Department of Biology, Faculty of Sciences, Karadeniz Technical University, 61080, Trabzon, Turkey.
J Microbiol. 2011 Oct;49(5):782-90. doi: 10.1007/s12275-011-1136-8. Epub 2011 Nov 9.
This study was conducted to investigate the applicability of rpoB, which encodes the β subunit of RNA polymerase, to be used as an alternative to 16S rRNA gene sequence similarity analysis in the thermophilic genus Anoxybacillus. Partial rpoB sequences were generated for the 14 type strains of Anoxybacillus species and 6 other strains of four Anoxybacillus species. The sequences and the phylogenetic tree of rpoB were compared with those obtained from 16S rRNA gene analysis. The rpoB gene was found to provide a better resolution for Anoxybacillus species, with lower interspecies sequence similarities. The rpoB sequence similarity analysis permitted a more accurate discrimination of the species within the Anoxybacillus genus than the more commonly used 16S rRNA gene. Furthermore, rapid and reproducible repetitive extragenic palindromic fingerprinting techniques (REP-, ERIC-, and BOX-PCR) were employed for the specimens of genus Anoxybacillus. Through comparison of the three methods, it was found that the BOX-PCR method generated more informative results than REP-PCR for the studied strains; BOX-PCR profiles were more distinct for the different strains, including a higher number of bands. Rapid and reproducible repetitive extragenic palindromic fingerprinting techniques (rep-PCR) constitute a suitable molecular approach for the validation and maintenance of taxonomy within the Anoxybacillus genus. The results of this study show that rpoB and rep-PCR provide rapid and reliable methods for molecular typing of Anoxybacillus species.
本研究旨在探讨 rpoB（编码 RNA 聚合酶β亚基）在嗜热属 Anoxybacillus 中的应用，作为 16S rRNA 基因序列相似性分析的替代方法。对 14 株 Anoxybacillus 种的模式株和 6 株其他 4 种 Anoxybacillus 种的菌株进行了部分 rpoB 序列生成。比较了 rpoB 序列和系统发育树与 16S rRNA 基因分析获得的序列和系统发育树。rpoB 基因对 Anoxybacillus 种的分辨率更高，种间序列相似度更低。rpoB 序列分析比更常用的 16S rRNA 基因更能准确地区分 Anoxybacillus 属内的种。此外，还对 Anoxybacillus 属的标本进行了快速且可重复的重复外回文重复单位指纹图谱技术（REP-PCR、ERIC-PCR 和 BOX-PCR）。通过比较这三种方法，发现 BOX-PCR 方法比 REP-PCR 更能为研究菌株提供更具信息量的结果；BOX-PCR 图谱对不同菌株更为独特，包括更多的条带。快速且可重复的重复外回文重复单位指纹图谱技术（rep-PCR）构成了验证和维护 Anoxybacillus 属分类学的合适分子方法。本研究结果表明，rpoB 和 rep-PCR 为 Anoxybacillus 种的分子分型提供了快速可靠的方法。
