Fukushima Masao, Kakinuma Kenichi, Kawaguchi Ryuji
Genomics Research Institute, SRL, Inc., 5-6-50 Shinmachi, Hino-shi, Tokyo 191-0002, Japan.
J Clin Microbiol. 2002 Aug;40(8):2779-85. doi: 10.1128/JCM.40.8.2779-2785.2002.
Phylogenetic analysis of about 200 strains of Salmonella, Shigella, and Escherichia coli was carried out using the nucleotide sequence of the gene for DNA gyrase B (gyrB), which was determined by directly sequencing PCR fragments. The results establish a new phylogenetic tree for the classification of Salmonella, Shigella, and Escherichia coli in which Salmonella forms a cluster separate from but closely related to Shigella and E. coli. In comparison with 16S rRNA analysis, the gyrB sequences indicated a greater evolutionary divergence for the bacteria. Thus, in screening for the presence of bacteria, the gyrB gene might be a useful tool for differentiating between closely related species of bacteria such as Shigella spp. and E. coli. At present, 16S rRNA sequence analysis is an accurate and rapid method for identifying most unknown bacteria to the genus level because the highly conserved 16S rRNA region is easy to amplify; however, analysis of the more variable gyrB sequence region can identify unknown bacteria to the species level. In summary, we have shown that gyrB sequence analysis is a useful alternative to 16S rRNA analysis for constructing the phylogenetic relationships of bacteria, in particular for the classification of closely related bacterial species.
利用通过直接对聚合酶链反应(PCR)片段进行测序而确定的DNA促旋酶B(gyrB)基因的核苷酸序列,对约200株沙门氏菌、志贺氏菌和大肠杆菌进行了系统发育分析。结果建立了一个用于沙门氏菌、志贺氏菌和大肠杆菌分类的新系统发育树,其中沙门氏菌形成了一个与志贺氏菌和大肠杆菌分开但密切相关的簇。与16S核糖体RNA(rRNA)分析相比,gyrB序列表明这些细菌的进化差异更大。因此,在筛查细菌的存在时,gyrB基因可能是区分志贺氏菌属和大肠杆菌等密切相关细菌物种的有用工具。目前,16S rRNA序列分析是将大多数未知细菌鉴定到属水平的准确且快速的方法,因为高度保守的16S rRNA区域易于扩增;然而,对更具变异性的gyrB序列区域进行分析可以将未知细菌鉴定到种水平。总之,我们已经表明,gyrB序列分析是构建细菌系统发育关系的16S rRNA分析的有用替代方法,特别是对于密切相关细菌物种的分类。