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Natural Transformation of Helicobacter pylori Involves the Integration of Short DNA Fragments Interrupted by Gaps of Variable Size

机译:幽门螺杆菌的自然转化涉及短DNA片段的整合由可变大小的间隙打断。

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摘要

Helicobacter pylori are gram-negative bacteria notable for their high level of genetic diversity and plasticity, features that may play a key role in the organism's ability to colonize the human stomach. Homeologous natural transformation, a key contributor to genomic diversification, has been well-described for H. pylori. To examine the mechanisms involved, we performed restriction analysis and sequencing of recombination products to characterize the length, fragmentation, and position of DNA imported via natural transformation. Our analysis revealed DNA imports of small size (1,300 bp, 95% confidence limits 950–1850 bp) with instances of substantial asymmetry in relation to selectable antibiotic-resistance markers. We also observed clustering of imported DNA endpoints, suggesting a possible role for restriction endonucleases in limiting recombination length. Additionally, we observed gaps in integrated DNA and found evidence suggesting that these gaps are the result of two or more separate strand invasions. Taken together, these observations support a system of highly efficient short-fragment recombination involving multiple recombination events within a single locus.
机译:幽门螺杆菌是革兰氏阴性细菌,以其高水平的遗传多样性和可塑性而著称,这些特征可能在生物体在人胃中的定殖能力中发挥关键作用。同源自然转化是基因组多样化的关键因素,对于幽门螺杆菌已有很好的描述。为了检查涉及的机制,我们进行了限制性分析和重组产物测序,以表征通过自然转化导入的DNA的长度,片段化和位置。我们的分析显示,DNA导入量小(1,300 bp,95%置信度限制950–1850 bp),并且与可选的抗生素抗性标记物存在明显的不对称性。我们还观察到了进口DNA端点的聚类,表明限制性核酸内切酶在限制重组长度方面可能发挥作用。此外,我们观察到了整合DNA中的缺口,并发现证据表明这些缺口是两个或多个分开的链入侵的结果。综上所述,这些观察结果支持了涉及单个位点内多个重组事件的高效短片段重组系统。

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