哈维弧菌16S rRNA基因拷贝数的种内变异  

Intra-species variation of 16S rRNA gene copy number of Vibrio harveyi

作　　者：郑嘉来 阎永伟[1] 唐姝[1] 杨坤杰[1] 李长红[1] 王凯[1] 张德民[1] 

机构地区：[1]宁波大学应用海洋生物技术教育部重点实验室,宁波315211

出　　处：《生物学杂志》2015年第6期6-11,共6页Journal of Biology

基　　金：国家自然科学基金项目(30970104);国家863项目(2012AA092000);宁波市科技计划项目(2010C10013)

摘　　要：利用荧光定量PCR法测定哈维弧菌（Vibrio harveyi）基因组内16S rRNA基因拷贝数。基于16S rDNA、pyrH、rpo A和rec A4个基因的系统发育学分析鉴定弧菌菌株,然后通过重叠PCR构建包含哈维弧菌模式菌株16S rDNA和gyrB基因片段的标准质粒,建立了定量PCR测定细胞内16S rRNA基因拷贝数方法,并测定了6株测试菌株和哈维弧菌模式株的16S rRNA基因拷贝数,同时以已知拷贝数的菌株ATCC BAA-1116做对照。6个测试菌株均为哈维弧菌。菌株ATCC BAA-1116的16S rRNA基因拷贝数为11个,与已知值相符。模式菌株MCCC 1H00031-T和6个测试菌株SXB-C、SCB-4、NBV00029、NBV00035、NBV00039和NBV00269的拷贝数分别为9、12、13、12、11、13和9个。利用荧光定量PCR法测定哈维弧菌基因组内16S rRNA基因拷贝数的方法简单可行。哈维弧菌16S rRNA基因拷贝数的种内变异可能是其适应近岸多变生境的原因之一。This study aims to determine intra-genomic 16S rRNA gene copy number of Vibrio harveyi strains by a method of fluorescent quantitative PCR. Vibrio strains were identified by multilocussequence analysis based on 16S rDNA, pyrH, rpoA and recA. A standard plasmid, harboring fragments of 16S rDNA and the single copy gene gyrB from V. harveyi type strain, was constructed. A fluorescent quantitative method was established and used to determine the intra-genomic 16S rRNA gene copy number of the tested 6 strains, taken the type strains of V. harveyi and V. harvei ATCC BAA-1116 as reference, whose 16S rRNA gene copy number is well known. All the 6 tested Vibrio strains were identified as V. harveyi. The copy number of 16S rRNA gene in strain ATCC BAA-1116 was 11, consistent with the known values. The copy number of strain MCCC 1H00031T, SXB-C, SCB-4, NBV00029, NBV00035, NBV00039 and NBV00269 was 9, 12, 13, 12, 11, 13 and 9, respectively. Fluorescent quantitative PCR method is simple and reliable for determi-ning 16S rRNA gene copy number within genome. Intra-species variation of 16S rRNA gene copy number of V. harveyi might be of eco-logical significance for their adaptation to fluctuant coastal niches.

关 键 词：哈维弧菌 16S RRNA基因 基因拷贝数 定量PCR 

分 类 号：Q93-331[生物学—微生物学] Q93-31