大连海湾仿刺参肠道细菌多样性的16S rDNA分析

采用构建16SrDNA克隆文库的方法对大连湾仿刺参肠道中的细菌进行了多样性研究,NCBI数据库中序列的Blast比对结果表明,大连湾仿刺参肠道细菌具有高度多样性。挑取149个阳性克隆子,经HinfⅠ和HaeⅢ限制性内切酶谱型分析后得到36个可操作分类单元(OTUs)进行序列测定,克隆文库库容覆盖率为75.8%,结果是大连湾仿刺参肠道细菌分属于14种(属),主要为弧菌(Vibiro),优势菌群为Proteobacteria,占86.1%。用CLUSTAL X软件对36个OTUs序列和NCBI基因库中最相似的序列进行分析,应用MEGA 4软件中neighbor-joining方法构建系统发育树,确定大连湾仿刺参肠道细菌与已知菌之间基因系统进化关系。

Bacterial diversity in the intestine of Apostichopus japonicus in Dalian Bay

Bacterial diversity in the intestine of Apostichopus japonicus in Dalian Bay has been analyzed by constructing a 16SrDNA clone library.Results indicated the high bacterial diversity in the intestine of A.japonicus in Dalian Bay.A total of 149positive clones of 16SrDNA clone library were selected and 36operational taxonomy units(OTUs)were sequenced after digested by Hinf Ⅰ and Hae Ⅲ,and coverage of the library was 75.8%.The result shows that Vibiro sp.was the predominant bacteria of 14bacterial groups in the intestine of A.japonicus and the predominant group with approximately 86.1%of the sequences affiliated with Proteobacteria.The identities of the 36full-length sequences were confirmed by BLAST in NCBI databases,and the best matching sequences were retrieved from the database and analyzed with CLUSTAL X.The phylogenetic tree was constructed by neighbor-joining approach implemented in MEGA 4.