Vibrio alginolyticus gyrB sequence analysis and gyrB-targeted PCR identification in environmental isolates
Luo, P; Hu, CQ; cqhu@scsio.ac.cn
2008
发表期刊DISEASES OF AQUATIC ORGANISMS
ISSN0177-5103
卷号82期号:3页码:209-216
摘要gyrB fragments (about 1.2 kb) of 9 Vibrio alginolyticus strains were sequenced, and their phylogenetic relationship with other closely related Vibrio species was analyzed. All the V alginolyticus strains grouped into one strongly supported cluster in the phylogenetic tree. There were 54 base variations among the 1167 bp mutual gyrB regions of 11 V alginolyticus strains; all the V alginolyticus strains shared the same amino acid sequence except V alginolyticus ATCC 17749. Based on the gyrB sequences, we designed 2 primers for specific PCR identification of V alginolyticus. Fifty-two bacterial strains from 12 genera were used to test the PCR specificity, and only V alginolyticus strains produced the predicted 568 bp amplification fragment. In addition, PCR screening of 50 randomly selected environmental strains, grown on thiosulfate citrate bile salts-sucrose (TCBS) medium, gave rise to a positive amplification result for V alginolyticus from 37 of them. To further confirm accuracy of PCR identification, biochemical identification of the 50 strains was carried out. Strains giving positive PCR amplification were biochemically identified as V alginolyticus, while strains that gave negative results were biochemically identified as other Vibrio or non-Vibrio species. Using the basic local alignment search tool (BLAST), gyrB sequences obtained from 2 randomly selected strains (YJ0666 and YJ16713) of the 37 PCR-positive strains showed highest identity values with V. alginolyticus strains (>96%). Thus, our results demonstrated that gyrB is a good marker for molecular identification of V alginolyticus, and a gyrB-based PCR method was successfully developed.
部门归属[Luo, Peng; Hu, Chaoqun] Chinese Acad Sci, S China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China
关键词Gyrb Vibrio Alginolyticus Pcr Identification
学科领域Fisheries ; Veterinary Sciences
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收录类别sci
资助项目LMB
WOS记录号WOS:000262839900005
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被引频次:18[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/4932
专题中科院海洋生物资源可持续利用重点实验室
通讯作者cqhu@scsio.ac.cn
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Luo, P,Hu, CQ,cqhu@scsio.ac.cn. Vibrio alginolyticus gyrB sequence analysis and gyrB-targeted PCR identification in environmental isolates[J]. DISEASES OF AQUATIC ORGANISMS,2008,82(3):209-216.
APA Luo, P,Hu, CQ,&cqhu@scsio.ac.cn.(2008).Vibrio alginolyticus gyrB sequence analysis and gyrB-targeted PCR identification in environmental isolates.DISEASES OF AQUATIC ORGANISMS,82(3),209-216.
MLA Luo, P,et al."Vibrio alginolyticus gyrB sequence analysis and gyrB-targeted PCR identification in environmental isolates".DISEASES OF AQUATIC ORGANISMS 82.3(2008):209-216.
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