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961.
Predictable and reproducible production of transgenic rats from a standardized input of egg donors and egg recipients is essential for routine rat model production. In the course of establishing a transgenic rat service, transgenic founders were produced from three transgenes in outbred Sprague-Dawley (SD) rats and four transgenes in inbred Fischer 344 (F344) rats. Key parameters that affect transgenesis efficiency were assessed, including superovulation treatments, methods to prepare pseudopregnant recipients, and microinjection technique. Five superovulation regimens were compared and treatment with 20 IU PMSG and 30 IU HCG was selected for routine use. Four methods to prepare pseudopregnant egg recipients were compared and estrus synchronization with LHRHa and mating to vasectomized males was selected as most effective. More than 80% of eggs survived microinjection when modified pronuclear microinjection needles and DNA buffers were used. The efficiencies of transgenic production in rats and C57BL/6J (B6J) mice were compared to provide a context for assessing the difficulty of transgenic rat production. Compared to B6J mice, SD rat transgenesis required fewer egg donors per founder, fewer pseudopregnant egg recipients per founder, and produced more founders per eggs microinjected. Similar numbers of injection days were required to produce founders. These results suggest that SD rat transgenesis can be more efficient than B6J mouse transgenesis with the appropriate technical refinements. Advances in transgenic rat production have the potential to increase access to rat models. 相似文献
962.
Pathogenomic sequence analysis of Bacillus cereus and Bacillus thuringiensis isolates closely related to Bacillus anthracis 总被引:6,自引:0,他引:6 下载免费PDF全文
Han CS Xie G Challacombe JF Altherr MR Bhotika SS Brown N Bruce D Campbell CS Campbell ML Chen J Chertkov O Cleland C Dimitrijevic M Doggett NA Fawcett JJ Glavina T Goodwin LA Green LD Hill KK Hitchcock P Jackson PJ Keim P Kewalramani AR Longmire J Lucas S Malfatti S McMurry K Meincke LJ Misra M Moseman BL Mundt M Munk AC Okinaka RT Parson-Quintana B Reilly LP Richardson P Robinson DL Rubin E Saunders E Tapia R Tesmer JG Thayer N Thompson LS Tice H Ticknor LO Wills PL Brettin TS Gilna P 《Journal of bacteriology》2006,188(9):3382-3390
Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis are closely related gram-positive, spore-forming bacteria of the B. cereus sensu lato group. While independently derived strains of B. anthracis reveal conspicuous sequence homogeneity, environmental isolates of B. cereus and B. thuringiensis exhibit extensive genetic diversity. Here we report the sequencing and comparative analysis of the genomes of two members of the B. cereus group, B. thuringiensis 97-27 subsp. konkukian serotype H34, isolated from a necrotic human wound, and B. cereus E33L, which was isolated from a swab of a zebra carcass in Namibia. These two strains, when analyzed by amplified fragment length polymorphism within a collection of over 300 of B. cereus, B. thuringiensis, and B. anthracis isolates, appear closely related to B. anthracis. The B. cereus E33L isolate appears to be the nearest relative to B. anthracis identified thus far. Whole-genome sequencing of B. thuringiensis 97-27and B. cereus E33L was undertaken to identify shared and unique genes among these isolates in comparison to the genomes of pathogenic strains B. anthracis Ames and B. cereus G9241 and nonpathogenic strains B. cereus ATCC 10987 and B. cereus ATCC 14579. Comparison of these genomes revealed differences in terms of virulence, metabolic competence, structural components, and regulatory mechanisms. 相似文献
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964.
Shilpa Jain Xiaoling Zhang Preeti J. Khandelwal Aleister J. Saunders Brian S. Cummings Peter Oelkers 《Journal of lipid research》2009,50(8):1563-1570
Esterifying lysophospholipids may serve a variety of functions, including phospholipid remodeling and limiting the abundance of bioactive lipids. Recently, a yeast enzyme, Lpt1p, that esterifies an array of lysophospholipids was identified. Described here is the characterization of a human homolog of LPT1 that we have called lysophosphatidylcholine acyltransferase 3 (LPCAT3). Expression of LPCAT3 in Sf9 insect cells conferred robust esterification of lysophosphatidylcholine in vitro. Kinetic analysis found apparent cooperativity with a saturated acyl-CoA having the lowest K0.5 (5 μM), a monounsaturated acyl-CoA having the highest apparent Vmax (759 nmol/min/mg), and two polyunsaturated acyl-CoAs showing intermediate values. Lysophosphatidylethanolamine and lysophosphatidylserine were also utilized as substrates. Electrospray ionization mass spectrometric analysis of phospholipids in Sf9 cells expressing LPCAT3 showed a relative increase in phosphatidylcholine containing saturated acyl chains and a decrease in phosphatidylcholine containing unsaturated acyl chains. Targeted reduction of LPCAT3 expression in HEK293 cells had essentially an opposite effect, resulting in decreased abundance of saturated phospholipid species and more unsaturated species. Reduced LPCAT3 expression resulted in more apoptosis and distinctly fewer lamellipodia, suggesting a necessary role for lysophospholipid esterification in maintaining cellular function and structure. 相似文献
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966.
967.
Bjork BC Fujiwara Y Davis SW Qiu H Saunders TL Sandy P Orkin S Camper SA Beier DR 《PloS one》2010,5(12):e14375
RNA interference (RNAi) is a powerful strategy for studying the phenotypic consequences of reduced gene expression levels in model systems. To develop a method for the rapid characterization of the developmental consequences of gene dysregulation, we tested the use of RNAi for "transient transgenic" knockdown of mRNA in mouse embryos. These methods included lentiviral infection as well as transposition using the Sleeping Beauty (SB) and PiggyBac (PB) transposable element systems. This approach can be useful for phenotypic validation of putative mutant loci, as we demonstrate by confirming that knockdown of Prdm16 phenocopies the ENU-induced cleft palate (CP) mutant, csp1. This strategy is attractive as an alternative to gene targeting in embryonic stem cells, as it is simple and yields phenotypic information in a matter of weeks. Of the three methodologies tested, the PB transposon system produced high numbers of transgenic embryos with the expected phenotype, demonstrating its utility as a screening method. 相似文献
968.
Tellew JE Lanier M Moorjani M Lin E Luo Z Slee DH Zhang X Hoare SR Grigoriadis DE St Denis Y Di Fabio R Di Modugno E Saunders J Williams JP 《Bioorganic & medicinal chemistry letters》2010,20(24):7259-7264
Antagonists of the corticotropin-releasing factor (CRF) neuropeptide may prove effective in treating stress and anxiety related disorders. In an effort to identify antagonists with improved physico-chemical properties a new series of CRF(1) antagonists were designed to substitute the propyl groups at the C7 position of the pyrazolo[1,5-a]pyrimidine core of 1 with heterocycles. Compound (S)-8d was identified as a high affinity ligand with a pK(i) value of 8.2 and a functional CRF(1) antagonist with pIC(50) value of 7.0 in the in vitro CRF ACTH production assay. 相似文献
969.
Louise E. Phillips Hang‐Gu Choi Gary W. Saunders Gerald T. Kraft 《Journal of phycology》2000,36(1):199-219
A morphological, anatomical, and molecular study of the two genera (Heterocladia and Trigenea) and three species of the tribe Heterocladieae (Rhodomelaceae, Ceramiales) is presented. First collections of male and female gametophytes of Heterocladia australis Decaisne and Trigenea umbellata J. Agardh and of tetrasporophytes of the type species of Trigenea, T. australis Sonder, have allowed a much clearer assessment of these taxa from a classical morphological standpoint. Reproductive and vegetative characters of the two Trigenea species are shown to be virtually identical to those of Heterocladia, which differs from Trigenea principally in having both flattened and terete lateral branches, as opposed to exclusively terete axes throughout. As a consequence, we propose to transfer the Trigenea species to the earlier‐named genus Heterocladia as H. caudata L. Phillips, H.‐G. Choi, G.W. Saunders et Kraft, nom. nov. and H. umbellata ( J. Agardh) L. Phillips, H.‐G. Choi, G.W. Saunders et Kraft, comb. nov. The close relationship of the three species is supported by molecular data, as nucleotide sequences of the 18S rRNA gene from each are nearly identical. The same sequences from species of eight other rhodomelaceous genera plus those from five outgroup taxa are analyzed to provide grounds for preliminary phylogenetic inferences about the position of the Heterocladieae in the Rhodomelaceae. Both the Heterocladieae and the Rhodomelaceae are monophyletic taxa in our analyses, the Heterocladieae grouping weakly with the Bostrychieae and the problematic Australian endemic genus Sonderella, the latter yet to be assigned to a tribe. Representatives of groups with which Heterocladia has been associated previously, such as the Lophothalieae and Brongniartelleae, appear to be only distantly related, although many more taxa need to be analyzed before the systematic position of the genus becomes clear. 相似文献
970.