Lin Edward A, Zhang Xue-Song, Levine Steven M, Gill Steven R, Falush Daniel, Blaser Martin J
Department of Medicine, New York University School of Medicine, New York, New York, United States of America.
PLoS Pathog. 2009 Mar;5(3):e1000337. doi: 10.1371/journal.ppat.1000337. Epub 2009 Mar 13.
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.
幽门螺杆菌是革兰氏阴性菌,以其高度的遗传多样性和可塑性而闻名,这些特性可能在该生物体定殖于人类胃部的能力中起关键作用。同源自然转化是基因组多样化的一个关键因素,幽门螺杆菌的同源自然转化已得到充分描述。为了研究其中涉及的机制,我们对重组产物进行了限制性分析和测序,以表征通过自然转化导入的DNA的长度、片段化和位置。我们的分析揭示了小尺寸的DNA导入(1300 bp,95%置信区间为950 - 1850 bp),且与可选择的抗生素抗性标记相比存在明显的不对称情况。我们还观察到导入DNA端点的聚集,这表明限制性内切酶在限制重组长度方面可能发挥作用。此外,我们在整合的DNA中观察到间隙,并发现证据表明这些间隙是两个或更多单独的链侵入的结果。综上所述,这些观察结果支持了一个高效的短片段重组系统,该系统涉及单个基因座内的多个重组事件。