Ki Jang-Seu, Zhang Rui, Zhang Wen, Huang Yi-Li, Qian Pei-Yuan
Department of Biology, Coastal Marine Laboratory, Hong Kong University of Science and Technology, Clearwater Bay, Kowloon, Hong Kong SAR, China.
Microb Ecol. 2009 Nov;58(4):679-91. doi: 10.1007/s00248-009-9519-7. Epub 2009 May 7.
In the present study, we sequenced the RNA polymerase beta subunit (rpoB) gene of marine Vibrio species and assessed its discriminative power in identifying vibrios. Both the rpoB and 16S rRNA sequences of 29 phenotypically different Vibrio strains isolated from coastal waters were determined. Molecular and phylogenetic comparisons of the sequences of these two genes classified the 29 strains into 11 different species. The resolution of the Vibrio spp. on the rpoB phylogenetic tree was approximately three times greater than that on the 16S rRNA phylogenetic tree. Moreover, by comparing the rpoB sequences of 98 marine gamma-Proteobacteria, including 38 marine Vibrio species, Vibrio-specific primers were developed to amplify a 730-bp fragment of the rpoB gene. Using these primers, we successfully detected Vibrio signals in environmental samples and determined their relative abundances via comparisons with known standards. This rpoB-targeting polymerase chain reaction assay can be used efficiently to monitor relative Vibrio abundance in marine waters.
在本研究中，我们对海洋弧菌属物种的RNA聚合酶β亚基（rpoB）基因进行了测序，并评估了其在鉴定弧菌方面的鉴别能力。测定了从沿海水域分离出的29株表型不同的弧菌菌株的rpoB和16S rRNA序列。对这两个基因序列进行的分子和系统发育比较将这29株菌株分为11个不同的物种。弧菌属在rpoB系统发育树上的分辨率大约是在16S rRNA系统发育树上的三倍。此外，通过比较98种海洋γ-变形菌（包括38种海洋弧菌属物种）的rpoB序列，开发了弧菌属特异性引物，以扩增rpoB基因的一个730bp片段。使用这些引物，我们成功地在环境样品中检测到弧菌信号，并通过与已知标准进行比较确定了它们的相对丰度。这种针对rpoB的聚合酶链反应测定法可有效地用于监测海水中弧菌的相对丰度。
