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绵羊夏柏特线虫和叶氏夏柏特线虫线粒体nad5基因和cytb基因的序列分析

ISSN号：1673-4696
期刊名称：《中国兽医科学》
时间：0
分类：S852.731[农业科学—基础兽医学;农业科学—兽医学;农业科学—畜牧兽医]
作者机构：[1]吉林农业大学动物科技学院,吉林长春130118, [2]中国农业科学院兰州兽医研究所家畜疫病病原.生物学国家重点实验室,甘肃兰州730046, [3]湖南农业大学动物医学院,湖南长沙410128, [4]西北农林科技大学动物医学院,陕西杨凌712100, [5]青海省畜牧兽医科学院,青海西宁810016
相关基金：公益性行业（农业）科研专项（201303037）;甘肃省创新研究群体计划项目（1210RJIA006）

作者：赵雷[1,2], 江海海[2], 刘国华[2,3], 赵光辉[4], 蔡进忠[5], 周东辉[2], 朱兴全[2], 钱爱东[1]

关键词：绵羊夏柏特线虫, 叶氏夏柏特线虫, 线粒体DNA, nad5基因, Cytb基因, 种系发育分析, Chabertia ovina Chabertia erschowi mitochondrial DNA, nad5 gene cytb gene, phylo-genetic relationship

中文摘要：

本研究的目的是通过对绵羊夏柏特线虫和叶氏夏柏特线虫线粒体细胞色素b基因（cytb）部分序列（pcytb）和烟酰胺嘌呤二核苷酸（NADH）脱氢酶亚单位5基因（nad5）部分序列（pnad5）的克隆和遗传变异分析，首次为在分子水平上区分绵羊夏柏特线虫和叶氏夏柏特线虫提供理论依据。采用PCR方法分别克隆了6条绵羊夏柏特线虫陕西分离株和9条叶氏夏柏特线虫青海分离株的pcytb和pnad5。将测序结果应用ClustalX1．81程序进行比对，然后用PAUP4．0程序中的MP法将pcytb和pnad5串联后构建系统发育树。结果显示，获得的pnad5和pcytb序列的长度分别为388bp和659bp，其变异率在绵羊夏柏特线虫分离株中分别为0．3％～1．3％和0．2％～2．6％，在叶氏夏柏特线虫分离株中分别为0～1．8％和0．2％～1．2％，较为保守。但pnad5和pcytb序列在两种间差异率较大，分别为14．7％～16．0％和15．0％～17．39／6。用pcytb和pnad5串联后的序列进行的种系发育分析表明，绵羊夏柏特线虫和叶氏夏柏特线虫分别位于2个分支，支持叶氏夏柏特线虫为独立种；pcytb和pnad5序列可作为夏柏特线虫种间鉴定的遗传标记。

英文摘要：

The present study aimed tO examine sequence variations in cytochrome b（cytb） and NADH dehydrogenase submit 5（nad5） genes among Chabertia ovina isolates from goats in Shaanxi Province and Chabertia erschowi isolates from yaks in Qinghai Province, and to study the phylogenetic relationship of the two species based on two genes. The partial cytb（pcytb） and nadS（pnadS） were amplified from individual samples and then sequenced. The obtained sequences were aligned using the ClustalX 1.81. Then MP tree was constructed using PAUP 4. 0. The lengths of all the pcytb and pnad5 sequences were 388 bp and 659 bp,respectively. The intra-specific sequence differences were 0.3 % to 1.3 % for pnad5 and 0.2 % to 2.6 % for pcytb among the examined C. ovina isolates, and were 0 to 1.8% for pnad5 and 0.2% to 1.2% for pcytb among the examined C. erschowi isolates. The inter-specific sequence variations between the two spe- cies were 14.7% to 16.0% for pnad5 and 15.0% to 17.3% for pcytb,which were significantly higher than intra-specific sequence variations. Phylogenetic analysis showed that C. ovina and C. erschovoi were distinct nematode species. The mitochondrial cytb and had5 sequences can be used as genetic markers for the diffe- rentiation of Chabertia spp.

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