猬迭宫绦虫线粒体cox3基因的克隆及序列分析

Cloning and Sequence Analysis of Mitochondrial Cytochrome c Oxidase Subunit 3 of Spirometra erinaceieuropaei Isolates from Hunan Province

本研究旨在阐明猬迭宫绦虫湖南分离株线粒体细胞色素c氧化酶第Ⅲ亚基(cox3)基因部分序列(pcox3)的遗传变异情况。利用聚合酶链反应(PCR)扩增猬迭宫绦虫的pcox3,应用ClustalX 1.81程序对序列进行比对,同时利用DNAStar5.0中的Megalign程序进行同源性分析,再用Phylip 3.67程序MP法绘制系统发育树,并与GenBank中已知猬迭宫绦虫相应基因序列进行比对分析。结果显示,所获得的pcox3序列长度一致,均为264bp,湖南分离株与已知猬迭宫绦虫位于同一分支,来自湖南省的猬迭宫绦虫pcox3序列有微小差异。本研究结果为猬迭宫绦虫进一步的分类、鉴定和遗传变异研究奠定了基础。

The objectives of the present study was to examine sequence variation in the mitochondrial cytochrome c oxidase subunit 3（cox3） gene among Spirometra erinaceieuropaei isolates from Hunan province.The partial cox3（pcox3） was amplified from each S.erinaceieuropaei sample by PCR,and pcox3 sequences were aligned using the ClustalX 1.81.MP tree was constructed using the software Phylip 3.67 version 4.0.Sequence homology analysis was performed using the Megalign program of the software DNAStar 5.0 by comparing with the corresponding sequence of S.erinaceieuropaei available in GenBank.The results showed that the lengths of pcox3 were 264 bp.The phylogenetic tree showed that the Hunan isolates and the S.erinaceieuropaei available in GenBank were clustered in the same clade.The pcox3 sequences of S.erinaceieuropaei isolates from Hunan province were slight different.The results provided a foundation for further studies of molecular identification and molecular genetics of S.erinaceieuropaei.