斯氏副柔线虫线粒体CO1基因的克隆与序列分析  

Cloning and sequence analysis of the partial CO1 gene within mitochondrial DNA of Parabronema skrjabini

作　　者：李文生[1] 杨莲茹[1] 杨晓野[1] 赵治国 赵晓慧[1] 李鑫 王瑞[1] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]内蒙古出入境检验检疫局,内蒙古呼和浩特010020 [3]内蒙古巴彦淖尔市乌拉特后旗潮格尔镇兽医工作站,内蒙古巴彦淖尔015500

出　　处：《畜牧与兽医》2011年第5期25-30,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(30660124);内蒙古自治区自然科学基金项目(200607010401)

摘　　要：为研究斯氏副柔线虫(Parabronema skrjabini)种间遗传标记特点,从内蒙古地区骆驼皱胃中采集斯氏副柔线虫成虫,提取虫体DNA,利用线虫通用引物扩增细胞色素氧化酶1(CO1)基因,将其克隆到pMD19-T载体后,用PCR技术鉴定阳性菌落并进行测序,运用DNAStar 5.0和MEGA 4.0软件将测序结果与GenBank公布的相关线虫CO1序列进行比对分析。结果显示:斯氏副柔线虫DNA扩增出的CO1基因序列片段长度为689 bp。同其他相关属线虫CO1基因序列比对,斯氏副柔线虫与小口柔线虫(Habronema microstoma)、蝇柔线虫(Habronema muscae)的同源性最高,为86.7%,遗传距离为0.143;与加利西亚光丝虫(Litomosoides galizai)的同源性最低,为80.1%,遗传距离为0.229。通过两种不同方法建立的系统发育树比较,证实斯氏副柔线虫与柔线属线虫均位于同一分支,自展值都在47%以上。表明CO1基因不仅可以作为斯氏副柔线虫理想的种间遗传标记,也可为该线虫进一步的分子分类学研究提供重要基础。To investigate the inter-specific genetic marker of Parabronema skrjabini, the genomic DNA of P. skrjabini collected from the in; fected camels in Inner Mongolia were extracted. The mitochondrial cytochrome oxidase subunit 1 （ CO1 ） gene was amplified by PCR using universal primers, and then the PCR product was cloned into pMD19-T vectors. The insert was sequenced successfully and compared with other nematode sequences by DNAStar 5.0 and MEGA 4.0. The results showed that the PCR product was 689 bp in length. Compared with other CO1 gene sequences of related nematode, the homology among P. skrjabini, Habronema microstoma, and H. muscae is 86. 7% and the nu＇ cleotide pairwise distance is 0. 143. Conversely, the homology between P. skrjabini and Litomosoides galizai is 80. 1% and the nucleotide pairwise distance is 0. 229. It suggested that the CO1 gene of P. skrjabini can be not only used as a genetic marker, but also, used for further study in molecular classification.

关 键 词：斯氏副柔线虫 CO1基因 克隆 序列分析 

分 类 号：S852.731[农业科学—基础兽医学]