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33.
Baar C d'Abbadie M Vaisman A Arana ME Hofreiter M Woodgate R Kunkel TA Holliger P 《Nucleic acids research》2011,39(8):e51
Potent inhibitors limit the use of PCR assays in a wide spectrum of specimens. Here, we describe the engineering of polymerases with a broad resistance to complex environmental inhibitors using molecular breeding of eight different polymerase orthologues from the genus Thermus and directed evolution by CSR in the presence of inhibitors. Selecting for resistance to the inhibitory effects of Neomylodon bone powder, we isolated 2D9, a chimeric polymerase comprising sequence elements derived from DNA polymerases from Thermus aquaticus, Thermus oshimai, Thermus thermophilus and Thermus brockianus. 2D9 displayed a striking resistance to a broad spectrum of complex inhibitors of highly divergent composition including humic acid, bone dust, coprolite, peat extract, clay-rich soil, cave sediment and tar. The selected polymerase promises to have utility in PCR-based applications in a wide range of fields including palaeobiology, archaeology, conservation biology, forensic and historic medicine. 相似文献
34.
Blood biochemical polymorphisms as markers for genetic characteristics of wild Spanish and domestic rabbits 总被引:1,自引:0,他引:1
Seventeen blood proteins were studied in a sample of 412 Spanish wild rabbits and in 598 domestic rabbits belonging to various breeds. The wild rabbit populations showed a high level of genetic polymorphism. Six loci were monomorphic, while the remaining ten loci were segregating for at least two alleles. Two of the loci that were polymorphic in the wild rabbits were monomorphic in the domestic ones. Wright's inbreeding coefficient in the total Spanish wild rabbit population was F=5.66, indicating subdivision of the total population. Inbreeding coefficients, estimated by Kidd et al.'s method (Anim. Blood Grps, Biochem. Genet. 11: 21–38), differed significantly from zero, being 15.62%, in wild rabbits and 6–12% in domestic breeds, indicating consanguinity.Genetic distances between wild rabbit populations showed that factors other than geographic distance (e.g., bottlenecks, barriers such as rivers, mountains, etc.) may explain the result that a northern population forms a cluster with two central populations whereas the northeastern populations form a different cluster with another central population. Populations of the first cluster are more closely related to the captive populations than others.There are three population clusters of domestic rabbits, namely (1) New Zealand White and a hybrid combination; (2) Spanish Common, Butterfly, Burgundy, and Californian; and (3) Spanish Giant. 相似文献
35.
Using the method of gene transfection with liposome, we obtained the mouse erythroleukemia cell line MEL-TF19, which stably carries TFAR19, a novel apoptosis-related gene. The expression of TFAR19 was detected by Western blot. Growth curve and flow cytometry analysis showed that after being transfected with TFAR19 gene, the growth of MEL-TF19 is suppressed and its apoptosis is accelerated because of the serum deprivation. Our biorheological study indicated that in the apoptotic process, compared with MEL cells, MEL-TF19 cells exhibit larger osmotic fragility, lower cell surface charge density, increased elastic modulus K1 which is inversely proportional to cells' maximal deformation ability, obviouslydiminished surface viscosity μ, with elastic modulus K2 having no distinct changes. The above results provided some bases for recognizing the function of TFAR19 completely from the viewpoint of biorheology. 相似文献
36.
Human DNA polymerase nu (pol nu) is one of three A family polymerases conserved in vertebrates. Although its biological functions are unknown, pol nu has been implicated in DNA repair and in translesion DNA synthesis (TLS). Pol nu lacks intrinsic exonucleolytic proofreading activity and discriminates poorly against misinsertion of dNTP opposite template thymine or guanine, implying that it should copy DNA with low base substitution fidelity. To test this prediction and to comprehensively examine pol nu DNA synthesis fidelity as a clue to its function, here we describe human pol nu error rates for all 12 single base-base mismatches and for insertion and deletion errors during synthesis to copy the lacZ alpha-complementation sequence in M13mp2 DNA. Pol nu copies this DNA with average single-base insertion and deletion error rates of 7 x 10(-5) and 17 x 10(-5), respectively. This accuracy is comparable to that of replicative polymerases in the B family, lower than that of its A family homolog, human pol gamma, and much higher than that of Y family TLS polymerases. In contrast, the average single-base substitution error rate of human pol nu is 3.5 x 10(-3), which is inaccurate compared to the replicative polymerases and comparable to Y family polymerases. Interestingly, the vast majority of errors made by pol nu reflect stable misincorporation of dTMP opposite template G, at average rates that are much higher than for homologous A family members. This pol nu error is especially prevalent in sequence contexts wherein the template G is preceded by a C-G or G-C base pair, where error rates can exceed 10%. Amino acid sequence alignments based on the structures of more accurate A family polymerases suggest substantial differences in the O-helix of pol nu that could contribute to this unique error signature. 相似文献
37.
石羊河下游民勤绿洲退耕地植被自然演替特征及物种多样性分析 总被引:3,自引:0,他引:3
采用重要值、Margalef物种丰富度指数、Shannon-wiener和Simpson多样性指数、Pielou和Alatalo均匀度指数对石羊河下游民勤绿洲不同年限退耕地自然恢复的植物群落进行了研究。结果表明:(1)石羊河下游民勤绿洲退耕地在50a自然恢复过程中共出现植物34种,14科,其中灌木6种,草本28种,分别占总物种数的17.65%和82.35%;藜科(8种)、禾本科(6种)、蒺藜科(4种)植物占总物种数的52.94%。(2)研究区退耕后50a植物群落演替经历为:田旋花→藜→苦苣菜→骆驼蒿→骆驼蓬→盐生草→黑果枸杞→红砂→盐爪爪的演替过程。(3)群落物种丰富度和多样性指数均随退耕时间的延长而呈现出波动式下降的变化趋势,而均匀度指数则呈现出在退耕初期(1~5a)先下降然后呈波动式上升的变化趋势。研究表明,随着退耕年限的增加,群落物种组成逐渐减少,植物群落演替向前发展的大致经历分为4个阶段:退耕1~5a为一年生草本和宿根植物迅速恢复阶段,退耕5~15a为一年生草本植物向多年生草本演替阶段,退耕15~30a为多年生草本向多年生灌木演替阶段,退耕30~50a为多年生灌木植物稳定阶段。 相似文献
38.
Aurora Fernández-Astorga María Joseé Hijarrubia María Hernández Ines Arana Ester Suñen 《Letters in applied microbiology》1995,20(5):308-311
A total of 330 strains of psychrotrophic non-fermenting Gram-negative bacteria isolated from vegetables were studied. In spite of the wide range of antibiotic resistance occurring, less than 10% showed resistance patterns which included mezclocillin-ticarcillin-gentamicin or ceftizoxime-norfloxacin. Reductions of > 5 log10 in the numbers of cfu were found when these strains were exposed for 30 min to a quaternary ammonium compound (1% w/v). 相似文献
39.
Borda Enri Camusso Juan Jose Perez Leiros Claudia Bacman Sandra Hubscher Osvaldo Arana Roberto Sterin-Borda Leonor 《Molecular and cellular biochemistry》1996,163(1):335-341
Isolated congenital heart block may be associated with Primary Sjogren's Syndrome. In this work we demonstrated that IgG present in the sera ofpatients with Primary Sjogren's Syndrome (PSS) could bind and activate muscarinic acetylcholine receptors of rat neonatal atria. These antibodies were able to inhibit in a irreversible manner the binding of 3H-QNB to muscarinic cholinergic receptors of purified rat atria membranes. Moreover, IgG from PSS individuals could modify biological effects mediated by muscarinic cholinoceptors activation, i.e. decrease contractility and cAMP and increase phosphoinositide turnover and cGMP. Atropine blocked all of these effects and carbachol mimicked them; confirming muscarinic cholinergic receptors-mediated PSS IgG action. Neither binding nor biological effect were obtained using adult instead of neonatal rat atria. IgG from sera of normal women were not effective in the studied system. The prevalence of cholinergic antibody was 100% in PSS and was independent of Ro/SS-A and La/SS-B antibodies. It could be concluded that antibody against muscarinic cholinergic receptors may be another serum factor to be considered in the pathophysiology of the development of congenital heart block. 相似文献
40.
Siddhivinayak Hirve Marleen Boelaert Greg Matlashewski Dinesh Mondal Byron Arana Axel Kroeger Piero Olliaro 《PLoS neglected tropical diseases》2016,10(8)
BackgroundAs Bangladesh, India and Nepal progress towards visceral leishmaniasis (VL) elimination, it is important to understand the role of asymptomatic Leishmania infection (ALI), VL treatment relapse and post kala-azar dermal leishmaniasis (PKDL) in transmission.