猪流行性腹泻病毒贵州株部分S基因原核表达质粒的构建  

Plasmid Construction of Prokaryotic Expression of S Gene in Guizhou Porcine Epidemic Diarrhea Virus Strain

作　　者：何玉侠 毛黎红 周碧君[1,2] 王开功[1,2] 文明[1,2] 程振涛[1,2] 温贵兰[1,2] 冯旭芳[1] 潘廷云 张艳[1] 刘长素 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病与兽医公共卫生重点实验室,贵州贵阳550025 [3]贵州省贵安新区党武镇农业服务中心,贵州贵安550025

出　　处：《贵州农业科学》2015年第8期189-191,195,共4页Guizhou Agricultural Sciences

基　　金：贵州省农业科技攻关项目"贵州省主要人畜共患病综合防控技术研究"[黔科合NY字(2011)3061];贵州省农业委员会科技计划项目"贵州省2013年重大动物疫病省级定点监测"(2013-06);"2014年贵州省重大动物疫病省级定点监测"(2014-06);大学生创新创业训练计划项目"猪流行性腹泻病毒贵州株部分S基因的克隆与原核载体构建"(201410657016);贵州省生猪质量安全工程技术研究中心建设项目"生猪健康养殖实验室建设与技术研发"[黔科合农C字(2011)4022]

摘　　要：为给贵州猪流行性腹泻(PED)的诊断及防治提供科学依据,同时为猪流行性腹泻病毒(PEDV)部分S蛋白进一步表达建立相应检测方法奠定理论基础,利用贵州PEDV分离株,参考GenBank登录PEDV CV777株的S基因序列,选取主要抗原区域(COE)设计1对PEDV COE基因特异性引物,进行部分保护性抗原COE基因的PCR扩增,扩增产物与pMD19-T载体链接,鉴定正确后再亚克隆至pET32a位置,结果成功构建了贵州PEDV分离株COE基因的原核表达质粒,分离株COE基因与HBQX株的序列同源性达99.1%,与经典毒株CV777株的同源性为93.4%。The prokaryotic expression plasmid of COE gene of Guizhou PEDV isolated strain was successfully established through amplifying part protective antigens COE gene of PEDV COE specific primer of Guizhou PEDV strain by using the selected main antigen domain （COE） design 1 according to S gene sequence of pEDV CV777 strain registered in GenBank and then the link between amplification products and PMD19-T vector was sub-cloned on pET32a to provide the scientific basis for diagnosis and prevention of porcine epidemic diarrhea （PED） and lays the theoretical foundation for establishing the corresponding detection method of PEDV-S gene expression in Guizhou. The sequence homology between COE gene of Guizhou PEDV isolated strain and HBQX strain reaches 99.1 % and the sequence homology between COE ~ene of Guizhou PEDV isolated strain and classical strain CV777 is uo to 93.4%.

关 键 词：猪流行性腹泻病毒 COE基因 原核表达 质粒构建 贵州 

分 类 号：S852.65[农业科学—基础兽医学]