01-01

137

Yeast mutants with increased bacterial transposon Tn5 excision.  

PubMed

Five complementing recessive mutations that exhibit increased bacterial transposon Tn5 precise excision in yeast Saccharomyces cerevisiae were obtained by ethylmethanesulfonate treatment. One of these mutations (tex1) was submitted to extensive genetic analysis. tex1 is a recessive temperature-sensitive mutation resulting in a 20-100-fold increase in Tn5 excision. It also has increased frequencies of ochre mutation reversion, of forward mutation to canavanine resistance, and loss of chromosome III or its right arm. The possible mechanism of tex1 effects is discussed. PMID:1850571

Gordenin, D A; Proscyavichus, Y Y; Malkova, A L; Trofimova, M V; Peterzen, A

1991-01-01

138

Demonstration of criticality safety for the modified TN-REG and TN-BRP transport/storage casks  

SciTech Connect

An inability to model the structural performance of borated steel baskets under accident conditions forced the specially designed TN-BRP and TN-REG casks to be modified for half-loaded shipments. This paper discusses the approach used to demonstrate that the half-loaded casks would remain safely subcritical even if no credit were taken for the borated basket. Normal and accident configurations were analyzed with the KENO V.a code. The strategy conceived and the analyses performed to demonstrate an acceptable margin of safety are discussed. 5 refs., 3 figs., 2 tabs.

Parks, C.V.; Fox, P.B. (Oak Ridge National Lab., TN (USA))

1989-01-01

139

Genetic manipulations in Rhizobium meliloti utilizing two new transposon Tn5 derivatives.  

PubMed

Two derivatives of the prokaryotic transposon Tn5 were constructed in vitro. In Tn5-233, the central area of Tn5, which carries resistance to kanamycin/neomycin, bleomycin and streptomycin, is replaced by a fragment carrying resistance to the aminocyclitol antibiotics gentamycin/kanamycin and streptomycin/spectinomycin. In Tn5-235, the Escherichia coli beta-galactosidase gene is inserted within the streptomycin resistance gene of Tn5, and constitutively expressed from a Tn5 promoter. Both constructs transpose with about the same frequency as Tn5 in Escherichia coli and Rhizobium meliloti. When a Tn5-derivative is introduced into an R. meliloti strain which already contains a different Tn5-derivative, in situ transposon replacement is obtained at high frequency, presumably by a pair of crossovers between the IS50 sequences at the ends of the incoming and resident transposons. In this way we converted a previously isolated recA::Tn5 mutant into the corresponding recA::Tn5-233 strain, which can now be used as a genetic background in the study of complementation of other Tn5-induced mutations. We also replaced the drug markers of several Tn5-induced exo mutants, which we were then able to map relative to each other by transduction with phage phi M12. In a strain carrying Tn5-235 located near Tn5-233, we were able to isolate deletions of the intervening markers, presumably resulting from general recombination between the two transposons, by screening for loss of the Lac+ phenotype. Unlike Tn5 itself, resident Tn5-233 does not appear to suppress transposition of another incoming Tn5-derivative. PMID:3020382

De Vos, G F; Walker, G C; Signer, E R

1986-09-01

140

Cytochrome mutants of bradyrhizobium induced by transposon tn5.  

PubMed

Transposon Tn5 was used to mutate Bradyrhizobium japonicum USDA 61N. From over 5000 clones containing Tn5, 12 were selected and purified using a chemical reaction to identify oxidase-deficient clones. Four classes of mutants were identified based on the alterations in cytochromes. Most of the mutants had alterations in more than one cytochrome. Southern hybridization analysis of restricted genomic DNA of a representative strain of each class demonstrated that each mutant had a single Tn5 insert. Thus a single Tn5 insert produced pleiotropic effects on cytochromes. One class, which was totally deficient in cytochromes aa(3) and c, produced ineffective nodules on soybeans. Most of the strains representing the other classes produced effective nodules but exceptions were observed in each class. Bacteroids of the wild-type strain contained cytochrome aa(3). Bacteroids from one class of mutants were totally devoid of cytochrome aa(3). Several of these strains produced effective symbioses indicating that cytochrome aa(3) is not required for an effective symbiosis in this DNA homology group II strain which normally has this terminal oxidase in bacteroids. PMID:16666807

Nautiyal, C S; van Berkum, P; Sadowsky, M J; Keister, D L

1989-06-01

141

Insertion site specificity of the transposon Tn3.  

PubMed

The Tn3-deletion method [Davies and Hutchison, Nucleic Acids Res. 19, 5731-5738, (1991)] was used to sequence a 9.4 kb DNA fragment. Transpositional 'warm' spots were not a limiting factor but a 935 bp 'cold' spot was completed using a synthetic oligonucleotide primer. Two hundred and twenty three miniTn3 insertion sites from three sequencing projects were aligned and a 19 bp asymmetric consensus site was identified. There is no absolute sequence requirement at any position in this consensus, so insertion occurs promiscuously (approximately 37% of sites are potential targets). In our sequencing projects, multiply targeted sites always closely matched the consensus, although not all close matches were targeted frequently. The 935 bp cold spot showed no unusual features when analysed with the consensus sequence. The consensus can be used to accurately predict likely insertion sites in a new sequence. Synthetic oligonucleotides based on the consensus and a known hot spot for Tn3 were mutagenised. These sequences were not hot spots in our vectors, suggesting that the primary sequence alone is not sufficient to create an insertional hot spot. We conclude that some other factor, such as DNA secondary structure, also plays an important role in target site selection for the transposon Tn3. PMID:7885847

Davies, C J; Hutchison, C A

1995-02-11

142

Effect of predeformational basin geometry in the kinematic evolution of a thin-skinned orogenic wedge: Insights from three-dimensional finite element modeling of the Provo salient, Sevier fold-thrust belt, Utah  

Microsoft Academic Search

In fold-thrust belts, sedimentary cover rocks are detached from undeformed basement and undergo crustal-scale shortening and internal deformation. We have investigated a three-dimensional (3-D), nonlinear, elastic-plastic finite element model using the restored Provo salient of the Sevier belt as our initial configuration. In the model the deformed sedimentary prism displays large-scale geometries that are seen in many natural fold-thrust belts

Sanghoon Kwon; Gautam Mitra; Renato Perucchio

2007-01-01

143

Combining paleomagnetism and structural geology to better understand complex orogenic systems: development of a robust kinematic model for Sevier fold-thrust belt and Laramide foreland of Wyoming, U.S.A.  

NASA Astrophysics Data System (ADS)

Varying patterns of layer-parallel shortening (LPS) and vertical-axis rotations from the thin-skin Sevier fold-thrust belt to the thick-skin Laramide foreland of Wyoming are quantified from integrated structural, anisotropy of magnetic susceptibility (AMS), and paleomagnetic analyses. Within the Sevier belt, widespread early LPS was accommodated by spaced cleavage, fracture sets, minor folds, and minor faults. LPS directions are subperpendicular to structural trends of systematically curved thrust sheets of the Wyoming salient, reflecting a combination of primary dispersion and secondary rotation during thrusting. Within the Laramide foreland, limited LPS was accommodated mostly by minor faults with conjugate wedge and strike-slip geometries. LPS directions in gentler fold limbs vary from perpendicular to acute with structural trends of variably oriented, anastomosing basement-cored arches. Steep forelimbs display more complex relations, including younger fault sets that developed during evolving stress states and localized vertical-axis rotations. Although internal strain is limited, weak AMS lineations defined by kinked and rotated phyllosilicates are widely developed and consistently oriented perpendicular to measured LPS directions. Palinspastically restored LPS directions, corrected for paleomagnetically determined vertical-axis rotations, vary on average from W-E in the Sevier belt to WSW-ENE in the Laramide foreland. In detail, LPS directions display deflections related to primary sedimentary wedge geometry and basement fabrics. LPS in the Sevier belt is interpreted to partly reflect stress transmitted from the hinterland through the growing orogenic wedge and topographic stress along the front of the wedge. LPS in the Laramide foreland is interpreted to partly reflect basal traction during flat-slab subduction beneath thick cratonic lithosphere, with spatial-temporal variations in stress trajectories related to basement heterogeneities and evolving fault systems. Evidence for significant late N-S Laramide shortening, as proposed by multi-stage shortening models, is not observed.

Weil, Arlo; Yonkee, Adolph

2013-04-01

144

Failure of normal red blood cells to become Tn-activated when transfused into a patient with Tn-polyagglutination.  

PubMed

A Group O patient with Tn-polyagglutinable RBCs was transfused with two units of Group O packed RBCs. Patient and transfused RBCs were separated by differential agglutination in blood samples drawn over a period of two months after transfusion. The transfused RBCs did not become polyagglutinable. PMID:7416196

Reid, M E; Ellisor, S S; Swanson, J; Edwards, G; Stanik, F

1980-09-01

145

75 FR 27846 - Tennessee Disaster Number TN-00039  

Federal Register 2010, 2011, 2012, 2013

...Smith, Stewart, Trousdale, Wayne, Wilson. Contiguous Counties: (Economic Injury Loans Only): Tennessee: Overton, Pickett, Putnam, Warren, White. Alabama: Lauderdale. Kentucky: Calloway, Clinton, Cumberland, Logan, Monroe, Trigg....

2010-05-18

146

[Transposon Tn5 and its derivatives effectively transpose over a broad temperature range].  

PubMed

It was shown that the 1st class composite transposon Tn5 (5.8 Kb) and its synthetic derivatives--TnV (Tn5-ReppSC101; 6.1 Kb) and Tn5-MobRP4 (about 7.7 Kb) transpose in Escherichia coli K-12 cells (RecA strain HB101) with similar efficiency both at 28 and at 42 degrees C as well as at 37 degrees C. This property of Tn5-like elements distinguishes them from the class II transposons (such as Tn3, Tn21 etc.), whose transposition, as is well known, is strongly suppressed even at 37 degrees C. It was also demonstrated that transposition frequency of Tn5-derived elements depends on their copy number. PMID:1964141

Borovok, I A

1990-09-01

147

From Source to Sink: Exhumation of the North America Cordillera Revealed by Multi-dating of Detrital Minerals from Upper Jurassic-Upper Cretaceous Sevier Foreland Basin  

NASA Astrophysics Data System (ADS)

We sampled twenty-two Late Jurassic to Late Cretaceous syn-orogenic conglomerate clasts in proximal units in the Sevier fold-thrust belt and their distal sandstone equivalents up to 300 km east of the thrust front, in Utah, Colorado, Wyoming, and South Dakota. To better constrain depositional ages, these samples were analyzed using detrital zircon U-Pb (DZ U-Pb) geochronology. To identify a thermochronometer that measures source exhumation in the North America Cordillera, both zircon (U-Th)/He and apatite fission track (AFT) thermochronology was utilized, on both the conglomerate cobbles and sandstone (detrital) samples. Eleven samples were analyzed with zircon (U-Th)/He; however, discordant ages in the conglomerate cobble samples suggest that this system was not fully reset and never experienced T> ~180 °C in the source stratigraphy during the Sevier orogeny. Eleven other samples are analyzed using apatite fission track thermochronology (AFT); AFT ages are generally similar or older than depositional ages indicating that the detrital ages record source exhumation signals, and that exhumation depth corresponds to T>~120 °C. In order to test whether or not the youngest cooling AFT age population represents a source exhumation signal or a co-magmatic signal we here performed double dating of the detrital AFT samples using apatite U-Pb thermochronology. Maximum depositional ages using DZ U-Pb match existing age controls on basin stratigraphy. Our study shows that AFT is an effective thermochronometer to detect source exhumation for Cretaceous foreland stratigraphy in the western U.S.A. Lag-times (i.e. the difference between the source exhumation age and depositional age) are ~0 to 5 Myr with relatively steady-state to slightly increasing exhumation rates suggesting orogenic growth at this time. The very short lag times also indicate limited to no storage time between source and sink. The AFT lag time of the Early Cretaceous Kelvin Formation is ~5 Myr and represents a time when the Canyon Range Thrust was active to the southwest. Upper Cretaceous units have an approximate lag time of 0 Myr, during a time when the Willard, Crawford, Medicine Butte and Absaroka Thrust systems were active to the west. These lag-time measurements correspond to exhumation rates of ~0.8 mm/yr to >>1 mm/yr.

Painter, C. S.; Carrapa, B.; DeCelles, P. G.; Gehrels, G. E.; Thomson, S. N.

2013-12-01

148

Host-dependent transposon Tn5-mediated streptomycin resistance.  

PubMed

Transposon Tn5 encodes streptomycin resistance in addition to kanamycin-neomycin resistance. This resistance was not detectable in Escherichia coli but was efficiently expressed in Rhizobium meliloti and certain other strains. By analysis of cloned Tn5 restriction endonuclease fragments, the streptomycin resistance (str) gene was located in the right-hand side of the central region as the transposon is conventionally drawn. Transcription of str appeared to originate at pL, the promoter for the neo gene (neomycin phosphotransferase type II). Expression of streptomycin resistance in E. coli was obtained after cloning of the neo-str region downstream of a strong E. coli promoter. A construct in which PL was deleted also showed differential expression of streptomycin resistance. PMID:6330042

De Vos, G F; Finan, T M; Signer, E R; Walker, G C

1984-07-01

149

The transposon Tn5 carries a bleomycin-resistance determinant.  

PubMed

Transposon Tn5 carries a determinant for resistance to bleomycin (Bm). Deletion mapping and cloning experiments have shown that this determinant, gene ble, is located between the determinant for kanamycin (Km) and neomycin resistance (gene neo) and the determinant for streptomycin resistance (gene str). Genes neo, ble, and str belong to an operon controlled by the common promoter. The Mr of the ble product, as determined by polyacrylamide gel electrophoresis, is 12000 to 13000. PMID:6085062

Genilloud, O; Garrido, M C; Moreno, F

1984-12-01

150

Dissecting Tn5 transposition using HIV-1 integrase diketoacid inhibitors.  

PubMed

Diketoacid (DKA) compounds have been shown to inhibit HIV-1 integrase by a mechanism that involves sequestration of the active site metals. Because HIV-1 integrase and Tn5 transposase have similar active site architectures and catalytic mechanisms, we investigated whether DKA analogues would inhibit Tn5 transposase activity and provide a model system to explore the mechanisms of action of these inhibitors. A screen of several hundred DKA analogues identified several with activity against Tn5 Tnp. Six DKA inhibitors used in this study manifested a variety of effects on different transposition steps suggesting that different analogues may have different binding contacts with transposase. All DKA compounds inhibited paired end complex (PEC) formation in which the nucleoprotein complex required for catalysis is assembled. Dissociation of PECs by some DKA compounds indicates that these inhibitors can decrease PEC stability. Four DKA compounds inhibited the two cleavage steps releasing transposon DNA from flanking DNA, and one of these four compounds preferentially inhibited the second cleavage step. The differential effect of this inhibitor on the second cleavage event indicates that cleavage of the two transposon-donor DNA boundaries is a sequential process requiring a conformational change. The requirement for a conformational change between cleavage events was also demonstrated by the inability of transposase to perform second cleavage at 25 degrees C. Finally, all six compounds inhibit strand transfer, the final step of Tn5 transposition. Two of the compounds that inhibited strand transfer have no effect on DNA cleavage. The strand transfer inhibition properties of various DKA compounds was sensitive to the structure of the 5'-non-transferred strand, suggesting that these compounds bind in or near the transposase active site. Other results that probe compound binding sites include the effects of active site mutations and donor DNA on DKA compound inhibition activities. Thus, DKA inhibitors will provide an important set of tools to investigate the mechanism of action of transposases and integrases. PMID:17725323

Czyz, Agata; Stillmock, Kara A; Hazuda, Daria J; Reznikoff, William S

2007-09-25

151

Tn4399, a conjugal mobilizing transposon of Bacteroides fragilis.  

PubMed Central

Conjugal transposons play an important role in the dissemination of antibiotic resistance determinants in the streptococci and have been postulated to exist in Bacteroides fragilis. To investigate the presence of conjugal transposons in B. fragilis, we employed a Tra- derivative of the transfer factor pBFTM10 contained in the chimeric plasmid pGAT400 delta BglII. We attempted to restore transferability to this plasmid from a series of transconjugants generated by crossing B. fragilis TMP230 containing the TET transfer factor with B. fragilis TM4000, a standard recipient. Transconjugant TM4.2321 transferred pGAT400 delta BglII to Escherichia coli HB101 at almost the same frequency as did the Tra+ parental plasmid, pGAT400. Analysis of the transferred plasmids revealed the presence of 9.6 kilobases of additional DNA in every case but at different positions in independent isolates. The presence of this DNA, designated Tn4399, allowed the pGAT400 delta BglII derivatives to retransfer from the TM4000 background to B. fragilis or E. coli recipients. DNA hybridization studies demonstrated the presence of one copy of Tn4399 in TMP230 and three copies at new sites in TM4.2321. Tn4399 is a new B. fragilis transposon with unique transfer properties that may play a role in the dissemination of drug resistance genes. It differs from previously described conjugal transposons by its ability to mobilize nonconjugal plasmids in cis. Images

Hecht, D W; Malamy, M H

1989-01-01

152

Transposons Tn1696 and Tn21 and Their Integrons In4 and In2 Have Independent Origins  

Microsoft Academic Search

The first 13.6 kb of the mercury and multidrug resistance transposon Tn1696, which includes the class 1 integron In4, has been sequenced. In4 is 8.33 kb long and contains the 5*-conserved segment (5*-CS) and 2.24 kb of the 3*-conserved segment (3*-CS) flanking four integrated cassettes. The 3*-CS region is followed by one full copy and an adjacent partial copy of

SALLY R. PARTRIDGE; HEIDI J. BROWN; H. W. Stokes; RUTH M. HALL

2001-01-01

153

75 FR 29591 - Tennessee Disaster Number TN-00038  

Federal Register 2010, 2011, 2012, 2013

...include the following areas as adversely affected by the disaster. Primary Counties: Benton, Cannon, Giles, Marshall, Pickett, Sumner. All other information in the original declaration remains unchanged. (Catalog of Federal Domestic Assistance...

2010-05-26

154

IL17A-producing neutrophil-regulatory Tn lymphocytes  

Microsoft Academic Search

The proinflammatory cytokine IL-17A, mainly produced by specialized T cells, plays an important homeostatic role in regulating\\u000a neutrophil production and blood neutrophil counts. This review will assemble and discuss the evidence for this function of\\u000a IL-17A-producing cells, which are collectively called neutrophil-regulatory T cells or Tn cells. IL-17A-producing lymphocytes\\u000a are most abundant in the mesenteric lymph node, where they account

Klaus Ley; Emily Smith; Matthew A. Stark

2006-01-01

155

Tn5 transposase with an altered specificity for transposon ends.  

PubMed

Tn5 is a composite bacterial transposon that encodes a protein, transposase (Tnp), required for movement of the transposon. The initial step in the transposition pathway involves specific binding of Tnp to 19-bp end recognition sequences. Tn5 contains two different specific end sequences, termed outside end (OE) and inside end (IE). In Escherichia coli, IE is methylated by Dam methylase (IE(ME)). This methylation greatly inhibits recognition by Tnp and greatly reduces the ability of transposase to facilitate movement of IE defined transposons. Through use of a combinatorial random mutagenesis technique (DNA shuffling), we have isolated an IE(ME)-specific hyperactive form of Tnp, Tnp sC7v.2.0, that is able to promote high levels of transposition of IE(ME) defined transposons in vivo and in vitro while functioning at wild-type levels with OE transposons. This protein contains a critical glutamate-to-valine mutation at amino acid 58 that is responsible for this change in end specificity. PMID:11741865

Naumann, Todd A; Reznikoff, William S

2002-01-01

156

Percent Uninsured by County  

NSDL National Science Digital Library

This is a county by county visualization of the percentage of residents that are uninsured. The data are from a set available here: http://manyeyes.alphaworks.ibm.com/manyeyes/datasets/percent-uninsured-by-county/versions/1

Manyeyes

157

Characterization of Tn1546, a Tn3-related transposon conferring glycopeptide resistance by synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147.  

PubMed Central

Sequence determination of the flanking regions of the vancomycin resistance van gene cluster carried by pIP816 in Enterococcus faecium BM4147 revealed similarity to transposons of the Tn3 family. Imperfect inverted repeats (36 of 38 bp) delineated a 10,851-bp element designated Tn1546. The 4-kb region located upstream from the vanR gene contained two open reading frames (ORF) transcribed in opposite directions. The deduced amino acid sequence of ORF1 (988 residues) displayed, respectively, 56 and 42% identity to those of the transposases of Tn4430 from Bacillus thuringiensis and of Tn917 from Enterococcus faecalis. The product of ORF2 (191 residues) was related to the resolvase of Tn917 (33% amino acid identity) and to the Res protein (48%) of plasmid pIP404 from Clostridium perfringens. Tn1546 transposed consecutively in Escherichia coli from plasmid pUC18 into pOX38 and from pOX38 into various sites of pBR329. Transposition was replicative, led to the formation of cointegrates, and produced a 5-bp duplication at the target site. Southern hybridization and DNA amplification revealed the presence of Tn1546-related elements in enterococci highly resistant to glycopeptides. Analysis of sequences surrounding these elements indicated that transposition plays a role in dissemination of the van gene cluster among replicons of human clinical isolates of E. faecium.

Arthur, M; Molinas, C; Depardieu, F; Courvalin, P

1993-01-01

158

BWR - Spent Fuel Transport and Storage with the TN{sup TM}9/4 and TN{sup TM}24BH Casks  

SciTech Connect

The Swiss Nuclear Utilities have started in 2001 to store spent fuel in dry metallic dual-purpose casks at ZWILAG, the Swiss interim storage facility. BKW FMB Energy Ltd., the Muehleberg Nuclear Power Plant owner, is involved in this process and has elected to store its BWR spent fuel in a new high capacity dual-purpose cask, the TNeTeM24BH from the COGEMA Logistics/TRANSNUCLEAR TN{sup TM}24 family. The Muehleberg BWR spent fuels are transported by road in a medium size shuttle transport cask and then transferred to a heavy transport/storage cask (dry transfer) in the hot cell of ZWILAG site. For that purpose, COGEMA Logistics designed and supplied: - Two shuttle casks, TN{sup TM}9/4, mainly devoted to transport of spent fuel from Muehleberg NPP to ZWILAG. Licensed according to IAEA 1996, the TN{sup TM}9/4 is a 40 ton transport cask, for 7 BWR high bum-up spent fuel assemblies. - A series of new high capacity dual-purpose casks, TN{sup TM}24BH, holding 69 BWR spent fuels. Two transport campaigns took place in 2003 and 2004. For each campaign, ten TN{sup TM}9/4 round trips are performed, and one TN{sup TM}24BH is loaded. 5 additional TN{sup TM}24BH are being manufactured for BKW, and the next transport campaigns are scheduled from 2006. The TN{sup TM}24BH high capacity dual purpose cask and the TN{sup TM}9/4 transport cask characteristics and capabilities will then be detailed. (authors)

Wattez, L. [COGEMA LOGISTICS - AREVA Group (France); Marguerat, Y. [BKW FMB Energy Ltd (Switzerland); Hoesli, C. [ZWILAG Zwischenlager Wuerenlingen AG (Switzerland)

2006-07-01

159

Occurrence of Tn4371-Related Mobile Elements and Sequences in (Chloro)biphenyl-Degrading Bacteria  

PubMed Central

Tn4371, a 55-kb transposable element involved in the degradation and biphenyl or 4-chlorobiphenyl identified in Ralstonia eutropha A5, displays a modular structure including a phage-like integrase gene (int), a Pseudomonas-like (chloro)biphenyl catabolic gene cluster (bph), and RP4- and Ti-plasmid-like transfer genes (trb) (C. Merlin, D. Springael, and A. Toussaint, Plasmid 41:40–54, 1999). Southern blot hybridization was used to examine the presence of different regions of Tn4371 in a collection of (chloro)biphenyl-degrading bacteria originating from different habitats and belonging to different bacterial genera. Tn4371-related sequences were never detected on endogenous plasmids. Although the gene probes containing only bph sequences hybridized to genomic DNA from most strains tested, a limited selection of strains, all ?-proteobacteria, displayed hybridization patterns similar to the Tn4371 bph cluster. Homology between Tn4371 and DNA of two of those strains, originating from the same area as strain A5, extended outside the catabolic genes and covered the putative transfer region of Tn4371. On the other hand, none of the (chloro)biphenyl degraders hybridized with the outer left part of Tn4371 containing the int gene. The bph catabolic determinant of the two strains displaying homology to the Tn4371 transfer genes and a third strain isolated from the A5 area could be mobilized to a R. eutropha recipient, after insertion into an endogenous or introduced IncP1 plasmid. The mobilized DNA of those strains included all Tn4371 homologous sequences previously identified in their genome. Our observations show that the bph genes present on Tn4371 are highly conserved between different (chloro)biphenyl-degrading hosts, isolated globally but belonging mainly to the ?-proteobacteria. On the other hand, Tn4371-related mobile elements carrying bph genes are apparently only found in isolates from the environment that provided the Tn4371-bearing isolate A5.

Springael, Dirk; Ryngaert, Annemie; Merlin, Christophe; Toussaint, Ariane; Mergeay, Max

2001-01-01

160

Biochemical characterization of soluble Tn glycoproteins from malignant effusions of patients with carcinomas.  

PubMed

The Tn determinant (GalNAc-O-Ser/Thr) is one of the most specific human tumor markers. In normal cells Tn is a cryptic structure in the peptide core of mucin type O-glycoproteins, and it is detected in an unmasked form in most human carcinomas evaluated by immunohistochemistry. Scarce data are available regarding the characteristics of soluble Tn bearing glycoproteins. We herein report the first comparative characterization of soluble Tn glycoproteins derived from different kinds of human tumors (breast, colon, gastric, ovarian and liver). Considerable heterogeneity was observed in the physicochemical properties of Tn soluble glycoproteins from all the tumor-associated effusions evaluated. In SDS-PAGE analysis Tn glycoproteins from liver and colon effusions migrated as a broad single major component (>500 kDa), while several components of >200 kDa were identified in samples from breast, ovarian, and gastric cancer. The results of perchloric acid (PCA) treatment and CsCl gradient ultracentrifugation indicated that the Tn glycoproteins in effusion fluids correspond predominantly to mucin-like glycoproteins. However, in samples from patients with colon and liver cancer, a fraction of Tn glycoproteins formed part of the immune complexes that precipitated in PCA, suggesting that the anti-Tn immune response in vivo could modify their physicochemical properties. The four apomucins evaluated (MUC1, MUC2, MUC5AC and MUC6) carried Tn epitopes in each of the effusions, indicating that soluble apomucin detection may reflect the abnormal expression of MUC genes inherent to these tumors. Taking together, these results indicate that apomucin expression profile is responsible, at least in part, for the high heterogeneity of soluble Tn glycoproteins, and suggest that the identification of Tn determinant on the different soluble apomucins could be useful for the development of new diagnostic tools as well as to evaluate the anti-tumor immune response in patients with cancer. PMID:12883744

Freire, Teresa; Medeiros, Andrea; Reis, Celso A; Real, Francisco X; Osinaga, Eduardo

2003-01-01

161

Synthesis of Double C-Glycoside Analogue of sTn  

PubMed Central

A sTn double C-glycoside, sTn analogue 2, was synthesized using samarium chemistry developed in our laboratory. Complications in the oxidation reaction affording aldehyde acceptor were overcome by double protection of amide and the use of a room-temperature ionic liquid as solvent. Studies are underway to conjugate the sTn double C-glycoside hapten 2 to KLH carrier protein for biological evaluation as a vaccine.

Ress, Dino K.; Baytas, Sultan N.; Wang, Qun; Munoz, Eva M.; Tokuzoki, Kazuo; Tomiyama, Hiroshi; Linhardt, Robert J.

2014-01-01

162

TN approximation to neutron transport equation and application to critical slab problem  

NASA Astrophysics Data System (ADS)

The critical slab problem has been studied in one-speed neutron transport equation with isotropic scattering by using the TN method. TN moment criticality solutions are obtained for the uniform finite slab using Mark and Marshak type vacuum boundary conditions. Results obtained by TN method, using the two type boundary conditions mentioned above, were presented in the Tables and also the Tables included the results obtained by PN method for the comparisons.

Anli, Fikret; Ya?a, Faruk; Güngör, Süleyman; Öztürk, Hakan

2006-09-01

163

Transfer of Tn5385, a Composite, Multiresistance Chromosomal Element from Enterococcus faecalis  

Microsoft Academic Search

Tn5385 is a ca. 65-kb element integrated into the chromosomes of clinical Enterococcus faecalis strains CH19 and CH116. It confers resistance to erythromycin, gentamicin, mercuric chloride, streptomycin, tetracycline- minocycline, and penicillin via b-lactamase production. Tn5385 is a composite structure containing regions previously found in staphylococcal and enterococcal plasmids. Several transposons and transposon-like ele- ments within Tn5385 have been identified, including

LOUIS B. RICE; LENORE L. CARIAS

1998-01-01

164

DRAFT LANDSAT DATA MOSAIC: MONTGOMERY COUNTY, TEXAS; HARRIS COUNTY, TEXAS; FORT BEND COUNTY, TEXAS; BRAZORIA COUNTY, TEXAS; GALVESTON COUNTY, TEXAS  

EPA Science Inventory

This is a draft Landsat Data Mosaic, which contains remote sensing information for Montgomery County, Texas Harris County, Texas Fort Bend County, Texas Brazoria County, Texas Galveston County, and Texas Imagery dates on the following dates: October 6, 1999 and September 29, 200...

165

Nonlinear pressure dependence of TN in almost multiferroic EuTiO3  

NASA Astrophysics Data System (ADS)

The antiferromagnetic (AFM) phase transition temperature TN of EuTiO3 has been studied as a function of pressure p. The data reveal a nonlinear dependence of TN on p with TN increasing with increasing pressure. The exchange interactions exhibit an analogous dependence on p as TN (if the absolute value of the nearest neighbor interaction is considered) and there is evidence that the AFM transition is robust with increasing pressure. The corresponding Weiss temperature ?W remains anomalous since it always exhibits positive values. The data are analyzed within the Bloch power law model and provide excellent agreement with experiment.

Guguchia, Z.; Caslin, K.; Kremer, R. K.; Keller, H.; Shengelaya, A.; Maisuradze, A.; Bettis, J. L., Jr.; Köhler, J.; Bussmann-Holder, A.; Whangbo, M.-H.

2013-09-01

166

Nonlinear pressure dependence of TN in almost multiferroic EuTiO3.  

PubMed

The antiferromagnetic (AFM) phase transition temperature TN of EuTiO3 has been studied as a function of pressure p. The data reveal a nonlinear dependence of TN on p with TN increasing with increasing pressure. The exchange interactions exhibit an analogous dependence on p as TN (if the absolute value of the nearest neighbor interaction is considered) and there is evidence that the AFM transition is robust with increasing pressure. The corresponding Weiss temperature ?W remains anomalous since it always exhibits positive values. The data are analyzed within the Bloch power law model and provide excellent agreement with experiment. PMID:23963024

Guguchia, Z; Caslin, K; Kremer, R K; Keller, H; Shengelaya, A; Maisuradze, A; Bettis, J L; Köhler, J; Bussmann-Holder, A; Whangbo, M-H

2013-09-18

167

Specific DNA cleavage mediated by the integrase of conjugative transposon Tn916.  

PubMed Central

The conjugative transposon Tn916 encodes a protein called INT(Tn916) which, based on DNA sequence comparisons, is a member of the integrase family of site-specific recombinases. Integrase proteins such as INT(lambda), FLP, and XERC/D that promote site-specific recombination use characteristic, conserved amino acid residues to catalyze the cleavage and ligation of DNA substrates during recombination. The reaction proceeds by a two-step transesterification reaction requiring the formation of a covalent protein-DNA intermediate. Different requirements for homology between recombining DNA sites during integrase-mediated site-specific recombination and Tn916 transposition suggest that INT(Tn916) may use a reaction mechanism different from that used by other integrase recombinases. We show that purified INT(Tn916) mediates specific cleavage of duplex DNA substrates containing the Tn916 transposon ends and adjacent bacterial sequences. Staggered cleavages occur at both ends of the transposon, resulting in 5' hydroxyl protruding ends containing coupling sequences. These are sequences that are transferred with the transposon from donor to recipient during conjugative transposition. The nature of the cleavage products suggests that a covalent protein-DNA linkage occurs via a residue of INT(Tn916) and the 3'-phosphate group of the DNA. INT(Tn916) alone is capable of executing the strand cleavage step required for recombination during Tn916 transposition, and this reaction probably occurs by a mechanism similar to that of other integrase family site-specific recombinases.

Taylor, K L; Churchward, G

1997-01-01

168

Chiral fluctuations in triangular antiferromagnets at T? TN  

NASA Astrophysics Data System (ADS)

Chiral fluctuations in triangular antiferromagnets (TAFs) at T?TN are studied theoretically. The case of a ferromagnetic interaction along the c axis (which is directed perpendicular to the plane of the lattice), Dzyaloshinskii-Moriya interaction D?c and a weak magnetic field H?c , is considered in detail. Previously, this model has been proposed to describe quantum TAF CsCuCl3 . Expressions for dynamical chirality (DC) are derived within the linear spin-wave approximation. In contrast to nonfrustrated antiferromagnets, DC is found to be nonzero even at D,H=0 in a one-domain sample. We argue that this unusual behavior stems from the fact that a ground state of XY and Heisenberg TAFs is characterized by an axial vector along which DC is directed.

Syromyatnikov, A. V.

2005-04-01

169

Interaction of Tn5 transposase with the transposon termini.  

PubMed

Transposition of Tn5 requires the binding of the transposase protein to the transposon outside end (OE) DNA sequences. Transposase mutants that increase the transposition frequency result in the formation of two distinct transposase/OE DNA complexes, observed by gel retardation analysis. The slower migrating complex I, also formed by wild-type transposase, contains protein oligomers of transposase and transposase related proteins. The faster migrating, novel complex II is caused by the binding of monomeric, proteolytic transposase fragments gamma and delta that have lost the carboxy-terminus of the protein. Transposase gamma and delta bind OE DNA with a high apparent affinity but are unable to promote transposition in vivo. We propose that the transposase protein is functionally unstable and can undergo a conformational change that reduces the activity but protects the protein from proteolysis. The transposase mutants favor the more active but proteolytically hypersensitive protein conformation. PMID:8289277

Wiegand, T W; Reznikoff, W S

1994-01-14

170

8. DETAIL SHOT OF POST AND RAIL AND CUT AWAY ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. DETAIL SHOT OF POST AND RAIL AND CUT AWAY VIEW OF ABUTMENT SUPPORT INTO THE GROUND - Lost Creek Road Bridge, Spanning Sevier River at County Road 243, 2 miles east of Aurora, Aurora, Sevier County, UT

171

Precise excision and instability of the transposon Tn5 in Pseudomonas aeruginosa.  

PubMed

Excision of the transposon Tn5 from sites of insertion in plasmid DNA was shown to occur at high frequency in Pseudomonas aeruginosa. Plasmids with Tn5 insertions conjugated poorly into P. aeruginosa and adversely affected growth compared to the respective parental plasmids. The kanamycin-resistance phenotype of Tn5 was expressed poorly in P. aeruginosa and kanamycin-sensitive strains were common during the manipulation of the P. aeruginosa transconjugants. Examination of plasmid DNA isolated from kanamycin-sensitive P. aeruginosa transconjugants revealed excision of Tn5 sequences. A plasmid containing a selectable marker (mercury resistance) inactivated by a Tn5 insertion was constructed, and Tn5 excised precisely, permitting the expression of the mercury-resistance marker at high frequency (10(-3) in P. aeruginosa and at the expected low frequency (10(-7] in Escherichia coli. The recombinational mechanism that promotes frequent Tn5 excision in P. aeruginosa operated in the absence of the P. aeruginosa recA gene product. Fragments of Tn5 were also examined for excision and instability in P. aeruginosa. A plasmid containing the terminal 485 bp of inverted repeat sequences from Tn5, but lacking the transposase or kanamycin-resistance genes, also showed precise excision of Tn5 DNA at high frequency (10(-2] in P. aeruginosa. Unlike plasmids containing a complete Tn5 insertion, this plasmid transferred to P. aeruginosa at normal frequencies and growth of the host was not severely impaired. In contrast, plasmids containing either IS50 element transferred to P. aeruginosa at greatly reduced frequencies, and transconjugants containing the IS50R element (which contains the active transposase gene) were small and especially difficult to maintain. P. aeruginosa transconjugants harbouring a plasmid containing only the DNA between the IS50 elements (which included the kanamycin-resistance gene) were of normal size and stably maintained.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2166130

Goldberg, J B; Won, J; Ohman, D E

1990-05-01

172

TN approximation to reflected slab and computation of the critical half thicknesses  

NASA Astrophysics Data System (ADS)

The criticality solution to one-speed neutron transport equation using the TN approximation is described for reflected slab. In the solution, Marshak type boundary condition is used. The critical half thicknesses are computed for different values of c and reflection coefficients. Computations are made by using the both TN and PN approximation for the comparison.

Anli, F.; Güngör, S.; Ya?a, F.; Öztürk, H.

2006-09-01

173

Bleomycin increases amikacin and streptomycin resistance in Escherichia coli harboring transposon Tn5.  

PubMed

The antitumor antibiotic bleomycin acts as a transcriptional inducer of the neo-ble-str operon of the transposon Tn5, increasing the resistance level to streptomycin and amikacin in Tn5-containing Escherichia coli. The mechanism may involve a recA-independent induction mediated by DNA damage. PMID:7694544

Blazquez, J; Martinez, J L; Baquero, F

1993-09-01

174

Transcriptional activity of the transposable element Tn10 in the Salmonella typhimurium ilvGEDA operon.  

PubMed Central

Polarity of Tn10 insertion mutations in the Salmonella typhimurium ilvGEDA operon depends on both the location and the orientation of the Tn10 element. One orientation of Tn10 insertions in ilvG and ilvE permits low-level expression of the downstream ilvEDA and ilvDA genes, respectively. Our analysis of Salmonella ilv recombinant plasmids shows that this residual ilv expression must result from Tn10-directed transcription and does not reflect the presence of internal promoters in the ilvGEDA operon, as was previously suggested. The opposite orientation of Tn10 insertion in ilvE prevents ilvDA expression, indicating that only one end of Tn10 is normally active in transcribing adjacent genes. Both orientations of Tn10 insertion in ilvD exert absolute polarity on ilvA expression. Expression of ilvA is known to be dependent on effective translation of ilvD, perhaps reflecting the lack of a ribosome binding site proximal to the ilvA sequence. Therefore, recognition of the ability of Tn10 to promote transcription of contiguous genes in the ilvGEDA operon apparently requires the presence of associated ribosome binding sites.

Blazey, D L; Burns, R O

1982-01-01

175

The Lower Sevier River Basin Crop Monitor and Forecast Decision Support System: Exploiting Landsat Imagery to Provide Continuous Information to Farmers and Water Managers  

NASA Astrophysics Data System (ADS)

The last century has seen a large number of innovations in agriculture such as better policies for water control and management, upgraded water conveyance, irrigation, distribution, and monitoring systems, and better weather forecasting products. In spite of this, irrigation management and irrigation water deliveries by farmers/water managers is still based on factors like water share amounts, tradition, and past experience on irrigation. These factors are not necessarily related to the actual crop water use; they are followed because of the absence of related information provided in a timely manner at an affordable cost. Thus, it is necessary to develop means to deliver continuous and personalized information about crop water requirements to water users/managers at the field and irrigation system levels so managers at these levels can better quantify the required versus available water for irrigation during the irrigation season. This study presents a new decision support system (DSS) platform that addresses the absence of information on actual crop water requirements and crop performance by providing continuous updated farm-based crop water use along with other farm performance indicators such as crop yield and farm management to irrigators and water managers. This DSS exploits the periodicity of the Landsat Satellite Mission (8 to 16 days, depending on the period of interest) to provide remote monitoring at the individual field and irrigation system levels. The Landsat satellite images are converted into information about crop water use, yield performance and field management through application of state-of-the-art semi-physical and statistical algorithms that provide this information at a pixel basis that are ultimately aggregated to field and irrigation system levels. A version of the DSS has been implemented for the agricultural lands in the Lower Sevier River, Utah, and has been operational since the beginning of the 2013 irrigation season. The main goal of this DSS implementation is to provide continuous and personalized information to farmers and water managers regarding crops in fields and the irrigation delivery system throughout the irrigation season so that decisions related to agricultural water use can result in water savings while not diminishing crop yields.

Torres-Rua, A. F.; Walker, W. R.; McKee, M.

2013-12-01

176

Control of Neogene Extension in Idaho and Utah by Thrust Ramps in the Sevier Orogenic Belt: Evidence From Gravity Modeling in Cache and Cottonwood Valleys  

NASA Astrophysics Data System (ADS)

The subsurface structure of Cache and Cottonwood Valleys is largely due to two episodes of deformation. The Cretaceous to Eocene Sevier orogeny caused compressional folding and faulting across the region presently occupied by the basins. Following this, Neogene extension occurred along the Bannock detachment fault and other normal faults in Cache and Cottonwood Valleys. See-saw slippage of the west-dipping East Cache Valley fault and east-dipping West Cache fault in Utah and Idaho created a graben which filled with sediments of the Salt Lake Formation. Gravity modeling shows the subsurface structure of the basin using layer densities constrained by well logs and previously published densities. The base of the Tertiary was also constrained by seismic interpretations. The gravity model shows the location and dip of the East Cache Valley fault, West Cache fault, and Wasatch fault. It also indicates a ramp in the Paris thrust fault beneath the East Cache Valley fault and the Bear River Range (called the Logan ramp in this study), which puts higher density Proterozoic to middle Paleozoic units over lower density upper Paleozoic to Mesozoic units. In Cottonwood Valley, which is northeast of Cache Valley in Idaho, gravity data indicate that Neogene extension created a series of smaller northeast-dipping half grabens above the proposed breakaway zone of the southwest-dipping Bannock detachment fault. Like the East Cache Valley fault, the gravity data also indicate that the breakaway zone of the Bannock detachment fault in Cottonwood Valley formed above a ramp in the Paris thrust (called the Treasureton ramp in this and a previous study) which also placed higher density units over lower density ones. Based on the gravity and geologic data in this study, we interpret the East Cache Valley fault to be the breakaway zone for the Bannock detachment fault in southern and central Cache Valley. It is also likely that this fault connects to the Bannock detachment fault breakaway zone in Cottonwood Valley. Furthermore, we believe that the Logan and Treasureton ramps in the Paris thrust are connected, and that this ramp, as well as overlying thrust faults, were somehow responsible for localizing the breakaway zone of the Bannock detachment fault system tens of millions of years after thrusting ceased.

Groze, C. L.; Kruger, J. M.; Eversaul, M. L.

2006-12-01

177

Low-temperature thermochronology of the Laramide Ranges and eastward translation of shortening in the Sevier Belt, Wyoming, Utah and Montana  

NASA Astrophysics Data System (ADS)

This dissertation contains two studies that use very different techniques to investigate the Mesozoic and Cenozoic tectonics of the western USA. The first study investigates shortening in the Sevier thrust belt of northeast Utah and southwest Wyoming using cross sections and seismic reflection data. The second study investigates the low-temperature thermochronology of the Laramide Ranges using apatite (U-Th)/He dating. We used cross sections and seismic reflection data to investigate bed length discrepancies within the hanging wall of the Absaroka thrust in the Sevier thrust belt of northeast Utah and southwest Wyoming. Restoration of cross sections suggests that there was ˜8-14 km of pre-Absaroka-thrust shortening above the Jurassic Preuss salt detachment, but not below it, in the hanging wall of the Absaroka thrust. Reflection seismic data over the hanging wall of the Crawford thrust show that the Crawford thrust is not offset along the Preuss salt detachment, indicating that the additional shortening on the Absaroka plate was transferred east before main movement on the Crawford thrust. Although early displacement on the Crawford thrust cannot be ruled out as the cause of the extra shortening, surface and subsurface geology suggests slip from the western thrust system (Willard and Lost Creek thrusts) was transferred several tens of kilometers east along the Jurassic Preuss salt detachment between ˜102-90 Ma, to the future location of the Absaroka thrust hanging wall. The lack of deformation of the Crawford thrust on the seismic data, along with shortening and extension estimates from cross sections, also indicate that the magnitude of Paleocene and post-early Eocene shortening on the Medicine Butte thrust was essentially offset by subsequent extension on the middle Eocene to late Oligocene Almy-Acocks normal-fault system. For the second study in this dissertation, we dated 91 borehole and surface samples from Laramide-age, basement-cored uplifts of the Rocky Mountain foreland (Wind River, Beartooth, Bighorn and Laramie Ranges) and the Uncompahgre Uplift using the apatite (U-Th)/He system. Apatite (U-Th)/He ages generally decrease with increasing subsurface depth (decreasing elevation) and most samples show age dispersion ranging from tens to hundreds of Myr. Additionally, several samples show correlations between apatite (U-Th)/He age and effective U concentration (eU = [U] + 0.235[Th]) of the crystal, indicating that radiation damage has affected He diffusivity, and hence (U-Th)/ He age. Many surface and near-surface samples have apatite (U-Th)/He ages that are older than corresponding apatite fission-track ages. Forward modeling of Laramide-type thermal histories using a radiation damage diffusion model showed that (U-Th)/He ages may be widely dispersed, and may be older than corresponding apatite fission-track ages within a fossil He partial retention zone. Most of our samples, however, do not show the correlation between (U-Th)/He age and eU predicted by radiation damage diffusion models. We investigated the influence of both grain size and eU content and show that the effects of grain size can obscure (U-Th)/ He age-eU correlations and, similarly, the effect of eU variation can obscure (U-Th)/ He age-grain size correlations. (U-Th)/He ages that are older than fission-track ages from high peaks in the Wind River Range, and from some samples from the Beartooth Range, are most likely the result of He implantation from high eU phases. Best-fit thermal histories from the inversion of age-eU pairs were extrapolated to other elevations to create model age-elevation profiles for a range of eU concentrations. These model profiles approximate our real data. Inverse modeling of (U-Th)/He age data suggests that rapid exhumation within the Laramide province likely began earlier in the Bighorn Mountains (before ˜71 Ma) than the Beartooth Range (before ˜58 Ma), and that the borehole at the northern end of the Laramie Range penetrated a fault sliver at depth.

Peyton, Sara Lynn

178

Spectroscopic and ITC study of the conformational change upon Ca{sup 2+}-binding in TnC C-lobe and TnI peptide complex from Akazara scallop striated muscle  

SciTech Connect

Akazara scallop (Chlamys nipponensis akazara) troponin C (TnC) of striated adductor muscle binds only one Ca{sup 2+} ion at the C-terminal EF-hand motif (Site IV), but it works as the Ca{sup 2+}-dependent regulator in adductor muscle contraction. In addition, the scallop troponin (Tn) has been thought to regulate muscle contraction via activating mechanisms that involve the region spanning from the TnC C-lobe (C-lobe) binding site to the inhibitory region of the TnI, and no alternative binding of the TnI C-terminal region to TnC because of no similarity between second TnC-binding regions of vertebrate and the scallop TnIs. To clarify the Ca{sup 2+}-regulatory mechanism of muscle contraction by scallop Tn, we have analyzed the Ca{sup 2+}-binding properties of the complex of TnC C-lobe and TnI peptide, and their interaction using isothermal titration microcalorimetry, nuclear magnetic resonance, circular dichroism, and gel filtration chromatography. The results showed that single Ca{sup 2+}-binding to the Site IV leads to a structural transition not only in Site IV but also Site III through the structural network in the C-lobe of scallop TnC. We therefore assumed that the effect of Ca{sup 2+}-binding must lead to a change in the interaction mode between the C-lobe of TnC and the TnI peptide. The change should be the first event of the transmission of Ca{sup 2+} signal to TnI in Tn ternary complex.

Yumoto, Fumiaki [Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan); Department of Physiology II, Jikei University School of Medicine, Tokyo (Japan); Tanaka, Hiroyuki [Laboratory of Biochemistry and Biotechnology, Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido (Japan); Nagata, Koji; Miyauchi, Yumiko; Miyakawa, Takuya [Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan); Ojima, Takao [Laboratory of Biochemistry and Biotechnology, Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido (Japan); Tanokura, Masaru [Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan)], E-mail: amtanok@mail.ecc.u-tokyo.ac.jp

2008-04-25

179

Tn5401, a new class II transposable element from Bacillus thuringiensis.  

PubMed Central

A new class II (Tn3-like) transposable element, designated Tn5401, was recovered from a sporulation-deficient variant of Bacillus thuringiensis subsp. morrisoni EG2158 following its insertion into a recombinant plasmid. Sequence analysis of the insert revealed a 4,837-bp transposon with two large open reading frames, in the same orientation, encoding proteins of 36 kDa (306 residues) and 116 kDa (1,005 residues) and 53-bp terminal inverted repeats. The deduced amino acid sequence for the 36-kDa protein shows 24% sequence identity with the TnpI recombinase of the B. thuringiensis transposon Tn4430, a member of the phage integrase family of site-specific recombinases. The deduced amino acid sequence for the 116-kDa protein shows 42% sequence identity with the transposase of Tn3 but only 28% identity with the TnpA transposase of Tn4430. Two small open reading frames of unknown function, designated orf1 (85 residues) and orf2 (74 residues), were also identified. Southern blot analysis indicated that Tn5401, in contrast to Tn4430, is not commonly found among different subspecies of B. thuringiensis and is not typically associated with known insecticidal crystal protein genes. Transposition was studied with B. thuringiensis by using plasmid pEG922, a temperature-sensitive shuttle vector containing Tn5401. Tn5401 transposed to both chromosomal and plasmid target sites but displayed an apparent preference for plasmid sites. Transposition was replicative and resulted in the generation of a 5-bp duplication at the target site. Transcriptional start sites within Tn5401 were mapped by primer extension analysis. Two promoters, designated PL and PR, direct the transcription of orf1-orf2 and tnpI-tnpA, respectively, and are negatively regulated by TnpI. Sequence comparison of the promoter regions of Tn5401 and Tn4430 suggests that the conserved sequence element ATGTCCRCTAAY mediates TnpI binding and cointegrate resolution. The same element is contained within the 53-bp terminal inverted repeats, thus accounting for their unusual lengths and suggesting an additional role for TnpI in regulating Tn5401 transposition. Images

Baum, J A

1994-01-01

180

The Topology of T-Duality for Tn-BUNDLES  

NASA Astrophysics Data System (ADS)

In string theory, the concept of T-duality between two principal Tn-bundles E and Ê over the same base space B, together with cohomology classes h ? H3(E,?) and ? ? H3(Ê,?), has been introduced. One of the main virtues of T-duality is that h-twisted K-theory of E is isomorphic to ?-twisted K-theory of Ê. In this paper, a new, very topological concept of T-duality is introduced. We construct a classifying space for pairs as above with additional "dualizing data", with a forgetful map to the classifying space for pairs (also constructed in the paper). On the first classifying space, we have an involution which corresponds to passage to the dual pair, i.e. to each pair with dualizing data exists a well defined dual pair (with dualizing data). We show that a pair (E, h) can be lifted to a pair with dualizing data if and only if h belongs to the second step of the Leray-Serre filtration of E (i.e. not always), and that in general many different lifts exist, with topologically different dual bundles. We establish several properties of the T-dual pairs. In particular, we prove a T-duality isomorphism of degree -n for twisted K-theory.

Bunke, Ulrich; Rumpf, Philipp; Schick, Thomas

181

The organization of the outside end of transposon Tn5.  

PubMed

The end sequences of the IS50 insertion sequence are known as the outside end (OE) and inside end. These complex ends are related but nonidentical 19-bp sequences that serve as substrates for the activity of the Tn5 transposase. Besides providing the binding site of the transposase, the end sequences of a transposon contain additional types of information necessary for transposition. These additional properties include but are not limited to host protein interaction sites and sites that program synapsis and cleavage events. In order to delineate the properties of the IS50 ends,the base pairs involved in the transposase binding site have been defined. This has been approached through performing a variety of in vitro analyses: a ++hydroxyl radical missing-nucleoside interference experiment, a dimethyl sulfate interference experiment, and an examination of the relative binding affinities of single-site end substitutions. These approaches have led to the conclusion that the transposase binds to two nonsymmetrical regions of the OE, including positions 6 to 9 and 13 to 19. Proper binding occurs along one face of the helix, over two major and minor grooves, and appears to result in a significant bending of the DNA centered approximately 3 bp from the donor DNA-OE junction. PMID:8626296

Jilk, R A; York, D; Reznikoff, W S

1996-03-01

182

78 FR 58489 - Proposed Amendment of Class E Airspace; McMinnville, TN  

Federal Register 2010, 2011, 2012, 2013

...Procedures have been developed at Warren County Memorial Airport. This action...Approach Procedures developed at Warren County Memorial Airport, McMinnville...would amend Class E airspace at Warren County Memorial Airport,...

2013-09-24

183

Tn6001, a Transposon-Like Element Containing the blaVIM-3-Harboring Integron In450?  

PubMed Central

We describe the structure of a transposon-like element named Tn6001, which contains a blaVIM-3-harboring integron In450, which was derived from a multidrug-resistant Pseudomonas aeruginosa clinical isolate in Taiwan. The transposon backbone structure is most closely related to those of Tn1404* and Tn1403. Tn6001 was inserted into the chromosome of the clinical isolate.

Tseng, Sung-Pin; Hsueh, Po-Ren; Tsai, Jui-Chang; Teng, Lee-Jene

2007-01-01

184

The development of Tn Nuc and its use for the isolation of novel secretion signals in Lactococcus lactis  

Microsoft Academic Search

We have previously used Tn917 for the identification and characterization of regulated promoters from Lactococcus lactis [Israelsen et al., Appl. Environ. Microbiol. 61 (1995) 2540–2547]. We describe here the construction of a new Tn917-transposon derivative, termed TnNuc, which includes the Staphylococcus aureus nuclease gene (nuc) as a reporter for secretion. Transposition of TnNuc into the L. lactis chromosome allows the

Peter Ravn; José Arnau; Søren M. Madsen; Astrid Vrang; Hans Israelsen

2000-01-01

185

Identification of Tn4430, a transposon of Bacillus thuringiensis functional in Escherichia coli.  

PubMed

The mobile genetic element Tn4430, originating from the gram-positive bacterium, Bacillus thuringiensis, and previously described as the Th-sequence, is the first transposon isolated from the genus Bacillus. In the present work a gene (APH-III) conferring resistance to kanamycin was inserted into this 4.2 kb transposon. Transposition experiments showed that Tn4430 omega APH-III could transpose in the gram-negative host Escherichia coli when its insertion functions were supplied by an intact copy of Tn4430. By transposing Tn4430 omega APH-III directly onto pBR322, it was possible to determine the nucleotide sequence of the terminal inverted repeats of Tn4430 and of the target DNA site. Identical 38 bp in inverted orientation are situated at each end of the transposon and there is a direct duplication of 5 bp at the insertion site. Thus, it is clear that Tn4430 is closely related to the transposons belonging to the Tn3 family (class II elements). PMID:3018445

Lereclus, D; Mahillon, J; Menou, G; Lecadet, M M

1986-07-01

186

Tn4556, a 6.8-kilobase-pair transposable element of Streptomyces fradiae.  

PubMed

A 6.8-kilobase-pair (kbp) transposable element (Tn4556) was found in a neomycin-producing strain of Streptomyces fradiae. This element was first observed in two 30.3-kbp plasmids (pUC1123 and pUC1124) which arose when a thiostrepton resistance gene (1 kbp) was ligated with the BclI-2 fragment (22.5 kbp) that contains the origin of replication of phage SF1. The Tn4556 segment was deleted when these plasmids were transduced into another S. fradiae host with phage SF1. These deletion plasmids (pUC1210 and pUC1211) had copy numbers of less than 1 per chromosome and were unstable. In contrast, pUC1123 and pUC1124, with copy numbers of 12 to 15 per chromosome, respectively, were relatively stable. When pUC1210 and pUC1211 were reintroduced into S. fradiae by protoplast transformation, the Tn4556 element transposed again to the plasmids at numerous new locations in either of two orientations. A copy of Tn4556 was found in the S. fradiae chromosome by hybridization studies. It appears that Tn4556 originated from the chromosome, transposed into unstable pUC1210 and pUC1211, and made stable plasmids. A temperature-sensitive hybrid plasmid carrying a viomycin resistance derivative of Tn4556 (pMT660::Tn4556::vph) was constructed. When Streptomyces lividans UC8390 containing the hybrid plasmid was grown at 39 degrees C, Tn4556::vph (Tn4560) transposed to random positions in the host chromosome. PMID:2820925

Chung, S T

1987-10-01

187

Epigenetic Silencing of the Chaperone Cosmc in Human Leukocytes Expressing Tn Antigen*  

PubMed Central

Cosmc is the specific molecular chaperone in the endoplasmic reticulum for T-synthase, a Golgi ?3-galactosyltransferase that generates the core 1 O-glycan, Gal?1–3GalNAc?-Ser/Thr, in glycoproteins. Dysfunctional Cosmc results in the formation of inactive T-synthase and consequent expression of the Tn antigen (GalNAc?1-Ser/Thr), which is associated with several human diseases. However, the molecular regulation of expression of Cosmc, which is encoded by a single gene on Xq24, is poorly understood. Here we show that epigenetic silencing of Cosmc through hypermethylation of its promoter leads to loss of Cosmc transcripts in Tn4 cells, an immortalized B cell line from a male patient with a Tn-syndrome-like phenotype. These cells lack T-synthase activity and express the Tn antigen. Treatment of cells with 5-aza-2?-deoxycytidine causes restoration of Cosmc transcripts, restores T-synthase activity, and reduces Tn antigen expression. Bisulfite sequencing shows that CG dinucleotides in the Cosmc core promoter are hypermethylated. Interestingly, several other X-linked genes associated with glycosylation are not silenced in Tn4 cells, and we observed no correlation of a particular DNA methyltransferase to aberrant methylation of Cosmc in these cells. Thus, hypermethylation of the Cosmc promoter in Tn4 cells is relatively specific. Epigenetic silencing of Cosmc provides another mechanism underlying the abnormal expression of the Tn antigen, which may be important in understanding aberrant Tn antigen expression in human diseases, including IgA nephropathy and cancer.

Mi, Rongjuan; Song, Lina; Wang, Yingchun; Ding, Xiaokun; Zeng, Junwei; Lehoux, Sylvain; Aryal, Rajindra P.; Wang, Jianmei; Crew, Vanja K.; van Die, Irma; Chapman, Arlene B.; Cummings, Richard D.; Ju, Tongzhong

2012-01-01

188

A clinical isolate of transposon Tn5 expressing streptomycin resistance in Escherichia coli.  

PubMed Central

The central region of transposon Tn5 carries three antibiotic resistance markers: neo, ble, and str. The str gene codes for a phosphotransferase that inactivates streptomycin. This activity is phenotypically expressed in several gram-negative bacteria but not in Escherichia coli. We identified a Tn5 variant in E. coli clinical isolates that express streptomycin resistance. This transposon carries a 6-base-pair deletion within the str gene, near the 3' end. The same kind of mutation had been previously obtained experimentally from Tn5. Images

Genilloud, O; Blazquez, J; Mazodier, P; Moreno, F

1988-01-01

189

A clinical isolate of transposon Tn5 expressing streptomycin resistance in Escherichia coli.  

PubMed

The central region of transposon Tn5 carries three antibiotic resistance markers: neo, ble, and str. The str gene codes for a phosphotransferase that inactivates streptomycin. This activity is phenotypically expressed in several gram-negative bacteria but not in Escherichia coli. We identified a Tn5 variant in E. coli clinical isolates that express streptomycin resistance. This transposon carries a 6-base-pair deletion within the str gene, near the 3' end. The same kind of mutation had been previously obtained experimentally from Tn5. PMID:2830233

Genilloud, O; Blázquez, J; Mazodier, P; Moreno, F

1988-03-01

190

Tn5supF, a 264-base-pair transposon derived from Tn5 for insertion mutagenesis and sequencing DNAs cloned in phage lambda.  

PubMed

We constructed a derivative of transposon Tn5 called Tn5supF for insertion mutagenesis and sequencing DNAs cloned in phage lambda. This element carries a supF amber-suppressor tRNA gene. Its insertion into lambda can be selected by plaque formation by using nonsuppressing (sup0) Escherichia coli for amber mutant lambda phage and sup0 dnaB-amber E. coli for nonamber lambda phage. Tn5supF is just 264 base pairs long. It transposes efficiently and inserts quasi-randomly into DNA targets. The unique sequences near its termini can be used as primer binding sites for dideoxynucleotide DNA sequencing, thus permitting the direct sequencing of DNAs cloned in phage lambda without subcloning. PMID:2548192

Phadnis, S H; Huang, H V; Berg, D E

1989-08-01

191

Polarization dependence of the magnetic fluctuations in Cr below TN  

NASA Astrophysics Data System (ADS)

We have investigated the magnetic excitations in single-Q, single-domain Cr below TN using polarization analysis of inelastically scattered neutrons for energy transfers up to 18 meV. The results show that the longitudinal (L) magnetic modes are enhanced below ~=8 meV with respect to the transverse (T) modes at low temperature. The latter are independent of the polarization with respect to the ordering vector Q'+/- in the transverse spin-density-wave phase, i.e., they are isotropic. The L and T modes are not affected by the spin-flop transition to the longitudinal spin-density-wave phase at Tsf=121 K indicating that the transition is due to anisotropies and that it is not an intrinsic feature of the spin-density wave. There are indications that the L modes have a less steep dispersion curve and are more damped than the T modes in accordance with the three-band model by Fishman and Liu. With increasing temperature the intensity from the T and L modes increases strongly at small E transfer and their E dependence becomes similar. In particular, the enhancement of the L modes with respect to the T modes vanishes. Significant inelastic scattering is present at the silent peak positions. It surpasses even the scattering at the allowed peak positions at large E transfers. We argue that the silent inelastic scattering is partly responsible for the ``commensurate'' scattering at the (100) position due to resolution effects and that the major cross section at (100) is due to the longitudinally polarized Fincher-Burke mode at 4.2 meV.

Böni, P.; Sternlieb, B. J.; Shirane, G.; Roessli, B.; Lorenzo, J. E.; Werner, S. A.

1998-01-01

192

Percent Uninsured by County  

NSDL National Science Digital Library

This is a county by county visualization of the percentage of residents that are uninsured. The data are from a set available here: http://www-958.ibm.com/software/data/cognos/manyeyes/datasets/percent-uninsured-by-county/versions/1

Manyeyes

193

Reviewing County Extension Programs  

ERIC Educational Resources Information Center

A review of the effectiveness of extension education programs in Douglas County, Kansas is presented. Data were collected through a review of program data, interviews, and questionnaires sent to county residents. The results indicate a general knowledge and acceptance of the county's extension services among the respondents. (EC)

Prawl, Warren L.; Jorns, William J.

1976-01-01

194

Direct interaction between the TnsA and TnsB subunits controls the heteromeric Tn7 transposase  

PubMed Central

The transposon Tn7 transposase that recognizes the transposon ends and mediates breakage and joining is heteromeric. It contains the Tn7-encoded proteins TnsB, which binds specifically to the transposon ends and carries out breakage and joining at the 3? ends, and TnsA, which carries out breakage at the 5? ends of Tn7. TnsA apparently does not bind specifically to DNA, and we have hypothesized that it is recruited to the ends by interaction with TnsB. In this work, we show that TnsA and TnsB interact directly and identify several TnsA and TnsB amino acids involved in this interaction. We also show that TnsA can stimulate two key activities of TnsB, specific binding to the ends and pairing of the Tn7 ends. The ends of Tn7 are structurally asymmetric (i.e., contain different numbers of TnsB-binding sites), and Tn7 also is functionally asymmetric, inserting into its specific target site, attachment site attTn7 (attTn7) in a single orientation. Moreover, Tn7 elements containing two Tn7 right ends can transpose, but elements with two Tn7 left ends cannot. We show here that TnsA + TnsB are unable to pair the ends of a Tn7 element containing two Tn7 left ends. This pairing defect likely contributes to the inability of Tn7 elements with two Tn7 left ends to transpose.

Choi, Ki Young; Li, Ying; Sarnovsky, Robert; Craig, Nancy L.

2013-01-01

195

Tn4563 transposition in Streptomyces coelicolor and its application to isolation of new morphological mutants.  

PubMed Central

The Tn3-like transposon Tn4556 (and its derivatives Tn4560 and Tn4563) has been used for insertion mapping of genetic loci cloned on plasmids, but it has been difficult to obtain chromosomal insertions, largely because of the lack of a strong selection against transposon donor molecules. In this communication, we report two efficient selection techniques for transposition and their use in the isolation of chromosomal insertion mutations. A number of independent Streptomyces coelicolor morphological mutants (bld and whi) were obtained. Two of the bld mutations were mapped to locations on the chromosome by SCP1-mediated conjugation; at least one mutation, bld-5m1, appears to define a novel locus involved in control of S. coelicolor morphogenesis and antibiotic production. Images

Schauer, A T; Nelson, A D; Daniel, J B

1991-01-01

196

Inelastic neutron scattering in some ?-Mn alloys below and above TN  

NASA Astrophysics Data System (ADS)

Inelastic neutron scattering (INS) in the FCC Mn(37% Fe, 3% Cu) alloy and in the FCT ( c< a) Mn(12% Ge) and Mn(13.7% Ni) alloys was measured in a wide temperature range starting from few kelvins up to 1.3 TN. The low-temperature data are well explained in terms of neutron scattering on damped spin waves but essential changes start to appear already ca. 100-200 K below TN. The measurements in Mn(13.7% Ni) sample with unequal domain population allowed one to demonstrate the appearance of neutron scattering around the magnetic BZ boundary point, equivalent to the magnetic Brillouin zone center (MBZC) for differently oriented domains, in addition to the “normal” INS around the MBZC. The intensity distribution around this unfulfilled MBZC is identical with the distribution around the fulfilled MBZC at TN and above it, but differs below TN.

Mikke, Kazimierz; Jankowska-Kisielinska, Joanna; Hennion, Bernard

197

Tn7 transposition: a multigene process. Identification of a regulatory gene product.  

PubMed Central

Tn7 transposition is abolished by the deletion of a 2.2kb HindIII fragment from the central region of the transposon. Transposition is restored when the fragment is present in trans. When this fragment is present in trans with wild-type Tn7, transposition frequencies are stimulated 10-100-fold. The DNA sequence of this fragment has been determined and found to contain one long open reading frame coding for a protein of molecular weight 61 187. We have visualised this protein using a DNA-directed prokaryotic transcription-translation system. This gene may fill a regulatory role in the mechanism of Tn7 transposition. When present in trans the 2.2kb HindIII fragment alleviates a transcriptional block of promoter activity detected in Tn7. Images

Smith, G M; Jones, P

1986-01-01

198

76 FR 35370 - Proposed Establishment of Class E Airspace; Copperhill, TN  

Federal Register 2010, 2011, 2012, 2013

...Instrument Approach Procedures at Martin Campbell Field Airport. This action would enhance...instrument approach procedures for Martin Campbell Field Airport. Controlled airspace extending...establish Class E airspace at Martin Campbell Field Airport, Copperhill, TN....

2011-06-17

199

An alternative inverse PCR (IPCR) method to amplify DNA sequences flanking Tn5 transposon insertions.  

PubMed

We have developed an alternative method to amplify DNA sequences flanking Tn5 transposon insertions. This method relies on the identical sequences of inverted terminal repeats, located at the 5' and 3' ends of Tn5, to determine the location and orientation of a transposon insertion within a restriction endonuclease fragment. From this information, PCR primers can be designed to selectively amplify by inverse PCR the DNA flanking one side of the transposon. This method avoids the problem of amplifying or cloning long sequences flanking Tn5. To demonstrate the applicability of this method, we generated Tn5 transposon mutants of Pseudomonas abietaniphila BKME-9 which no longer grew on dehydroabietic acid (DhA). The flanking sequence of one of the mutant (strain BKME-941) which accumulated 7-oxoDhA, was amplified. PMID:10192049

Martin, V J; Mohn, W W

1999-03-01

200

The Levels and Distribution of TN, TP and TOC in the South China Sea  

NASA Astrophysics Data System (ADS)

The marine biogeochemistries of carbon, nitrogen and phosphorous have come under increased scrutiny because of their close involvement in climate change and coastal eutrophication. The South China Sea is unique in that located in a subtropical zone, and therefore represents an important regime for biogeochemical studies. However, to our knowledge, few data are available for total nitrogen (TN), total phosphorous (TP) and total organic carbon (TOC) in South Sea, China. The present study aims to contribute to the knowledge of their status through investigating the level and distribution of TN, TP and TOC in South China Sea. A total of 108 seawater samples of 11 sites in south sea, china were collected during August 29- September 4, 2006. An automated and simultaneous method for determination of TN and TOC was applied to all seawater samples. The combined system allowed simultaneous determination for TOC and TN in the same sample using a single injection and provided low detection limits and excellent linear ranges for both TOC and TN. The risk of contamination has been remarkably reduced due to the minimal sample manipulation and automated analyses. And quantitative analyses of TP in seawater were accomplished by a typical chemical method. Concentration ranges of TN and TP were 0.06-0.67, and 0.003-0.071 mg/L, respectively, as well as that of TOC were 0.23-2.51mg/L. The values of TN and TP showed that the status of nutrition is relatively better in south china sea than other marine areas. Moreover, the upright change trend of TN concentration level as well as TP and TOC according to the experimental results at the total 11 sites are simultaneity studied. The concentration of TN initial increases with the increasing of the depth, later the value becomes almost constant. In contrast, the concentration of TOC reduces with the increasing of the depth, later the value becomes almost unchangeable. Compared with the trend of TN and TOC, that of TP appears relatively stable. Thus, TP could be regarded as the key factor about eutrophication. This work should provide some useful information to better understand the environmental status of south china sea. Keywords: Concentration level, distribution, TN, TP, TOC, South China Sea. Acknowledgments The work was supported by The National Natural Science Foundation of China (No. 40976050), and the National Public Benefit (Ocean) Research Foundation of China (201105013).

Wang, H.; Han, D.

2012-04-01

201

Molecular Analysis of Tn1546 in Enterococcus faecium Isolated from Animals and Humans  

Microsoft Academic Search

The internal areas and the position of integration of the glycopeptide resistance element Tn1546 were char- acterized by using PCR fragment length polymorphism, sequencing, and DNA hybridization techniques with 38 high-level vancomycin-resistant Enterococcus faecium isolates of human and animal origins from Europe and the United States. Only minor variations in the coding regions within Tn1546 were found, suggesting high genetic

LARS BOGØ JENSEN; PETER AHRENS; LONE DONS; RONALD N. JONES; ANETTE M. HAMMERUM; FRANK MØLLER AARESTRUP

202

New mutations in C1GALT1C1 in individuals with Tn positive phenotype.  

PubMed

Tn polyagglutination results from inactivating mutations in C1GALT1C1, an X-borne gene encoding a core 1 beta3-galactosyltransferase-specific molecular chaperone (cosmc) required for the functioning of T-synthase (beta 1,3-galactosyltransferase), a glycosyltransferase essential for the correct biosynthesis of O-glycans. This study found novel inactivating mutations (Glu152Lys, Ser193Pro and Met1Ile) in the coding sequence of C1GALT1C1 in three Tn positive individuals and a complete lack of C1GALT1C1 cDNA expression was observed in an additional Tn positive individual. In addition, expression of ST6GALNAC1, which encodes (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1, 3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 1 and gives rise to sialyl-Tn antigen, was present at comparable levels in normal and Tn-positive human erythroblasts. Expression studies of wild-type and Tn positive C1GALT1C1 cDNA in the Jurkat cell line confirmed that the amino acid substitutions observed in Tn are inactivating. Analysis of the transcriptome of cultured normal and Tn positive erythroblasts revealed numerous differences in gene expression. Reduced transcript levels for fatty acid binding protein 5 (FABP5) and plexin D1 (PLXND1), and increased levels for aquaporin 3 (AQP3) were confirmed by quantitative real-time polymerase chain reaction. These data show that alteration of O-glycan structures resulting from T-synthase deficiency is accompanied by altered expression of a wide variety of genes in erythroid cells. PMID:18537974

Crew, Vanja Karamatic; Singleton, Belinda K; Green, Carole; Parsons, Stephen F; Daniels, Geoff; Anstee, David J

2008-08-01

203

Observation of two length scales above TN in a holmium thin film  

NASA Astrophysics Data System (ADS)

Two-axis neutron-scattering measurements on a 1 ?m thick holmium film reveal the presence of two magnetic correlation lengths above TN that are qualitatively similar to those recently observed in X-ray and neutron scattering studies of bulk holmium and terbium. The scattering profile is well described by a broad Lorentzian plus a narrow Lorentzian-squared line-shape with widths that, for temperatures sufficiently close to TN, differ by a factor often or more.

Gehring, P. M.; Vigliante, A.; McMorrow, D. F.; Gibbs, Doon; Majkrzak, C. F.; Helgesen, G.; Cowley, R. A./; Ward, R. C. C.; Wells, M. R.

1996-02-01

204

Soliton-magnon bound states in TMMC above and below TN  

NASA Astrophysics Data System (ADS)

The problem of soliton-magnon bound states occurring in the quasi-one-dimensional antiferromagnet TMMC [(CH 3) 4NMnCL 3] is studied. Above TN we found a bound state which is due to out-of-plane spin components when going beyond the sine-Gordon limit. Below TN we found a bound state which originates from the pairing of ?-kinks described by a double-sine-Gordon equation.

Ho?yst, J. A.; Benner, H.

1995-02-01

205

Tn917 transposon mutagenesis and marker rescue of interrupted genes of Streptococcus mutans  

Microsoft Academic Search

In order to study virulence factors of the opportunistic oral pathogen Streptococcus mutans we have used Tn917 mutagenesis to generate mutants defective in specific phenotypic properties believed to be associated with virulence. This work describes the procedures required to use the temperature-sensitive transposon Tn917 delivery vector pTV1-OK, along with two techniques to recover inactivated genes. This vector has proven useful

Dennis G. Cvitkovitch; Juan A. Gutierrez; Paula J. Crowley; Laura Wojciechowski; Jeffrey D. Hillman; Arnold S. Bleiweis

1998-01-01

206

Rapid physical mapping by transposon Tn5 mutagenesis to localize the cloned yeast ILV2 gene.  

PubMed

A rapid method for Tn5 mutagenesis of cloned genes on multicopy plasmids was used to map a yeast ILV2 mutant allele encoding a sulfometuron methyl-resistant acetolactate synthase. Twenty-one of 40 independent Tn5 insertions were within the 5.6-kilobase-pair cloned segment. Of these, seven adjacent transposition events inactivated the sulfometuron methyl resistance determinant, localizing the ILV2 gene to a minimum 1.4-kilobase-pair region. PMID:3006586

Van Dyk, T K; Falco, S C; LaRossa, R A

1986-01-01

207

[Genetic mapping of transposon Tn5-induced mutation blocking threonine dehydrogenase in Escherichia coli K-12].  

PubMed

The paper deals with a mutant of Escherichia coli K-12 obtained by transposon Tn5 mutagenesis. Insertion of this transposon inactivated the gene for L-threonine dehydrogenase catalysing the first step of L-threonine degradation. The insertion of Tn5 was mapped by using conjugation as well as transduction by T4GT7 and P1. It is located at 81 min of the E. coli genetic map between mtl and pyrE genes. PMID:2831110

Shakalis, I O; Gustiatiner, M M; Zhdanova, N I

1987-12-01

208

Tn 5044 -Conferred Mercury Resistance Depends on Temperature: the Complexity of the Character of Thermosensitivity  

Microsoft Academic Search

Escherichia coli K12 containing the transposon Tn5044 mer operon (merR, T, P, C, and A genes) is resistant to mercuric chloride at 30°C but sensitive to this compound at 37–41.5°C. We have studied the mechanism underlying the temperature-sensitive nature of this mercury resistance phenotype, and found that the expression of the Tn5044 merA gene coding for mercuric reductase (MerA) is

Gennady Kholodii; Elena Bogdanova

2002-01-01

209

Identification of Tn 4430 , a transposon of Bacillus thuringiensis functional in Escherichia coli  

Microsoft Academic Search

The mobile genetic element Tn4430, originating from the gram-positive bacterium, Bacillus thuringiensis, and previously described as the Th-sequence, is the first transposon isolated from the genus Bacillus. In the present work a gene (APH-III) conferring resistance to kanamycin was inserted into this 4.2 kb transposon. Transposition experiments showed that Tn4430OAPH-III could transpose in the gram-negative host Escherichia coli when its

Didier Lereclus; Jacques Mahillon; Ghislaine Menou; Marguerite-M. Lecadet

1986-01-01

210

Site-Specific Nicking in Vitroat oriT by the DNA Relaxase of Tn 5252  

Microsoft Academic Search

Tn5252is a promiscuous streptococcal element capable of madiating horizontal spread of multiple antibiotic resistance. To begin understanding the functional role of a transfer-related region in Tn5252,its nucleotide sequence was determined. Sequence of this 3.3-kb DNA segment revealed the presence of six open reading frames. The predicted amino acid sequence of one of the open reading frames, ORF9, showed similarity to

Pravina Srinivas; Ali O. Kiliç; Moses N. Vijayakumar

1997-01-01

211

The electrokinetic behavior of red blood cells from a patient with Tn syndrome by Doppler electrophoretic light scattering analysis.  

PubMed

The Tn syndrome is an acquired form of persistent mixed-field polyagglutination displaying two distinct populations of Tn positive (Tn) and Tn negative (tn) red blood cells (RBCs). We investigated the electrophoretic behavior of RBCs showing polyagglutination from a patient with Tn syndrome by the doppler electrophoretic light scattering (D.E.L.S.) analysis. The mean of zeta potential of normal RBCs from ten individuals was -13.07 +/- 0.61 mV (mean +/- S.D.). The content of membrane-associated sialic acid equated with the zeta potential of RBCs. Among the proteases ficin was most effective on the zeta potential of RBCs. The zeta potential of the patient Tn RBCs and tn RBCs were -4.73 mV and -13.32 mV, respectively. Tn RBCs reduced 64.5% of zeta potential compared with tn RBCs and formed 48.8%. These results may provide some useful information for classification of Tn syndrome. PMID:7732519

Omi, T; Kajii, E; Ikemoto, S

1994-12-01

212

A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2  

PubMed Central

We report the characterization of a DNA primase/polymerase protein (PolpTN2) encoded by the pTN2 plasmid from Thermococcus nautilus. Sequence analysis revealed that this protein corresponds to a fusion between an N-terminal domain homologous to the small catalytic subunit PriS of heterodimeric archaeal and eukaryotic primases (AEP) and a C-terminal domain related to their large regulatory subunit PriL. This unique domain configuration is not found in other virus- and plasmid-encoded primases in which PriS-like domains are typically fused to different types of helicases. PolpTN2 exhibited primase, polymerase and nucleotidyl transferase activities and specifically incorporates dNTPs, to the exclusion of rNTPs. PolpTN2 could efficiently prime DNA synthesis by the T. nautilus PolB DNA polymerase, suggesting that it is used in vivo as a primase for pTN2 plasmid replication. The N-terminal PriS-like domain of PolpTN2 exhibited all activities of the full-length enzyme but was much less efficient in priming cellular DNA polymerases. Surprisingly, the N-terminal domain possesses reverse transcriptase activity. We speculate that this activity could reflect an ancestral function of AEP proteins in the transition from the RNA to the DNA world.

Gill, Sukhvinder; Krupovic, Mart; Desnoues, Nicole; Beguin, Pierre; Sezonov, Guennadi; Forterre, Patrick

2014-01-01

213

Transposon Tn5 as an identifiable marker in rhizobia: Survival and genetic stability of Tn5 mutant bean rhizobia under temperature stressed conditions in desert soils.  

PubMed

Five transposon Tn5 insertion mutants of a beanRhizobium strain (Rhizobium leguminosarum b. v.phaseoli) were used in an ecological study to evaluate the extent to which transposon Tn5 was stable to serve as an identifiable marker in rhizobia under a high temperature stress condition in two Sonoran Desert soils. All the mutants possessed single chromosomal insertions of the transposon. In both soils, under the temperature stress conditions that were employed (40°C), both wild type and mutant populations possessing functional transposable elements declined rapidly. After 12 days, mutant cells, when screened using the Tn5 coded antibiotic resistance markers, were significantly less in number than when they were screened using only their intrinsic antibiotic resistance markers. There were no significant differences in numbers between the mutant cell population and the wild type when the mutant cells were screened using only the intrinsic antibiotic resistance markers. DNA-DNA hybridizations using a probe indicated neither deletion nor transposition of the transposable element. The results indicate that transposon DNA sequences are present within cells under high temperature stress conditions, but kanamycin/neomycin resistance is not expressed by some of these cells, suggesting that Tn5 undergoes a possible functional inactivation under these conditions. The possible implications of these findings are discussed. PMID:24194199

Pillai, S D; Pepper, I L

1991-12-01

214

The Bacteroides mobilizable transposon Tn4555 integrates by a site-specific recombination mechanism similar to that of the gram-positive bacterial element Tn916.  

PubMed Central

The Bacteroides mobilizable transposon Tn4555 is a 12.2-kb molecule that encodes resistance to cefoxitin. Conjugal transposition is hypothesized to occur via a circular intermediate and is stimulated by coresident tetracycline resistance elements and low levels of tetracycline. In this work, the ends of the transposon were identified and found to consist of 12-bp imperfect inverted repeats, with an extra base at one end. In the circular form, the ends were separated by a 6-bp "coupling sequence" which was associated with either the left or the right transposon terminus when the transposon was inserted into the chromosome. Tn4555 does not duplicate its target site upon insertion. Using a conjugation-based transposition assay, we showed that the coupling sequence originated from 6 bases of genomic DNA flanking either side of the transposon prior to excision. Tn4555 preferentially transposed into a 589-bp genomic locus containing a 207-bp direct repeat. Integration occurred before or after the repeated sequence, with one integration site between the two repeats. These observations are consistent with a transposition model based on site-specific recombination. In the bacteriophage lambda model for site-specific recombination, the bacteriophage recombines with the Escherichia coli chromosome via a 7-bp "crossover" region. We propose that the coupling sequence of Tn4555 is analogous in function to the crossover region of lambda but that unlike the situation in lambda, recombination occurs between regions of nonhomologous DNA. This ability to recombine into divergent target sites is also a feature of the gram-positive bacterial transposon Tn916.

Tribble, G D; Parker, A C; Smith, C J

1997-01-01

215

The SF1 helicase encoded by the archaeal plasmid pTN2 of Thermococcus nautili.  

PubMed

We expressed, purified, and characterized the helicase encoded by ORF1 of the Thermococcus nautili pTN2 plasmid (Soler et al. Nucl Acids Res 38, 5088-5104, 2010). The enzyme, which belongs to the SF1 family of helicases, possesses NTPase activity, with a strong preference for ATP and GTP as compared to CTP and TTP; dATP was also a substrate. Triphosphatase activity was strongly stimulated by single-stranded DNA and, to a lesser extent, by double-stranded DNA. Unwinding of duplexes comprising a fluorescent oligonucleotide was monitored by fluorescence polarization spectroscopy and by polyacrylamide gel electrophoresis. As observed for enzymes of the same family, pTN2 helicase displays a strong preference for duplexes comprising a 3' single-stranded extension and proceeds from the 3' to the 5' end of the loading strand. Under the conditions of the in vitro assay, pTN2 helicase did not appear to be recycled, but stayed bound to single-stranded DNA, which explains why high concentrations of enzyme are required to unwind long stretches of duplex DNA. The helicase enhances the synthesis of double-stranded DNA by pTN2 primase and by T. nautili PolB polymerase primed by pTN2 primase but it did not enhance synthesis by Taq DNA polymerase. PMID:24889120

Béguin, Pierre; Baron, Bruno; Gill, Sukhvinder; Charpin, Nicole; Forterre, Patrick

2014-07-01

216

Transposon mutagenesis of Xylella fastidiosa by electroporation of Tn5 synaptic complexes.  

PubMed

Pierce's disease, a lethal disease of grapevine, is caused by Xylella fastidiosa, a gram-negative, xylem-limited bacterium that is transmitted from plant to plant by xylem-feeding insects. Strains of X. fastidiosa also have been associated with diseases that cause tremendous losses in many other economically important plants, including citrus. Although the complete genome sequence of X. fastidiosa has recently been determined, the inability to transform or produce transposon mutants of X. fastidiosa has been a major impediment to understanding pathogen-, plant-, and insect-vector interactions. We evaluated the ability of four different suicide vectors carrying either Tn5 or Tn10 transposons as well as a preformed Tn5 transposase-transposon synaptic complex (transposome) to transpose X. fastidiosa. The four suicide vectors failed to produce any detectable transposition events. Electroporation of transposomes, however, yielded 6 x 10(3) and 4 x 10(3) Tn5 mutants per microg of DNA in two different grapevine strains of X. fastidiosa. Molecular analysis showed that the transposition insertions were single, independent, stable events. Sequence analysis of the Tn5 insertion sites indicated that the transpositions occur randomly in the X. fastidiosa genome. Transposome-mediated mutagenesis should facilitate the identification of X. fastidiosa genes that mediate plant pathogenicity and insect transmission. PMID:11386365

Guilhabert, M R; Hoffman, L M; Mills, D A; Kirkpatrick, B C

2001-06-01

217

Insertional mutagenesis of Listeria monocytogenes with a novel Tn917 derivative that allows direct cloning of DNA flanking transposon insertions.  

PubMed Central

To carry out efficient insertional mutagenesis in Listeria monocytogenes and to facilitate the characterization of disrupted genes, two novel derivatives of Tn917 were constructed, Tn917-LTV1 and Tn917-LTV3. The derivatives (i) transpose at a significantly elevated frequency, (ii) generate transcriptional lacZ fusions when inserted into a chromosomal gene in the appropriate orientation, and (iii) allow the rapid cloning in Escherichia coli of chromosomal DNA flanking transposon insertions. The rapid cloning of DNA flanking insertions is possible because the transposon derivatives carry ColE1 replication functions, a cluster of polylinker cloning sites, and antibiotic resistance genes selectable in E. coli (bla in the case of Tn917-LTV1; neo and ble in the case of Tn917-LTV3). The enhanced transposition frequency of Tn917-LTV1 and Tn917-LTV3 (about 100-fold in Bacillus subtilis) is believed to be due to the fortuitous placement of vector-derived promoters upstream from the Tn917 transposase gene. In L. monocytogenes, Tn917-LTV3 transposed at a frequency of 8 x 10(-4) when introduced on a pE194Ts-derived vector and generated at least eight different auxotrophic mutations. Two nonhemolytic insertion mutants of L. monocytogenes were isolated, and DNA flanking the transposon insertions was cloned directly into E. coli, making use of the ColE1 rep functions and neo gene carried by Tn917-LTV3. Both insertions were shown to be within hlyA, the L. monocytogenes hemolysin structural gene. Although Tn917-LTV1 and Tn917-LTV3 were constructed specifically for genetic analysis of L. monocytogenes, their enhanced transposition frequency and convenience for cloning of DNA adjacent to sites of insertions make them the transposon derivatives of choice for insertional mutagenesis in any gram-positive bacteria that support replication of pE194Ts.

Camilli, A; Portnoy, A; Youngman, P

1990-01-01

218

High frequency mobilization of gram-negative bacterial replicons by the in vitro constructed Tn 5 Mob transposon  

Microsoft Academic Search

A DNA fragment of the broad host range plasmid RP4 carrying the cis-acting DNA recognition site for conjugative DNA transfer between bacterial cells (Mobsite) was cloned into the kanamycin-neomycin resistance transposon Tn5. Using conventrional transposon mutagenesis techniques the new transposon, called Tn5-Mob, can easily be inserted into the host DNA of gram-negative bacteria. A host replicon carrying Tn5-Mob is then

Reinhard Simon

1984-01-01

219

Mapping of chromosomal loci associated with lipopolysaccharide synthesis and serotype specificity in Vibrio cholerae 01 by transposon mutagenesis using Tn5 and Tn2680.  

PubMed

Vibrio cholerae strains of the 01 serotype have been classified into three subclasses, Ogawa, Inaba and Hikojima, which are associated with the O-antigen of the lipopolysaccharide (LPS). The DNA encoding the biosynthesis of the O-antigen, the rfb locus, has been cloned and analysed (Manning et al. 1986; Ward et al. 1987). Transposon mutagenesis of the Inaba and Ogawa strains of V. cholerae, using Tn5 or Tn2680 allowed the isolation of a series of independent mutants in each of these serotypes. Some of the insertions were mapped to the rfb region by Southern hybridization using the cloned rfb DNA as a probe, confirming this location to be responsible for both O-antigen production and serotype specificity. The other insertions allowed a second region to be identified which is involved in V. cholerae LPS biosynthesis. PMID:2550772

Ward, H M; Manning, P A

1989-08-01

220

Hancock County Awards Gala  

NASA Technical Reports Server (NTRS)

Gene Goldman (left), deputy director of NASA's John C. Stennis Space Center, accepts an Award of Excellence from Jack Zink, executive director of the Hancock County Port and Harbor Commission, during the 2008 Annual Hancock County Awards Gala. The Award of Excellence was presented to recognize Stennis Space Center's contribution to NASA's 50 years of excellence in space exploration.

2008-01-01

221

County Government Employment: 1990.  

National Technical Information Service (NTIS)

The report provides national statistics on county government employment and payrolls for the month of October 1990. Statistics are based on a mail canvass survey that includes a sample of 2,026 county governments. The survey measures the number of governm...

1991-01-01

222

Transposon Tn5 mutagenesis of pseudomonas fluorescens to isolate mutants deficient in antibacterial activity.  

PubMed

Pseudomonas fluorescens was subjected to insertion mutagenesis studies using the transposon Tn5-GM to generate mutants deficient in antibacterial activity minus mutants. The transposon located on the temperature-sensitive plasmid pCHR84 was conjugally transferred into the non-pathogenic pseudomonad using the triparental mating procedure. Random integration of Tn5-GM into the chromosome of P. fluorescens was achieved by heat treatment of the transformed cells at 42 degrees C. Approximately 2% of transconjugants revealed an auxotrophic phenotype indicating efficient integration of the employed transposon into the chromosome of P. fluorescens. One transposon insertion mutant was obtained showing an antibacterial activity minus phenotype. This mutant (MM-7) was found to be defective in the production of an unidentified antibacterial compound against B. subtilis. These results introduce Tn5 transposon mutagenesis as a new useful tool for the molecular analysis of P. fluorescens. PMID:8138133

Rajendran, N; Jahn, D; Jayaraman, K; Marahiel, M A

1994-01-15

223

Novel Tn4371-ICE like element in Ralstonia pickettii and Genome mining for comparative elements  

PubMed Central

Background Integrative Conjugative Elements (ICEs) are important factors in the plasticity of microbial genomes. An element related to the ICE Tn4371 was discovered during a bioinformatic search of the Ralstonia pickettii 12J genome. This element was analysed and further searches carried out for additional elements. A PCR method was designed to detect and characterise new elements of this type based on this scaffold and a culture collection of fifty-eight Ralstonia pickettii and Ralstonia insidiosa strains were analysed for the presence of the element. Results Comparative sequence analysis of bacterial genomes has revealed the presence of a number of uncharacterised Tn4371-like ICEs in the genomes of several ? and ?- Proteobacteria. These elements vary in size, GC content, putative function and have a mosaic-like structure of plasmid- and phage-like sequences which is typical of Tn4371-like ICEs. These elements were found after a through search of the GenBank database. The elements, which are found in Ralstonia, Delftia, Acidovorax, Bordetella, Comamonas, Acidovorax, Congregibacter, Shewanella, Pseudomonas Stenotrophomonas, Thioalkalivibrio sp. HL-EbGR7, Polaromonas, Burkholderia and Diaphorobacter sp. share a common scaffold. A PCR method was designed (based on the Tn4371- like element detected in the Ralstonia pickettii 12J genome) to detect and characterise new elements of this type. Conclusion All elements found in this study possess a common scaffold of core genes but contain different accessory genes. A new uniform nomenclature is suggested for ICEs of the Tn4371 family. Two novel Tn4371-like ICE were discovered and characterised, using the novel PCR method described in two different isolates of Ralstonia pickettii from laboratory purified water.

2009-01-01

224

Persistent Tn polyagglutination syndrome during febrile neutropenia: a case report and review of the literature  

PubMed Central

Introduction Tn polyagglutination syndrome is a rare disorder that has been reported on only a few occasions in the literature, and, to the best of our knowledge, never before in the context of febrile neutropenia. Case presentation We report the case of a 26-year-old Caucasian woman who presented to our emergency department complaining of a persistent fever over the previous three days. She had a history of long-standing refractory pancytopenia with multi-lineage dysplasia and severe neutropenia, but she had rarely experienced infection. The results of a physical examination and multiple laboratory tests were unremarkable. While investigating the possible causes of the refractory, long-standing pancytopenia, the possibility of a polyagglutinable state was suggested. Blood samples were sent to the laboratory for an analysis of mixed-field seed lectin agglutination assay. A serum lectin panel confirmed the final diagnosis of Tn-activation. Conclusions We should include Tn-activation in our differential whenever we encounter cases of refractory long-standing idiopathic cytopenias and inconclusive bone marrow results displaying multi-lineage dysplasia. Novel genetic techniques have recently revealed the interesting pathophysiology of this phenomenon. The recognition and inclusion of Tn polyagglutination syndrome in our differential diagnoses has important clinical implications, given its main associated features, such as severe thrombocytopenia and neutropenia, which are usually linked to a benign clinical course and prognosis. Increased awareness of the polyagglutinable disorders will potentially decrease the need for invasive and costly medical interventions and also raises the need for monitoring of this specific sub-set of patients. In addition, the study of the expression and implications of Tn, and other similar antigens, offers a fascinating perspective for the study of its role in the diagnosis, prognosis and immunotherapy of solid tumors and hematological malignancies. The infrequency with which Tn polyagglutination syndrome is encountered, its clinical features and its pathophysiology make it a formidable diagnostic challenge.

2011-01-01

225

High-pressure effects on a manganese hexacyanomanganate ferrimagnet with TN=29 K  

NASA Astrophysics Data System (ADS)

The effects of high pressure, up to 1 GPa, on a Mn-based Prussian blue ferrimagnet, Mn 3II[Mn III(CN) 6] 2·12H 2O·1.7(CH 3OH) with TN=29 K, were studied. The transition temperature gradually increases with an increase in pressure and becomes ˜40 K at 1 GPa. The pressure dependence of TN can be semi-quantitatively interpreted in terms of the shrinkage of the Mn-CN-Mn superexchange pathway.

Awaga, Kunio; Sekine, Taketoshi; Okawa, Masaki; Fujita, Wataru; Holmes, Stephen M.; Girolami, Gregory S.

1998-09-01

226

Tn4351 transposes in Bacteroides spp. and mediates the integration of plasmid R751 into the Bacteroides chromosome  

SciTech Connect

The gene for resistance to erythromycin and clindamycin, which is carried on the conjugative Bacteroides plasmid, pBF4, has been shown previously to be part of an element (Tn4351) that transposes in Escherichia coli. The authors have now introduced Tn4351 into Bacteroides uniformis 0061 on the following two suicide vectors: (i) the broad-host-range IncP plasmid R751 (R751::Tn4351) and (ii) pSS-2, a chimeric plasmid which contains 33 kilobases of pBF4 (including Tn4351) cloned into the IncQ plasmid RSF1010 and which is mobilized by R751. When E. coli HB101, carrying either R751::Tn4351 or R751 and pSS-2, was mated with B. uniformis under aerobic conditions, Em/sup r/ transconjugants were detected at a frequency of 10 /sup -6/ to 10/sup -5/ (R751::Tn4351) or 10/sup -8/ to 10/sup -6/ (R751 and pSS-2). In matings involving pSS-2, all Em/sup r/ transconjugants contained simple insertions of Tn4351 in the chromosome, whereas in matings involving R751::Tn4351, about half of the Em/sup r/ transconjugants had R751 cointegrated with Tn4351 in the chromosome. Of the Em/sup r/ transconjugants, 13% were auxotrophs. Bacteroides spp. which had R751 cointegrated with Tn4351 in the chromosome did not transfer R751 or Tn4351 to E. coli HB101 or to isogenic B. uniformis, nor did the integrated R751 mobilize pE5-2, an E. coli-Bacteroides shuttle vector that contains a transfer origin that is recognized by R751.

Shoemaker, N.B.; Getty, C.; Gardner, J.F.; Salyers, A.A.

1986-03-01

227

New Sr, Nd, and Pb isotopic data from plutons in the northern Great Basin: Implications for crustal structure and granite petrogenesis in the hinterland of the Sevier thrust belt  

SciTech Connect

The influence of tectonic setting and age on the variation of isotopic signatures of granitic plutons in the northern Great Basin has, in general, not been apparent from previous investigations. None of these studies revealed the remarkable correlation shown by this expanded Sr, Nd, and Pb isotopic data base. Jurassic-Early Cretaceous plutons in the northern Great Basin have a limited range of Sr and Nd isotopic values that cluster near bulk earth. Construction of Sr 0.706 and {epsilon}{sub Nd} = {minus}7 isotopic boundaries is virtually impossible for plutons of this age range. In contrast, Upper Cretaceous peraluminous granites east of the {epsilon}{sub Nd} = {minus}7 line have very negative {epsilon}{sub Nd} values and high initial Sr ratios, and they appear to represent essentially pure crustal melts. The data favor a model that equates generation of these plutons via crustal thickening associated with the Sevier thrust belt. Cenozoic plutons appear to be mixtures of mantle and crustal reservoirs, and their isotopic systematics, along with those of the Late Cretaceous age plutonic suite, define a previously unrecognized, approximately east-west-trending crustal boundary between predominantly Archean crust to the north and predominantly Proterozoic crust to the south. The isotopic data from the Jurassic-Early Cretaceous plutonic suite do not reflect the presence of this boundary, suggesting that the isotopic systematics of this plutonic suite may not have been controlled by the same variations in crustal and/or mantle lithospheric structure at depth.

Wright, J.E. (Rice Univ., Houston, TX (USA)); Wooden, J.L. (Geological Survey, Menlo Park, CA (USA))

1991-05-01

228

76 FR 12280 - Revisions to the California State Implementation Plan, for Imperial County, Kern County, and...  

Federal Register 2010, 2011, 2012, 2013

...County, and Ventura County; Air Pollution Control Districts AGENCY: Environmental...revisions to the Imperial County Air Pollution Control District (ICAPCD), Kern County Air Pollution Control District...

2011-03-07

229

76 FR 12306 - Revisions to the California State Implementation Plan, Imperial County, Kern County, and Ventura...  

Federal Register 2010, 2011, 2012, 2013

...County, and Ventura County; Air Pollution Control Districts AGENCY: Environmental...revisions to the Imperial County Air Pollution Control District (ICAPCD), Kern County Air Pollution Control District...

2011-03-07

230

Novel Tn4371ICE like element in Ralstonia pickettii and Genome mining for comparative elements  

Microsoft Academic Search

BACKGROUND: Integrative Conjugative Elements (ICEs) are important factors in the plasticity of microbial genomes. An element related to the ICE Tn4371 was discovered during a bioinformatic search of the Ralstonia pickettii 12J genome. This element was analysed and further searches carried out for additional elements. A PCR method was designed to detect and characterise new elements of this type based

Michael P Ryan; J Tony Pembroke; Catherine C Adley

2009-01-01

231

Defining characteristics of Tn5 Transposase non-specific DNA binding  

PubMed Central

While non-specific DNA plays a role in target localization for many recombinases, transcription factors and restriction enzymes, the importance of non-specific DNA interactions for transposases has not been investigated. Here, we discuss non-specific DNA-Tn5 Transposase (Tnp) interactions and suggest how they stabilize the Tnp and modulate Tnp localization of the 19 bp Tnp recognition end sequences (ESes). DNA protection assays indicate that full-length Tnp interacts efficiently with supercoiled DNA that does not contain ESes. These interactions significantly prolong the lifetime of Tnp, in vitro. The balance between non-specific DNA bound and free Tnp is affected by DNA topology, yet, intermolecular transfer of active Tnp occurs with both supercoiled and linear non-specific DNA. Experiments with substrates of varying lengths show that Tn5 Tnp can utilize non-specific DNA to facilitate localization of an intramolecular ES over distances less than 464 bp. Finally, synaptic complex formation is inhibited in the presence of increasing concentrations of supercoiled and linear pUC19. These experiments strongly suggest that Tn5 Tnp has a robust non-specific DNA binding activity, that non-specific DNA modulates ES sequence localization within the global DNA, most likely through a direct transfer mechanism, and that non-specific DNA binding may play a role in the cis bias manifested by Tn5 transposition.

Steiniger, Mindy; Adams, Christian D.; Marko, John F.; Reznikoff, William S.

2006-01-01

232

Preclinical class 1 integron with a complete Tn402-like transposition module.  

PubMed

The presence of integrons was assessed in gut bacteria isolated from wild-caught prawns. A pseudomonad was recovered that contained a Tn402-like class 1 integron with a complete transposition module and two gene cassettes. One cassette was identical to a previously described cassette from a chromosomal class 3 integron in Delftia tsuruhatensis. PMID:21037292

Sajjad, Ammara; Holley, Marita P; Labbate, Maurizio; Stokes, H W; Gillings, Michael R

2011-01-01

233

A "new" lectin for detection of T, Tn, and Th polyagglutination.  

PubMed

A lectin reacting with T, Tn, and Th polyagglutinable red blood cells was found in an extract from the seeds of Hibiscus sabdariffa (Sorrel). This lectin is thus a useful addition to those available for distinguishing various types of red blood cell polyagglutinability. PMID:7101429

Yang, E K; Spence, L R; Harding, R Y; Moore, B P

1982-01-01

234

Member expansion in quantum ( t,n) threshold secret sharing schemes  

NASA Astrophysics Data System (ADS)

A protocol for member expansion in quantum ( t,n) threshold secret sharing schemes was proposed. Without a trusted center and modifying the shares of old participants, the protocol needs that t ( t is the threshold) old participants cooperate to generate the new share. Compared with the previous secret sharing protocols, the proposed protocol has the advantage of joining new participants agilely.

Yang, Yu-Guang; Wang, Yuan; Chai, Hai-Ping; Teng, Yi-Wei; Zhang, Hua

2011-06-01

235

78 FR 35776 - Proposed Establishment of Class E Airspace; Cleveland, TN  

Federal Register 2010, 2011, 2012, 2013

A notice of proposed rulemaking published in the Federal Register of March 6, 2013, to establish Class E airspace at Cleveland Regional Jetport, Cleveland, TN, is being withdrawn. Upon review, the FAA found that, for clarity, combining this proposed rulemaking with another proposal to amend existing airspace is...

2013-06-14

236

Transposon mutagenesis of the anaerobic commensal, Bacteroides fragilis, using the EZ?TN5 transposome  

PubMed Central

Genetic analysis of Bacteroides fragilis (BF) is hindered due to the lack of efficient transposon mutagenesis methods. Here we describe a simple method for transposon mutagenesis using EZ?TN5, a commercially available system that we optimized for use in BF638R. The modified EZ?TN5 transposon contains an E. coli conditional origin of replication, a kanamycin resistance gene for E. coli, an erythromycin resistance gene for BF and 19 basepair transposase recognition sequences on either ends. Electroporation of the transposome (transposon-transposase complex) into BF638R yielded 3.2± 0.35×103 CFU/?g of transposon DNA. Modification of the transposon by the BF638R restriction/modification system increased transposition efficiency 6-fold. Electroporation of the EZ?TN5 transposome results in a single copy insertion of the transposon evenly distributed across the genome of BF638R and can be used to construct a BF638R transposon library. The transposon was also effective in mutating a BF clinical isolate and a strain of the related species, B. thetaiotaomicron. The EZ?TN5 based mutagenesis described here is more efficient than other transposon mutagenesis approaches previously reported for BF.

Veeranagouda, Yaligara; Husain, Fasahath; Wexler, Hannah M.

2012-01-01

237

Investigation of Escherichia coli fumarate reductase subunit function using transposon Tn5.  

PubMed

Seventy two Tn5 transposon insertions were isolated in the frd operon carried on the multicopy plasmid pFRD79. The polar nature of these mutations permitted examination of the expression and localization of the frd polypeptides in novel subunit combinations. The minimal catalytic unit is the FRDA plus B dimer. A transposon within frdB (frdB::Tn5) produces inactive, soluble FRDA polypeptide which has covalently attached 8 alpha(N3-histidyl)flavin adenine dinucleotide cofactor. A transposon mutation within frdC (frdC::Tn5) produces soluble, catalytically active dimer. An insertion in frdD (frdD::Tn5) produces both a soluble trimer composed of FRDABC, and a tetramer of FRDABC and truncated FRDD bound to the inner membrane. Eighty percent of the activity is in the soluble form. Using this mutant, the requirement for FRDD both for optimal activity of the catalytic domain and for proper anchorage in the cytoplasmic membrane was demonstrated. PMID:2821165

Latour, D J; Weiner, J H

1987-03-01

238

Mutagenesis of acetobacter methanolicus MB58 with the transposon Tn5.  

PubMed

Transposon mutagenesis was applied to the isolation of mutants of the facultatively methylotrophic Acetobacter methanolicus MB 58. The transposon Tn5 (pSU2011) was transferred from Escherichia coli SM 10 by means of conjugation to Acetobacter methanolicus MB 58. Four out of 1850 stable Km-resistant transconjugants were identified that were formaldehyde sensitive and failed to grow on methanol. PMID:2166787

Dobrowolski, P; Gründig, M W

1990-01-01

239

Construction and characterization of versatile kanamycin-resistance cassettes derived from the Tn5 transposon.  

PubMed

We have developed plasmids with the Tn5 kanamycin-resistance gene (kan) flanked either symmetrically or asymmetrically by several restriction sites. These can be used to provide a selectable genetic marker or to mobilize restriction sites and sense or nonsense codons into genes. The 1.3-kb kan cassette exhibits polarity effects in both directions. PMID:2559874

Musso, R E; Hodam, T

1989-12-21

240

[Transposon Tn5 and its derivatives used in genetic analysis of bacteria].  

PubMed

The review deals with the derivatives of transposon Tn5 carrying new genes of antibiotic resistance. The derivatives were constructed for mobilization of genetically labeled replicons, for direct selection of mutants having lost the marked plasmids, for obtaining the genes with the strong constitutive or regulated expression, for isolation of conditional mutations, for faster physical mapping of megaplasmids. PMID:8394506

Katsy, E I

1993-01-01

241

Elution of Nitrate at the NABIR Field Research Center, Oak Ridge Reservation, Oak Ridge, TN  

Microsoft Academic Search

As part of a bioremediation project for the in situ bioreduction of uranium at the Department of Energy Natural and Accelerated Bioremediation Research (NABIR) Field Research Center (FRC) in Oak Ridge, TN, aquifer and groundwater conditioning is required before conducting the remediation experiment. One step includes flushing of the aquifer with pH-adjusted fresh water in order to remove extremely high

M. N. Fienen; C. S. Criddle; P. M. Jardine; P. K. Kitanidis; T. L. Mehlhorn; D. B. Watson; W. Wu

2003-01-01

242

76 FR 4147 - Putnam-Cumberland, TN-Improve Power Supply  

Federal Register 2010, 2011, 2012, 2013

...Putnam-Cumberland, TN--Improve Power Supply AGENCY: Tennessee Valley Authority...of proposed electrical power supply improvements in the Putnam...power to meet increasing power demands in the project area. In its...of transmission lines. TVA supplies bulk electric power to...

2011-01-24

243

Lactobacillus plantarum TN8 exhibits protective effects on lipid, hepatic and renal profiles in obese rat.  

PubMed

This study aimed to first investigate the immuno-modulatory effects of six newly isolated lactic acid bacteria (LAB) on the peripheral blood mononuclear cells (PBMC) of Wistar rats. Except for Lactobacillus plantarum TN8, all the other strains were noted to induce high levels of pro-inflammatory cytokine IL-12 and low levels of anti-inflammatory cytokine IL-10. The strains also generated low ratios of IL-10/IL-12 cytokine. Strain TN8 was, on the other hand, noted to induce an increase in anti-inflammatory IL-10 cytokine secretion rates and a decrease in pro-inflammatory IL-12, IFN-? and TNF-? cytokine production. The oral administration of TN8 improved the hepatic and urinary functions of obese rats by inducing decreases (P < 0.05) in alanine amino transferase (ALAT), gamma glutamyl transferase (GGT), plasmatic triglycerides, total cholesterol concentrations, creatinine, urea, and body weight when compared to the control group of animals that underwent an increase in aspartate amino transferase (ASAT) and high density lipoprotein (HDL). Overall, the findings indicate that strain TN8 exhibited a number of attractive properties that might open new promising opportunities for the improvement of various parameters related to animal health performance and the avoidance of antibiotics and drugs as promoting factors. PMID:23891961

Ben Salah, Riadh; Trabelsi, Imen; Hamden, Khaled; Chouayekh, Hichem; Bejar, Samir

2013-10-01

244

IS10/Tn10 transposition efficiently accommodates diverse transposon end configurations.  

PubMed Central

Transposon Tn10 and its component insertion sequence IS10 move by non-replicative transposition. We have studied the array of reaction intermediates and products in a high efficiency in vitro IS10/Tn10 transposition reaction. Synapsis of two transposon ends, followed by cleavage and strand transfer, can occur very efficiently irrespective of the relative locations and orientations of the two ends. The two participating ends can occur in inverted or direct orientation on the same molecule or, most importantly, on two different molecules. This behavior contrasts sharply with that of Mu, in which transposition is strongly biased in favor of inverted repeat synapsis. Mechanistically, the absence of discrimination amongst various end configurations implies that the architecture within the IS10/Tn10 synaptic complex is relatively simple, i.e. lacking any significant intertwining of component DNA strands. Biologically these observations are important because they suggest that the IS10 insertion sequence module has considerable flexibility in the types of DNA rearrangements that it can promote. Most importantly, it now seems highly probable that a single non-replicative IS10 element can promote DNA rearrangements usually attributed to replicative transposition, i.e. adjacent deletions and cointegrates, by utilizing transposon ends on two sister chromosomes. Other events which probably also contribute to the diversity of IS10/Tn10-promoted rearrangements are discussed. Images

Chalmers, R M; Kleckner, N

1996-01-01

245

Tn tau sub E Comparisons of Tokamak Reactors of Different Aspect Ratios.  

National Technical Information Service (NTIS)

The ignition parameter Tn tau (sub E) of the International Tokamak Experimental Reactor (ITER) with an aspect ratio A = 2.6, a major radius R = 5.8 m, a plasma current I(sub p) = 22 MA, and an external toroidal field of B(sub t0) = 5 T at R; a High Field ...

Y. K. M. Peng

1989-01-01

246

Transposon mutagenesis of the anaerobic commensal, Bacteroides fragilis, using the EZ::TN5 transposome.  

PubMed

Genetic analysis of Bacteroides fragilis (BF) is hindered because of the lack of efficient transposon mutagenesis methods. Here, we describe a simple method for transposon mutagenesis using EZ::TN5, a commercially available system that we optimized for use in BF638R. The modified EZ::TN5 transposon contains an Escherichia coli conditional origin of replication, a kanamycin resistance gene for E. coli, an erythromycin resistance gene for BF , and 19 basepair transposase recognition sequences on either ends. Electroporation of the transposome (transposon-transposase complex) into BF638R yielded 3.2 ± 0.35 × 10(3) CFU ?g(-1) of transposon DNA. Modification of the transposon by the BF638R restriction/modification system increased transposition efficiency sixfold. Electroporation of the EZ::TN5 transposome results in a single-copy insertion of the transposon evenly distributed across the genome of BF638R and can be used to construct a BF638R transposon library. The transposon was also effective in mutating a BF clinical isolate and a strain of the related species, Bacteroides thetaiotaomicron. The EZ::TN5-based mutagenesis described here is more efficient than other transposon mutagenesis approaches previously reported for BF. PMID:22639975

Veeranagouda, Yaligara; Husain, Fasahath; Wexler, Hannah M

2012-08-01

247

Characterization of glucansucrase and dextran from Weissella sp. TN610 with potential as safe food additives.  

PubMed

Pear-derived Weissella sp. TN610 produced extracellular glycosyltransferase activity responsible for the synthesis of soluble exopolysaccharide from sucrose. Acid and dextranase-catalyzed hydrolysis revealed that the synthesized polymer was a glucan. According to (1)H and (13)C NMR analysis, the glucan produced by TN610 was a linear dextran made of 96% ?-(1?6) and 4% ?-(1?3) linkages. Zymogram analysis confirmed the presence of a unique glucansucrase of approximately 180 kDa in the cell-free supernatant from TN610. The crude enzyme, optimally active at 37°C and pH 5, has promising potential for application as a food additive since it catalyzes dextran synthesis in sucrose-supplemented milk, allowing its solidification. A 4257-bp product corresponding to the mature glucansucrase gene was amplified by PCR from TN610. It encoded a polypeptide of 1418 residues having a calculated molecular mass of 156.089 kDa and exhibiting 96% and 95% identity with glucansucrases from Lactobacillus fermentum Kg3 and Weissella cibaria CMU, respectively. PMID:23046923

Bejar, Wacim; Gabriel, Valérie; Amari, Myriam; Morel, Sandrine; Mezghani, Monia; Maguin, Emmanuelle; Fontagné-Faucher, Catherine; Bejar, Samir; Chouayekh, Hichem

2013-01-01

248

Complete nucleotide sequence of macrolide-lincosamide-streptogramin B-resistance transposon Tn917 in Streptococcus faecalis.  

PubMed Central

Streptococcus faecalis transposon Tn917 was cloned in Escherichia coli on plasmid vector pBR325. The erythromycin resistance determinant of Tn917 was not expressed in the E. coli background. The nucleotide sequence of Tn917 was determined and found to be 5,257 base pairs in length. Six open reading frames (ORFs) were identified and designated 1 through 6 (5' to 3'); all were on the same DNA strand. A region exhibiting strong homology with known promoters was identified upstream from ORF1. ORFs 1 to 3 were virtually identical to the previously sequenced erythromycin resistance determinant on Streptococcus sanguis plasmid pAM77. At the 3' point, where the homology between Tn917 and pAM77 ends, was a 20-base-pair region about 80% homologous with a component of the res site of Tn3. The amino acid sequence of ORF4 showed homology with other site-specific recombination enzymes, including approximately 30% homology with the resolvase of Tn3. Contained within Tn917 was a directly oriented 73-base-pair duplication of the left terminus. The Tn917 sequence revealed that antibiotic-enhanced transposition might be due to extension of transcription from the resistance-related genes (in ORFs 1 to 3) into transposition genes (in ORFs 4 to 6). Transcription analyses resulted in data consistent with this interpretation. Images

Shaw, J H; Clewell, D B

1985-01-01

249

Chromosomal integration of plasmid DNA by homologous recombination in Enterococcus faecalis and Lactococcus lactis subsp. lactis hosts harboring Tn919.  

PubMed Central

Integration of pCI192, a pBR322-derived vector plasmid containing homology to the chromosomally located conjugative transposon Tn919 was observed in two strains that harbor Tn919, namely, Enterococcus faecalis GF590 and Lactococcus lactis subsp. lactis CH919. Hybridization analysis indicated that single-copy integration of the plasmid had occurred at low frequency. The Tn919::plasmid structure was conjugated from an E. faecalis donor to a L. lactis recipient, although at lower frequencies than was Tn919. Segregation of the tetracycline and chloramphenicol resistance markers during conjugation was observed. The integration strategy described allows for DNA manipulations to be performed in an easily manipulated model host strain with the subsequent transfer of integrated structures by conjugation to any strain capable of receiving Tn919. The results indicate that homologous recombination events may be used to introduce plasmid-encoded genes to the lactococcal chromosome. Images

Casey, J; Daly, C; Fitzgerald, G F

1991-01-01

250

The novel conjugative transposon tn1207.3 carries the macrolide efflux gene mef(A) in Streptococcus pyogenes.  

PubMed

The macrolide efflux gene mef(A) of the Streptococcus pyogenes clinical strain 2812A was found to be carried by a 52-kb chromosomal genetic element that could be transferred by conjugation to the chromosome of other streptococcal species. The characteristics of this genetic element are typical of conjugative transposons and was named Tn1207.3. The size of Tn1207.3 was established by pulsed-field gel electrophoresis (PFGE), and DNA sequencing analysis showed that the 7,244 bp at the left end of Tn1207.3 were identical to those of the pneumococcal Tn1207.1 element. Tn1207.3-like genetic elements were found to be inserted at a single specific chromosomal site in 12 different clinical isolates S. pyogenes exhibiting the M phenotype of resistance to macrolides and carrying the mef(A) gene. Tn1207.3 was transferred from S. pyogenes 2812A to Streptococcus pneumoniae, and sequence analysis carried out on six independent transconjugants showed that insertion of Tn1207.3 in the pneumococcal genome always occurred at a single specific site as in Tn1207.1. Using MF2, a representative S. pneumoniae transconjugant, as a donor, Tn1207.3 was transferred again by conjugation to S. pyogenes and Streptococcus gordonii. The previously described nonconjugative element Tn1207.1 of S. pneumoniae appears to be a defective element, part of a longer conjugative transposon that carries mef(A) and is found in clinical isolates of S. pyogenes. PMID:12959402

Santagati, Maria; Iannelli, Francesco; Cascone, Carmela; Campanile, Floriana; Oggioni, Marco R; Stefani, Stefania; Pozzi, Gianni

2003-01-01

251

A Tn5051-like mer-containing transposon identified in a heavy metal tolerant strain Achromobacter sp. AO22  

PubMed Central

Background Achromobacter sp. AO22 (formerly Alcaligenes sp. AO22), a bacterial strain isolated from a lead-contaminated industrial site in Australia, was previously found to be resistant to moderate to high levels of mercury, copper and other heavy metals. However, the nature and location of the genetic basis for mercuric ion resistance in this strain, had not been previously identified. Findings Achromobacter sp. AO22 contains a functional mer operon with all four essential genes (merRTPA) and shows >99% DNA sequence identity to that of Tn501. The mer operon was present on a transposon, designated TnAO22, captured by introducing a broad-host-range IncP plasmid into Achromobacter sp. AO22 and subsequently transferring it to E. coli recipients. The transposition frequency of TnAO22 was 10-2 to 10-3 per target plasmid transferred. Analysis of TnAO22 sequence revealed it belonged to the Tn21 subgroup of the Tn3 superfamily of transposons, with the transposition module having >99% identity with Tn5051 of a Pseudomonas putida strain isolated from a water sample in New York. Conclusion TnAO22 is thus a new variant of Tn5051 of the Tn3 superfamily and the transposon and its associated mercury resistance system are among the few such systems reported in a soil bacterium. Achromobacter sp. AO22 can thus be exploited for applications such as in situ mercury bioremediation of contaminated sites, or the mobile unit and mer operon could be mobilized to other bacteria for similar purposes.

Ng, Shee Ping; Davis, Belinda; Palombo, Enzo A; Bhave, Mrinal

2009-01-01

252

Characterization of the conjugative transposon Tn6000 from Enterococcus casseliflavus 664.1H1 (formerly Enterococcus faecium 664.1H1).  

PubMed

Tn6000 (formerly EfcTn1) from Enterococcus casseliflavus strain 664.1H1 (previously Enterococcus faecium 664.1H1) is a tetracycline resistance-encoding conjugative transposon of the Tn916-like family of mobile genetic elements. Sequence analysis of Tn6000 shows that it has a novel modular structure, comprising fragments of diverse proven and putative mobile elements including plasmids, conjugative transposons and virulence and pathogenicity islands. PMID:20528943

Brouwer, Michael S M; Mullany, Peter; Roberts, Adam P

2010-08-01

253

An improved procedure and new vectors for transposon Tn5 mutagenesis of the phototrophic bacterium Rhodospirillum rubrum.  

PubMed

A detailed examination of vectors and procedures used for Tn5 mutagenesis of the phototrophic purple non-sulfur bacterium Rhodospirillum rubrum has been performed. The mobilizable Tn5 suicide vectors currently available show a frequency of Tn5 mutagenesis for R. rubrum of approx. 10(-7)-10(-8), approx. 100-1000-fold lower than observed for the related bacteria Rhodobacter capsulatus and Rhodobacter sphaeroides. Using the blue-to-red reversion of a blue-green mutant, R. rubrum ST6, containing a single Tn5 lesion in one of the early genes for carotenoid biosynthesis, we have shown that the frequency of precise excision of a chromosomally inserted Tn5 element, to restore the wild-type phenotype in the absence of selection, is 10(-6). We have constructed three new suicide vectors for Tn5 mutagenesis, where the transposase encoded by the IS50R element was placed in the same (pSUPEG11, pSUPEG21) or in the opposing (pSUPEG22) orientation from the weak promoter of the RK2-derived tetR gene. With the vector pSUPEG11, the frequency of Tn5 mutagenesis was increased to 10(-5), approx. 100-fold higher than observed previously. PMID:7959072

Ghosh, R; Elder, D J; Saegesser, R; Kelly, D J; Bachofen, R

1994-12-01

254

Transposon Tn5-259 mutagenesis of Pseudomonas cepacia to isolate mutants deficient in antifungal activity.  

PubMed

Transposon Tn5-259 was inserted into the chromosome of Pseudomonas cepacia by mating with an Escherichia coli strain harboring a self-mobilizable, temperature-sensitive plasmid, pME12. Data from Southern blots and auxotroph analyses indicated that a single copy of the transposon was inserted in several places into the chromosome of P. cepacia. Among 1500 Tn5-259 transconjugants, only one mutant was found to be defective in the production of an antifungal compound, pyrrolnitrin. In addition, this mutant lost its ability to antagonize fungal phytopathogens. Using flanking DNA of the mutated gene as a probe, we have isolated four overlapping cosmid clones from a genomic library of P. cepacia. However, we were unable to complement the mutant because of difficulty in mobilizing the cosmids from E. coli to P. cepacia. PMID:1377094

Jayaswal, R K; Fernandez, M A; Visintin, L; Upadhyay, R S

1992-04-01

255

Enhancement of polyunsaturated fatty acid production by Tn5 transposon in Shewanella baltica.  

PubMed

Transposon Tn5 mutagenesis was used to generate random mutations in Shewanella baltica MAC1, a polyunsaturated fatty acid (PUFA)-producing bacterium. Three mutants produced 3-5 times more eicosapentaenoic acid (EPA 20:5 n-3) compared to the wild type at 10 degrees C. One of the mutants produced 0.3 mg EPA g(-1) when grown at high temperature (30 degrees C). Moreover, 2 mg docosahexaenoic acid (DHA 22:6 n-3) g(-1) was produced by S. baltica mutants at 4 degrees C. Sequencing of insertion mutation(s) showed 96% homology to trimethylamine N-oxide (TMAO) reductase gene and 85% homology to rRNA operons of E. coli. Tn5 transposon mutagenesis therefore is a suitable technique to increase PUFA formation in bacteria. PMID:16799757

Amiri-Jami, Mitra; Wang, Haifeng; Kakuda, Yukio; Griffiths, Mansel W

2006-08-01

256

Transposon Tn5 target specificity: preference for insertion at G/C pairs.  

PubMed

The procaryotic transposon Tn5 inserts into many different sites within a single gene, but some sites (hotspots) are targeted repeatedly. Hotspots are not closely related in sequence, but most have G/C pairs at the ends of the nine base pairs duplicated by Tn5 insertion. In pBR322, the major hotspot coincides with the "-10 region" of the tet promoter. We mutated the G/C pairs at this hotspot and assayed for insertion into hotspot I, resistance to tetracycline, and plasmid supercoiling. We found that changing the G/C pairs to A/T pairs reduced the frequency of insertion into the hotspot by at least fivefold. The reduction in hotspot use caused by these G/C to A/T changes was not attributable to changes in plasmid supercoiling or tet promoter strength. PMID:2852135

Lodge, J K; Weston-Hafer, K; Berg, D E

1988-11-01

257

The TN-24P PWR spent-fuel storage cask: Testing and analyses: Interim report  

SciTech Connect

A performance test of a Transnuclear, Inc. TN-24P storage cask configured for pressurized water reactor (PWR) spent fuel was performed. The test was the second of a series of cask performance tests planned under a cooperative agreement between Virginia Power and the US Department of Energy. The performance test consisted of loading the TN-24P cask with 24 PWR spent fuel assemblies from Virginia Power's Surry reactor. Cask surface and fuel assembly guide tube temperatures were measured, as were cask surface gamma and neutron dose rates. Testing was performed with vacuum, nitrogen, and helium backfill environment in both vertical and horizontal cask orientations. Limited spent fuel integrity data were also obtained.

Creer, J.M.; Michener, T.E.; McKinnon, M.A.; Tanner, J.E.; Gilbert, E.R.; Dziadosz, D.A.; Moore, E.V.; Schoonen, D.H.; Jensen, M.F.

1987-04-01

258

Lactobacillus plantarum TN627 significantly reduces complications of alloxan-induced diabetes in rats.  

PubMed

This study aimed to assess the potential of the probiotic strain Lactobacillus plantarum TN627 for preventing alloxan-induced diabetes in rats. The oral administration of this probiotic was noted to significantly improve the immunological parameters, protect the pancreatic tissues, and reduce the pancreatic and plasmatic ?-amylase activities and level of plasma glucose in the treated as compared to the control group of rats. Furthermore, this probiotic treatment was observed to markedly reduce pancreatic and plasmatic lipase activities and serum triglyceride and LDL-cholesterol rates and to increase the level of HDL-Cholesterol. It also exerted efficient protective effects on the liver and kidney functions evidenced by significant decreases in serum aspartate transaminase, alanine transaminase, lactate dehydrogenase, and gamma-glutamyl transpeptidase activities, as well as creatinine and urea contents. Taken together, the findings indicate that L. plantarum TN627 exhibits attractive in vivo antidiabetic effects that may be helpful in preventing diabetic complications in adult rats. PMID:23999246

Bejar, Wacim; Hamden, Khaled; Ben Salah, Riadh; Chouayekh, Hichem

2013-12-01

259

Construction of Tn5 lac, a transposon that fuses lacZ expression to exogenous promoters, and its introduction into Myxococcus xanthus.  

PubMed

A promoterless trp-lac fusion fragment was inserted near one end of the bacterial transposon Tn5 in the correct orientation to fuse lacZ gene expression to promoters outside Tn5. The resulting transposon, Tn5 lac, retains the kanamycin-resistance gene of Tn5 and transposes in Escherichia coli at 6% the frequency of Tn5 to many different sites in a bacteriophage lambda target. Expression of beta-galactosidase, the product of the lacZ gene, from Tn5 lac insertions in phage lambda depends both on insertion into a transcription unit in the correct orientation and on the regulation of the promoter of the transcription unit, verifying that by transposition Tn5 lac can fuse lacZ expression to outside promoters. An insertion of Tn5 lac in bacteriophage P1 was isolated and used to introduce Tn5 lac into Myxococcus xanthus, a bacterium that undergoes multicellular development. Stable kanamycin-resistant transductants are obtained that contain no P1 DNA sequences but have Tn5 lac inserted at different sites in the Myxococcus chromosome. Individual transductants express different levels of beta-galactosidase. A chromogenic substrate of beta-galactosidase, 5-bromo-4-chloro-3-indolyl beta-D-galactoside, is toxic in Myxococcus when cleaved in large amounts. In principle, Tn5 lac could be used to assay transcription in any bacterium in which Tn5 can transpose and beta-galactosidase can be measured. PMID:6091110

Kroos, L; Kaiser, D

1984-09-01

260

County Waterford Image Archive  

NSDL National Science Digital Library

Using volunteer help and a locally written image management system, the Waterford County Museum in Ireland has created this collection of close to 3,000 historical images that document Waterford County and its parishes from the late 1890s to the present. The Web interface to the collection includes a number of helpful features - for those familiar with Waterford county and its place and family names, there is a search box, but for those less familiar there are several ways to browse the collection, by selecting photographer, location, image format, or subjects from drop down lists, as well as selecting parishes from a map to see the pictures associated with each. A photo of the week is posted on the site, and visitors can sign up to receive an RSS feed or, alternatively, an email message when new images are added. There are also easy ways to submit photographs for inclusion in the archive, and for locals to contribute identifying information.

261

Finite strain and strain variation analysis in the Sheeprock Thrust Sheet: an internal thrust sheet in the Provo salient of the Sevier Fold-and-Thrust belt, Central Utah  

NASA Astrophysics Data System (ADS)

The Sheeprock thrust sheet in west-central Utah is an internal thrust sheet in the Provo salient of the Sevier fold-and-thrust belt. We have measured finite strain in quartzites (the dominant lithology), sampled along a square grid within the thrust sheet, using the modified normalized Fry method (McNaught M.A. (1994) Modifying the normalized Fry method for aggregates of non-elliptical grains. Journal of Structural Geology16 493-503). The {X}/{Y} and {X}/{Z} axial ratios from unsampled locations within the sample area were estimated using the spatial statistics approach. The strain ellipsoids exhibit a variable three-dimensional orientation pattern resulting from modification of the initial layer parallel shortening (LPS) strain ellipsoid by fault parallel shear in conjunction with vertical flattening and/or horizontal stretching indicating that the thrust sheet did not undergo plane strain deformation in the transport plane. This suggests that the plane strain assumption used in drawing restorable balanced cross-sections breaks down for internal thrust sheets with more than one penetrative-strain producing deformation event. The {X}/{Z} strain axial ratios decrease away from the thrust towards the middle of the sheet. The {X}/{Y} strain axial ratios from interpolated image diagrams indicate transport-parallel stretching at the front end of the sheet and strike-parallel stretching at the back end of the sheet. The footwall and hanging wall finite strain patterns are similar indicating that most of the strain in the Sheeprock thrust sheet developed early in the deformation history of the thrust sheet before and perhaps during the growth of a large fault propagation fold pair.

Mukul, Malay; Mitra, Gautam

1998-04-01

262

Effect of predeformational basin geometry in the kinematic evolution of a thin-skinned orogenic wedge: Insights from three-dimensional finite element modeling of the Provo salient, Sevier fold-thrust belt, Utah  

NASA Astrophysics Data System (ADS)

In fold-thrust belts, sedimentary cover rocks are detached from undeformed basement and undergo crustal-scale shortening and internal deformation. We have investigated a three-dimensional (3-D), nonlinear, elastic-plastic finite element model using the restored Provo salient of the Sevier belt as our initial configuration. In the model the deformed sedimentary prism displays large-scale geometries that are seen in many natural fold-thrust belts (e.g., arcuate salient, wedge-shaped cross section) and kinematics that are compatible with observations in the internal and external portions of the Provo salient; these suggest that the model can be used to predict geologic information that is generally not available from detailed observational studies in natural fold-thrust belts (e.g., strain history, material displacements, stress conditions). The model results indicate symmetric, noncoaxial, plane strain paths with consistent stress and strain orientations and material displacement directions in the middle of the 3-D wedge, and fully 3-D, nonsymmetric, noncoaxial, nonplane strain paths with out-of-transport material displacements over the lateral boundaries. The results from test runs further suggest that oblique ramps with strike direction less than 20° from the regional transport direction behave like lateral ramps, and those with strike direction greater than 80° from the regional transport direction behave similar to frontal ramps. Oblique ramps with dips greater than 60° behave like tear faults. These variations in different parts of the wedge are caused mainly by interaction between the transport parallel motion of the moving wedge and the preexisting footwall template of ramps and flats that the wedge has to ride over during its evolution.

Kwon, Sanghoon; Mitra, Gautam; Perucchio, Renato

2007-02-01

263

Mucin-associated sialosyl-Tn antigen expression in gastric cancer correlates with an adverse outcome.  

PubMed Central

The expression of sialosyl-Tn (STn) antigen was evaluated by immunohistochemistry in primary gastric cancers. Twenty-one of 31 (68%) gastric cancers expressed STn, regardless of tumour location, stage or histological type. Eighty-one per cent of patients with STn-positive tumours died of their disease or had recurrent cancer, compared with 20% of patients with STn-negative tumours (P < 0.002). STn may be a useful prognostic marker in patients with gastric cancer. Images Figure 1

Werther, J. L.; Rivera-MacMurray, S.; Bruckner, H.; Tatematsu, M.; Itzkowitz, S. H.

1994-01-01

264

Characterization of a "mobilization cassette" in transposon Tn4399 from Bacteroides fragilis.  

PubMed Central

Derivatives of nonconjugal plasmids that carry Tn4399, a transposon isolated from Bacteroides fragilis, can be mobilized for transfer by the broad-host-range IncP plasmids pRK231 or R751 in Escherichia coli. To characterize regions of Tn4399 involved in mobilization, we have isolated and analyzed subcloned fragments of Tn4399 in E. coli, as well as mutations within the element. We have identified a "mobilization cassette" within a 2.8-kb region of Tn4399 which, when cloned into mobilization-deficient plasmids, allows these plasmids to be mobilized in trans by the IncP plasmids pRK231 and R751. The 2.8-kb region has been sequenced, and several open reading frames have been identified. Mutants defective in two genes, designated mocA and mocB, coding for deduced products of 36.4 and 16.4 kDa, respectively, cannot be mobilized by either IncP plasmid; these mutants can be complemented in the presence of the respective wild-type genes in trans. This suggests that the putative MocA and MocB proteins have a role in the mobilization process. The 36.4-kDa MocA protein contains a 14-amino-acid sequence which is closely related to a highly conserved motif within DNA relaxases encoded by a wide variety of conjugal or mobilizable plasmids. Subcloning experiments also lead to the localization of an oriT region within a 199-bp fragment, internal to the mobilization cassette.

Murphy, C G; Malamy, M H

1993-01-01

265

Persistent repression of a functional allele can be responsible for galactosyltransferase deficiency in Tn syndrome.  

PubMed Central

A human hematopoietic disorder designated as Tn syndrome or permanent mixed-field polyagglutinability has been ascribed to a stem cell mutation leading to a specific deficiency of UDP-Gal:GalNAc alpha 1-O-Ser/Thr beta 1-3 galactosyltransferase (beta 3 Gal-T) activity in affected cells. To test for the possibility that an allele of the beta 3Gal-T gene might be repressed instead of mutated, we have investigated whether 5-azacytidine or sodium n-butyrate, both inducers of gene expression, would reactivate expression of beta 3Gal-T in cloned enzyme-deficient T cells derived from a patient affected by the Tn syndrome. Flow cytometry revealed that a single treatment induced de novo expression of the Thomsen-Friedenreich antigen (Gal beta 1-3GalNAc-R), the product of beta 3Gal-T activity. In addition, a sialylated epitope on CD43 (leukosialin), which is present on normal but not on beta 3Gal-T-deficient T cells, was also reexpressed. Although no beta 3Gal-T activity was detectable in untreated Tn syndrome T cells, after exposure to 5-azaC,beta 3Gal-T activity reached nearly normal values. Both agents failed to reactivate beta 3Gal-T in Jurkat T leukemic cells, which also lack beta 3Gal-T activity. These data demonstrate that Tn syndrome T cells contain an intact beta 3Gal-T gene copy and that the enzyme deficiency in this patient is due to a persistent and complete but reversible repression of a functional allele. In contrast, the cause of beta 3Gal-T deficiency appears to be different in Jurkat T cells. Images

Thurnher, M; Rusconi, S; Berger, E G

1993-01-01

266

ROLE OF NA+ IN TRANSPORT OF HG2+ AND INDUCTION OF THE TN21 "MER" OPERON  

EPA Science Inventory

The effects of sodium ions on the uptake of Hg 2 + and induction of the TN21 mer operon were studied using E. coli HMS174 harboring the reporter plasmids pRB28 and pOS14. lasmid pRB28 carries merRT' and pOS14 carries merRTPC of the mer operon, both cloned upstream of a promoterle...

267

Polaron Effects on Superexchange Interaction: Isotope Shifts of TN Tc and T* in Layered Copper Oxides  

NASA Astrophysics Data System (ADS)

A compact expression has been obtained for the superexchange coupling of magnetic ions via intermediate anions with regard to polaron effects at both magnetic ions and intermediate anions. This expression is used to analyze the main features of the behavior of isotope shifts for temperatures of three types in layered cuprates: the Neel temperatures ($T_N$), critical temperatures of transitions to a superconducting state ($T_C$), and characteristic temperatures of the pseudogap in the normal state ($T^*$).

Eremin, M. V.; Eremin, I. M.; Larionov, I. A.; Terzi, A. V.

2002-04-01

268

The Sugar Creek zinc deposit, Jackson Co. TN -- Exploration history, geology and mineralization  

Microsoft Academic Search

During the 60's and 70's zinc exploration of central TN and KY was active. The Sugar Creek Project was one of several investigated by Exxon. The discovery hole, Cu 15, was drilled in early 1973. The Sugar Creek Zinc Deposit was acquired by Independence Mining Co. in 1986 and I.M.C. has subsequently completed additional drilling, both stepout and confirmation holes.

G. Reinbold; A. V. Moran; D. L. Stevens

1993-01-01

269

Family of Class 1 Integrons Related to In4 from Tn1696  

Microsoft Academic Search

The class 1 integron In28, found in the multidrug resistance transposon Tn1403, was found to be located in the res site of the backbone transposon and is flanked by a 5-bp direct duplication, indicating that it reached this position by transposition. In28 has a backbone structure related to that of In4, but has lost internal sequences, including the sul1 gene,

SALLY R. PARTRIDGE; GAVIN D. RECCHIA; H. W. Stokes; RUTH M. HALL

2001-01-01

270

Genechip-detecting mutations in exon 8 in cTnI gene associated with FHCM  

NASA Astrophysics Data System (ADS)

As the rate of gene discovery accelerates, more efficient methods are needed to analyze genes in human tissues. Genechip, a kind of new device, is composed of DNA probes immobilized on a solid substrate. With the advantage of the high throughput information, genechip has become one of the best solutions to detect and analyse the mutations in genes. Hypertrophic cardiomyopathy (HCM), the most common cause of the sudden death in the young, is one of the diseases damaging people health most badly. It is an autosomal dominant disease. More than 55% of the HCM patients are genetic. The mutations of exon 8 in the Cardiac troponin I (cTnI) gene are closely associated with Family Hypertrophic Cardiomyopathy (FHCM). Our purpose is to perform the assay of the mutations in exon 8 in cTnI gene based on the genechip theory and technology. Special probes were designed to fabricate the genechip to detect the mutations in cTnI gene simultaneously. We designed two oligonucleotide sequences 5"-end labeled with fluorescein, one simulating wild-type and the other simulating mutant. We mixed oligonucleotide I and II together to simulate heterozygote. After optimizing the hybridization protocols, the fabricated genechip can detect the mutations in exon 8 in cTnI gene with relative high sensitivity and specificity. When applying the fabricated genechip to detect the target DNA sequence, we found that the fully complementary probe gave a fluorescent signal almost 50% stronger than that of the one base mismatched one, which is in accordance with the result from theoretic estimate. It is believed that an applicable special genechip can be developed for investigating and diagnosing FHCM after further improvement.

Zhang, Yuanying; He, Nongyue; Guo, Huishi; Yang, Di; Wan, Wenhui; Bian, Zhiping; Zhang, Jinan

2005-01-01

271

Regulated expression of heterologous genes in Bacillus subtilis using the Tn 10 encoded tet regulatory elements  

Microsoft Academic Search

TheEscherichia coli-derivedtet regulatory elements from Tn10 have been used to construct vectors allowing the regulated, inducible, high-level expression of foreign genes inBacillus subtilis. While the wild-typetet promoters are inactive inB. subtilis, a synthetic mutanttet sequence with improved promoter consensus sequences and upstream poly A blocks shows activity inB. subtilis. The expression of an indicatorcat gene is inducible by sublethal amounts

Manfred Geissendiirfer; Wolfgang Hillen

1990-01-01

272

Defects in fruiting body development caused by Tn5 lac insertions in Myxococcus xanthus.  

PubMed Central

Mutations caused by insertions of Tn5 lac that block development are rare. At least six of the eight mutations examined appeared to be regulatory. Three of these were found to disrupt social motility, suggesting a particular importance for this function. One other occurred in a known cell-cell interaction gene, bsgA, and the remaining two were located in genes operative early in the developmental program. Images FIG. 1 FIG. 2

Kroos, L; Kuspa, A; Kaiser, D

1990-01-01

273

Defects in fruiting body development caused by Tn5 lac insertions in Myxococcus xanthus.  

PubMed

Mutations caused by insertions of Tn5 lac that block development are rare. At least six of the eight mutations examined appeared to be regulatory. Three of these were found to disrupt social motility, suggesting a particular importance for this function. One other occurred in a known cell-cell interaction gene, bsgA, and the remaining two were located in genes operative early in the developmental program. PMID:2152913

Kroos, L; Kuspa, A; Kaiser, D

1990-01-01

274

TN method for the critical thickness of one-speed neutrons in a slab with forward and backward scattering  

NASA Astrophysics Data System (ADS)

The critical slab problem in the case of combination of forward and backward scattering with usual isotropic scattering is studied for one-speed neutrons in a uniform finite slab by using TN method based on Chebyshev polynomial approximation and Marshak boundary conditions. It is shown that TN method gives accurate results in one-dimensional geometry and is very efficient both in derivation of equations and rapid convergence. Numerical results obtained by TN method are compared against the PN method in tabular form, which agreed quite well.

Öztürk, Hakan; Anli, Fikret; Güngör, Süleyman

2007-06-01

275

Purification and biochemical analyses of a monomeric form of Tn5 transposase.  

PubMed Central

The binding of transposase (Tnp) to the specific Tn5 end sequences is the first dedicated reaction during transposition. In this study, comparative DNA-binding analyses were performed using purified full-length Tnp and a C-terminal deletion variant (delta369) that lacks the putative dimerization domain. The shape of the binding curve of full-length Tnp is sigmoidal in contrast to the hyperbolic-shaped binding curve of delta369. This observation is consistent with previous observations as well as a rate of binding study presented here, which suggest that the full-length Tnp-end interaction, unlike that of the truncated protein, is a complex time-dependent reaction possibly involving a subunit exchange. Circular permutation assay results indicate that both proteins are capable of distorting the Tn5end sequences upon binding. Molecular weight determinations based on the migratory patterns of complexed DNA in polyacrylamide gels has shown that delta369 specifically binds the Tn5 end sequences as a monomer while full-length Tnp in complex represents a heterodimer.

York, D; Reznikoff, W S

1996-01-01

276

TnAraOut, a transposon-based approach to identify and characterize essential bacterial genes.  

PubMed

Identification of genes that encode essential products provides a promising approach to validation of new antibacterial drug targets. We have developed a mariner-based transposon, TnAraOut, that allows efficient identification and characterization of essential genes by transcriptionally fusing them to an outward-facing, arabinose-inducible promoter, PBAD, located at one end of the transposon. In the absence of arabinose, such TnAraOut fusion strains display pronounced growth defects. Of a total of 16 arabinose-dependent TnAraOut mutants characterized in Vibrio cholerae, four were found to carry insertions upstream of known essential genes (gyrB, proRS, ileRS, and aspRS) whereas the other strains carried insertions upstream of known and hypothetical genes not previously shown to encode essential gene products. One of the essential genes identified by this analysis appears to be unique to V. cholerae and thus may represent an example of a species-specific drug target. PMID:10888841

Judson, N; Mekalanos, J J

2000-07-01

277

Origin of the second length scale found above TN in UO2  

NASA Astrophysics Data System (ADS)

We present the results of x-ray- and neutron-scattering studies of the temperature dependence of the magnetic scattering exhibited by the type-I, triple-Q antiferromagnet UO2. Our neutron-scattering results are consistent with those of earlier studies, including the observation of short-ranged magnetic correlations at temperatures near and above TN. However, it is found by x-ray diffraction that a second, longer length scale is induced near TN when the near-surface volume of the sample is mechanically roughened. The longitudinal and transverse widths of the additional scattering increase continuously with increasing temperature above TN, similar to that which has been observed near the magnetic ordering transitions of Ho, Tb, and NpAs and near the tetragonal-to-cubic transitions of various perovskites. Another unusual feature of the present results for UO2 involves the apparent shift with temperature of the magnetic scattering along the surface normal direction at the (1,1,0) reflection, but not at the (2,1,0) reflection. To our knowledge, this is the first observation of a second length scale near a first-order transition.

Watson, G. M.; Gaulin, B. D.; Gibbs, Doon; Thurston, T. R.; Simpson, P. J.; Shapiro, S. M.; Lander, G. H.; Matzke, Hj.; Wang, S.; Dudley, M.

1996-01-01

278

Melting of the antiferromagnetic structure in UNiAL near TN  

NASA Astrophysics Data System (ADS)

UNiAl has the hexagonal ZrNiAl type of structure and below TN=19.3 K exhibits antiferromagnetic ordering of U moments with the propagation vector q=(0.1 0.1 0.5). In a sufficiently high magnetic field Bc applied along the c-axis ( Bc=11.4 T at 2 K), UNiAl undergoes a metamagnetic transition from an antiferromagnetic structure towards a ferromagnetic structure. In fields above ?8 T (<11.4 T), a new antiferromagnetic structure is established in UNiAl with the in-plane modulation direction tilted to the [1 0 0] direction. The propagation vector of the new antiferromagnetic structure has the same length as in the original and also the direction of U moments along the c-axis is preserved. In this paper we show that the competition between the two indicated antiferromagnetic structures takes place even in zero field in the vicinity of TN. In a small temperature range around TN a diffuse ring of intensity can be observed, indicating that angular coherence is lost and the modulation of U moments exists in several directions.

Prokeš, K.; Brück, E.; Sechovský, V.

2006-11-01

279

Exchange bias in a ferromagnet/antiferromagnet system with TC<<TN  

NASA Astrophysics Data System (ADS)

This paper reports experimental results obtained on an unconventional exchange bias (EB) system where the ferromagnetic layer, Ni0.75Cu0.25, has lower ordering temperature (TC) than that (TN) of the antiferromagnetic one, NiO, with emphasis on modifying EB through either magnetic annealing or light-ion irradiation. Samples were cooled from temperatures higher than TN or in between TC and TN to room temperature with magnetic field applied in different in-plane directions. Upon ion irradiation, magnetic fields, parallel or antiparallel to the orientation of the field present during the films deposition, were applied to explore different effects on EB. We found that the EB direction can be completely reversed by means of either annealing or ion bombardment; however, both postdeposition treatments provide very little variation of the EB field value over that produced during the film's growth. The importance of the annealing field strength was also discussed. The results were interpreted based on a mechanism which assumes that the interfacial moments adjacent to the antiferromagnetic layer are responsible for establishing the exchange biasing in the paramagnetic state.

Sossmeier, K. D.; Pereira, L. G.; Schmidt, J. E.; Geshev, J.

2011-04-01

280

A functional origin of transfer (oriT) on the conjugative transposon Tn916.  

PubMed Central

The origin of transfer (oriT) of the 18-kb conjugative transposon Tn916 has been localized to a 466-bp region which spans nucleotides 15215 to 15681 on the transposon map. The oriT lies within an intercistronic region between open reading frames ORF20 and ORF21 that contains six sets of inverted repeats ranging from 10 to 20 bp in size. The segment contains three sequences showing identity in 9 of 12 bp to the consensus nicking site (nic) of the IncP family of conjugative plasmids found in gram-negative bacteria. Overlapping one of these sequences is a region similar to the nic site of the F plasmid. Functionality was based on the ability of the oriT-containing sequence to provide a cis-acting mobilization of chimeras involving the shuttle vector pWM401 in response to activation in trans by an intact chromosome-borne transposon Tn916 delta E. Cloned segments of 466 or 376 nucleotides resulted in unselected cotransfer of the plasmid at levels of about 40% when selection was for Tn916 delta E, whereas a 110-bp segment resulted in cotransfer at a frequency of about 7%. Mobilization was specific in that gram-positive plasmids, such as pAD1 and pAM beta 1, and the gram-negative plasmids pOX38 (a derivative of F) and RP1 did not mobilize oriT-containing chimeras.

Jaworski, D D; Clewell, D B

1995-01-01

281

Alteration of surface properties in a Tn5 mutant strain of Rhizobium trifolii 0403.  

PubMed Central

A symbiotically defective mutant strain of Rhizobium trifolii, UR251, was obtained by transposon Tn5 mutagenesis of R. trifolii 0403 rif and recognized by its partially ineffective (Fix +/-) phenotype on white clover plants. UR251 had a single Tn5 insertion in plasmid DNA, a wild-type plasmid pattern, and no detectable Mu DNA sequences originally present in the vector used for Tn5 mutagenesis. Agglutination by the clover lectin trifoliin A and attachment to clover root hairs was higher with UR251 than with the wild-type strain. The capsular polysaccharide (CPS) of UR251 was altered, as shown by a slower rate of CPS depolymerization with a CPS beta-lyase, PD-I; more pyruvate and less acetate and 3-hydroxybutanoate noncarbohydrate substitutions as quantitated by 1H nuclear magnetic resonance; and a higher pyruvyl transferase activity (enzymatic pyruvylation of lipid-bound saccharides). The site of increased pyruvylation in the CPS of UR251 was on the terminal galactose of the branch of the repeating oligosaccharide unit. These results show that the level of noncarbohydrate substitutions of the CPS as well as pyruvyl transferase activity are altered in R. trifolii UR251 and that trifoliin A-binding ability and clover root hair attachment are improved in this mutant strain of R. trifolii 0403 rif. Images

Gardiol, A E; Hollingsworth, R I; Dazzo, F B

1987-01-01

282

Pierce County Hospital Development Guide.  

National Technical Information Service (NTIS)

A hospital development guide that was developed for Pierce County, Washington is presented. Planning for communitywide hospital development in Pierce County was initiated in 1971, based on a draft report and passage of State certificate of need legislatio...

1974-01-01

283

Simple mucin-type carbohydrate antigens (Tn, sialosyl-Tn, T and sialosyl-T) and gp 230 mucin-like glycoprotein are candidate markers for neoplastic transformation of the human cervix  

Microsoft Academic Search

Mucins and simple mucin-type carbohydrates are cancer-associated antigens in several human tumors. Expression of Tn, sialosyl-Tn,\\u000a Thomsen-Friedenreich (T), sialosyl-T and of a recently identified mucin-like glycoprotein (gp230) has not yet been thoroughly\\u000a investigated in human cervix carcinogenesis. In the present study sections from normal cervix (n=10), CIN III lesions (n=10), and invasive carcinomas (n=47) were evaluated immunohistochemically using monoclonal antibodies.

Carla Carrilho; Martha Cantel; Patricia Gouveia; Leonor David

2000-01-01

284

Iron oxide superparamagnetic nanoparticles conjugated with a conformationally blocked ?-Tn antigen mimetic for macrophage activation  

NASA Astrophysics Data System (ADS)

Among new therapies to fight tumors, immunotherapy is still one of the most promising and intriguing. Thanks to the ongoing structural elucidation of several tumor antigens and the development of innovative antigen carriers, immunotherapy is in constant evolution and it is largely used either alone or in synergy with chemotherapy/radiotherapy. With the aim to develop fully synthetic immunostimulants we have recently developed a mimetic of the ?-Tn mucin antigen, a relevant tumor antigen. The 4C1 blocked mimetic 1, unique example of an ?-Tn mimetic antigen, was functionalized with an ?-phosphonate linker and used to decorate iron oxide superparamagnetic nanoparticles (MNPs), employed as multivalent carriers. MNPs, largely exploited for supporting and carrying biomolecules, like antibodies, drugs or antigens, consent to combine in the same nanometric system the main features of an inorganic magnetic core with a bioactive organic coating. The superparamagnetic glyconanoparticles obtained, named GMNPs, are indeed biocompatible and immunoactive, and they preserve suitable characteristics for use as heat mediators in the magnetic fluid hyperthermia (MFH) treatment of tumors. All together these properties make GMNPs attracting devices for innovative tumor treatment.Among new therapies to fight tumors, immunotherapy is still one of the most promising and intriguing. Thanks to the ongoing structural elucidation of several tumor antigens and the development of innovative antigen carriers, immunotherapy is in constant evolution and it is largely used either alone or in synergy with chemotherapy/radiotherapy. With the aim to develop fully synthetic immunostimulants we have recently developed a mimetic of the ?-Tn mucin antigen, a relevant tumor antigen. The 4C1 blocked mimetic 1, unique example of an ?-Tn mimetic antigen, was functionalized with an ?-phosphonate linker and used to decorate iron oxide superparamagnetic nanoparticles (MNPs), employed as multivalent carriers. MNPs, largely exploited for supporting and carrying biomolecules, like antibodies, drugs or antigens, consent to combine in the same nanometric system the main features of an inorganic magnetic core with a bioactive organic coating. The superparamagnetic glyconanoparticles obtained, named GMNPs, are indeed biocompatible and immunoactive, and they preserve suitable characteristics for use as heat mediators in the magnetic fluid hyperthermia (MFH) treatment of tumors. All together these properties make GMNPs attracting devices for innovative tumor treatment. Electronic supplementary information (ESI) available: Optical rotation and spectroscopic data for all compounds; selected TEM images of MNPs, CMNPs and GMNPs; XRD patterns and FTIR spectra of MNPs coated with surfactants and with the ?-Tn mimetic; MNP magnetic properties. See DOI: 10.1039/c4nr00070f

Manuelli, Massimo; Fallarini, Silvia; Lombardi, Grazia; Sangregorio, Claudio; Nativi, Cristina; Richichi, Barbara

2014-06-01

285

Housing: Dutchess County.  

National Technical Information Service (NTIS)

One of the most crucial responsibilities of any fast-growing area is that of providing housing for all segments of its population. As pressures for growth are increasingly exerted on Dutchess County, it will be all the more important for the area to know ...

1972-01-01

286

Reviewing County Extension Programs  

ERIC Educational Resources Information Center

An evaluation of the Douglas County (Kansas) Extension Program to determine clientele's level of awareness and acceptance of the program and program strengths and weaknesses, to establish new program directions and a base line for future evaluation efforts, and to develop methodology and materials to be used in evaluation. Procedures, findings,…

Prawl, Warren L.; Jorns, William J.

1976-01-01

287

Dielectric anomaly at TN in LaMnO3 as a signature of coupling between spin and orbital degrees of freedom  

NASA Astrophysics Data System (ADS)

We observe a distinct anomaly in dielectric permittivity (?') as well as relaxation time (?0) versus temperature (T) pattern at the antiferromagnetic transition point TN in a single crystal of LaMnO3 . The equivalent circuit analysis of the impedance spectra across TN reveals a clear anomaly in the capacitive component C0 at TN . Since no structural transition takes place across TN , the anomaly in ?0 and C0 at TN possibly signifies multiferroicity stemming from coupling between orbital and spin order in LaMnO3 .

Mondal, Parthasarathi; Bhattacharya, Dipten; Choudhury, Pranab; Mandal, Prabhat

2007-11-01

288

The entirely carbohydrate immunogen Tn-PS A1 induces a cancer cell selective immune response and cytokine IL-17.  

PubMed

The tumor-associated carbohydrate antigen/hapten Thomsen-nouveau (Tn; a-D-GalpNAc-ONH2) was conjugated to a zwitterionic capsular polysaccharide, PS A1, from commensal anaerobe Bacteroides fragilis ATCC 25285/NCTC 9343 for the development of an entirely carbohydrate cancer vaccine construct and probed for immunogenicity. This communication discloses that murine anti-Tn IgG3 antibodies both bind to and recognize human tumor cells that display the Tn hapten. Furthermore, the sera from immunization of mice with Tn-PS A1 contain cytokine interleukin 17 (IL-17A), which is known to possess anti-tumor function and represents a striking difference to an IL-2, and IL-6 profile obtained with anti-PS A1 sera. PMID:22297566

De Silva, Ravindra A; Appulage, Dananjaya K; Pietraszkiewicz, Halina; Bobbitt, Kevin R; Media, Joe; Shaw, JiaJiu; Valeriote, Fred A; Andreana, Peter R

2012-04-01

289

Immunological response from an entirely carbohydrate antigen: design of synthetic vaccines based on Tn-PS A1 conjugates.  

PubMed

An entirely carbohydrate-based immunogen consisting of a zwitterionic polysaccharide (ZPS) PS A1 and the well-known tumor antigen Tn has been designed, synthesized, and studied for immunological effects. The PS A1 motif was included to act as an MHCII elicitor for a T-cell-dependent immune response with increased immunogenicity against tumor-associated carbohydrate antigens, providing an alternative to carrier proteins. Through the use of C57BL/6 mice, it has been shown that chemical modification of PS A1 does not alter the recognition sequence responsible for an MHCII-mediated, T-cell-dependent immune response. The Tn-PS A1 conjugate construct confers specificity toward the Tn antigen alone, and specific carbohydrate immunoglobulins, namely, IgG3, are generated from intraperitoneal immunizations with or without adjuvant. The properties of the vaccine candidate are attributed to a site-specific linking strategy that incurs significant incorporation of Tn antigen. PMID:19552408

De Silva, Ravindra A; Wang, Qianli; Chidley, Tristan; Appulage, Dananjaya K; Andreana, Peter R

2009-07-22

290

The entirely carbohydrate immunogen Tn-PS A1 induces a cancer cell selective immune response and cytokine IL-17  

PubMed Central

The tumor-associated carbohydrate antigen/hapten Thomsen-nouveau (Tn; ?-D-GalpNAc-ONH2) was conjugated to a zwitterionic capsular polysaccharide, PS A1, from commensal anaerobe Bacteroides fragilis ATCC 25285/NCTC 9343 for the development of an entirely carbohydrate cancer vaccine construct and probed for immunogenicity. This communication discloses that murine anti-Tn IgG3 antibodies both bind to and recognize human tumor cells that display the Tn hapten. Furthermore, the sera from immunization of mice with Tn-PS A1 contain cytokine interleukin 17 (IL-17A), which is known to possess anti-tumor function and represents a striking difference to an IL-2, and IL-6 profile obtained with anti-PS A1 sera.

De Silva, Ravindra A.; Appulage, Dananjaya K.; Pietraszkiewicz, Halina; Bobbitt, Kevin R.; Media, Joe; Shaw, JiaJiu; Valeriote, Fred A.

2012-01-01

291

Polarized neutron scattering study of the magnetic response across TN in a single crystal of UO2  

NASA Astrophysics Data System (ADS)

We present the results of an inelastic neutron scattering experiment with polarization analysis performed on a single crystal of UO2. A magnetic inelastic response is observed above TN in the energy region between 3 and 10 meV, at least up to 6.5TN. This result provides evidence of the fact that in the time scale of our experiment the uranium triplet ground state is split into three singlets because of a dynamical Jahn-Teller distortion of the oxygen cage, reducing the point symmetry at the uranium site. The position of the peaks and their dispersion are compatible with monoclinic 1-k distortions along the three directions of the <100> star. This suggests a picture in which as TN is approached, a correlation builds up between the phases of the corresponding vibrations until, eventually, a static 3-k distortion is obtained below TN.

Amoretti, G.; Caciuffo, R.; Santini, P.; Lander, G. H.; Kulda, J.; de v. Du Plessis, P.

1999-04-01

292

Dissemination of TnMERI1-like mercury resistance transposons among Bacillus isolated from worldwide environmental samples.  

PubMed

Fifty-six mercury-resistant (Hg(R)) Bacillus strains were isolated from natural environments at various sites of the world. Southern hybridisation and polymerase chain reaction (PCR) analysis showed that 21 of the 56 isolates have closely related or identical mer operons to that of Bacillus megaterium MB1. These 21 isolates displayed a broad-spectrum mercury resistance and volatilised Hg(0). PCR amplification with a single primer and restriction fragment length polymorphism analysis showed that these 21 isolates had TnMERI1-like class II transposons. These transposons can be classified into Tn5084, Tn5085, or TnMERI1. From these results, at least three types of class II mercury resistance transposons exist in Hg(R)Bacillus and these transposons may contribute the worldwide distribution and horizontal dissemination of the mer operons among Bacillus strains in natural environments. PMID:19712430

Narita, Masaru; Matsui, Kazuaki; Huang, Chieh-Chen; Kawabata, Zen'ichiro; Endo, Ginro

2004-04-01

293

Isolation of Salmonella typhimurium Tn5 mutants defective for survival on egg shell surface using transposon footprinting.  

PubMed

The objective of this study was to screen the genome of Salmonella typhimurium for genes potentially required for survival on egg shell surfaces using a novel transposon footprinting method based on a previously developed Tn5 mutagenesis technique. Since the amplified DNA fragment length should usually be unique for each mutant, the polymerase chain reaction products separated on an agarose gel generate a transposon footprint with each band in the footprint representing the corresponding Tn5 mutant. By comparing the footprints from the pools of S. typhimurium Tn5 mutants before and after inoculation on egg shell surfaces, Tn5 mutants not recovered after selection were rapidly identified for potential isolation and genetic analysis. PMID:12602827

Kwon, Y M; Kubena, L F; Nisbet, D J; Ricke, S C

2003-01-01

294

Fate of Tn5 Mutants of Root Growth-Inhibiting 'Pseudomonas' sp. in Intact Soil-Core Microcosms.  

National Technical Information Service (NTIS)

Transposon Tn5 mutants of a wheat root growth-inhibiting nonfluorescent Pseudomonas sp. were inoculated into intact soil-core microcosms to determine the utility of intact soil cores for evaluating the fate and transport of microorganisms in agricultural ...

J. M. Fredrickson S. A. Bentjen H. Bolton S. W. Li P. Van Voris

1989-01-01

295

Ca(2+)-regulatory function of the inhibitory peptide region of cardiac troponin I is aided by the C-terminus of cardiac troponin T: Effects of familial hypertrophic cardiomyopathy mutations cTnI R145G and cTnT R278C, alone and in combination, on filament sliding.  

PubMed

Investigations of cardiomyopathy mutations in Ca(2+) regulatory proteins troponin and tropomyosin provide crucial information about cardiac disease mechanisms, and also provide insights into functional domains in the affected polypeptides. Hypertrophic cardiomyopathy-associated mutations TnI R145G, located within the inhibitory peptide (Ip) of human cardiac troponin I (hcTnI), and TnT R278C, located immediately C-terminal to the IT arm in human cardiac troponin T (hcTnT), share some remarkable features: structurally, biochemically, and pathologically. Using bioinformatics, we find compelling evidence that TnI and TnT, and more specifically the affected regions of hcTnI and hcTnT, may be related not just structurally but also evolutionarily. To test for functional interactions of these mutations on Ca(2+)-regulation, we generated and characterized Tn complexes containing either mutation alone, or both mutations simultaneously. The most important results from in vitro motility assays (varying [Ca(2+)], temperature or HMM density) show that the TnT mutant "rescued" some deleterious effects of the TnI mutant at high Ca(2+), but exacerbated the loss of function, i.e., switching off the actomyosin interaction, at low Ca(2+). Taken together, our experimental results suggest that the C-terminus of cTnT aids Ca(2+)-regulatory function of cTnI Ip within the troponin complex. PMID:24418317

Brunet, Nicolas M; Chase, P Bryant; Mihajlovi?, Goran; Schoffstall, Brenda

2014-06-15

296

Mixtures of glyphosate and surfactant TN20 accelerate cell death via mitochondrial damage-induced apoptosis and necrosis.  

PubMed

Glyphosate, a common herbicide, is not toxic under normal exposure circumstances. However, this chemical, when combined with a surfactant, is cytotoxic. In this study, the mechanism of the additive effect of glyphosate and TN-20, a common surfactant in glyphosate herbicides, was investigated. After exposure of rat H9c2 cells to glyphosate and TN-20 mixtures, following assays were performed: flow cytometry to determine the proportion of cells that underwent apoptosis and necrosis; western blotting to determine expression of mitochondrial proteins (Bcl-2 and Bax); immunological methods to evaluate translocation of cytochrome C; luminometric measurements to determine activity of caspases 3/7 and 9; and tetramethyl rhodamine methyl ester assay to measure mitochondrial membrane potentials. Bcl-1 intensity decreased while Bax intensity increased with exposure to increasing TN-20 and/or glyphosate concentrations. Caspase activity increased and mitochondrial membrane potential decreased only when the cells were exposed to a mixture of both TN-20 and glyphosate, but not after exposure to either one of these compounds. The results support the possibility that mixtures of glyphosate and TN-20 aggravate mitochondrial damage and induce apoptosis and necrosis. Throughout this process, TN-20 seems to disrupt the integrity of the cellular barrier to glyphosate uptake, promoting glyphosate-mediated toxicity. PMID:23099315

Kim, Young-hee; Hong, Jung-rak; Gil, Hyo-wook; Song, Ho-yeon; Hong, Sae-yong

2013-02-01

297

75 FR 52367 - Notice of Inventory Completion: Memphis Pink Palace Museum, Memphis, TN  

Federal Register 2010, 2011, 2012, 2013

...associated funerary objects are present. In 1972, human remains representing a minimum of one individual were removed from the Togo site (3CS24), Cross County, AR, during amateur excavations. The human remains were donated to the museum by Ms....

2010-08-25

298

Systematic sequencing of cDNA clones using the transposon Tn5.  

PubMed

In parallel with the production of genomic sequence data, attention is being focused on the generation of comprehensive cDNA-sequence resources. Such efforts are increasingly emphasizing the production of high-accuracy sequence corresponding to the entire insert of cDNA clones, especially those presumed to reflect the full-length mRNA. The complete sequencing of cDNA clones on a large scale presents unique challenges because of the generally small, yet heterogeneous, sizes of the cloned inserts. We have developed a strategy for high-throughput sequencing of cDNA clones using the transposon Tn5. This approach has been tailored for implementation within an existing large-scale 'shotgun-style' sequencing program, although it could be readily adapted for use in virtually any sequencing environment. In addition, we have developed a modified version of our strategy that can be applied to cDNA clones with large cloning vectors, thereby overcoming a potential limitation of transposon-based approaches. Here we describe the details of our cDNA-sequencing pipeline, including a summary of the experience in sequencing more than 4200 cDNA clones to produce more than 8 million base pairs of high-accuracy cDNA sequence. These data provide both convincing evidence that the insertion of Tn5 into cDNA clones is sufficiently random for its effective use in large-scale cDNA sequencing as well as interesting insight about the sequence context preferred for insertion by Tn5. PMID:12034835

Shevchenko, Yuriy; Bouffard, Gerard G; Butterfield, Yaron S N; Blakesley, Robert W; Hartley, James L; Young, Alice C; Marra, Marco A; Jones, Steven J M; Touchman, Jeffrey W; Green, Eric D

2002-06-01

299

Insertional transposon mutagenesis by electroporation of released Tn5 transposition complexes.  

PubMed

DNA transposition is an important biological phenomenon that mediates genome rearrangements, inheritance of antibiotic resistance determinants, and integration of retroviral DNA. Transposition has also become a powerful tool in genetic analysis, with applications in creating insertional knockout mutations, generating gene-operon fusions to reporter functions, providing physical or genetic landmarks for the cloning of adjacent DNAs, and locating primer binding sites for DNA sequence analysis. DNA transposition studies to date usually have involved strictly in vivo approaches, in which the transposon of choice and the gene encoding the transposase responsible for catalyzing the transposition have to be introduced into the cell to be studied (microbial systems and applications are reviewed in ref. 1). However, all in vivo systems have a number of technical limitations. For instance, the transposase must be expressed in the target host, the transposon must be introduced into the host on a suicide vector, and the transposase usually is expressed in subsequent generations, resulting in potential genetic instability. A number of in vitro transposition systems (for Tn5, Tn7, Mu, Himar1, and Ty1) have been described, which bypass many limitations of in vivo systems. For this purpose, we have developed a technique for transposition that involves the formation in vitro of released Tn5 transposition complexes (TransposomesTM) followed by introduction of the complexes into the target cell of choice by electroporation. In this report, we show that this simple, robust technology can generate high-efficiency transposition in all tested bacterial species (Escherichia coli, Salmonella typhimurium, and Proteus vulgaris) We also isolated transposition events in the yeast Saccharomyces cerevisiae. PMID:10625401

Goryshin, I Y; Jendrisak, J; Hoffman, L M; Meis, R; Reznikoff, W S

2000-01-01

300

Tn916-generated, lipooligosaccharide mutants of Neisseria meningitidis and Neisseria gonorrhoeae.  

PubMed Central

A library of Tn916-generated, tetracycline-resistant (Tc) mutants of the group B Neisseri meningitidis strain NMB was screened by using monoclonal antibodies (MAbs) that recognize structural differences in neisserial lipooligosaccharide (LOS). The LOS of parental strain NMB had a relative molecular mass of 4.5 kDa, reacted with MAbs 3F11 and 6B4 but not with MAb 4C4 or 6E4, and contained a lacto-N-neotetrose unit. Two phenotypically stable mutants, SS3 and R6, altered in LOS, were identified by colony immunoblots, electrophoresis, and Western immunoblots. The LOS of mutant SS3 was 3.4 kDa and reacted with MAbs 4C4 and 6E4 but not MAb 3E11 or 6B4. The LOS of mutant R6 was 3.1 to 3.2 kDa and reacted with MAb 6E4 but not MAb 3F11, 6B4, or 4C4. Thus, the LOSs of the R6 and SS3 mutants were predicted to contain different truncations of the core oligosaccharide. The LOS phenotype of each mutant was linked to Tc(r), as determined by transformation of the parent strain with DNA from the mutant. Southern hybridizations and single-specific-primer PCR revealed in each mutant a single truncated tn916 insertion which had lost genes required for mobilization. Tn916 mutagenesis was used to identify two distinct genetic sites in the meningococcal chromosome involved in biosynthesis of the oligosaccharide chain of LOS and to create genetically defined LOS mutants of N. meningitidis and Neisseria gonorrhoeae. Images

Stephens, D S; McAllister, C F; Zhou, D; Lee, F K; Apicella, M A

1994-01-01

301

County Business Patterns 1995  

NSDL National Science Digital Library

The US Census Bureau has made available the state publications for the 1995 version of its annual County Business Patterns 1995 series (Adobe Acrobat [.pdf] format only). Data available include employment, payroll, and number of establishments by Standard Industrial Code category. Establishments are further broken down by nine employment size classes ranging from 1-4 to 1,000 or more. The US volume is forthcoming. Complete 1993 and 1994 (also .pdf format) series are available at the site.

1997-01-01

302

Characterization of a transposon Tn916-generated mutant of Haemophilus ducreyi 35000 defective in lipooligosaccharide biosynthesis.  

PubMed Central

To define the role of the surface lipooligosaccharide (LOS) of Haemophilus ducreyi in the pathogenesis of chancroid, Tn916 mutants of H. ducreyi 35000 defective in expression of the murine monoclonal antibody (MAb) 3F11 epitope on H. ducreyi LOS were identified by immunologic screening. One mutant, designated 1381, has an LOS which lacks the MAb 3F11 epitope and migrates with an increased mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gene disrupted by the Tn916 element in strain 1381 was identified by cloning the sequences flanking the Tn916 element. The sequences were then used to probe a lambda DASHII genomic library. In strain 1381, Tn916 interrupts a gene which encodes an open reading frame (ORF) with an Mr of 40,246. This ORF has homology to the product of the rfaK gene of Escherichia coli. The major LOS glycoform produced by strain 1381 was analyzed by using a combination of mass spectrometry, linkage and composition analysis, and 1H nuclear magnetic resonance spectroscopy. The major LOS species was found to terminate in a single glucose attached to the heptose (L-glycero-D-manno-heptose, or Hep) trisaccharide core. In the wild-type strain 35000, glucose serves as the acceptor for the addition of the D-glycero-D-manno-heptose (or DDHep), which extends to form the mature branch of the H. ducreyi LOS. This mature oligosaccharide is in turn partially capped by the addition of sialic acid (NeuAc), i.e., NeuAc2 alpha-->3Gal beta1-->4GlcNAc beta1-->3Gal beta1-->4DDHep alpha1-->6Glc beta1 (W. Melaugh et al., Biochemistry 33:13070-13078, 1994). Since this LOS terminates prior to the addition of the branch DD-heptose, this gene is likely to encode the D-glycero-D-manno-heptosyltransferase. Strain 1381 exhibits a significant reduction in adherence to and invasion of primary human keratinocytes. This defect was complemented by the cloned heptosyltransferase gene, indicating that the terminal portion of the LOS oligosaccharide plays an important role in adherence to human keratinocytes.

Gibson, B W; Campagnari, A A; Melaugh, W; Phillips, N J; Apicella, M A; Grass, S; Wang, J; Palmer, K L; Munson, R S

1997-01-01

303

The bleomycin resistance gene of transposon Tn5 is an excellent marker for transformation of corynebacteria.  

PubMed

Corynebacteria are highly sensitive to the glycopeptide antibiotic bleomycin. The bleomycin resistance gene of transposon Tn5 is expressed very efficiently in Brevibacterium lactofermentum. This gene constitutes an excellent marker for selection of transformants of corynebacteria. The bleomycin resistance gene is expressed from the same promoter as the neomycin resistance gene, which is already used as marker in many vectors of corynebacteria. The promoter of the neo-ble cluster is expressed in a variety of Gram-negative and Gram-positive microorganisms and eucaryotic organisms. PMID:1373065

Guerrero, C; Mateos, L M; Malumbres, M; Martín, J F

1992-03-01

304

Transposon insertion mutagenesis of Pseudomonas aeruginosa with a Tn5 derivative: application to physical mapping of the arc gene cluster.  

PubMed

For insertional mutagenesis of Pseudomonas aeruginosa, a derivative of the kanamycin-resistance (KmR) transposon Tn5 was constructed (Tn5-751) that carried the trimethoprim-resistance (TpR) determinant from plasmid R751 as an additional marker. Double selection for KmR and TpR avoided the isolation of spontaneous aminoglycoside-resistant mutants which occur at high frequencies in P. aeruginosa. As a delivery system for the recombinant transposon, plasmid pME305, a derivative of the broad-host-range plasma RP1, proved effective; pME305 is temperature-sensitive at 43 degrees C for maintenance in Escherichia coli and P. aeruginosa and deleted for IS21 and the KmR and primase genes. In matings with an E. coli donor carrying pME9(= pME305::Tn5-751), transposon insertion mutants of P. aeruginosa PAO were recovered at approx. 5 X 10(-7)/donor at 43 degrees C. Among Tn5-751 insertional mutants 0.9% were auxotrophs. A thr::Tn5-751 mutation near the recA-like locus rec-102 is useful for the construction of recombination-deficient strains. Several arc::Tn5-751 mutants could be isolated that were defective in anaerobic utilization of arginine as an energy source. From three of these mutants the arc gene region was cloned into an E. coli vector plasmid. Since Tn5-751 has a single EcoRI site between the TpR and KmR genes, EcoRI-generated fragments carrying either resistance determinant plus adjacent chromosomal DNA could be selected separately in E. coli. Thus, a restriction map of the arc region was constructed and verified by hybridization experiments. The arc genes were tightly clustered, confirming earlier genetic evidence. PMID:2989092

Rella, M; Mercenier, A; Haas, D

1985-01-01

305

The Evaluation of Diagnostic Role of Cardiac Troponin T (cTnT) in Newborns with Heart Defects  

PubMed Central

Heart diseases are a significant cause of morbidity and mortality in newborns. Diagnostic methods are often not sufficient or, in many cases, cannot be used. There is a great advance in medical knowledge concerning biomarkers in the diagnosis of circulatory system in adult patients. Among them, cardiac troponins play the main role. In current literature, there is not enough data concerning the possibility of using them in neonatal cardiac diagnostics. Aim of the Study. To evaluate diagnostic usefulness of cTnT in correlation with other markers of circulatory failure and myocardial damage in newborns with heart defects. Patients and Methods. The study involved 83 newborns up to 46 weeks of postmenstrual age. The exclusion criteria were severe perinatal asphyxia and presence of severe noncardiac diseases. Patients were divided into 2 main groups: group I—54 patients with congenital heart defects (CHDs), and group II (control)—29 healthy neonates. All patients underwent detailed examination of circulatory system. Cardiac troponin T (cTnT) concentrations were evaluated by Roche CARDIAC T Quantitive test. Results. Performed studies revealed that cTnT levels in newborns with heart pathology were significantly higher than in healthy ones. However, cTnT concentrations in patients with CHD did not correlate with clinical symptoms of heart failure, nor with echocardiographic markers of LV function. Type of heart defect did not influence cTnT levels as well. Only hemodynamic significance evaluated by echocardiography influenced the cTnT levels with statistical significance. Conclusions. (1) Statistically significant differences in cTnT levels between newborns with heart defects and healthy subjects were shown. (2) CTnT levels in newborns with heart defects refer only to hemodynamic significance of the defect.

Tarkowska, Agata; Furmaga-Jablonska, Wanda

2012-01-01

306

The Tn7 transposition regulator TnsC interacts with the transposase subunit TnsB and target selector TnsD  

PubMed Central

The excision of transposon Tn7 from a donor site and its insertion into its preferred target site, attachment site attTn7, is mediated by four Tn7-encoded transposition proteins: TnsA, TnsB, TnsC, and TnsD. Transposition requires the assembly of a nucleoprotein complex containing all four Tns proteins and the DNA substrates, the donor site containing Tn7, and the preferred target site attTn7. TnsA and TnsB together form the heteromeric Tn7 transposase, and TnsD is a target-selecting protein that binds specifically to attTn7. TnsC is the key regulator of transposition, interacting with both the TnsAB transposase and TnsD-attTn7. We show here that TnsC interacts directly with TnsB, and identify the specific region of TnsC involved in the TnsB–TnsC interaction during transposition. We also show that a TnsC mutant defective in interaction with TnsB is defective for Tn7 transposition both in vitro and in vivo. Tn7 displays cis-acting target immunity, which blocks Tn7 insertion into a target DNA that already contains Tn7. We provide evidence that the direct TnsB–TnsC interaction that we have identified also mediates cis-acting Tn7 target immunity. We also show that TnsC interacts directly with the target selector protein TnsD.

Choi, Ki Young; Spencer, Jeanelle M.; Craig, Nancy L.

2014-01-01

307

Characterization of the Tn916 conjugative transposon in a food-borne strain of Lactobacillus paracasei.  

PubMed

Food-borne antibiotic-resistant lactic acid bacteria have received growing attention in the past few years. We have recently identified tetracycline-resistant Lactobacillus paracasei in samples of milk and natural whey starter cultures employed in the manufacturing process of a typical Italian fermented dairy product, Mozzarella di Bufala Campana. In the present study, we have characterized at the molecular level the genetic context of tetracycline resistance determinants in these natural strains, which we have identified as tet(M). This gene was present in 21 independent isolates, whose fingerprinting profiles were distributed into eight different repetitive extragenic palindromic groups by cluster analysis. We provide evidence that the gene is associated with the broad-host, conjugative transposon Tn916, which had never before been described to occur in L. paracasei. PCR analysis of four independent isolates by use of specifically designed primer pairs detected the presence of a circular intermediate form of the transposon, carrying a coupling sequence (GGCAAA) located between the two termini of Tn916. This novel coupling sequence conferred low conjugation frequency in mating experiments with the recipient strain JH2-2 of Enterococcus faecalis. PMID:19395574

Devirgiliis, Chiara; Coppola, Doriana; Barile, Simona; Colonna, Bianca; Perozzi, Giuditta

2009-06-01

308

Isolation and characterization of transposon Tn5-induced symbiotic mutants of Rhizobium loti.  

PubMed

Rhizobium loti NZP2037 and NZP2213, each cured of its single large indigenous plasmid, formed effective nodules on Lotus spp., suggesting that the symbiotic genes are carried on the chromosome of these strains. By using pSUP1011 as a vector for introducing transposon Tn5 into R. loti NZP2037, symbiotic mutants blocked in hair curling (Hac), nodule initiation (Noi), bacterial release (Bar), and nitrogen fixation (Nif/Cof) on Lotus pedunculatus were isolated. Cosmids complementing the Hac, Noi, and Bar mutants were isolated from a pLAFR1 gene library of NZP2037 DNA by in planta complementation and found to contain EcoRI fragments of identical sizes to those into which Tn5 had inserted in the mutants. The cosmids that complemented the mutants of these phenotypic classes did not share common fragments, nor did cosmids that complemented four mutants within the Noi class, suggesting that these symbiotically important regions are not tightly linked on the R. loti chromosome. PMID:2984178

Chua, K Y; Pankhurst, C E; Macdonald, P E; Hopcroft, D H; Jarvis, B D; Scott, D B

1985-04-01

309

Isolation and characterization of transposon Tn5-induced symbiotic mutants of Rhizobium loti.  

PubMed Central

Rhizobium loti NZP2037 and NZP2213, each cured of its single large indigenous plasmid, formed effective nodules on Lotus spp., suggesting that the symbiotic genes are carried on the chromosome of these strains. By using pSUP1011 as a vector for introducing transposon Tn5 into R. loti NZP2037, symbiotic mutants blocked in hair curling (Hac), nodule initiation (Noi), bacterial release (Bar), and nitrogen fixation (Nif/Cof) on Lotus pedunculatus were isolated. Cosmids complementing the Hac, Noi, and Bar mutants were isolated from a pLAFR1 gene library of NZP2037 DNA by in planta complementation and found to contain EcoRI fragments of identical sizes to those into which Tn5 had inserted in the mutants. The cosmids that complemented the mutants of these phenotypic classes did not share common fragments, nor did cosmids that complemented four mutants within the Noi class, suggesting that these symbiotically important regions are not tightly linked on the R. loti chromosome. Images

Chua, K Y; Pankhurst, C E; Macdonald, P E; Hopcroft, D H; Jarvis, B D; Scott, D B

1985-01-01

310

Tn5-induced mutations affecting sulfur-oxidizing ability (Sox) of Thiosphaera pantotropha  

SciTech Connect

Mutants of Thiosphaera pantotropha defective in chemolithoautotrophic growth were obtained by transpositional mutagenesis with Tn5 coding for kanamycin resistance. The suicide vehicle for introducing Tn5 to T. pantotropha was pSUP5011 harbored by Escherichia coli. Kanamycin-resistant isolates were screened for the inability to grow with reduced sulfur compounds (Sox/sup -/). Four classes of Sox/sup -/ mutants were obtained. Three were of different pleiotropic phenotypes: (i) unable to grow with formate, nitrate, and xanthine; (this class strongly suggested the involvement of a molybdenum cofactor in inorganic sulfur-oxidizing ability); (ii) no growth with hydrogen; (iii) slight growth with hydrogen and formate. Two plasmids, pHG41 (about 450 kilobase pairs) and pHG42 (110 kilobases), were identified in lysates of T. pantotropha. In one Sox/sup -/ mutant pHG41 could not be detected. Revertant analysis suggested that pHG41 and pHG42 were not involved in the Sox character.

Chandra, T.S.; Friedrich, C.G.

1986-05-01

311

Purification and characterization of cis-aconitic acid decarboxylase from Aspergillus terreus TN484-M1.  

PubMed

cis-Aconitic acid decarboxylase (CAD) was assumed to be a key enzyme in the production of itaconic acid by comparing the activity of CAD from Aspergillus terreus TN484-M1 with that of CAD from the low-itaconate yielding strain Aspergillus terreus CM85J. The constitutive CAD was purified to homogeneity from A. terreus TN484-M1 by ammonium sulfate fractionation, and column chromatography on DEAE-toyopearl, Butyl-toyopearl, and Sephacryl S200HR, and then characterized. A molecular mass of 55 kDa for the native enzyme was determined by SDS-PAGE. The enzymic activity was optimal at a pH of 6.2 and temperature of 45 degrees C. The K(m) value for cis-aconitic acid was determined as 2.45 mM (pH 6.2, 37 degrees C). The enzyme was completely inactivated by Hg+, Cu2+, Zn2+, p-chloromercuribenzoate, and 5,5'-dithio-bis(2-nitrobenzoate). PMID:16233265

Dwiarti, Lies; Yamane, Ken; Yamatani, Hitoshi; Kahar, Prihardi; Okabe, Mitsuyasu

2002-01-01

312

Isolation and symbiotic characterization of transposon Tn5-induced arginine auxotrophs of Sinorhizobium meliloti.  

PubMed

Seventeen arginine auxotrophic mutants of Sinorhizobium meliloti Rmd201 were isolated by random transposon Tn5 mutagenesis using Tn5 delivery vector pGS9. Based on intermediate feeding studies, these mutants were designated as argA/argB/argC/argD/argE (ornithine auxotrophs), argF/argI, argG and argH mutants. The ornithine auxotrophs induced ineffective nodules whereas all other arginine auxotrophs induced fully effective nodules on alfalfa plants. In comparison to the parental strain induced nodule, only a few nodule cells infected with rhizobia were seen in the nitrogen fixation zone of the nodule induced by the ornithine auxotroph. TEM studies showed that the bacteroids in the nitrogen fixation zone of ornithine auxotroph induced nodule were mostly spherical or oval unlike the elongated bacteroids in the nitrogen fixation zone of the parental strain induced nodule. These results indicate that ornithine or an intermediate of ornithine biosynthesis, or a chemical factor derived from one of these compounds is required for the normal development of nitrogen fixation zone and transformation of rhizobial bacteria into bacteroids during symbiosis of S. meliloti with alfalfa plants. PMID:15242285

Kumar, Anvita; Vij, Neeraj; Randhawa, Gursharn S

2003-10-01

313

The Sugar Creek zinc deposit, Jackson Co. TN -- Exploration history, geology and mineralization  

SciTech Connect

During the 60's and 70's zinc exploration of central TN and KY was active. The Sugar Creek Project was one of several investigated by Exxon. The discovery hole, Cu 15, was drilled in early 1973. The Sugar Creek Zinc Deposit was acquired by Independence Mining Co. in 1986 and I.M.C. has subsequently completed additional drilling, both stepout and confirmation holes. A total of 137 holes for 300,833 ft have been drilled. The Sugar Creek deposit is a typical Tennessee zinc deposit (Mississippi Valley Type) which occurs in solution collapse breccias in the Lower Ordovician, Knox Dolomite. The Knox consists of fine grained dolomite with interlayered limestones and crystalline dolomite. Only scattered residual limestone is found in the Sugar Creek area. Collapse breccias have formed which control zinc deposition and are similar to other TN Zn. deposits. At Sugar Creek the types of breccias include: a vertically exaggerated glory hole breakthrough breccia which extends to within 137 ft. of the Knox unconformity, has 500 ft. of zinc mineralization with 8 significant zinc intervals; holes with stacked zinc intervals interpreted to be sides of breakthrough breccia; and single zinc intervals in laterally positioned bedded mineral zones. A total of 99 holes were drilled in the more intense mineralized areas. The ratio of ore to non ore holes is nearly 1 to 1. The mineralization is typical M.V.T. with predominantly sphalerite and only minor occurrences of galena, fluorite, pyrite, etc.

Reinbold, G.; Moran, A.V.; Stevens, D.L. (Independence Mining Co. Inc., Reno, NV (United States))

1993-03-01

314

Tn5044-conferred mercury resistance depends on temperature: the complexity of the character of thermosensitivity.  

PubMed

Escherichia coli K12 containing the transposon Tn5044 mer operon (merR, T, P, C, and A genes) is resistant to mercuric chloride at 30 degrees C but sensitive to this compound at 37-41.5 degrees C. We have studied the mechanism underlying the temperature-sensitive nature of this mercury resistance phenotype, and found that the expression of the Tn5044 merA gene coding for mercuric reductase (MerA) is severely inhibited at non-permissive temperatures. Additionally, MerA showed a considerably reduced functional activity in vivo at non-permissive temperatures. However, the temperature-sensitive character of the functioning of this enzyme in cell extracts, where it interacted with one of the low-molecular weight SH compounds rather than with the transport protein MerT (as is the case in vivo), was not apparent. These data suggest that the temperature-sensitive mercury resistance phenotype should stay under control at two stages: when the merA gene is expressed and when its product interacts with MerT to accept the mercuric ion. PMID:12403178

Kholodii, Gennady; Bogdanova, Elena

2002-06-01

315

Bioactive secondary metabolites from a new terrestrial Streptomyces sp. TN262.  

PubMed

During our search for Streptomyces spp. as new producers of bioactive secondary metabolites, the ethyl acetate extract of the new terrestrial Streptomyces isolate TN262 delivered eight antimicrobially active compounds. They were identified as 1-acetyl-beta-carboline (1), tryptophol (2), cineromycin B (3), 2,3-dihydrocineromycin B (4), cyclo-(tyrosylprolyl) (5), 3-(hydroxyacetyl)-indole (6), brevianamide F (7), and cis-cyclo-(L-prolyl-L-leucyl) (8). Three further metabolites were detected in the unpolar fractions using GC-MS and tentatively assigned as benzophenone (9), N-butyl-benzenesulfonamide (10), and hexanedioic acid-bis-(2-ethylhexyl) ester (11). This last compound is known as plasticizer derivatives, but it has never been described from natural sources. In this article, we describe the identification of the new Streptomyces sp. isolate TN262 using its cultural characteristics, the nucleotide sequence of the corresponding 16S rRNA gene and the phylogenetic analysis, followed by optimization, large-scale fermentation, isolation of the bioactive constituents, and determination of their structures. The biological activity of compounds (2), (3), (4), and those of the unpolar fractions was addressed as well. PMID:19842066

Elleuch, Lobna; Shaaban, Mohamed; Smaoui, Slim; Mellouli, Lotfi; Karray-Rebai, Ines; Fourati-Ben Fguira, Lilia; Shaaban, Khaled A; Laatsch, Hartmut

2010-09-01

316

Molecular Analysis of Tn1546 in Enterococcus faecium Isolated from Animals and Humans  

PubMed Central

The internal areas and the position of integration of the glycopeptide resistance element Tn1546 were characterized by using PCR fragment length polymorphism, sequencing, and DNA hybridization techniques with 38 high-level vancomycin-resistant Enterococcus faecium isolates of human and animal origins from Europe and the United States. Only minor variations in the coding regions within Tn1546 were found, suggesting high genetic stability. The isolates originated from broilers (n = 5), a chicken (n = 1), a duck (n = 1), a turkey (n = 1), pigs (n = 8), a pony (n = 1), and humans (n = 23). A total of 13 different types were defined based on a single-nucleotide difference in the vanX gene, the presence of insertion sequences, and hybridization patterns. For some types more than one isolate were found. For type 1, 10 isolates of both human and animal origins were 1found. All were indistinguishable from the reference strain, BM4147. For type 2, 11 isolates of human and animal origins were found. Six human isolates from England were all of type 3. Two human isolates from the United States, indistinguishable from each other, were type 9. These results showed that vancomycin-resistant E. faecium of animal and human origins can contain indistinguishable genetic elements coding for vancomycin resistance, indicating either horizontal gene transfer between E. faecium organisms of human and animal origins or the existence of a common reservoir for glycopeptide resistance.

Jensen, Lars Bog?; Ahrens, Peter; Dons, Lone; Jones, Ronald N.; Hammerum, Anette M.; Aarestrup, Frank M?ller

1998-01-01

317

A novel dihydrofolate reductase cassette inserted in an integron borne on a Tn21-like element.  

PubMed Central

In this study, a 498-bp dhfrXII gene coding for trimethoprim resistance was found inserted in a cassette-like manner in the recombinationally active locus, the integron, borne on a transposon Tn21-like element. The dhfrXII cassette is distinct from those cassettes earlier observed in integrons and was found here upstream of two similarly inserted cassettes. The second one carried the new unidentified orfF, which is 85% identical to the orfD cassette in R46. The third cassette contained the aadA2 gene mediating spectinomycin resistance. The plasmid carrying this Tn21-like element was originally isolated from a trimethoprim-resistant urinary tract pathogen, Escherichia coli, from Turku City Hospital, Turku, Finland. By colony hybridization and polymerase chain reaction, this group of three cassettes, including dhfrXII, was detected in four additional E. coli strains of similar origin and in four Shigella strains isolated in Finland but originating from Asia. The dihydrofolate reductase produced from dhfrXII showed an unusual drug resistance in that 50% of the enzymatic activity remained at a trimethoprim concentration of 1 mM.

Heikkila, E; Skurnik, M; Sundstrom, L; Huovinen, P

1993-01-01

318

Iron oxide superparamagnetic nanoparticles conjugated with a conformationally blocked ?-Tn antigen mimetic for macrophage activation.  

PubMed

Among new therapies to fight tumors, immunotherapy is still one of the most promising and intriguing. Thanks to the ongoing structural elucidation of several tumor antigens and the development of innovative antigen carriers, immunotherapy is in constant evolution and it is largely used either alone or in synergy with chemotherapy/radiotherapy. With the aim to develop fully synthetic immunostimulants we have recently developed a mimetic of the ?-Tn mucin antigen, a relevant tumor antigen. The (4)C1 blocked mimetic , unique example of an ?-Tn mimetic antigen, was functionalized with an ?-phosphonate linker and used to decorate iron oxide superparamagnetic nanoparticles (MNPs), employed as multivalent carriers. MNPs, largely exploited for supporting and carrying biomolecules, like antibodies, drugs or antigens, consent to combine in the same nanometric system the main features of an inorganic magnetic core with a bioactive organic coating. The superparamagnetic glyconanoparticles obtained, named GMNPs, are indeed biocompatible and immunoactive, and they preserve suitable characteristics for use as heat mediators in the magnetic fluid hyperthermia (MFH) treatment of tumors. All together these properties make GMNPs attracting devices for innovative tumor treatment. PMID:24898009

Manuelli, Massimo; Fallarini, Silvia; Lombardi, Grazia; Sangregorio, Claudio; Nativi, Cristina; Richichi, Barbara

2014-06-12

319

Hydrology of Lake County, Florida  

USGS Publications Warehouse

Lake County includes a 1,150 square-mile area consisting of ridges, uplands, and valleys in central-peninsular Florida. About 32 percent of the county is covered by lakes, swamps, and marshes. Water requirements in 1970 averaged about 54 million gallons per day. About 85 percent of the water was obtained from wells; about 15 percent from lakes. The Floridan aquifer supplies almost all the ground water used in Lake County. Annual recharge to the Floridan aquifer averages about 7 inches over the county; runoff average 8.5 inches. The quality of ground and surface water in Lake County is in general good enough for most uses; however, the poor quality of Floridan-aquifer water in the St. John River Valley probably results from the upward movement of saline water along a fault zone. Surface water in Lake County is usually less mineralized than ground water but is more turbid and colored. (Woodard-USGS)

Knochenmus, Darwin D.; Hughes, G. H.

1976-01-01

320

77 FR 72968 - Revisions to the California State Implementation Plan, for Imperial County, Placer County and...  

Federal Register 2010, 2011, 2012, 2013

...Placer County and Ventura County Air Pollution Control Districts AGENCY: Environmental...revisions to the Imperial County Air Pollution Control District (ICAPCD), Placer County Air Pollution Control District (PCAPCD)...

2012-12-07

321

77 FR 73005 - Revisions to the California State Implementation Plan, Imperial County, Placer County, and...  

Federal Register 2010, 2011, 2012, 2013

...Placer County, and Ventura County Air Pollution Control Districts AGENCY: Environmental...revisions to the Imperial County Air Pollution Control District (ICAPCD), Placer County Air Pollution Control District (PCAPCD)...

2012-12-07

322

Identification of a novel composite transposable element, Tn5280, carrying chlorobenzene dioxygenase genes of Pseudomonas sp. strain P51.  

PubMed

Analysis of one of the regions of catabolic plasmid pP51 which encode chlorobenzene metabolism of Pseudomonas sp. strain P51 revealed that the tcbA and tcbB genes for chlorobenzene dioxygenase and dehydrogenase are located on a transposable element, Tn5280. Tn5280 showed the features of a composite bacterial transposon with iso-insertion elements (IS1066 and IS1067) at each end of the transposon oriented in an inverted position. When a 12-kb HindIII fragment of pP51 containing Tn5280 was cloned in the suicide donor plasmid pSUP202, marked with a kanamycin resistance gene, and introduced into Pseudomonas putida donor plasmid pSUP202, marked with a kanamycin resistance gene, and introduced into Pseudomonas putida KT2442, Tn5280 was found to transpose into the genome at random and in single copy. The insertion elements IS1066 and IS1067 differed in a single base apir located in the inner inverted repeat and were found to be highly homologous to a class of repetitive elements of Bradyrhizobium japonicum and distantly related to IS630 of Shigella sonnei. The presence of the catabolic genes tcbA and tcbB on Tn5280 suggests a mechanism by which gene clusters can be mobilized as gene cassettes and joined with others to form novel catabolic pathways. PMID:1657878

van der Meer, J R; Zehnder, A J; de Vos, W M

1991-11-01

323

Integrative gene cloning and expression system for Streptomyces sp. US 24 and Streptomyces sp. TN 58 bioactive molecule producing strains.  

PubMed

Streptomyces sp. US 24 and Streptomyces sp. TN 58, two strains producing interesting bioactive molecules, were successfully transformed using E. coli ET12567 (pUZ8002), as a conjugal donor, carrying the integrative plasmid pSET152. For the Streptomyces sp. US 24 strain, two copies of this plasmid were tandemly integrated in the chromosome, whereas for Streptomyces sp. TN 58, the integration was in single copy at the attB site. Plasmid pSET152 was inherited every time for all analysed Streptomyces sp. US 24 and Streptomyces sp. TN 58 exconjugants under nonselective conditions. The growth, morphological differentiation, and active molecules production of all studied pSET152 integrated exconjugants were identical to those of wild type strains. Consequently, conjugal transfer using pSET152 integration system is a suitable means of genes transfer and expression for both studied strains. To validate the above gene transfer system, the glucose isomerase gene (xylA) from Streptomyces sp. SK was expressed in strain Streptomyces sp. TN 58. Obtained results indicated that heterologous glucose isomerase could be expressed and folded effectively. Glucose isomerase activity of the constructed TN 58 recombinant strain is of about eighteenfold higher than that of the Streptomyces sp. SK strain. Such results are certainly of importance due to the potential use of improved strains in biotechnological process for the production of high-fructose syrup from starch. PMID:19547659

Sioud, Samiha; Aigle, Bertrand; Karray-Rebai, Ines; Smaoui, Slim; Bejar, Samir; Mellouli, Lotfi

2009-01-01

324

Participation of the recA determinant in the transposition of class II transposon mini-TnMERI1.  

PubMed

As an initial step to understand the mobile nature of class II mercury resistance transposon TnMERI1, the effect of the recA gene on translocation of mini-TnMERI1 was evaluated. A higher transposition frequency in the LE392 strain (2.4+/-1.2x10(-5)) than in the recA-deficient DH1 strain (1.2+/-0.8x10(-6)) indicated participation of the recA gene in mini-TnMERI1 transposition. Introduction of the recA gene into the DH1 strain complemented the transposition frequency at the same level as in LE392 and confirmed participation of the recA gene in transposition. However, treatment of cells by stress agents, including irradiation of up to 3000 Jm(-2) UV doses, did not alter the transposition frequency and suggested independence of RecA from the SOS stress response. Further analysis of transconjugants indicated participation of RecA in the resolution of the cointegrate structure of the transposon. These results suggested that RecA is a constitutive cellular factor that increases translocation of mini-TnMERI1 and may participate in dissemination of TnMERI1-like transposons. PMID:16243449

Matsui, Kazuaki; Narita, Masaru; Ishii, Hidenori; Endo, Ginro

2005-12-15

325

Fast electro-optic switching of frequency modulation TN-LCDs fabricated by doping nanoparticles and their mechanism  

NASA Astrophysics Data System (ADS)

Electro-optic characteristics of a twisted nematic (TN) liquid crystal display (LCD) fabricated by doping Ag nanoparticles protected with NLC, 5CB (K-15, Merck) molecules has been investigated and it is shown that the device exhibits a unique electrooptic response characteristics that is sensitive to the high frequency component of the operating voltage together with the conventional root-mean-square response to the operating voltage. We call this device frequency modulation (FM) TN-LCD. The FM TN-LCD is switched by switching the frequency of the operating voltage while its amplitude is kept unchanged. As an example, by switching the frequency from 20Hz to 500Hz or 50Hz to 2kHz, and the switching is done with the time constant of several ms. This device shows a peculiar response to a burst AC square voltage waveforms, where the device exhibits bipolar response with the time constant of 500 ms and the envelope with the time constants shorter than those of an undoped TN-LCD by from 30% to 80%. The mechanism of the FM-TN-LCD is investigated through the study of its dielectric properties.

Miyama, Tomohiro; Kundu, Sudarshan; Shiraki, Hiroyuki; Sakai, Yoshio; Shiraishi, Yukihide; Toshima, Naoki; Kobayashi, Shunsuke

2004-05-01

326

Digital Atlas of Texas Counties  

NSDL National Science Digital Library

Created by the Center for Geosptial Technology at Texas Tech University, this digital atlas is a fantastic find. It brings together information about all of Texas's counties, including satellite relief maps and data sets about the roads, rivers, lakes, and other features in each area. Visitors can use the interactive map to click on a county of interest, or use the drop down menu to select a region. Once users select a county, they can download information about it for future use. The site also contains a number of Featured Links to resources from the U.S. Census Bureau, the Texas Association of Counties, and several tourism agencies.

327

75 FR 49016 - County of Greenville, S.C.-Acquisition Exemption-Greenville County Economic Development Corporation  

Federal Register 2010, 2011, 2012, 2013

...Exemption--Greenville County Economic Development Corporation The County of...acquire from Greenville County Economic Development Corporation (GCEDC) approximately...See Greenville County Economic Development...

2010-08-12

328

73 FR 46416 - Medicare Program; Prospective Payment System and Consolidated Billing for Skilled Nursing...  

Federal Register 2010, 2011, 2012, 2013

...1.0548 MD. Frederick County, MD Montgomery County, MD 13740...8155 Radford, VA. Giles County, VA Montgomery County, VA Pulaski County, VA Radford...Christian County, KY Trigg County, KY Montgomery County, TN Stewart County, TN...

2008-08-08

329

Cloning and functional analysis of the sequences flanking mini-Tn5 in the magnetosomes deleted mutant NM4 of Magnetospirillum gryphiswaldense MSR1  

Microsoft Academic Search

A magnetosome deleted mutant NM4 of Magnetospirillum gryphiswaldense MSR-1 was generated by mini-Tn5 transposon mutagenesis, and a 5045-bp fragment flanking mini-Tn5 in NM4 was cloned by Anchored\\u000a PCR. Sequencing analysis showed that this fragment involved six putative open reading frames (ORFs); the mini-Tn5 was inserted\\u000a into ORF4. Functional complementary test indicated that the 5045-bp fragment was required for biosynthesis of

Feng Li; Ying Li; Wei Jiang; Zhenfang Wang; Jilun Li

2005-01-01

330

Establishment of an Arbitrary PCR for Rapid Identification of Tn917 Insertion Sites in Staphylococcus epidermidis: Characterization of Biofilm-Negative and Nonmucoid Mutants  

Microsoft Academic Search

Transposon mutagenesis with the Enterococcus faecalis transposon Tn917 is a genetic approach frequently used to identify genes related with specific phenotypes in gram-positive bacteria. We established an arbitrary PCR for the rapid and easy identification of Tn917 insertion sites in Staphylococcus epidermidis with six independent, well-characterized biofilm-negative Tn917 transposon mutants, which were clustered in the icaADBC gene locus or harbor

Johannes K.-M. Knobloch; Max Nedelmann; Kathrin Kiel; Katrin Bartscht; Matthias A. Horstkotte; Sabine Dobinsky; Holger Rohde; Dietrich Mack

2003-01-01

331

Bifunctional gfp-and gusA-containing mini-Tn 5 transposon derivatives for combined gene expression and bacterial localization studies  

Microsoft Academic Search

The gfp gene, encoding the green fluorescent protein, was combined with the gusA gene, coding for the ?-glucuronidase enzyme, in mini-Tn5 transposon derivatives for use in Gram-negative bacteria. These mini-Tn5 elements allow simultaneously monitoring of gene expression and localization of the marked bacteria. Introduction of the resultant mini-Tn5 transposons into Rhizobium etli, Azospirillum brasilense and Pseudomonas stutzeri allowed us to

Chuanwu Xi; Mark Lambrecht; Jos Vanderleyden; Jan Michiels

1999-01-01

332

Sequencing and expression of the 6'-N-acetyltransferase gene of transposon Tn1331 from Klebsiella pneumoniae.  

PubMed Central

Plasmid-mediated amikacin resistance in Klebsiella pneumoniae resides on a 1.5-kilobase BamHI fragment which is part of the Tn3-related multiresistance transposon Tn1331. In this work, we present the complete nucleotide sequence of the amikacin resistance gene and the neighboring sequences. Maxicell experiments detected only one polypeptide of 23 kilodaltons, the product of one of the open reading frames identified as ORF I. Comparison of the complete sequence with that of Tn3 indicated that 396 base pairs located just upstream from ORF I are identical to a region between the end of the tnpR gene and the first six amino acids of the beta-lactamase transcript. Sequences which may act as hot spots for recombination were identified. One was located just after amino acid 6 of beta-lactamase, and the other was located at the end of the amikacin resistance gene. Images

Nobuta, K; Tolmasky, M E; Crosa, L M; Crosa, J H

1988-01-01

333

Hydrostatic pressure effects on TN for single-crystal U(Pt0.98Pd0.02)3  

NASA Astrophysics Data System (ADS)

Substitution of Pd for Pt in the heavy-fermion superconductor UPt3 is known to suppress superconductivity and induce conventional antiferromagnetic order for x >= 0.006. It has been postulated that the primary effect of Pd is to exert a negative pressure, and that there is an antiferromagnetic quantum critical point (QCP) at x = 0.006. We are testing this through application of hydrostatic pressure on a single crystal sample of U(Pt0.98Pd0.02)3 with ordering temperature TN = 3.30 K by crossing the QCP from the ordered state. Our first measurements for temperatures above 2 K indicate that the pressure required to suppress the TN to 0 K is approximately 6 +/- 1 kbar. We are now extending our measurements to lower temperatures (a) to refine this estimate, and (b) to look for signatures of quantum critical behavior in the low-temperature transport when TN = 0 K.

Cuff, Diana; Graf, Michael

2007-03-01

334

Bleomycin-kanamycin resistance as a marker of the presence of transposon Tn5 in clinical strains of Escherichia coli.  

PubMed

The aminoglycoside modifying enzyme aminoglycoside 3'-phosphotransferase II (APH(3')II) is encoded for on transposon Tn5 by the aphA gene, in the same operon as the ble gene determining bleomycin resistance. To document this linkage 82 kanamycin-resistant Escherichia coli strains of clinical origin were studied; all 18 isolates presenting bleomycin-kanamycin resistance were shown by an enzymatic assay to produce APH(3')II, and the presence of Tn5 was demonstrated by gene hybridization. Similarly, bleomycin-kanamycin resistance was shown to be linked to APH(3')II production in Salmonella spp. The epidemiology of strains with Tn5-encoded APH(3')II may thus be studied, at least in Escherichia coli, by a simple diffusion test using bleomycin and kanamycin discs. PMID:2480898

Baquero, F; Saldaña, M A; Blazquez, J; Palacios, R G; Aguiar, J M; Martinez, J L; Vicente, M F; Rubio, C; Gómez-Lus, R

1989-11-01

335

Inducible transfer of conjugative transposon Tn1545 from Enterococcus faecalis to Listeria monocytogenes in the digestive tracts of gnotobiotic mice.  

PubMed

Transfer of conjugative transposon Tn1545 from Enterococcus faecalis to Listeria monocytogenes was studied in vitro and in vivo. Tn1545 transferred following filter mating at a frequency of 2.5 x 10(-7) transconjugants per donor colony. A 20-fold increase in transfer frequency was observed when matings were performed in the presence of a subinhibitory concentration of tetracycline. The frequency of in vivo transfer of Tn1545, expressed as the number of transconjugants per donor cell extracted from the intestines of the gnotobiotic mice after 35 days of experiment, was 1.1 x 10(-8). Presence of a low concentration of tetracycline in the drinking water increased this frequency 10-fold. PMID:1849709

Doucet-Populaire, F; Trieu-Cuot, P; Dosbaa, I; Andremont, A; Courvalin, P

1991-01-01

336

Glycan elongation beyond the mucin associated Tn antigen protects tumor cells from immune-mediated killing.  

PubMed

Membrane bound mucins are up-regulated and aberrantly glycosylated during malignant transformation in many cancer cells. This results in a negatively charged glycoprotein coat which may protect cancer cells from immune surveillance. However, only limited data have so far demonstrated the critical steps in glycan elongation that make aberrantly glycosylated mucins affect the interaction between cancer cells and cytotoxic effector cells of the immune system. Tn (GalNAc-Ser/Thr), STn (NeuAc?2-6GalNAc-Ser/Thr), T (Gal?1-3GalNAc-Ser/Thr), and ST (NeuAc?2-6Gal?1-3GalNAc-Ser/Thr) antigens are recognized as cancer associated truncated glycans, and are expressed in many adenocarcinomas, e.g. breast- and pancreatic cancer cells. To investigate the role of the cancer associated glycan truncations in immune-mediated killing we created glyco-engineered breast- and pancreatic cancer cells expressing only the shortest possible mucin-like glycans (Tn and STn). Glyco-engineering was performed by zinc finger nuclease (ZFN) knockout (KO) of the Core 1 enzyme chaperone COSMC, thereby preventing glycan elongation beyond the initial GalNAc residue in O-linked glycans. We find that COSMC KO in the breast and pancreatic cancer cell lines T47D and Capan-1 increases sensitivity to both NK cell mediated antibody-dependent cellular-cytotoxicity (ADCC) and cytotoxic T lymphocyte (CTL)-mediated killing. In addition, we investigated the association between total cell surface expression of MUC1/MUC16 and NK or CTL mediated killing, and observed an inverse correlation between MUC16/MUC1 expression and the sensitivity to ADCC and CTL-mediated killing. Together, these data suggest that up-regulation of membrane bound mucins protects cells from immune mediated killing, and that particular glycosylation steps, as demonstrated for glycan elongation beyond Tn and STn, can be important for fine tuning of the immune escape mechanisms in cancer cells. PMID:24039759

Madsen, Caroline B; Lavrsen, Kirstine; Steentoft, Catharina; Vester-Christensen, Malene B; Clausen, Henrik; Wandall, Hans H; Pedersen, Anders Elm

2013-01-01

337

Glycan Elongation Beyond the Mucin Associated Tn Antigen Protects Tumor Cells from Immune-Mediated Killing  

PubMed Central

Membrane bound mucins are up-regulated and aberrantly glycosylated during malignant transformation in many cancer cells. This results in a negatively charged glycoprotein coat which may protect cancer cells from immune surveillance. However, only limited data have so far demonstrated the critical steps in glycan elongation that make aberrantly glycosylated mucins affect the interaction between cancer cells and cytotoxic effector cells of the immune system. Tn (GalNAc-Ser/Thr), STn (NeuAc?2-6GalNAc-Ser/Thr), T (Gal?1–3GalNAc-Ser/Thr), and ST (NeuAc?2-6Gal?1–3GalNAc-Ser/Thr) antigens are recognized as cancer associated truncated glycans, and are expressed in many adenocarcinomas, e.g. breast- and pancreatic cancer cells. To investigate the role of the cancer associated glycan truncations in immune-mediated killing we created glyco-engineered breast- and pancreatic cancer cells expressing only the shortest possible mucin-like glycans (Tn and STn). Glyco-engineering was performed by zinc finger nuclease (ZFN) knockout (KO) of the Core 1 enzyme chaperone COSMC, thereby preventing glycan elongation beyond the initial GalNAc residue in O-linked glycans. We find that COSMC KO in the breast and pancreatic cancer cell lines T47D and Capan-1 increases sensitivity to both NK cell mediated antibody-dependent cellular-cytotoxicity (ADCC) and cytotoxic T lymphocyte (CTL)-mediated killing. In addition, we investigated the association between total cell surface expression of MUC1/MUC16 and NK or CTL mediated killing, and observed an inverse correlation between MUC16/MUC1 expression and the sensitivity to ADCC and CTL-mediated killing. Together, these data suggest that up-regulation of membrane bound mucins protects cells from immune mediated killing, and that particular glycosylation steps, as demonstrated for glycan elongation beyond Tn and STn, can be important for fine tuning of the immune escape mechanisms in cancer cells.

Madsen, Caroline B.; Lavrsen, Kirstine; Steentoft, Catharina; Vester-Christensen, Malene B.; Clausen, Henrik

2013-01-01

338

Edgecombe County, North Carolina, Land Development Plan.  

National Technical Information Service (NTIS)

The Edgecombe County land development plan has been designed to utilize all possible areas of physical, social and economic potential within the county. The plan is essentially a long range program for the physical development of the county. (Author)

D. L. Pugh

1970-01-01

339

Magnetic Studies of Ca_2CuO_2Cl2 Above and Below T_N  

NASA Astrophysics Data System (ADS)

Single crystal neutron diffraction and magnetization studies of the spin-1/2 square lattice quantum Heisenberg antiferromagnet Ca_2CuO_2Cl2 are reported. The results indicate a Néel temperature of TN ~= 246 K with an in-plane antiferromagnetic spin ordering at lower temperatures. We present a model of the stacking of the spin planes which is unexpectedly different from Sr_2CuO_2Cl2 and related cuprates. Above TN we present evidence for the existence of quasistatic correlated domains consistent with a spin-dimensionality crossover from the Heisenberg to an XY-like or Ising-like model.

Miller, Lance L.; Vaknin, David; Zarestky, Jerel L.

1997-03-01

340

Conjugal transfer of bacterial chromosomes mediated by the RK2 plasmid transfer origin cloned into transposon Tn5.  

PubMed

We report here a novel system for the conjugal transfer of bacterial chromosomes which utilizes the transfer origin (oriT) of plasmid RK2 cloned into transposon Tn5. Tn5 with oriT was inserted by transposition into the chromosomes of Escherichia coli and Rhizobium meliloti. The oriT sequence then served as the origin of high-frequency chromosome transfer when a helper RK2 plasmid was present in the same cell. The broad host range features of RK2 make this system of oriented chromosome mobilization applicable to most gram-negative bacteria. PMID:6090433

Yakobson, E A; Guiney, D G

1984-10-01

341

Physical mapping of transposon Tn5 insertions defines a gene cluster functional in nitrous oxide respiration by Pseudomonas stutzeri.  

PubMed

By transposon Tn5 mutagenesis, 19 strains of Pseudomonas stutzeri were acquired that had defects in nitrous oxide respiration (Nos- phenotype). A physical map of the mutants showed nearly random Tn5 insertions into genomic DNA within a single region ca. 8 kilobases long. Mutants were characterized immunochemically, enzymatically, and chemically. Several functions related to the synthesis and regulation of nitrous oxide reductase were associated with this DNA region, indicating that in P. stutzeri part of the genetic information necessary to respire nitrous oxide is clustered. PMID:2820935

Viebrock, A; Zumft, W G

1987-10-01

342

The MerE protein encoded by transposon Tn 21 is a broad mercury transporter in Escherichia coli  

Microsoft Academic Search

In order to clarify the physiological role of the merE gene of transposon Tn21, a pE4 plasmid that contained the merR gene of plasmid pMR26 from Pseudomonas strain K-62, and the merE gene of Tn21 from the Shigella flexneri plasmid NR1 (R100) was constructed. Bacteria with plasmid pE4 (merR-o\\/p-merE) were more hypersensitive to CH3Hg(I) and Hg(II), and took up significantly

Masako Kiyono; Yuka Sone; Ryosuke Nakamura; Hidemitsu Pan-Hou; Kou Sakabe

2009-01-01

343

Equity and County College Financing.  

ERIC Educational Resources Information Center

An analysis by the Commission on Financial Postsecondary Education of the system by which New Jersey supports county colleges revealed disparities and inequities among the New Jersey counties in wealth and effort per full-time equivalent student, and current state financing practices that aggravate this problem. In response, alternative state…

New Jersey State Commission on Financing Postsecondary Education, Trenton.

344

Housing Cuyahoga County: Housing Sales.  

National Technical Information Service (NTIS)

The report is one of four which, together, constitute the second year work program of the Cuyahoga County Housing Study. It presents and analyzes data on sales prices of one-family houses in the county, both in aggregate and by small area, in terms of loc...

1971-01-01

345

Menominee: Wisconsin's 72nd County.  

ERIC Educational Resources Information Center

Menominee Indian Reservation became a county after the 1960 Census of Population; therefore, data for the county as a unit were distributed throughout the census publication and appeared as civil division tabulation. This 1963 report attempts to compile these data, as well as data from previous census publications, and present them in easily…

Weidemann, Wayne H.; Fuguitt, Glenn V.

346

Stereoselective total synthesis of glycopeptides bearing the dimeric and trimeric sialosyl-Tn epitope.  

PubMed

The dimeric and trimeric sialosyl-Tn epitopes, [alpha-D-Neup5Ac-(2----6)-alpha-D-GalpNAc-(1----3)-L-Ser]n-L-Val (n = 2 and 3), which represent part of a clustered carbohydrate region of glycophorin A, a human erythrocyte glycoprotein, have been synthesised stereoselectively. 2-Azido-3-O-benzyl-4,6-O-benzylidene-2-deoxy-D-galactopyranosyl fluoride (GalpNAc unit), Fmoc-L-serine phenacyl ester (Ser unit), and benzyl 5-acetamido-4,7,8,9-tetra-O-benzyl-5-deoxy-3-S-phenyl-3-thio-D-erythro-L - gluco-2-nonulopyranosylonate bromide (Neup5Ac unit) were the key intermediates for stereoselective glycosylation. 2-Ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline-promoted elongation of the peptide chain and then hydrogenolysis afforded the title compounds. PMID:1724631

Nakahara, Y; Iijima, H; Shibayama, S; Ogawa, T

1991-09-01

347

Radiative capture of polarized neutrons by polarized protons at Tn=183 MeV  

NASA Astrophysics Data System (ADS)

In order to provide a quantitative test of theoretical calculations incorporating meson-exchange currents and intermediate ? resonances, we measure the normal-component spin correlation coefficient CNN, the differential cross section d?/d?, and the neutron and proton analyzing powers An and Ap, each as a function of angle, for n-->p-->-->d? at Tn=183 MeV. Our n-->p-->-->d? results, combined with the previous cross section and photon asymmetry data collected in the past decade, place quite strong constraints on model calculations. Our data are in excellent agreement with theoretical predictions by Jaus and Woolcock that incorporate meson-exchange and isobar current effects and relativistic corrections, signifying great recent progress in our understanding of these effects in the nucleon-nucleon system.

Xu, G.; Pate, S. F.; Bloch, C.; Vigdor, S. E.; Bowyer, S. M.; Bowyer, T. W.; Jacobs, W. W.; Meyer, H. O.; Pierce, E.; Sowinski, J.; Whiddon, C.; Wissink, S. W.; Jolivette, P. L.; Pickar, M. A.

1995-12-01

348

Weak Ferromagnetism above TN in Gd_2CuO_4  

NASA Astrophysics Data System (ADS)

In this work we report magnetization and Raman experiments performed in single crystals of Gd_2CuO_4. We show the presence of weak ferromagnetism (WF) up to T ~ 650 K, well above T_N ~ 280 K. We have studied the temperature dependence of the intensity of the two forbidden Raman modes, B_1g^* and B_2g^* , and the WF component between 2 K and 800 K. The WF component follows the temperature evolution of the relative intensity of the two forbidden Raman modes and both vanishing for T>650 K. Recent neutron diffraction experiments showed an orthorhombic-tetragonal transition at ~ 650 K. Thus, our results may be interpreted in terms of structural distortions within the CuO2 planes, that disappear at the transition temperature.

Martinho, H.; Sanjurjo, J. A.; Rettori, C.; Martin, A. A.; Oseroff, S. B.; Fisk, Z.

2000-03-01

349

2D ferromagnetic fluctuation above TN in orbital-ordered LaMnO 3  

NASA Astrophysics Data System (ADS)

Onset of two-dimensional (2D) ferromagnetic (FM) short-range order with T 2 D?150 K above A-type antiferromagnetic (AF) Néel temperature T N=139 K was observed in 100 G low field for orbital-ordered LaMnO 3. The A-type AF long-range order is stabilized through strong 2D basal plane FM superexchange coupling of orbital-ordered e g electrons and weak c-axis AF coupling. Both T2D and TN are field dependent, and merge in 1 T high field around 130 K. Mn K-edge X-ray absorption near-edge spectrum for orbital-ordered LaMnO 3 at room temperature indicates a pre-edge splitting of 2.0 eV due to weakly allowed 1s-3d dipole transition.

Chen, C. T.; Wu, H. H.; Lin, B. N.; Hsu, Y. Y.; Ku, H. C.

2003-05-01

350

Localized transposon Tn5 mutagenesis of the photosynthetic gene cluster of Rhodobacter sphaeroides.  

PubMed

Four genes essential for bacteriochlorophyll biosynthesis were known to be encoded within a 45 kb region of the Rhodobacter sphaeroides genome, the boundaries of which are defined by puh and puf genes for reaction-centre and light-harvesting LH1 complexes. The cluster is represented by eight overlapping inserts cloned in the mobilizable vector pSUP202. We have used localized transposon Tn5 mutagenesis to characterize this cluster further; a total of 87 independent insertions were generated which identify nine genes for bacteriochlorophyll biosynthesis, six for carotenoid biosynthesis, and puhA encoding the reaction-centre H subunit. This work provides an essential framework for a detailed study of the structure and expression of genes for photosynthesis in this bacterium. PMID:2170816

Coomber, S A; Chaudhri, M; Connor, A; Britton, G; Hunter, C N

1990-06-01

351

Protein fusions with the kanamycin resistance gene from transposon Tn5.  

PubMed

The gene for the neomycin phosphotransferase II (NPT II) from transposon Tn5 was fused at the amino or carboxy terminus to foreign DNA sequences coding for 3-300 amino acids and the properties of the fused proteins were investigated. All amino-terminal fusions examined conferred kanamycin resistance to their host cell, but profound differences in their enzymatic activity and stability were detected. Short additions to the amino terminus of the NPT II resulted in highly enzymatically active fusion proteins whereas long amino-terminal fusions often had to be proteolytically degraded to release active proteins. Fusions at the carboxy-terminal end of the NPT II protein did not always induce kanamycin resistance and their enzymatic activity depended more stringently on the nature of the junction sequence. PMID:6098471

Reiss, B; Sprengel, R; Schaller, H

1984-12-20

352

A double diastereoselective Michael-type addition as an entry to conformationally restricted tn antigen mimics.  

PubMed

A totally stereocontrolled C-Michael addition of serine-equivalent C-nucleophiles to tri-O-benzyl-2-nitro-d-galactal was used as the key step to synthesize several pyrano[3,2-b]pyrrole structures. These scaffolds could be regarded as conformationally restricted Tn antigen mimics, as we have demonstrated by biological assays. The pyranose rings retain their (4)C1 chair conformation, as shown by molecular modeling and NMR spectroscopy. The expected bioactivity was established by a competition-tailored enzyme-linked lectin assay using both soybean and Vicia villosa agglutinins as model lectins. The facile described synthetic route and the strategic combination of computational and experimental techniques to reveal conformational features and bioactivity demonstrate the prepared glycomimics to be promising candidates for further exploitation of this scaffold to give glycans for lectin blocking and vaccination. PMID:24083620

Aydillo, Carlos; Navo, Claudio D; Busto, Jesús H; Corzana, Francisco; Zurbano, María M; Avenoza, Alberto; Peregrina, Jesús M

2013-11-01

353

Using transposon Tn5 insertions to sequence bacteriophage T4 gene 11.  

PubMed

A simple and rapid method of creating an overlapping set of deletions in cloned DNA in preparation for sequencing has been developed. The method is based on a positive selection for Tn5 transposition into the cloned DNA fragment on a high-copy-number filamid, resolution of potential filamid dimers by filamentous phage infection, and the use of Tn5 both as a "portable" restriction enzyme site for in vitro DNA deletion and a "portable" sequencing primer binding site to initiate DNA sequencing reactions using a custom primer complementary to the outside ends of IS50. This new method has been utilized to sequence bacteriophage T4 gene 11, encoding the T4 baseplate protein gp11. The coding sequence of gene 11 is 657 bp in length, and predicts a primary structure of 219 amino acids that agrees well with the biochemical data previously obtained. DNA sequence around gene 11 suggests that the expression of genes 10, 11, and 12 of phage T4 are translationally coupled. Plasmids carrying deletions generated using this method have been used to map genetically five amber alleles of gene 11. These amber alleles were sequenced to confirm the proposed reading frame. The five amber alleles actually represent two different mutational changes at either codon 206 or 207, changing these adjacent glutamine codons to amber. The position of these amber alleles lends support to earlier studies identifying the carboxyl terminus of gp11 as essential in the interaction of P11 with baseplate protein P10 (Plishker and Berget, 1984). PMID:2548819

Barrett, B K; Berget, P B

1989-05-01

354

TN-68 Spent Fuel Transport Cask Analytical Evaluation for Drop Events  

SciTech Connect

The U.S. Nuclear Regulatory Commission (NRC) is responsible for licensing commercial spent nuclear fuel transported in casks certified by NRC under the Code of Federal Regulations (10 CFR), Title 10, Part 71 [1]. Both the International Atomic Energy Agency regulations for transporting radioactive materials [2, paragraph 727], and 10 CFR 71.73 require casks to be evaluated for hypothetical accident conditions, which includes a 9-meter (m) (30-ft) drop-impact event onto a flat, essentially unyielding, horizontal surface, in the most damaging orientation. This paper examines the behavior of one of the NRC certified transportation casks, the TN-68 [3], for drop-impact events. The specific area examined is the behavior of the bolted connections in the cask body and the closure lid, which are significantly loaded during the hypothetical drop-impact event. Analytical work to evaluate the NRC-certified TN-68 spent fuel transport cask [3] for a 9-m (30-ft) drop-impact event on a flat, unyielding, horizontal surface, was performed using the ANSYS® [4] and LS DYNA™ [5] finite-element analysis codes. The models were sufficiently detailed, in the areas of bolt closure interfaces and containment boundaries, to evaluate the structural integrity of the bolted connections under 9-m (30-ft) free-drop hypothetical accident conditions, as specified in 10 CFR 71.73. Evaluation of the cask for puncture, caused by a free drop through a distance of 1-m (40-in.) onto a mild steel bar mounted on a flat, essentially unyielding, horizontal surface, required by 10 CFR 71.73, was not included in the current work, and will have to be addressed in the future. Based on the analyses performed to date, it is concluded that, even though brief separation of the flange and the lid surfaces may occur under some conditions, the seals would close at the end of the drop events, because the materials remain elastic during the duration of the event.

Shah, M. J.; Klymyshyn, Nicholas A.; Adkins, Harold E.; Koeppel, Brian J.

2007-03-30

355

Hydrology of Polk County, Florida  

USGS Publications Warehouse

Local water managers usually rely on information produced at the State and regional scale to make water-resource management decisions. Current assessments of hydrologic and water-quality conditions in Polk County, Florida, commonly end at the boundaries of two water management districts (South Florida Water Management District and the Southwest Florida Water Management District), which makes it difficult for managers to determine conditions throughout the county. The last comprehensive water-resources assessment of Polk County was published almost 40 years ago. To address the need for current countywide information, the U.S. Geological Survey began a 3?-year study in 2002 to update information about hydrologic and water-quality conditions in Polk County and identify changes that have occurred. Ground-water use in Polk County has decreased substantially since 1965. In 1965, total ground-water withdrawals in the county were about 350 million gallons per day. In 2002, withdrawals totaled about 285 million gallons per day, of which nearly 95 percent was from the Floridan aquifer system. Water-conservation practices mainly related to the phosphate-mining industry as well as the decrease in the number of mines in operation in Polk County have reduced total water use by about 65 million gallons per day since 1965. Polk County is underlain by three principal hydrogeologic units. The uppermost water-bearing unit is the surficial aquifer system, which is unconfined and composed primarily of clastic deposits. The surficial aquifer system is underlain by the intermediate confining unit, which grades into the intermediate aquifer system and consists of up to two water-bearing zones composed of interbedded clastic and carbonate rocks. The lowermost hydrogeologic unit is the Floridan aquifer system. The Floridan aquifer system, a thick sequence of permeable limestone and dolostone, consists of the Upper Floridan aquifer, a middle semiconfining unit, a middle confining unit, and the Lower Floridan aquifer. The Upper Floridan aquifer provides most of the water required to meet demand in Polk County. Data from about 300 geophysical and geologic logs were used to construct hydrogeologic maps showing the tops and thicknesses of the aquifers and confining units within Polk County. Thickness of the surficial aquifer system ranges from several feet thick or less in the extreme northwestern part of the county and along parts of the Peace River south of Bartow to more than 200 feet along the southern part of the Lake Wales Ridge in eastern Polk County. Thickness of the intermediate aquifer system/intermediate confining unit is highly variable throughout the county because of past erosional processes and sinkhole formation. Thickness of the unit ranges from less than 25 feet in the extreme northwestern part of the county to more than 300 feet in southwestern Polk County. The altitude of the top of the Upper Floridan aquifer in the county ranges from about 50 feet above National Geodetic Vertical Datum of 1929 (NGVD 29) in the northwestern part to more than 250 feet below NGVD 29 in the southern part. Water levels in the Upper Floridan aquifer fluctuate seasonally, increasing during the wet season (June through September) and decreasing during the rest of the year. Water levels in the Upper Floridan aquifer also can change from year to year, depending on such factors as pumpage and climatic variations. In the southwestern part of the county, fluctuations in water use related to phosphate mining have had a major impact on ground-water levels. Hydrographs of selected wells in southwestern Polk County show a general decline in water levels that ended in the mid-1970s. This water-level decline coincides with an increase in water use associated with phosphate mining. A substantial increase in water levels that began in the mid-1970s coincides with a period of decreasing water use in the county. Despite reductions in water use since 1970, howev

Spechler, Rick M.; Kroening, Sharon E.

2007-01-01

356

Modular Evolution of TnGBSs, a New Family of Integrative and Conjugative Elements Associating Insertion Sequence Transposition, Plasmid Replication, and Conjugation for Their Spreading  

PubMed Central

Integrative and conjugative elements (ICEs) have a major impact on gene flow and genome dynamics in bacteria. The ICEs TnGBS1 and TnGBS2, first identified in Streptococcus agalactiae, use a DDE transposase, unlike most characterized ICEs, which depend on a phage-like integrase for their mobility. Here we identified 56 additional TnGBS-related ICEs by systematic genome analysis. Interestingly, all except one are inserted in streptococcal genomes. Sequence comparison of the proteins conserved among these ICEs defined two subtypes related to TnGBS1 or TnGBS2. We showed that both types encode different conjugation modules: a type IV secretion system, a VirD4 coupling protein, and a relaxase and its cognate oriT site, shared with distinct lineages of conjugative elements of Firmicutes. Phylogenetic analysis suggested that TnGBSs evolved from two conjugative elements of different origins by the successive recruitment of a transposition module derived from insertion sequences (ISs). Furthermore, TnGBSs share replication modules with different plasmids. Mutational analyses and conjugation experiments showed that TnGBS1 and TnGBS2 combine replication and transposition upstream promoters for their transfer and stabilization. Despite an evolutionarily successful horizontal dissemination within the genus Streptococcus, these ICEs have a restricted host range. However, we reveal that for TnGBS1 and TnGBS2, this host restriction is not due to a transfer incompatibility linked to the conjugation machineries but most likely to their ability for transient maintenance through replication after their transfer.

Guerillot, Romain; Da Cunha, Violette; Sauvage, Elisabeth; Bouchier, Christiane

2013-01-01

357

Synthetic and Immunological Studies of 5?-N-phenylacetyl sTn to Develop Carbohydrate-Based Cancer Vaccines and to Explore the Impacts of Linkage between Carbohydrate Antigens and Carrier Proteins  

PubMed Central

5?-N-Phenylacetyl sTn (sTnNPhAc), an unnatural derivative of sTn antigen expressed by many tumors, and its ?-linked protein conjugates were prepared and investigated to explore glycoconjugate cancer vaccines. sTnNPhAc?-KLH elicited a robust T cell-dependent immunity. The antiserum derived from sTnNPhAc?- or sTnNPhAc?-KLH-inoculated mice was similarly reactive to sTnNPhAc? and sTnNPhAc? but showed very little reactivity to sTn, NeuNPhAc? (2,3)GalNAc – a regioisomer of sTnNPhAc, isolated phenylacetyl group, and the linker employed to conjugate sTnNPhAc and carrier protein. It was concluded that the sTnNPhAc-elicited immunity was specific for the whole antigen rather than the phenylacetyl group or other partial structures of sTnNPhAc and that the reducing end configuration or linkage of sTnNPhAc did not affect its immunological identity. It was also concluded that a new linker designed to conjugate carbohydrates and proteins did not provoke any immune reaction and that the linker, as well as the associated new and convenient coupling strategy, can be safely used for the development of glycoconjugate vaccines.

Wang, Qianli; Ekanayaka, Sandamali Amarasingha; Wu, Jian; Zhang, Junping; Guo, Zhongwu

2009-01-01

358

Capability Description for NASA's F/A-18 TN 853 as a Testbed for the Integrated Resilient Aircraft Control Project.  

National Technical Information Service (NTIS)

The NASA F/A-18 tail number (TN) 853 full-scale Integrated Resilient Aircraft Control (IRAC) testbed has been designed with a full array of capabilities in support of the Aviation Safety Program. Highlights of the system's capabilities include: 1) a quad-...

C. Hanson

2009-01-01

359

Erythrocyte membrane modification in malignant diseases of myeloid and lymphoreticular tissues. I. Tn-polyagglutination in acute myelocytic leukaemia.  

PubMed

Tn polyagglutination (persistent mixed-field polyagglutination) occurring in a patient with an acute myelocytic leukemia is described. The association of this RBC membrane change, thought to be due to somatic mutation at stem cell level, with a myeloproliferative disorder, has not been previously reported. PMID:1063594

Bird, G W; Wingham, J; Pippard, M J; Hoult, J G; Melikian, V

1976-06-01

360

Establishing exchange bias below TN with polycrystalline Ni0.52 Co0.48 O/Co bilayers  

NASA Astrophysics Data System (ADS)

Exchange-coupled bilayers of polycrystalline ferromagnetic (FM) Co on antiferromagnetic (AFM) Ni0.52 Co0.48 O were investigated with emphasis on the issue of establishing an exchange-bias field, HE , below the AFM ordering temperature, TN . It was found that field-cooling the bilayers through TN provided very little, if any, increase in HE over that produced by deposition of the Co at temperatures far below TN . Further significant aspects of this issue were also examined. The biasing field, HB , needed to be applied only during the deposition of a small fraction (1 nm) of the FM film below TN of the AFM to achieve the maximum HE ; if HB was reversed at any time during the FM deposition, HE was determined by the final direction of HB ; the direction of HE could be reversed after it had been established by applying a reversed field during a post-deposition latent period. These and other findings are considered with respect to clarification of the mechanisms for establishing HE .

Berkowitz, A. E.; Hansen, M. F.; Kodama, R. H.; Tang, Y. J.; Hong, J. I.; Smith, David J.

2005-10-01

361

Relationship between summarizing chemical parameters like AOX, TOC, TN b , and toxicity tests for effluents from the chemical production  

Microsoft Academic Search

Therefore this study was undertaken to investigate, whether there are correlations between summarizing parameters, which potentially indicate hazardous water components, like AOX (adsorbable organic halogen), TOC (total organic carbon) and TN b (total bound nitrogen) and biological effects, observed from different bioassays conducted in the laboratory. The toxic effects were investigated on luminescent bacteria, microcrustaceans and algae in accordance with

G. Gellert

2000-01-01

362

RADIOCARBON MEASUREMENT OF THE BIOGENIC CONTRIBUTION TO SUMMERTIME PM 2.5 AMBIENT AEROSOL IN NASHVILLE, TN  

EPA Science Inventory

Radiocarbon (14C) measurements performed on PM-2.5 samples collected near Nashville, TN from June 21 to July 13, 1999, showed high levels of modern carbon, ranging from 56 to 80% of the total carbon in the samples. Radiocarbon measurements performed on dichloromethane extracts of...

363

Complete DNA sequence, specific Tn5 insertion map, and gene assignment of the carotenoid biosynthesis pathway of Rhodobacter sphaeroides.  

PubMed

The carotenoid biosynthesis genes form a cluster within the genome of Rhodobacter sphaeroides, lying in the middle of a larger cluster and 45 kb in length, which contains genes for bacteriochlorophyll biosynthesis and for the reaction center and light-harvesting apoproteins. The positions and approximate limits of the carotenoid genes were determined previously by localized transposon Tn5 mutagenesis and by comparison with the closely related Rhodobacter capsulatus carotenoid gene cluster. In this report, analysis of the DNA and deduced amino acid sequences of the carotenoid genes in R. sphaeroides are presented. Twenty-five Tn5 insertion mutants were used to produce a base-specific Tn5 insertion map of this region, and carotenoid gene assignment was supported by spectroscopic, ultrastructural, and high-pressure liquid chromatography analyses of these mutants. A region in the 3' end of crtD which affects bacteriochlorophyll biosynthesis was discovered, and CrtA was found to possess a proline-rich C-terminal region containing a repeated (Ala-Pro)n motif. CrtF also showed a high degree of sequence conservation with eukaryotic O-methyltransferases. This study provides gene sequences and assignments based upon a comprehensive structural, spectroscopic, and biochemical analysis of a range of carotenoid biosynthetic mutants; in each mutation, the point of Tn5 insertion is determined accurate to 1 bp on the gene cluster. PMID:7721699

Lang, H P; Cogdell, R J; Takaichi, S; Hunter, C N

1995-04-01

364

Complete DNA sequence, specific Tn5 insertion map, and gene assignment of the carotenoid biosynthesis pathway of Rhodobacter sphaeroides.  

PubMed Central

The carotenoid biosynthesis genes form a cluster within the genome of Rhodobacter sphaeroides, lying in the middle of a larger cluster and 45 kb in length, which contains genes for bacteriochlorophyll biosynthesis and for the reaction center and light-harvesting apoproteins. The positions and approximate limits of the carotenoid genes were determined previously by localized transposon Tn5 mutagenesis and by comparison with the closely related Rhodobacter capsulatus carotenoid gene cluster. In this report, analysis of the DNA and deduced amino acid sequences of the carotenoid genes in R. sphaeroides are presented. Twenty-five Tn5 insertion mutants were used to produce a base-specific Tn5 insertion map of this region, and carotenoid gene assignment was supported by spectroscopic, ultrastructural, and high-pressure liquid chromatography analyses of these mutants. A region in the 3' end of crtD which affects bacteriochlorophyll biosynthesis was discovered, and CrtA was found to possess a proline-rich C-terminal region containing a repeated (Ala-Pro)n motif. CrtF also showed a high degree of sequence conservation with eukaryotic O-methyltransferases. This study provides gene sequences and assignments based upon a comprehensive structural, spectroscopic, and biochemical analysis of a range of carotenoid biosynthetic mutants; in each mutation, the point of Tn5 insertion is determined accurate to 1 bp on the gene cluster.

Lang, H P; Cogdell, R J; Takaichi, S; Hunter, C N

1995-01-01

365

Testing and analyses of the TN24P PWR spent-fuel dry storage cask loaded with consolidated fuel  

Microsoft Academic Search

A performance test of a Transnuclear, Inc. TN-24P storage cask configured for pressurized water reactor (PWR) spent fuel was performed. The work was performed by the Pacific Northwest Laboratory (PNL) and Idaho National Engineering Laboratory (INEL) for the US Department of Energy Office of Civilian Radioactive Waste Management (OCRWM) and the Electric Power Research Institute. The performance test consisted of

M. A. McKinnon; T. E. Michener; M. F. Jensen; G. R. Rodman

1989-01-01

366

The Effects of TN:TP Ratios on the Phytoplankton and Colonial Cyanobacteria in Eutrophic Shallow Lakes  

Microsoft Academic Search

The aim of this study was to assess the effects of nutrient and other environmental factors on phytoplankton community composition and the density of colonial cyanobacteria along the gradients of total nitrogen (TN) to total phosphorus (TP) ratios. Fifteen urban eutrophic lakes with different nutrients concentrations were sampled on a bimonthly basis for two years. In total, the absolutely dominance

Lv Jin; Wu Hongjuan

2010-01-01

367

Establishment of an arbitrary PCR for rapid identification of Tn917 insertion sites in Staphylococcus epidermidis: characterization of biofilm-negative and nonmucoid mutants.  

PubMed

Transposon mutagenesis with the Enterococcus faecalis transposon Tn917 is a genetic approach frequently used to identify genes related with specific phenotypes in gram-positive bacteria. We established an arbitrary PCR for the rapid and easy identification of Tn917 insertion sites in Staphylococcus epidermidis with six independent, well-characterized biofilm-negative Tn917 transposon mutants, which were clustered in the icaADBC gene locus or harbor Tn917 in the regulatory gene rsbU. For all six of these mutants, short chromosomal DNA fragments flanking both transposon ends could be amplified. All fragments were sufficient to correctly identify the Tn917 insertion sites in the published S. epidermidis genomes. By using this technique, the Tn917 insertion sites of three not-yet-characterized biofilm-negative or nonmucoid mutants were identified. In the biofilm-negative and nonmucoid mutant M12, Tn917 is inserted into a gene homologous to the regulatory gene purR of Bacillus subtilis and Staphylococcus aureus. The Tn917 insertions of the nonmucoid but biofilm-positive mutants M16 and M20 are located in genes homologous to components of the phosphoenolpyruvate-sugar phosphotransferase system (PTS) of B. subtilis, S. aureus, and Staphylococcus carnosus, indicating an influence of the PTS on the mucoid phenotype in S. epidermidis. PMID:14532029

Knobloch, Johannes K-M; Nedelmann, Max; Kiel, Kathrin; Bartscht, Katrin; Horstkotte, Matthias A; Dobinsky, Sabine; Rohde, Holger; Mack, Dietrich

2003-10-01

368

Solid-phase synthesis of a pentavalent GalNAc-containing glycopeptide (Tn antigen) representing the nephropathy-associated IgA hinge region.  

PubMed

Incomplete or aberrant glycosylation leading to Tn antigen (GalNAcalpha1-Ser/Thr) expression on human glycoproteins is strongly associated with human pathological conditions, including tumors, certain autoimmune diseases, such as the idiopathic IgA nephropathy, and may modulate immune homeostasis. In addition, the Tn antigen is highly expressed by certain pathogens and plays a role in host-pathogen interactions. To enable experimental approaches to study interactions of the Tn antigen with the immune system and analyze anti-Tn antibody responses in infection or disorders, we generated a Tn-expressing resource that can be used for high-throughput screening. In consideration of IgA nephropathy in which the hinge region is incompletely glycosylated, we used this hinge sequence that encodes five potential glycosylation sites as the ideal template for the synthesis of a Tn antigen-expressing glycopeptide. Inclusion of an N-terminal biotin in the peptide enabled binding to streptavidin-coated ELISA plates as monitored using Helix pomatia agglutinin or anti-Tn monoclonal antibody. We also found that the biotinylated IgA-Tn peptide is a functional acceptor for beta1-3-galactosylation using recombinant T-synthase (beta1-3-galactosyltransferase). Besides its immunochemical functionality as a possible diagnostic tool for IgA nephropathy, the peptide is an excellent substrate for glycan elongation and represents a novel template applicable for glycan-antigen-associated diseases. PMID:20719305

Bolscher, Jan G M; Brevoord, Judith; Nazmi, Kamran; Ju, Tongzhong; Veerman, Enno C I; van Wijk, Joanna A E; Cummings, Richard D; van Die, Irma

2010-09-23

369

Construction of Mobilizable Mini-Tn7 Vectors for Bioluminescent Detection of Gram-Negative Bacteria and Single-Copy Promoter lux Reporter Analysis  

PubMed Central

We describe the construction of mini-Tn7-based broad-host-range vectors encoding lux genes as bioluminescent reporters. These constructs can be mobilized into the desired host(s) by conjugation for chromosomal mini-Tn7-lux integration and are useful for localization of bacteria during infections or for characterizing regulation of promoters of interest in Gram-negative bacteria.

Damron, F. Heath; McKenney, Elizabeth S.; Barbier, Mariette; Liechti, George W.; Schweizer, Herbert P.

2013-01-01

370

The Tn7 transposase is a heteromeric complex in which DNA breakage and joining activities are distributed between different gene products.  

PubMed Central

The bacterial transposon Tn7 translocates by a cut and paste mechanism: excision from the donor site results from double-strand breaks at each end of Tn7 and target insertion results from joining of the exposed 3' Tn7 tips to the target DNA. Through site-directed mutagenesis of the Tn7-encoded transposition proteins TnsA and TnsB, we demonstrate that the Tn7 transposase is a heteromeric complex of these proteins, each protein executing different DNA processing reactions. TnsA mediates DNA cleavage reactions at the 5' ends of Tn7, and TnsB mediates DNA breakage and joining reactions at the 3' ends of Tn7. Thus the double-strand breaks that underlie Tn7 excision result from a collaboration between two active sites, one in TnsA and one in TnsB; the same (or a closely related) active site in TnsB also mediates the subsequent joining of the 3' ends to the target. Both TnsA and TnsB appear to be members of the retroviral integrase superfamily: mutation of their putative DD(35)E motifs blocks catalytic activity. Recombinases of this class require a divalent metal cofactor that is thought to interact with these acidic residues. Through analysis of the metal ion specificity of a TnsA mutant containing a sulfur (cysteine) substitution, we provide evidence that a divalent metal actually interacts with these acidic amino acids. Images

Sarnovsky, R J; May, E W; Craig, N L

1996-01-01

371

Functional Tn5393-Like Transposon in the R Plasmid pRAS2 from the Fish Pathogen Aeromonas salmonicida subspecies salmonicida Isolated in Norway  

PubMed Central

Tn5393c containing strA-strB was identified as part of R plasmid pRAS2 from the fish pathogen Aeromonas salmonicida subsp. salmonicida. This is the first time an intact and active transposon in the Tn5393 family has been reported in an ecological niche other than an agricultural habitat.

L'Abee-Lund, Trine M.; S?rum, Henning

2000-01-01

372

Mush Creek Watershed, Dallas County and Lowndes County, Alabama.  

National Technical Information Service (NTIS)

The proposal is for the development of a watershed project in Dallas and Lowndes Counties, AL. This project includes an accelerated conservation land treatment program and the installation of floodwater retarding structures. The planned combination of con...

1973-01-01

373

Robust rat pulmonary radioprotection by a lipophilic Mn N-alkylpyridylporphyrin, MnTnHex-2-PyP(5+).  

PubMed

With the goal to enhance the distribution of cationic Mn porphyrins within mitochondria, the lipophilic Mn(III)meso-tetrakis(N-n-hexylpyridinium-2-yl)porphyrin, MnTnHex-2-PyP(5+) has been synthesized and tested in several different model of diseases, where it shows remarkable efficacy at as low as 50 µg/kg single or multiple doses. Yet, in a rat lung radioprotection study, at higher 0.6-1 mg/kg doses, due to its high accumulation and micellar character, it became toxic. To avoid the toxicity, herein the pulmonary radioprotection of MnTnHex-2-PyP(5+) was assessed at 50 µg/kg. Fischer rats were irradiated to their right hemithorax (28 Gy) and treated with 0.05 mg/kg/day of MnTnHex-2-PyP(5+) for 2 weeks by subcutaneously-implanted osmotic pumps, starting at 2 h post-radiation. The body weights and breathing frequencies were followed for 10 weeks post-radiation, when the histopathology and immunohistochemistry were assessed. Impact of MnTnHex-2-PyP(5+) on macrophage recruitment (ED-1), DNA oxidative damage (8-OHdG), TGF-?1, VEGF(A) and HIF-1? were measured. MnTnHex-2-PyP(5+) significantly decreased radiation-induced lung histopathological (H&E staining) and functional damage (breathing frequencies), suppressed oxidative stress directly (8-OHdG), or indirectly, affecting TGF-?1, VEGF (A) and HIF-1? pathways. The magnitude of the therapeutic effects is similar to the effects demonstrated under same experimental conditions with 120-fold higher dose of ~5000-fold less lipophilic Mn(III)meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin, MnTE-2-PyP(5+). PMID:24624330

Gauter-Fleckenstein, Benjamin; Reboucas, Julio S; Fleckenstein, Katharina; Tovmasyan, Artak; Owzar, Kouros; Jiang, Chen; Batinic-Haberle, Ines; Vujaskovic, Zeljko

2014-01-01

374

Identification of a circular intermediate in the transfer and transposition of Tn4555, a mobilizable transposon from Bacteroides spp.  

PubMed Central

Transmissible cefoxitin (FX) resistance in Bacteroides vulgatus CLA341 was associated with the 12.5-kb, mobilizable transposon, Tn4555, which encoded the beta-lactamase gene cfxA. Transfer occurred by a conjugation-like mechanism, was stimulated by growth of donor cells with tetracycline (TC), and required the presence of a Bacteroides chromosomal Tcr element. Transconjugants resistant to either FX, TC, or both drugs were obtained, but only Fxr Tcr isolates could act as donors of Fxr in subsequent matings. Transfer of Fxr could be restored in Fxr Tcs strains by the introduction of a conjugal Tcr element from Bacteroides fragilis V479-1. A covalently closed circular DNA form of Tn4555 was observed in donor cells by Southern hybridization, and the levels of this circular transposon increased significantly in cells grown with TC. Both the cfxA gene and the Tn4555 mobilization region hybridized to the circular DNA, suggesting that this was a structurally intact transposon unit. Circular transposon DNA purified by CsCl-ethidium bromide density gradient centrifugation was used to transform Tcs B. fragilis 638, and Fxr transformants were obtained. Both the circular form and the integrated Tn4555 were observed in transformants, but the circular form was present at less than one copy per chromosomal equivalent. Examination of genomic DNA from Fxr transformants and transconjugants revealed that Tn4555 could insert at a wide variety of chromosomal sites. Multiple transposon insertions were present in many of the transconjugants, indicating that there was no specific barrier to the introduction of a second transposon copy. Images

Smith, C J; Parker, A C

1993-01-01

375

Use of the promoter fusion transposon Tn5 lac to identify mutations in Bordetella pertussis vir-regulated genes.  

PubMed

Mutants of Bordetella pertussis deficient in virulence-associated factors were identified by using the transposon Tn5 lac. Tn5 lac is a derivative of Tn5 which generates promoter fusions for beta-galactosidase. Tn5 lac insertions in the vir-regulated genes of B. pertussis were identified by selecting for kanamycin-resistant mutants that expressed beta-galactosidase when the vir-regulated genes were expressed but not when the vir-regulated genes were turned off. Fourteen different mutations in vir-regulated genes were identified. Two mutants were deficient in the production of the filamentous hemagglutinin, two mutants were deficient in the production of adenylate cyclase toxin and hemolysin, and one mutant was deficient in the production of dermonecrotic toxin. One insertion mapped adjacent to the pertussis toxin gene, but the mutant produced pertussis toxin. The phenotypes of the remaining eight mutants were not determined, but the mutants did not appear to be deficient in the production of the 69,000-dalton outer membrane protein (agglutinogen 3) or the capsule. Screening for mutations in either of the fimbrial genes proved to be problematic since the parental strain was found to switch from a fimbriated to a nonfimbriated state at a high frequency, which was suggestive of the metastable expression of pili in other bacteria. We used Southern blot analysis with a 30-mer specific for the fimbrial sequences. No bands with the predicted increase in size due to the 12 kilobases from Tn5 lac were observed, which suggests that none of these genes were mutated. Southern blot analysis also revealed that seven of the eight unidentified mutations mapped to different restriction fragments, which suggests that they could be deficient in as many as seven different genes. PMID:2569447

Weiss, A A; Melton, A R; Walker, K E; Andraos-Selim, C; Meidl, J J

1989-09-01

376

Transposon Tn5 excision in yeast: influence of DNA polymerases alpha, delta, and epsilon and repair genes.  

PubMed

Interaction between short repeats may be a source of genomic rearrangements and deletions. We investigated possible interactions between short (9 base pairs) direct repeats in yeast by using our previously described system for analyzing bacterial transposon Tn5 excision in yeast. Mutations of either POL3 or POL1, the proposed structural genes for polymerases delta and alpha, respectively, yield high levels of excision at semipermissive temperatures. pol2 (corresponding to polymerase epsilon) and pol2 pol3 double mutants do not exhibit enhanced excision. A majority of excision events involve direct repeats and are precise; the remaining imprecise excisions occur within or in the vicinity of the repeats. The three DNA repair pathways identified by rad1, rad6 and rad18, rad50 and rad52 mutations were examined for their possible role in Tn5 excision; no enhancement was observed in mutants. However, the pol3-stimulated Tn5 excision was reduced in rad52 and rad50 mutants. This suggests the potential for interaction between the systems for DNA double-strand break/recombinational repair and DNA synthesis. Based on the suggestion of Morrison et al. [Morrison, A., Araki, H., Clark, A. B., Hamatake, R. H. & Sugino, A. (1990) Cell 62, 1143-1151] that polymerases delta and alpha are responsible for lagging-strand synthesis and that polymerase epsilon is responsible for leading-strand synthesis, we suggest that Tn5 excision is stimulated under conditions of altered lagging-strand synthesis, possibly due to extended opportunities for single-strand interactions between the inverted insertion sequence I550 repeats of Tn5. PMID:1315039

Gordenin, D A; Malkova, A L; Peterzen, A; Kulikov, V N; Pavlov, Y I; Perkins, E; Resnick, M A

1992-05-01

377

Stable expression of sialyl-Tn antigen in T47-D cells induces a decrease of cell adhesion and an increase of cell migration.  

PubMed

Sialyl-Tn is a carbohydrate antigen overexpressed in several epithelial cancers including breast cancer, and usually associated with poor prognosis. Sialyl-Tn is synthesized by a CMP-Neu5Ac: GalNAc alpha2,6-sialyltransferase: ST6GalNAc I, which catalyzes the transfer of a sialic acid residue in alpha2,6-linkage to the GalNAcalpha1-O-Ser/Thr structure. The resulting disaccharide (Neu5Acalpha2-6GalNAcalpha1-O-Ser/Thr) cannot be further elongated and sialyl-Tn expression results therefore in a shortening of the O-glycan chains. However, usual breast cancer cell lines express neither ST6GalNAc I nor sialyl-Tn antigen. We have previously shown that stable transfection of MDA-MB-231 cells with the hST6GalNAc I cDNA induces the sialyl-Tn antigen expression at the cell surface and leads to a decreased cell growth and an increased cell migration. We describe herein the generation of new T47-D clones expressing sialyl-Tn antigen after hST6GalNAc I cDNA stable transfection. sialyl-Tn antigen is carried by several high molecular weight membrane bound O-glycoproteins, including MUC1. We show that sialyl-Tn expression induces a decrease of cell growth and adhesion, and an increase of cell migration in sialyl-Tn positive clones compared to mock transfected cells. These observations show that the alteration of the O-glycans pattern is sufficient to modify the biological features of cancer cells. These T47-D sialyl-Tn expressing clones might allow further in vivo investigation to determine precisely the impact of such O-glycosylation modifications on breast cancer development. PMID:15770530

Julien, Sylvain; Lagadec, Chann; Krzewinski-Recchi, Marie-Ange; Courtand, Gilles; Le Bourhis, Xuefen; Delannoy, Philippe

2005-03-01

378

Expression of sialosyl-Tn in colony-forming unit-erythroid, erythroblasts, B cells, and a subset of CD4+ cells.  

PubMed

The epitopes Tn and sialosyl-Tn are expressed on erythrocytes of individuals with a very rare blood group, who often suffer from "Tn syndrome." We surveyed expression of Tn and sialosyl-Tn in normal blood cells, malignant transformed cells, and progenitor stem cells from bone marrow (BM). An anti-Tn antibody, IE3, and an anti-sialosyl-Tn antibody, TKH2, were used in this study. TKH2 reacted with erythroblasts, B cells, and a subset of CD4+ cells; but not with erythrocytes. Erythroblastic cell lines (K562, HEL, and UT7/EPO) and B-cell lines (Daudi, Raji, and B-cell lines transformed by Epstein-Barr virus) showed reactivity to TKH2. Similar results from the reactivity of TKH2 with transformed cells from leukemia patients and lymphoma patients were obtained; TKH2 reacted with blasts from erythroleukemia (M6; for 4 of 4 cases) and with lymphocytes from B-cell chronic lymphocytic leukemia (3 of 3), B-cell lymphoma (5 of 5), and CD4+ adult T-cell leukemia (4 of 4), but did not react with blasts from acute myeloid leukemia (M0 to M5; 0 of 22) or acute lymphoid leukemia (B-lymphoid leukemia, 0 of 11; T-lymphoid leukemia, 0 of 2; undifferentiated leukemia, 0 of 1). IE3 did not react with all of the tested cells. CD2-CD19-TKH2+ normal BM cells (BMC) contained blasts and various maturation stages of erythroblasts. The TKH2+ cells produced a large number of colony-forming unit-erythroid (CFU-E) colonies, whereas they produced a small number of burst-forming unit-erythroid colonies and CFU-granulocyte-macrophage colonies. CD34+ normal BMC did not express Tn and sialosyl-Tn. These findings suggest that sialosyl-Tn expresses in CFU-E to erythroblasts. PMID:7903875

Muroi, K; Suda, T; Nakamura, M; Okada, S; Nojiri, H; Amemiya, Y; Miura, Y; Hakomori, S

1994-01-01

379

Bolivar County, Mississippi: Subdivision Regulations.  

National Technical Information Service (NTIS)

The report creates and establishes suggested general rules and regulations for the subdivision of land in Bolivar County, MS. It sets forth transportation and thoroughfare design standards within subdivisions; and outlines administrative enforcement, pena...

1973-01-01

380

County Business Patterns, 1997: Massachusetts.  

National Technical Information Service (NTIS)

County Business Patterns is an annual series that provides subnational economic data by industry. The series is useful for studying the economic activity of small areas; analyzing economic changes over time; and as a benchmark for statistical series, surv...

1999-01-01

381

County Business Patterns, 2004: Massachusetts.  

National Technical Information Service (NTIS)

County Business Patterns is an annual series that provides subnational economic data by industry. The series is useful for studying the economic activity of small areas; analyzing economic changes over time; and as a benchmark for statistical series, surv...

2006-01-01

382

County Business Patterns, 1996: Massachusetts.  

National Technical Information Service (NTIS)

County Business Patterns is an annual series that provides subnational economic data by industry. The series is useful for studying the economic activity of small areas; analyzing economic changes over time; and as a benchmark for statistical series, surv...

1998-01-01

383

County Business Patterns, 2003: Massachusetts.  

National Technical Information Service (NTIS)

County Business Patterns is an annual series that provides subnational economic data by industry. The series is useful for studying the economic activity of small areas; analyzing economic changes over time; and as a benchmark for statistical series, surv...

2005-01-01

384

County Business Patterns: Massachusetts, 2002.  

National Technical Information Service (NTIS)

County Business Patterns is an annual series that provides subnational economic data by industry. The series is useful for studying the economic activity of small areas; analyzing economic changes over time; and as a benchmark for statistical series, surv...

2004-01-01

385

VERMONT COUNTY HEALTH DATA2  

EPA Science Inventory

This datalayer contains Vermont Population and Health data describing public health (1986-2000), by county, extracted from various sources, such as; the Vermont Department of Health, the Vermont Center for Justice Research, the National Institute on Alcohol Abuse & Alcoholis...

386

Dauphin County Opportunity Industrialization Center.  

National Technical Information Service (NTIS)

The goals of the OIC organization in Dauphin County include providing hope for impoverished and hard-core unemployed in the form of job preparation by providing adult education, pre-vocational training, skills training, counseling, job development, placem...

1968-01-01

387

Geothermal development plan: Yuma county  

SciTech Connect

One hot spring and 33 wells drilled in the county discharge water at temperatures sufficient for direct-use geothermal applications such as process heat and space heating and cooling. Currently, one industry within the county has been identified which may be able to use geothermal energy for its process heat requirements. Also, a computer simulation model was used to predict geothermal energy on line as a function of time under both private and city-owned utility development of the resource.

White, D.H.

1981-01-01

388

Geothermal Development Plan: Pima County  

SciTech Connect

Pima County is located entirely within the Basin and Range physiographic province in which geothermal resources are known to occur. Continued growth as indicated by such factors as population growth, employment and income will require large amounts of energy. It is believed that geothermal energy could provide some of the energy that will be needed. Potential users of geothermal energy within the county are identified.

White, D.H.

1981-01-01

389

San Bernardino County Museum  

NSDL National Science Digital Library

The San Bernardino County Museum is a regional museum with exhibits and collections in cultural and natural history. Special exhibits, the Exploration Station live animal discovery center, extensive research collections, and public programs for adults, families, students, and children are all part of the museum experience. The Museum is surrounded by citrus groves, and orange blossoms perfume the air. The Zimmerman Citrus Kiosk explores citrus agricultural history in Southern California. Death Valley Alive! is a new traveling exhibit, available for rent, where visitors can explore the natural history and culture of the Death Valley region from its geologic beginnings 1.3 billion years ago. Online exhibits include The Importance of Museum Collections, The Barstow Fossil Beds, Motherlode of the Miocene, The Etiwanda Fan, and for kids, Mimicry, a study in camouflage and adaptation. Teacher Resources include professional development and workshops, and Trading Places, where 21 hours of time spent volunteering in the Museum earns a free program for your class. Sample opportunities include writing activities for study kits, developing post-visit activities, creating a museum gallery guide, translating student materials into Spanish, or developing an independent idea. There are programs for youth, Scouts, and adults, as well as various publications available in anthropology, archeology, biology, geology and history.

390

Atlas of Historical County Boundaries  

NSDL National Science Digital Library

So you'd like to know the boundaries of Washington's King County in the early 20th century? Or perhaps you'd like to know more about the shape of Nassau County in New York back in the 19th century? The Atlas of Historical County Boundaries can provide you such details. The project is housed at the William M. Scholl Center for American History and Culture at The Newberry Library in Chicago and it was completed in 2010. The Atlas includes such features as all boundary changes in states and countries, non-county areas, separate map or polygon files for every different county configuration, and other helpful features. Users can get started by clicking on different states of interest and just exploring as they see fit. It's also useful to read over the Using the Atlas area for additional assistance. Finally, visitors can also look at the Publications section for more information on the source material for these county boundaries.

391

Insertions of mini-Tn10 transposon T-POP in Salmonella enterica sv. typhi.  

PubMed Central

We have mutagenized a clinical strain of Salmonella enterica sv. typhi with mini-transposon Tn10dTet (T-POP) to obtain conditional lethal (tetracycline-dependent) mutants with T-POP insertions upstream of essential genes. Generalized transducing phage P22 was used to introduce T-POP from a S. typhimurium donor into a S. typhi recipient. Chromosomal DNA was purified from the mutagenized donor strains, fragmented, and then electroporated into S. typhi to backcross the original T-POP insertions. Four tetracycline-dependent mutants with two distinct terminal phenotypes were found among 1700 mutants with T-POP insertions. When grown in the absence of tetracycline, two of the four tetracycline-dependent mutants arrest at a late stage in the cell cycle, can be rescued by outgrowth in media with tetracycline, and define a reversible checkpoint late in the cell cycle. One of these insertions creates an operon fusion with a gene, yqgF, that is conserved among gram-negative bacteria and likely encodes an essential Holliday junction resolvase. T-POP insertions can be used not only to identify essential S. typhi genes but also to reveal novel phenotypes resulting from the depletion of their products.

Hidalgo, Alejandro A; Trombert, A Nicole; Castro-Alonso, J C; Santiviago, Carlos A; Tesser, Bruno R; Youderian, Philip; Mora, Guido C

2004-01-01

392

Determination of the stereostructure of the product of Tn3 resolvase by a general method.  

PubMed Central

A method has been developed for determination of the absolute structure of DNA catenanes. The catenated DNA is partially denatured before being thickened with a coating of RecA protein and spread for electron microscopy. This treatment allows visualization of the orientation of each ring as well as identification of the overlying and underlying DNAs at crossing points. These determinations define the topology of a catenane, providing a powerful means for testing mechanisms of catenane-producing enzymes in DNA recombination and replication. The technique was used to show that the single interlock of the catenated products of site-specific recombination mediated by Tn3 resolvase is exclusively of negative sign. The unique topology of the products indicates that resolvase fixes the sum of the number of supercoils between recombination sites at synapsis and the number of such supercoils lost or gained during strand exchange. The data strongly suggest that there are in fact three negative supercoils between synapsed sites; one supercoil is dissolved in the cross-over mechanism, whereas the other two are metamorphosed into the unique catenane interlock. Images

Wasserman, S A; Cozzarelli, N R

1985-01-01

393

Antiferromagnetic pinning of a phase-like mode below TN in NdMnO3  

NASA Astrophysics Data System (ADS)

We report on reflectivity and emission far infrared spectra of NdMnO3 between4K and its dissociation temperature. Phonon bands at 300K are in agreement with orthorhombic Pbnm space group assignments. In addition, a broad strong band. reminiscent to a phase-mode in quasi-one dimension metals, is found at very low frequencies that it is understood originating in charge fluctuations in d-orbitals. There is no distinctive behaviors between 1073 K and 1173 K, where orthorhombic O and O' coexist. Beyond ˜700 K a mid-infrared polaron band turns into a Drude tail suggesting hopping conductivity double exchange due to air heating oxidation Mn^3+-> Mn^4+ +1e^-. Below 300 K phonons are better defined and the low frequency giant dipole acquires strength. Few degrees above the antiferromagnetic transition it broadens as electrons loosing coherence. At TN˜76 K we find strong phonon magnetostriction while the band turns asymmetric locking-in to the underlying magnetic order. Preliminary measurements of hexagonal TmMnO3, show that asymmetry split due to lower symmetry and the triangular magnetic lattice two exchange integrals J1 and J2 in the a-b plane. Similar to a soft mode those two bands and a lower frequency resonance undergo strong hardening down to 4 K.

Massa, Nestor; Del Campo, Leire; de Sousa Meneses, Domingos; Echegut, Patrick; Martínez-Lope, Maria Jesus; Alonso, Jose Antonio

2012-02-01

394

Correlation of thermodynamic and genetic properties in the Tn10 encoded TET gene control region.  

PubMed Central

The thermal stability of the Tn10 encoded tetracycline resistance (TET) gene control region is investigated by melting studies using purified DNA restriction fragments containing various amounts of flanking sequences. In order to study the thermodynamic properties of this control region under conditions, where enough flanking DNA is present to mimic the situation in the chromosome, the five step melting process of a 1450-bp DNA fragment is analyzed. Because most of the sequence of this DNA is not known, the assignment of the melting transitions to segments of the DNA is done by an experimental method. This employs the preparation of subfragments from the 1450-bp DNA and comparison of their denaturation profiles with the one of the intact sequence. This approach results in the complete assignment of the five denaturation steps. Rather than from the ends, the unwinding starts from the TET gene control region in the middle of the 1450-bp sequence. A clear correlation between the thermodynamic and genetic properties of this DNA is observed. The regulatory sequence forms a small cooperative unit with the lowest stability in the entire fragment. The thermal denaturation of the TET repressor. TET operator complex reveals, that the TET repressor specifically recognizes the double stranded TET operator DNA and stabilizes this structure by 2.4 degrees C. This results is also discussed as an example of the possible action of denaturing or stabilizing proteins on this genetic control region.

Hillen, W; Unger, B

1982-01-01

395

Correlation between sialyl Tn antigen and lymphatic metastasis in patients with Borrmann type IV gastric carcinoma.  

PubMed Central

The expression of sialyl Tn (STn) antigen in 180 patients with Borrmann type IV gastric carcinomas was examined immunohistochemically. The rate of positive STn staining was 32% (57/180) for the primary tumours, and this positive staining correlated well with tumour extension, lymph node metastasis (P < 0.05) and peritoneal dissemination (P < 0.01). One-third (5/15) of patients with positive STn-staining cancer cells had a high level of serum STn. Lesions with positive STn staining were related to a lower survival rate for the patients (P < 0.05). Proliferative activity of the tumour, as measured by proliferating nuclear antigen (PCNA) labelling percentage and argyrophilic nucleolar organiser region (AgNOR) count, was significantly higher (41.5 +/- 13.0%, 3.78 +/- 0.98) in the STn-positive group than in the STn-negative group (34.2 +/- 13.2%, 3.48 +/- 0.85) (P < 0.01, P < 0.05 respectively). Estimating STn antigen may be useful for predicting the likelihood of lymph node metastasis or peritoneal dissemination and the clinical prognosis for patients with Borrmann type IV gastric carcinoma. Images Figure 1

Kakeji, Y.; Maehara, Y.; Morita, M.; Matsukuma, A.; Furusawa, M.; Takahashi, I.; Kusumoto, T.; Ohno, S.; Sugimachi, K.

1995-01-01

396

Sialyl-Tn in Cancer: (How) Did We Miss the Target?  

PubMed Central

Sialyl-Tn antigen (STn) is a short O-glycan containing a sialic acid residue ?2,6-linked to GalNAc?-O-Ser/Thr. The biosynthesis of STn is mediated by a specific sialyltransferase termed ST6GalNAc I, which competes with O-glycans elongating glycosyltransferases and prevents cancer cells from exhibiting longer O-glycans. While weakly expressed by fetal and normal adult tissues, STn is expressed by more than 80% of human carcinomas and in all cases, STn detection is associated with adverse outcome and decreased overall survival for the patients. Because of its pan-carcinoma expression associated with an adverse outcome, an anti-cancer vaccine, named Theratope, has been designed towards the STn epitope. In spite of the great enthusiasm around this immunotherapy, Theratope failed on Phase III clinical trial. However, in lieu of missing this target, one should consider to revise the Theratope design and the actual facts. In this review, we highlight the many lessons that can be learned from this failure from the immunological standpoint, as well as from the drug design and formulation and patient selection. Moreover, an irrefutable knowledge is arising from novel immunotherapies targeting other carbohydrate antigens and STn carrier proteins, such as MUC1, that will warrantee the future development of more successful anti-STn immunotherapy strategies.

Julien, Sylvain; Videira, Paula A.; Delannoy, Philippe

2012-01-01

397

Transposon Tn5-Generated Bradyrhizobium japonicum Mutants Unable To Grow Chemoautotrophically with H(2).  

PubMed

Twelve Tn5-induced mutants of Bradyrhizobium japonicum unable to grow chemoautotrophically with CO(2) and H(2) (Aut) were isolated. Five Aut mutants lacked hydrogen uptake activity (Hup). The other seven Aut mutants possessed wild-type levels of hydrogen uptake activity (Hup), both in free-living culture and symbiotically. Three of the Hup mutants lacked hydrogenase activity both in free-living culture and as nodule bacteroids. The other two mutants were Hup only in free-living culture. The latter two mutants appeared to be hypersensitive to repression by oxygen, since Hup activity could be derepressed under 0.4% O(2). All five Hup mutants expressed both ex planta and symbiotic nitrogenase activities. Two of the seven Aut Hup mutants expressed no free-living nitrogenase activity, but they did express it symbiotically. These two strains, plus one other Aut Hup mutant, had CO(2) fixation activities 20 to 32% of the wild-type level. The cosmid pSH22, which was shown previously to contain hydrogenase-related genes of B. japonicum, was conjugated into each Aut mutant. The Aut Hup mutants that were Hup both in free-living culture and symbiotically were complemented by the cosmid. None of the other mutants was complemented by pSH22. Individual subcloned fragments of pSH22 were used to complement two of the Hup mutants. PMID:16347549

Hom, S S; Novak, P D; Maier, R J

1988-02-01

398

Crystal structures of the transposon Tn5-carried bleomycin resistance determinant uncomplexed and complexed with bleomycin.  

PubMed

The transposon Tn5 carries a gene designated ble that confers resistance to bleomycin (Bm). In this study, we determined the x-ray crystal structures of the ble gene product, designated BLMT, uncomplexed and complexed with Bm at 1.7 and 2.5 A resolution, respectively. The structure of BLMT is a dimer with two Bm-binding pockets composed of two large concavities and two long grooves. This crystal structure of BLMT complexed with Bm gives a precise mode for binding of the antibiotic to BLMT. The conformational change of BLMT generated by binding to Bm occurs at a beta-turn composed of the residues from Gln(97) to Thr(102). Crystallographic analysis of Bm bound to BLMT shows that two thiazolium rings of the bithiazole moiety are in the trans conformation. The axial ligand, which binds a metal ion, seems to be the primary amine in the beta-aminoalanine moiety. This report, which is the first with regard to the x-ray crystal structure of Bm, shows that the bithiazole moiety of Bm is far from the metal-binding domain. That is, Bm complexed with BLMT takes a more extended form than the drug complexed with DNA. PMID:11134052

Maruyama, M; Kumagai, T; Matoba, Y; Hayashida, M; Fujii, T; Hata, Y; Sugiyama, M

2001-03-30

399

Tn5-Mob transposon mediated transfer of salt tolerance and symbiotic characteristics between Rhizobia genera.  

PubMed

Rhizobium meliloti 042B is a fast-growing, salt-tolerant and high efficiency nitrogen-fixing symbiont with alfalfa. Bradyrhizobium japonicum USDA110 grows slowly, and cannot grow in YMA medium containing 0.1M NaCl, but nodulates and fixed nitrogen efficiently with soybean. Eighty-six transconjugants, called SR, were obtained by inserting Tn5-Mob randomly into genomes of 042B using pSUP5011 and helper plasmid RP4. Selecting 4 SR strains randomly and introducing DNA fragment of SR into USDA110 with helper plasmid R68.45 by triparental mating, 106 transconjugants, called BSR, were constructed. Most of BSR strains had the fast-growing phenotype and could tolerate 0.3-0.5M NaCl generally. Some of them produced melanine. When soybean and alfalfa were inoculated with these transconjugants BSR, 47 out of 90 BSR were found to nodulate in both of these plants, but no nitrogenase activity was observed with alfalfa; 26 strains could only nodulate and fix nitrogen in soybean; 13 strains could nodulate in alfalfa but did not fix nitrogen; 4 strains failed to nodulate in either soybean or alfalfa. Among them, 4 transconjugants which tolerated and fixed nitrogen efficiently in soybean were constructed. PMID:8049344

Yang, S; Wu, Z; Gao, W; Li, J

1993-01-01

400

Mutational analysis of insertion sequence 50 (IS50) and transposon 5 (Tn5) ends.  

PubMed

Insertion sequence 50 (IS50) transposition utilizes a 19-base-pair "outside" end and a 19-base-pair "inside" end in inverted orientation relative to each other, whereas transposon 5 (Tn5) transposition utilizes two inverted outside ends. The frequency of transposition events that involve an inside end is regulated 1000-fold by the host dam methylase system. The end sequence requirements for transposition and its regulation by dam methylase were analyzed in Escherichia coli by generating random single base pair mutations in either an IS50 inside end or outside end placed in inverted orientation with respect to an unmutagenized outside end. The mutations were then isolated, assayed for transposition phenotype, and sequenced. Mutations were isolated at 15 of the 19 sites in the outside end. All of these mutations except those at position 4 decreased transposition. Mutations at position 4 (which is the only nonidentical base pair in a region of homology between the outside and inside ends) had no effect on transposition. Mutations were isolated at 11 of the 19 sites in the inside end. All of these mutations, including one at position 4, decreased transposition in dam- cells. Mutations at position 10 (within a dam recognition sequence) and 2 (not within a dam recognition sequence) reduced the magnitude of dam regulation. A mutation within a dam recognition sequence adjacent to the required 19 base pairs of the inside end did not reduce the magnitude of dam regulation. PMID:2832849

Makris, J C; Nordmann, P L; Reznikoff, W S

1988-04-01

401

Characterization of the bleomycin resistance determinant encoded on the transposon Tn5.  

PubMed

The transposon Tn5 carries a gene, ble, which confers resistance to bleomycin (Bm) and gives a survival advantage to its host cell. We found that the ble gene product, designated BLMT, is a binding protein with a strong affinity for Bm. BLMT quenched both the antibacterial and DNA-cleaving activities of Bm, when incubated with the antibiotic. An electron spin resonance spin-trapping analysis showed that BLMT inhibits the generation of Bm-induced hydroxyl radical, by trapping Bm but not the hydroxyl radical. Western blot analysis using an anti-BLMT monoclonal antibody revealed that BLMT is immunologically distinct from Bm-binding proteins from Streptomyces verticillus, Staphylococcus aureus and Streptoalloteichus hindustanus. Escherichia coli, transformed with a mutant ble having leucine instead of proline at N-terminal amino acid position 7, lost resistance to Bm, although the cell maintained the survival benefit. This suggests that the Bm resistance mediated by ble is independent of its ability to give a survival advantage to the host bacterium. PMID:9923599

Kumagai, T; Nakano, T; Maruyama, M; Mochizuki, H; Sugiyama, M

1999-01-01

402

Identification of multiple mercury sources to stream sediments near Oak Ridge, TN, USA.  

PubMed

Sediments were analyzed for total Hg concentration (THg) and isotopic composition from streams and rivers in the vicinity of the Y-12 National Security Complex (Y12) in Oak Ridge, TN (USA). In the stream directly draining Y12, where industrial releases of mercury (Hg) have been documented, high THg (3.26 to 60.1 ?g/g) sediments had a distinct Hg isotopic composition (?(202)Hg of 0.02 ± 0.15‰ and ?(199)Hg of -0.07 ± 0.03‰; mean ± 1SD, n = 12) compared to sediments from relatively uncontaminated streams in the region (?(202)Hg = -1.40 ± 0.06‰ and ?(199)Hg of -0.26 ± 0.03‰; mean ± 1SD, n = 6). Additionally, several streams that are nearby but do not drain Y12 had sediments with intermediate THg (0.06 to 0.21 ?g/g) and anomalous ?(202)Hg (as low as -5.07‰). We suggest that the low ?(202)Hg values in these sediments provide evidence for the contribution of an additional Hg source to sediments, possibly derived from atmospheric deposition. In sediments directly downstream of Y12 this third Hg source is not discernible, and the Hg isotopic composition can be largely explained by the mixing of low THg sediments with high THg sediments contaminated by Y12 discharges. PMID:24588770

Donovan, Patrick M; Blum, Joel D; Demers, Jason D; Gu, Baohua; Brooks, Scott C; Peryam, John

2014-04-01

403

Water Quality Management Plan: Campbell County, Johnson County, Sheridan County (Wyoming).  

National Technical Information Service (NTIS)

The Powder River Areawide Planning Organization (PRAPO) was designated by the Governor of Wyoming to administer a federal grant for areawide waste treatment planning. The major study area covered Campbell, Johnson and Sheridan Counties in Wyoming. Primary...

1978-01-01

404

76 FR 33401 - Environmental Impact Statement: Will and Kankakee Counties, Illinois and Lake County, IN  

Federal Register 2010, 2011, 2012, 2013

...Administration Environmental Impact Statement: Will and Kankakee Counties, Illinois and Lake County...that a Tier One Environmental Impact Statement will be prepared for the Illiana Corridor Project in Will and Kankakee Counties, Illinois and Lake...

2011-06-08

405

75 FR 25308 - Environmental Impact Statement: Winnebago County, IL and Rock County, WI  

Federal Register 2010, 2011, 2012, 2013

...Administration Environmental Impact Statement: Winnebago County, IL and Rock County, WI AGENCY: Federal Highway Administration (FHWA...Wisconsin Route 213 and Nye School Road northwest of Beloit, Rock County, Wisconsin to the interchange of Rockton Road and...

2010-05-07

406

The novel tetramethylpyrazine bis-nitrone (TN-2) protects against MPTP/MPP+-induced neurotoxicity via inhibition of mitochondrial-dependent apoptosis.  

PubMed

Mitochondrial-dependent apoptosis plays an important role in the degeneration of dopaminergic neurons in Parkinson's disease (PD). Methyl-4-phenyl-1,2,3,6-tetra- hydropyridine (MPTP), the most widely used neurotoxin to simulate PD, is converted to 1-methyl-4-phenylpyridinium (MPP(+)) in vivo. MPP(+) induces excessive intracellular reactive oxygen species (ROS), leading to mitochondrial-dependent apoptosis via sequentially opening mitochondria permeability transition pore (mPTP) to release cytochrome c from mitochondria into cytoplasm and activate pro-apoptotic caspase proteins. We have previously synthesized 2,5-[[(1,1-dimethylethyl)oxidoimino]methyl]-3,6-trimethylpyrazine (TN-2), a novel derivative of the Chinese herb medicine tetramethylpyrazine (TMP). TN-2 is armed with two powerful free radical-scavenging nitrone moieties. TN-2 significantly reversed the loss of dopaminergic neurons in the substantia nigra and the decrease in dopamine level in the striatum induced by MPTP in mice. TN-2 ameliorated the MPTP-induced decrease of brain superoxide dismutase activity and glutathione concentration and increase of brain malondialdehyde. In addition, TN-2 inhibited MPP(+)-induced neuronal damage/apoptosis in primary cerebellum granular neurons (CGNs) and SH-SY5Y cells. TN-2 decreased excessive intracellular ROS, prevented the loss of mitochondrial membrane potential, blocked the release of mitochondrial cytochrome c and inhibited the activation of caspase-3 and caspase-9. Moreover, TN-2 treatment increased the mRNA expression of mitochondrial biogenesis factors peroxisome proliferator-activated receptor (PPAR)-? coactivator-1 (PGC- 1? and ?) and mitochondrial transcription factor A (Tfam) in SH-SY5Y cells and CGNs. These results suggest that TN-2 protects dopaminergic neurons against MPTP/MPP(+)-induced neurotoxicity via the inhibition of mitochondrial-dependent apoptosis and possibly via the activation of mitochondrial biogenesis, indicating that TN-2 is a potential new treatment for PD. PMID:24233519

Xu, Daping; Duan, Hongwei; Zhang, Zaijun; Cui, Wei; Wang, Liang; Sun, Yewei; Lang, Ming; Hoi, Pui Man; Han, Yifan; Wang, Yuqiang; Lee, Simon MingYuen

2014-03-01

407

Primary Breast Cancer Tumours Contain High Amounts of IgA1 Immunoglobulin: An Immunohistochemical Analysis of a Possible Carrier of the Tumour-Associated Tn Antigen  

PubMed Central

The Tn antigen (GalNAc alpha-O-Ser/Thr) as defined by the binding of the lectin, helix pomatia agglutinin (HPA) or anti-Tn monoclonal antibodies, is known to be exposed in a majority of cancers, and it has also been shown to correlate positively with the metastatic capacity in breast carcinoma. The short O-glycan that forms the antigen is carried by a number of different proteins. One potential carrier of the Tn antigen is immunoglobulin A1 (IgA1), which we surprisingly found in tumour cells of the invasive parts of primary breast carcinoma. Conventional immunohistochemical analysis of paraffin-embedded sections from primary breast cancers showed IgA1 to be present in the cytoplasm and plasma membrane of 35 out of 36 individual primary tumours. The immunohistochemical staining of HPA and anti-Tn antibody (GOD3-2C4) did to some extent overlap with the presence of IgA1 in the tumours, but differences were seen in the percentage of stained cells and in the staining pattern in the different breast cancers analysed. Anti-Tn antibody and HPA were also shown to specifically bind to a number of possible constellations of the Tn antigen in the hinge region of IgA1. Both reagents could also detect the presence of Tn positive IgA in serum. On average 51% of the tumour cells in the individual breast cancer tumour sections showed staining for IgA1. The overall amount of staining in the invasive part of the tumour with the anti Tn antibody was 67%, and 93% with HPA. The intra-expression or uptake of IgA1 in breast cancer makes it a new potential carrier of the tumour associated and immunogenic Tn antigen.

Welinder, Charlotte; Baldetorp, Bo; Blixt, Ola; Grabau, Dorthe; Jansson, Bo

2013-01-01

408

Bleomycin-resistance gene derived from the transposon Tn5 confers selective advantage to Escherichia coli K-12.  

PubMed

The plasmid pRAB2 contains a silent operon derived from the transposon Tn5 and carrying the gene neo for neomycin-kanamycin resistance and a truncated ble gene (ble333) for bleomycin resistance. Spontaneous mutants that express the two resistances provide Escherichia coli cells an improved fitness during the phase of decline in the absence of the antibiotics. It is shown that the ble333 gene product is responsible for this better fitness. These results can explain a previously described selective advantage attributed to the presence of Tn5. The improved fitness of bleomycin-resistant bacteria is proposed to relate to DNA repair by the ble gene product. The consequences of the presence of an accessory gene improving fitness are discussed in terms of evolutionary stable strategy of a transposon in populations of E. coli. PMID:1717996

Blot, M; Meyer, J; Arber, W

1991-10-15

409

Isolation and characterization of transposon Tn5-induced mutants of Pseudomonas perfectomarina defective in nitrous oxide respiration.  

PubMed

Transposon (Tn5) mutagenesis of Pseudomonas perfectomarina with the plasmid pSUP2021 [(pBR325-Mob(RP4))::Tn5] and the chromosomally integrated RP4 plasmid in Escherichia coli as the donor, produced three distinct groups of mutants that were defective in nitrous oxide respiration. One group of mutants lacked the structural protein of N2O reductase, the second synthesized a copper-free apoprotein; and a third group expressed a low level of intact enzyme. The mutants provided evidence for N2O being the immediate precursor of dinitrogen in denitrification and documented the essentiality of the copper enzyme. Synthesis of N2O reductase depended strongly on the growth conditions, with N2O-grown cells expressing the lowest level of enzyme. Regulatory responses of mutants elicited by nitrate or oxygen were unaltered when compared with wild-type behavior. PMID:2993252

Zumft, W G; Döhler, K; Körner, H

1985-09-01

410

Molecular Description and Industrial Potential of Tn6098 Conjugative Transfer Conferring Alpha-Galactoside Metabolism in Lactococcus lactis? †  

PubMed Central

A novel 51-kb conjugative transposon of Lactococcus lactis, designated Tn6098, encoding the capacity to utilize ?-galactosides such as raffinose and stachyose, was identified and characterized. Alpha-galactosides are a dominant carbon source in many plant-derived foods. Most dairy lactococcus strains are unable to use ?-galactosides as a growth substrate, yet many of these strains are known to have beneficial industrial traits. Conjugal transfer of Tn6098 was demonstrated from the plant-derived donor strain L. lactis KF147 to the recipient L. lactis NZ4501, a derivative of the dairy model strain L. lactis MG1363. The integration of Tn6098 into the genome of the recipient strain was confirmed by Illumina sequencing of the transconjugant L. lactis NIZO3921. The molecular structure of the integration site was confirmed by a PCR product spanning the insertion site. A 15-bp direct repeat sequence (TTATACCATAATTAC) is present on either side of Tn6098 in the chromosome of L. lactis KF147. One copy of this sequence is also present in the L. lactis MG1363 chromosome and represents the sole integration site. Phenotypic characterization of all strains showed that the transconjugant has not only acquired the ability to grow well in soy milk, a substrate rich in ?-galactosides, but also has retained the flavor-forming capabilities of the recipient strain L. lactis MG1363. This study demonstrates how (induced) conjugation can be used to exploit the beneficial industrial traits of industrial dairy lactic acid bacteria in fermentation of plant-derived substrates.

Machielsen, Ronnie; Siezen, Roland J.; van Hijum, Sacha A. F. T.; van Hylckama Vlieg, Johan E. T.

2011-01-01

411

Tn6060, a Transposon from a Genomic Island in a Pseudomonas aeruginosa Clinical Isolate That Includes Two Class 1 Integrons?  

PubMed Central

A 25,441-bp transposon was recovered from a Pseudomonas aeruginosa clinical isolate. While the transposition module was >99% identical to sequence of Tn1403, the element had been subject to rearrangements, with two In70.2-like class 1 integrons inserted into it in an unusual “tail-to-tail” configuration. One cassette array was the same as that in In70.2; however, the second was different, generating a transposon that collectively includes six resistance cassettes.

Chowdhury, Piklu Roy; Merlino, John; Labbate, Maurizio; Cheong, Elaine Y.-L.; Gottlieb, Thomas; Stokes, H. W.

2009-01-01

412

Molecular and biochemical characterization of a new alkaline ?-propeller phytase from the insect symbiotic bacterium Janthinobacterium sp. TN115  

Microsoft Academic Search

A phytase-encoding gene (phyA115) was cloned from Janthinobacterium sp. TN115, a symbiotic bacterial strain isolated from the gut contents of Batocera horsfieldi larvae (Coleoptera: Cerambycidae), and expressed in Escherichia coli. The 1,884-bp full-length gene encodes a 28-residue putative signal peptide and a 599-residue mature protein with a calculated\\u000a mass of 64 kDa. The deduced PhyA115 shares low identity with known sequences

Rui Zhang; Peilong Yang; Huoqing Huang; Tiezheng Yuan; Pengjun Shi; Kun Meng; Bin Yao

413

Surface modifications in the platelets of a patient with alpha-N-acetyl-D-galactosamine residues, the Tn-syndrome.  

PubMed

The Tn-syndrome is an acquired disorder characterized by the polyagglutination of blood cells and the pathological exposure of alpha-N-acetyl-D-galactosamine residues (Tn-antigen) at the cell surface. We now report studies on the platelet of a patient (Ba.) of which 81% reacted positively with a fluorescein conjugate of Helix pomatia agglutinin (HPA). The surface proteins of Ba. platelets were labeled with 125I by the lactoperoxidase-catalyzed procedure; single and two-dimensional electrophoresis on sodium dodecyl sulfate (SDS)-polyacrylamide gels was followed by autoradiography that revealed normal 125I-labeling of the major membrane glycoproteins (GP) but that GP Ib had a faster than normal migration. the abnormal GP Ib of Ba. platelets was strongly labeled when platelet suspensions were treated sequentially with neuraminidase, galactose oxidase, and sodium [3H]borohydride. Unlike the GP Ib of normal human platelets, it was also strongly labeled when Ba. platelets were treated with galactose oxidase and sodium [3H]borohydride alone. Both the alloantigen, PlA1, and quinidine-dependent antibody receptor activity were normally expressed by Ba. platelets, which also bound a monoclonal antibody (AN51) to GP Ib. Analysis of Ba. platelets by crossed immunoelectrophoresis using a rabbit anti-human platelet antibody preparation revealed the presence of an immunoprecipitate in the GP Ib position that had an abnormal appearance and migration in the second dimension. An altered position of the precipitate given by Factor VIIIR:Ag was also noted. Incorporation of HPA into the agarose gel during the first dimension electrophoresis resulted in the specific precipitation of the abnormal GP Ib of Ba. platelets. Our studies show that circulating Tn-platelets contain GP Ib with a modified oligosaccharide chain structure responsible for the platelet expression of Tn-antigen activity. PMID:7174794

Nurden, A T; Dupuis, D; Pidard, D; Kieffer, N; Kunicki, T J; Cartron, J P

1982-12-01

414

Surface modifications in the platelets of a patient with alpha-N-acetyl-D-galactosamine residues, the Tn-syndrome.  

PubMed Central

The Tn-syndrome is an acquired disorder characterized by the polyagglutination of blood cells and the pathological exposure of alpha-N-acetyl-D-galactosamine residues (Tn-antigen) at the cell surface. We now report studies on the platelet of a patient (Ba.) of which 81% reacted positively with a fluorescein conjugate of Helix pomatia agglutinin (HPA). The surface proteins of Ba. platelets were labeled with 125I by the lactoperoxidase-catalyzed procedure; single and two-dimensional electrophoresis on sodium dodecyl sulfate (SDS)-polyacrylamide gels was followed by autoradiography that revealed normal 125I-labeling of the major membrane glycoproteins (GP) but that GP Ib had a faster than normal migration. the abnormal GP Ib of Ba. platelets was strongly labeled when platelet suspensions were treated sequentially with neuraminidase, galactose oxidase, and sodium [3H]borohydride. Unlike the GP Ib of normal human platelets, it was also strongly labeled when Ba. platelets were treated with galactose oxidase and sodium [3H]borohydride alone. Both the alloantigen, PlA1, and quinidine-dependent antibody receptor activity were normally expressed by Ba. platelets, which also bound a monoclonal antibody (AN51) to GP Ib. Analysis of Ba. platelets by crossed immunoelectrophoresis using a rabbit anti-human platelet antibody preparation revealed the presence of an immunoprecipitate in the GP Ib position that had an abnormal appearance and migration in the second dimension. An altered position of the precipitate given by Factor VIIIR:Ag was also noted. Incorporation of HPA into the agarose gel during the first dimension electrophoresis resulted in the specific precipitation of the abnormal GP Ib of Ba. platelets. Our studies show that circulating Tn-platelets contain GP Ib with a modified oligosaccharide chain structure responsible for the platelet expression of Tn-antigen activity. Images

Nurden, A T; Dupuis, D; Pidard, D; Kieffer, N; Kunicki, T J; Cartron, J P

1982-01-01

415

Sialyl-Tn antigen as a marker of colon cancer risk in ulcerative colitis: Relation to dysplasia and DNA aneuploidy  

Microsoft Academic Search

Background & Aims: Expression of the mucin-associated carbohydrate antigen sialyl-Tn (STn) and DNA aneuploidy has each been shown to correlate with malignant transformation in patients with sporadic colon cancer and in those with ulcerative colitis (UC). This study aimed to determine how STn expression topographically and temporally relates to aneuploidy and neoplasia in patients with long-standing UC. Methods: Twenty-six patients

Per Karlén; Eric Young; Olle Broström; Robert Löfberg; Bernhard Tribukait; Åke Öst; Carol Bodian; Steven Itzkowitz

1998-01-01

416

A high-performance short-channel bottom-contact OTFT and its application to AM-TN-LCD  

Microsoft Academic Search

We have demonstrated an organic thin-film-transistor (OTFT)-driven active-matrix twisted-nematic liquid crystal display (AM-TN-LCD) on a glass substrate, with a resolution of 160×120 pixels, 79 ppi. Substrate temperature was kept below the plastic-compatible temperature of 180°C throughout the fabrication process. In order to realize an OTFT-driven display with fine resolution, we employed short-channel bottom-contact (BC) pentacene OTFTs. It has been known

Kazumasa Nomoto; Nobukazu Hirai; Nobuhide Yoneya; Noriyuki Kawashima; Makoto Noda; Masaru Wada; Jiro Kasahara

2005-01-01

417

A new ?-galactosidase from symbiotic Flavobacterium sp. TN17 reveals four residues essential for ?-galactosidase activity of gastrointestinal bacteria  

Microsoft Academic Search

The ?-galactosidase gene, galA17, was cloned from Flavobacterium sp. TN17, a symbiotic bacterium isolated from the gut of Batocera horsfieldi larvae. The 2,205-bp full-length gene encodes a 734-residue polypeptide (GalA17) containing a putative 28-residue signal\\u000a peptide and a catalytic domain belonging to glycosyl hydrolase family 36 (GH 36). The deduced amino acid sequence of galA17 was most similar to a

Junpei Zhou; Pengjun Shi; Huoqing Huang; Yanan Cao; Kun Meng; Peilong Yang; Rui Zhang; Xiaoyan Chen; Bin Yao

2010-01-01

418

Particle size dependence of magnetization and phase transition near TN in multiferroic BiFeO3  

NASA Astrophysics Data System (ADS)

We report results of a comprehensive study of the phase transition at TN (~643 K) as a function of particle size in multiferroic BiFeO3 system. We employed electrical, thermal, and temperature dependent x-ray diffraction studies in order to characterize the transition in a host of samples. We also carried out detailed magnetic measurements over a temperature regime of 2-300 K under a magnetic field of 100-10 000 Oe both on bulk and nanocrystalline systems. While in the bulk system a sharp endothermic peak at TN together with a broad feature, ranging over nearly ~100 K (?T), could be observed in calorimetry, the nanoscale systems exhibit only the broad feature. The characteristic dielectric anomaly, expected at TN, is found to occur both at TO and TN across ?T in the bulk sample. The Maxwell-Wagner component due to interfaces between heterogenous regions with different conductivities is also present. The magnetic properties, measured at lower temperature, corroborate our observations in calorimetry. The metastability increases in the nanoscale BiFeO3 with divergence between zero-field cooled and field cooled magnetizations below ~100 K and faster magnetic relaxation. Interestingly, in nanoscale BiFeO3 one also observes finite coercivity at lower temperature, which points out that suitable design of particle size and shape may induce ferromagnetism. The inhomogeneous distribution of Bi/Fe ions and/or oxygen nonstoichiometry seems to be giving rise to broad features in thermal, magnetic as well as electrical responses.

Mazumder, R.; Ghosh, S.; Mondal, P.; Bhattacharya, Dipten; Dasgupta, S.; Das, N.; Sen, A.; Tyagi, A. K.; Sivakumar, M.; Takami, T.; Ikuta, H.

2006-08-01

419

A series of Tn5 variants with various drug-resistance markers and suicide vector for transposon mutagenesis.  

PubMed

A series of variants of transposon Tn5 were constructed by replacement of the 2.7-kb central segment which encodes kanamycin resistance with various other resistance-coding genes: tetracycline, chloramphenicol, gentamicin, trimethoprim, streptomycin or ampicillin. A thermosensitive replication mutant of the broad-host-range transmissible plasmid R388 was also constructed for use as a suicide vector for the delivery of transposable elements. PMID:2824292

Sasakawa, C; Yoshikawa, M

1987-01-01

420

Particle size dependence of magnetization and phase transition near TN in multiferroic BiFeO3  

Microsoft Academic Search

We report results of a comprehensive study of the phase transition at TN (~643 K) as a function of particle size in multiferroic BiFeO3 system. We employed electrical, thermal, and temperature dependent x-ray diffraction studies in order to characterize the transition in a host of samples. We also carried out detailed magnetic measurements over a temperature regime of 2-300 K

R. Mazumder; S. Ghosh; P. Mondal; Dipten Bhattacharya; S. Dasgupta; N. Das; A. Sen; A. K. Tyagi; M. Sivakumar; T. Takami; H. Ikuta

2006-01-01

421

Clonal Expression of the Tn Antigen in Erythroid and Granulocyte Colonies and Its Application to Determination of the Clonality of the Human Megakaryocyte Colony Assay  

PubMed Central

To evaluate whether exposure of Tn determinants at the surface of human erythrocytes, platelets, and granulocytes could arise from a somatic mutation in a hemopoietic stem cell, burst-forming unit erythroid (BFU-E) colonies, colony-forming unit granulocyte-macrophage (CFU-GM), and colony-forming unit-eosinophil (CFU-Eo) were grown from a blood group O patient with a typical Tn syndrome displaying two distinct populations (Tn+ and Tn-) of platelets, granulocytes, and erythrocytes. A large number of colonies was observed. Individual colonies were studied with a fluorescent conjugate of Helix pomatia agglutinin (HPA). A sizeable fraction of each of the erythroid and granulocytic colonies appeared to consist exclusively of either HPA-positive or HPA-negative cells, thereby demonstrating the clonal origin of those exhibiting the Tn marker. Similar results were obtained from a second patient. These findings establish that the HPA labeling of Tn cells is an accurate marker permitting assessment of the clonality of the human megakaryocyte (MK) colony assay. For the study of MK cultures a double-staining procedure using the HPA lectin and a monoclonal antiplatelet antibody (J-15) was applied in situ to identify all MK constituting a colony. Our results, obtained in studies of 133 MK colonies, provide definitive evidence that the human MK colony assay is clonal because all MK colonies were exclusively composed of Tn+ and Tn- MK. Furthermore, the distribution of MK within a single colony was shown to be seminormal with a mean at 6 MK, isolated MK typically being absent in culture. Comparison of the proportion of mature Tn+ cells in blood with their respective Tn+ progenitors has also shown that no proliferative advantage occurs after the commitment; because Tn polyagglutinability is an acquired disorder, then the expansion of the Tn+ clone must occur either during the proliferative stage of the pluripotent stem cell or during the commitment itself. This study therefore affords evidence that a blood group antigen plays a role in the differentiation of a pluripotent stem cell. Images

Vainchenker, William; Testa, Ugo; Deschamps, Jeanne Francoise; Henri, Annie; Titeux, Monique; Breton-Gorius, Janine; Rochant, Henri; Lee, Douglas; Cartron, Jean-Pierre

1982-01-01

422

Mobilization and transfer of Azospirillum lipoferum plasmid by the Tn5-Mob transposon into a plasmid-free Agrobacterium tumefaciens strain.  

PubMed

Azospirillum lipoferum 4B harbors five cryptic plasmids. Several suicide plasmids were used to transfer Tn5-Mob to A. lipoferum 4B. Tn5-Mob insertion mutations of this strain could be obtained at frequencies of 10(-8)-10(-7) per recipient cell. One hundred Tn5-Mob A. lipoferum 4B mutants were used in bacterial matings with a plasmid-free Agrobacterium tumefaciens recipient strain. This is the first report of mobilization, transfer, and replication of an Azospirillum plasmid in Agrobacterium tumefaciens. One transconjugant was found which had lost an indigenous plasmid. PMID:2852995

Bally, R; Givaudan, A

1988-12-01

423

Analysis for prevalence and physical linkages amongst integrons, ISEcp1, ISCR1, Tn21 and Tn7 encountered in Escherichia coli strains from hospitalized and non-hospitalized patients in Kenya during a 19-year period (1992-2011)  

PubMed Central

Background We determined the prevalence and evidence for physical linkage amongst integrons, insertion sequences, Tn21 and Tn7 transposons in a collection of 1327 E. coli obtained over a 19-year period from patients in Kenya. Results The prevalence of class 1 integrons was 35%, class 2 integrons were detected in 3 isolates but no isolate contained a class 3 integron. Integron lacking the 3’-CS or those linked to sul3 gene or IS26 or those containing the ISCR1 were only detected in multidrug resistant (MDR) strains. The dfrAs were the most common cassettes and their prevalence was: - dfrA1(28%), dfrA12(20%), dfA17(9%), dfrA7(9%), and dfrA16(5%). The aadA were the second most abundant cassettes and their prevalence was: - aadA1(25%), aadA2(21%), and aadA5(14%). Other cassettes occurred in lower prevalence of below 5%. Prevalence of Tn21, ISEcp1, ISCR1 and IS26 was 22%, 10%, 15%, and 7% respectively. Majority of Tn21 containing integrons carried a complete set of transposition genes while class 2 integrons were borne on Tn7 transposon. The qnrA genes were detected in 34(3%) isolates while 19(1%) carried qnrB. All qnr genes were in MDR strains carrying integrons containing the ISCR1. Close to 88% of blaTEM-52 were linked to IS26 while???80% of blaCTX-Ms and blaCMYs were linked to ISEcp1. Only a few studies have identified a blaCTX-M-9 containing an ISEcp1 element as reported in this study. Multiple genetic elements, especially those borne on incIl, incFII, and incL/M plasmids, and their associated resistance genes were transferrable en bloc to E. coli strain J53 in mating experiments. Conclusions This is the first detailed study on the prevalence of selected elements implicated in evolution of resistance determinants in a large collection of clinical E. coli in Africa. Proliferation of such strains carrying multiple resistance elements is likely to compromise the use of affordable and available treatment options for majority of poor patients in Africa. There is therefore a need to monitor the spread of these highly resistant strains in developing countries through proper infection control and appropriate use of antimicrobials.

2013-01-01

424

Validation of a Tn5 transposon mutagenesis system for Gluconacetobacter diazotrophicus through characterization of a flagellar mutant.  

PubMed

Gluconacetobacter diazotrophicus is a nitrogen-fixing bacterium, which was originally isolated from the interior of sugarcane plants. The genome of strain PAL5 of G. diazotrophicus has been completely sequenced and a next step is the functional characterization of its genes. The aim of this study was to establish an efficient mutagenesis method, using the commercial Tn5 transposon EZ::Tn5Tnp Transposome (Epicentre). Up to 1 x 10(6) mutants per microgram of transposome were generated in a single electroporation experiment. Insertion-site flanking sequences were amplified by inverse PCR and sequenced for 31 mutants. For ten of these mutants, both insertion flanks could be identified, confirming the 9 bp duplication that is typical for Tn5 transposition. Insertions occurred in a random fashion and were genetically stable for at least 50 generations. One mutant had an insertion in a homolog of the flagellar gene flgA, and was therefore predicted to be affected in flagella-dependent traits and used to validate the applied mutagenesis methodology. This mutant lacked flagella and was non-motile on soft agar. Interestingly, it was also strongly affected in the ability to form biofilm on glass wool. PMID:18060666

Rouws, Luc F M; Simões-Araújo, Jean L; Hemerly, Adriana S; Baldani, José I

2008-04-01

425

Testing and analyses of the TN-24P PWR spent-fuel dry storage cask loaded with consolidated fuel  

SciTech Connect

A performance test of a Transnuclear, Inc. TN-24P storage cask configured for pressurized water reactor (PWR) spent fuel was performed. The work was performed by the Pacific Northwest Laboratory (PNL) and Idaho National Engineering Laboratory (INEL) for the US Department of Energy Office of Civilian Radioactive Waste Management (OCRWM) and the Electric Power Research Institute. The performance test consisted of loading the TN-24P cask with 24 canisters of consolidated PWR spent fuel from Virginia Power's Surry and Florida Power and Light's Turkey Point reactors. Cask surface and fuel canister guide tube temperatures were measured, as were cask surface gamma and neutron dose rates. Testing was performed with vacuum, nitrogen, and helium backfill environments in both vertical and horizontal cask orientations. Transnuclear, Inc., arranged to have a partially insulated run added to the end of the test to simulate impact limiters. Limited spent fuel integrity data were also obtained. From both heat transfer and shielding perspectives, the TN-24P cask with minor refinements can be effectively implemented at reactor sites and central storage facilities for safe storage of unconsolidated and consolidated spent fuel. 35 refs., 93 figs., 17 tabs.

McKinnon, M A; Michener, T E; Jensen, M F; Rodman, G R

1989-02-01

426

Phenotypic Screening of Escherichia coli K-12 Tn5 Insertion Libraries, Using Whole-Genome Oligonucleotide Microarrays  

PubMed Central

Complete genome sequences in combination with global screening methods allow parallel analysis of multiple mutant loci to determine the requirement for specific genes in different environments. In this paper we describe a high-definition microarray approach for investigating the growth effects of Tn5 insertions in Escherichia coli K-12. Libraries of insertion mutants generated by a unique Tn5 mutagenesis system were grown competitively in defined media. Biotin-labeled runoff RNA transcripts were generated in vitro from transposon insertions in each population of mutants. These transcripts were then hybridized to custom-designed oligonucleotide microarrays to detect the presence of each mutant in the population. By using this approach, the signal associated with 25 auxotrophic insertions in a 50-mutant pool was not detectable following nine generations of growth in glucose M9 minimal medium. It was found that individual insertion sites could be mapped to within 50 bp of their genomic locations, and 340 dispensable regions in the E. coli chromosome were identified. Tn5 insertions were detected in 15 genes for which no previous insertions have been reported. Other applications of this method are discussed.

Winterberg, Kelly M.; Luecke, John; Bruegl, Amanda S.; Reznikoff, William S.

2005-01-01

427

Mn-oxidizing Bacteria in Oak Ridge, TN and the Potential for Mercury Remediation  

NASA Astrophysics Data System (ADS)

East Fork Poplar Creek (EFPC) in Oak Ridge, TN was highly contaminated with elemental mercury in the 1950 and 1960. The area is still experiencing the effects of mercury contamination, and researchers are searching for ways to remediate the EFPC. One possible mechanism for bioremediation is the use of biogenic Mn oxides to remove heavy metals from water systems. Six native Pseudomonas bacteria species were isolated from the EFPC in order to examine biogenic Mn oxides production and bioremediation of Oak Ridge slurries. To investigate the biochemical interactions of Pseudomonas and the native microbial communities with Hg, Mn, Fe, S, six different slurry treatment groups were compared using inductively coupled plasma-atomic emission spectrometry (ICP-AES) and cold vapor atomic absorption spectrometry (CVAAS). Oak Ridge slurries were autoclaved to inhibit microbial growth (group 1), autoclaved and amended with HgS (group 2), autoclaved and amended with Pseudomonas isolates and additional HgS (group 3), untreated slurry (group 4), normal slurry amended with HgS (group 5), and normal slurry amended with Pseudomonas isolates and additional HgS (group 6). The comparison of the autoclaved groups with the counterpart untreated and normal Oak Ridge slurries highlighted important microbial interactions. Also, the Pseudomonas isolates were grown separately in a MnSO4 media, and the individual bacteria were monitored for Mn-oxidization using ICP-AES and transmission electron microscopy (TEM). In the slurry sediments, the Pseudomonas isolates did produce Mn oxides which bound to mercury, and mercury bound to organic matter significantly decreased. However, after a significant decrease of dissolved mercury in the water, dissolved mercury was cycled back into the water system on day 10 of the study. Additionally, two individual native Oak Ridge Pseudomonas isolates demonstrated Mn-oxidization. Biogenic Mn oxides have the potential to decrease mercury cycling, however there is need for more in depth and long-term studies to confirm their sustained use as Hg bioremediators.

Wright, K. L.; McNeal, K. S.; Han, F. X.

2012-12-01

428

[Variation in the genome of CTXphi prophage of Vibrio cholerae biovar El Tor caused by Tn5-Mob transposon].  

PubMed

A key pathogenicity factor of the cholera etiologic agent is cholera toxin (CT) whose synthesis is encoded by the ctxAB operon forming apart of the CTXphi ptophage. Alterations in the virulent properties of the cholera vibrios are based on the variability of the CTXphi prophage containing the genes for ctxAB, zot, ace, cep, orfU, and psh in its core region. At the same time, the mechanism of the porophage genome reorganization needs further and more profound analysis. The goal of this work was to demonstrate that transposon Tn5-Mob (Kmr), when introduced into the chromosome of the V. cholera model strain MAK757 El Tor biovar containing two copies of the CTXphi prophage provoked a reorganization in the CTXphi prophage consisting in the deletion of zot, ace, cep, orfU genes. The level of the CT biosynthesis in the insertion mutants MAK757 chr::Tn5-Mob still retaining only the ctxAB operon, increased more than 2000 times as compared to that of the original strain. The enhanced CT production was shown to be associated with the altered structure of the chromosomal DNA region containing one copy of the ctxAB operon encoding this protein biosynthesis. The mutation in the CTXphi genome induced by Tn5-Mob was unstable. Among 600 isolated colonies obtained after dissemination of the MAK757 chr::Tn5-Mob transposant capable of CT overproduction in the full medium with no antibiotics, 5.8% gave clones that in parallel to the loss of Kmr marker, appeared to be deprived of the ctxAB operon thus becoming non-toxinogenic. The observed formation of the V. cholerae insertion mutants both capable of CT overproduction and non-toxinogenic ones, may be indicative of an important role played in the evolution of the cholera pathogen by the CTXphi genome variability induced by Tn elements. The plasmidless V. cholerae El Tor strain characterized by type II CT hyperproduction thus obtained in our experiments could be used for the production of this protein routinely applied to construct efficient cholera diagnostic and prophylactic preparations. PMID:18756818

Shchelkanova, E Iu; Goriaev, A A; Smirnova, N I

2008-01-01

429

Bi-functional gfp- and gusA-containing mini-Tn5 transposon derivatives for combined gene expression and bacterial localization studies.  

PubMed

The gfp gene, encoding the green fluorescent protein, was combined with the gusA gene, coding for the beta-glucuronidase enzyme, in mini-Tn5 transposon derivatives for use in Gram-negative bacteria. These mini-Tn5 elements allow simultaneously monitoring of gene expression and localization of the marked bacteria. Introduction of the resultant mini-Tn5 transposons into Rhizobium etli, Azospirillum brasilense and Pseudomonas stutzeri allowed us to visualise the interaction of these bacteria with their host plant. The dual-marker mini-Tn5 transposons constitute a powerful new tool for studying gene expression and ecology of bacteria in the environment and during the interaction with plants. PMID:10076635

Xi, C; Lambrecht, M; Vanderleyden, J; Michiels, J

1999-02-01

430

Construction of a modified mini-Tn5 luxCDABE transposon for the development of bacterial biosensors for ecotoxicity testing.  

PubMed

A mini-Tn5 transposon was modified to introduce a promoterless luxCDABE cassette from Vibrio fischeri into environmentally relevant bacterial strains in order to develop bioluminescence-based biosensors for toxicity testing. The mini-Tn5 luxCDABE transposon was chromosomally integrated downstream from an active promoter into two Pseudomonas strains (Pseudomonas fluorescens 8866 and Pseudomonas putida F1). Characterisation of the bioluminescent transconjugants demonstrated that the transposon integration was stable and had no effect on growth rate. Both P. fluorescens 8866 Tn5 luxCDABE and P. putida F1 Tn5 luxCDABE were used to assess the toxicity of standard solutions (Cu, Zn and 3,5-DCP) as well as Cu- and 3,5-DCP-spiked groundwater samples. They were successfully used for bioluminescence-based bioassays and the potential value of using different bacterial biosensors for ecotoxicity testing was shown. PMID:11313129

Weitz, H J; Ritchie, J M; Bailey, D A; Horsburgh, A M; Killham, K; Glover, L A

2001-04-13

431

Prototype Planning Study: Middlesex County, New Jersey.  

National Technical Information Service (NTIS)

Middlesex County, New Jersey is experiencing transportation problems. To help solve escalating traffic problems on the county roads, they explored the potential of transportation systems management (TSM) techniques. The strategies they investigated fell i...

1979-01-01

432

Cherokee County Kansas Lead Surveillance Program.  

National Technical Information Service (NTIS)

The Cherokee County Kansas Lead Surveillance Program was conducted as a two year screening program (1994-1996) by Kansas Department of Health and Environment and Cherokee County Health Department with funds made available by Agency for Toxic Substances an...

B. Fulghum D. Mulnix S. N. Paige

1998-01-01

433

Solid Waste Management Plan, Emmet County.  

National Technical Information Service (NTIS)

The Solid Waste Management Plan for Emmet County suggests economically feasible solid waste collection and disposal systems. Georgraphic factors related to County land area, outstanding natural resources, and soil conditions represent strong elements in t...

1973-01-01

434

The use of TN:TP and DIN:TP ratios as indicators for phytoplankton nutrient limitation in oligotrophic lakes affected by N deposition  

Microsoft Academic Search

The stoichiometric composition of lake water chemistry affects nutrient limitation among phytoplankton. I show how TN:TP and\\u000a DIN:TP ratios vary in oligotrophic lakes of Europe and the USA affected by different amounts of N deposition, and evaluate\\u000a whether the DIN:TP ratio is a better indicator than the TN:TP ratio for discriminating between N and P limitation of phytoplankton.\\u000a Data were

Ann-Kristin Bergström

2010-01-01

435

The malate dehydrogenase of Ralstonia eutropha and functionality of the C 3\\/C 4 metabolism in a Tn 5-induced mdh mutant  

Microsoft Academic Search

The Tn5-induced mutant VG12 of Ralstonia eutropha HF39, which was isolated in this study, revealed an interesting phenotype: it grew on fructose and pyruvate as well as autotrophically like the wild-type, whereas growth on tricarboxylic acid intermediates and glyoxylic acid was reduced, and no growth occurred if acetate, propionate or levulinate were provided as carbon source. Tn5 was mapped in

Christian O Brämer; Alexander Steinbüchel

2002-01-01

436

A hisT::Tn5 mutation affects production of microcins B17, C7, and H47 and colicin V.  

PubMed Central

A Tn5 insertion decreasing the production of microcin B17 was mapped to 50.2 min on the Escherichia coli chromosome map. Sequence analysis showed that the insertion disrupted hisT, the gene encoding pseudouridine synthase I, a tRNA-modifying enzyme. hisT::Tn5 mutant cells were also shown to be defective for the production of other antibiotic peptides, such as microcin C7, microcin H47, and colicin V.

Rodriguez-Sainz, M C; Hernandez-Chico, C; Moreno, F

1991-01-01

437

The conjugative transposon Tn 925 : enhancement of conjugal transfer by tetracycline in Enterococcus faecalis and mobilization of chromosomal genes in Bacillus subtilis and E. faecalis  

Microsoft Academic Search

The effects of tetracycline on transfer of the conjugative, tetracycline-resistance transposon, Tn925, as well as the ability of the transposon to promote the transfer of chromosomal genes was examined in Enterococcus faecalis and Bacillus subtilis. To test for chromosomal transfer, multiply-marked strains of each organism, each carrying a single chromosomal copy of Tn925, were mated on filters with suitable recipient

Olga R. Torres; Ruth Z. KormanZ; Stanley A. Zahler; Gary M. Dunny

1991-01-01

438

Underemployment Estimates by County, United States, 1960.  

ERIC Educational Resources Information Center

This is a collection of estimates, by county, of the unutilized labor due to underemployment in 1960. This technique of determining unemployment uses the national median income adjusted for each country's age-color mix, education status, and an employment factor, to provide a county standard against which the actual county median income can be…

Kampe, Ronald E.; Lindamood, William A.

439

Water Management in Cache County, Utah  

Microsoft Academic Search

Cache County is one of the biggest agricultural producers in Utah and over 70% of the county's water is used for irrigation. In this project, we use NASA's Terrestrial Observation and Prediction System (TOPS) and Moderate Resolution Imaging Spectroradiometer (MODIS) data to gain an understanding of the water cycle in Cache County by comparing the precipitation, snowpack, and runoff amounts

G. Zhang; K. Lowry; R. Nemani; C. Schmidt; J. Skiles

2008-01-01

440

Community Types and Mortality in Georgia Counties  

ERIC Educational Resources Information Center

Using an "ecological regional analysis" methodology for defining types of communities and their associated mortality rates, this study of Georgia's 159 counties finds that the suburban and town centered counties have low mortality while the city-centered type predicts low mortality for the whites. The military-centered counties do not predict. The…

Young, Frank W.