Iguchi Atsushi, Iyoda Sunao, Terajima Jun, Watanabe Haruo, Osawa Ro
Department of Bioscience, Graduate School of Science and Technology, Kobe University, 1-1 Rokko-dai, Kobe, Hyogo, 657-8501, Japan.
Gene. 2006 May 10;372:199-207. doi: 10.1016/j.gene.2006.01.005. Epub 2006 Mar 3.
It is known that XbaI-digested chromosomal DNAs of strains of Escherichia coli O157:H7 exhibit a wide variety in pulsed-field gel electrophoresis (PFGE) fragment patterns, which is used for epidemiological surveillance of this important pathogen. The variety in the restriction enzyme-digestion patterns suggests a wide genomic diversity, however, only a few studies have been conducted to investigate involvement of large-scale chromosomal rearrangements in development of the diversity. In this study, through rounds of subculturing E. coli O157:H7 strain EDL933, naturally occurring genome variation in the isolated derivatives was investigated. By comparing the PFGE patterns among clonally related derivatives, we found five types of large-scale inversions taking place within the chromosome. The five inversions found were across the replication axis and ranged from 250-kb to 1.4-Mb long, and all the corresponding recombination sites were associated with prophages or phage-like regions. Four inversions out of the five were resulted from recombination between pairs of lambda-like prophages disturbing the symmetry of the origin and terminus of the replication axis. These observations indicate that those prophage regions represent some of the hot spots for intrachromosomal recombination within the E. coli O157:H7 chromosome, where recombination between the prophage regions results not only in the large chromosomal inversions but might also in generation of chimeric phages.
已知大肠杆菌O157:H7菌株经XbaI酶切的染色体DNA在脉冲场凝胶电泳(PFGE)片段模式上表现出广泛的多样性,该技术用于对这种重要病原体进行流行病学监测。限制酶消化模式的多样性表明基因组存在广泛的差异,然而,仅有少数研究探讨了大规模染色体重排在这种差异形成过程中的作用。在本研究中,通过对大肠杆菌O157:H7菌株EDL933进行多轮传代培养,研究了分离出的衍生物中自然发生的基因组变异。通过比较克隆相关衍生物之间的PFGE模式,我们发现染色体内部发生了五种大规模倒位。所发现的这五种倒位跨越复制轴,长度从250 kb到1.4 Mb不等,所有相应的重组位点都与原噬菌体或类噬菌体区域相关。五种倒位中有四种是由成对的λ样原噬菌体之间的重组导致的,这破坏了复制轴起点和终点的对称性。这些观察结果表明,那些原噬菌体区域代表了大肠杆菌O157:H7染色体内部染色体间重组的一些热点区域,在这些区域,原噬菌体区域之间的重组不仅导致大规模染色体倒位,还可能产生嵌合噬菌体。