Desjardins P, Picard B, Kaltenböck B, Elion J, Denamur E
INSERM U 120, Hôpital Robert Debre, Paris, France.
J Mol Evol. 1995 Oct;41(4):440-8. doi: 10.1007/BF00160315.
Analysis of the Escherichia coli population by multilocus enzyme electrophoresis (MLEE) has established its clonal organization, but there is increasing evidence that horizontal DNA transfer occurs in E. coli. We have assessed the genetic structure of the species E. coli and determined the extent to which recombination can affect the clonal structure of bacteria. A panel of 72 E. coli strains from the ECOR collection was characterized by random amplified polymorphic DNA (RAPD) and restriction-fragment-length polymorphism (RFLP) of the ribosomal RNA gene (rrn) regions. These strains have been characterized by MLEE and are assumed to reflect the range of genotypic variation in the species as a whole. Statistical analysis, including factorial analysis of correspondence (FAC) and hierarchical classifications, established that the data obtained with the three genetic markers are mutually corroborative, thus providing compelling evidence that horizontal transfer does not disrupt the clonal organization of the population. However, there is a gradient of correlation between the different classifications which ranges from the highly clonal structure of B2 group strains causing extraintestinal infections in humans to the less-stringent structure of B1 group strains that came mainly from nonprimate mammals. This group (B1) appears to be the framework from which the remaining non-A group strains have emerged. These results indicate that RAPD analysis is well suited to intraspecies characterization of E. coli. Lastly, treating the RAPD data by FAC allowed description of subgroup-specific DNA fragments which can be used, in a strategy comparable to positional cloning, to isolate virulence genes.
通过多位点酶电泳(MLEE)对大肠杆菌群体进行分析已确定了其克隆组织,但越来越多的证据表明大肠杆菌中发生了水平DNA转移。我们评估了大肠杆菌物种的遗传结构,并确定了重组对细菌克隆结构的影响程度。利用随机扩增多态性DNA(RAPD)和核糖体RNA基因(rrn)区域的限制性片段长度多态性(RFLP)对来自ECOR菌库的72株大肠杆菌进行了特征分析。这些菌株已通过MLEE进行了特征分析,并被认为反映了整个物种的基因型变异范围。包括对应因子分析(FAC)和层次分类在内的统计分析表明,用这三种遗传标记获得的数据相互佐证,从而提供了令人信服的证据,证明水平转移不会破坏群体的克隆组织。然而,不同分类之间存在相关性梯度,范围从导致人类肠道外感染的B2组菌株的高度克隆结构到主要来自非灵长类哺乳动物的B1组菌株的不那么严格的结构。这一组(B1)似乎是其余非A组菌株出现的框架。这些结果表明,RAPD分析非常适合于大肠杆菌的种内特征分析。最后,通过FAC处理RAPD数据可以描述亚组特异性DNA片段,这些片段可用于一种类似于定位克隆的策略中,以分离毒力基因。