猪乙型脑炎病毒广西分离株FC792 E基因主要抗原域的原核表达及抗原性鉴定  

Prokaryotic Expression and Antigen Identification for Major Antigenic Domain of E Gene of Swine Japanese Encephalitis Virus Strain FC792 Isolated from Guangxi

作　　者：卢冰霞[1,2,3] 李斌[1,2] 秦毅斌[1,2] 赵武[1,2] 梁家幸[1,2] 韦超文 何颖[1,2] 苏乾莲[1,2] 梁保忠[1,2] 李莹莹[1,2,3] 姜佳佳[1,2,3] 杨盛斌 黄伟坚[3] 

机构地区：[1]广西兽医研究所,广西南宁530001 [2]广西畜禽疫苗新技术重点实验室,广西南宁530001 [3]广西大学动物科学技术学院,广西南宁530005 [4]广西田林县动物疫病预防控制中心,广西田林533300 [5]广西三江县林溪水产畜牧兽医站,广西三江545505

出　　处：《西南农业学报》2013年第3期1258-1263,共6页Southwest China Journal of Agricultural Sciences

基　　金：广西自然科学基金项目(2011GXNSFB018032);公益性行业(农业)科研专项(201103008);广西科技攻关项目(桂科攻1123007-3);广西畜禽疫苗新技术重点实验室系统性研究课题(12-071-28-A-5);广西基本科研业务费专项(桂科专项13-1)

摘　　要：对广西猪JEV FC792株的E基因主要抗原域Ⅰ、Ⅱ基因(Ea基因)进行原核表达及抗原性鉴定,为猪流行性乙型脑炎病毒(JEV)诊断抗原和基因工程疫苗的研制奠定基础。结果:成功构建广西猪JEV FC792株Ea基因的克隆重组质粒pMD18-T-Ea及表达重组质粒pET32a-Ea,经SDS-PAGE及Western blot鉴定表明,JEV FC792株Ea蛋白在Rosetta(DE3)菌中成功进行融合表达,表达出约58 KDa的融合蛋白;经抗原性鉴定表明,表达的融合蛋白Ea能与猪JEV阳性血清抗体发生特异性反应,融合蛋白Ea具有良好的JEV抗原性。In order prepare for the development of diagnostic antigens and genetic engineering vaccine of swine Japanese encephalitis virus（JEV）,prokaryotic expression and antigen identification for major antigenic domain I,Ⅱ gene（Ea gene） of E gene of swine JEV strain FC792 isolated from Guangxi were performed.Results： Guangxi swine JEV FC792 strain Ea gene cloning recombinant plasmid pMD18-T-Ea and expressing recombinant plasmid pET32a-Ea were successfully constructed.By SDS-PAGE and Western blot analysis showed that protein Ea of JEV FC792 strain was successfully fusion expressed in Rosetta（DE3） bacteria,and about 58 KDa fusion protein Ea was expressed.Antigenic identification showed that the expressed fusion protein Ea specifically react with swine JEV positive serum antibody,indicating the fusion protein Ea has good JEV antigenic.

关 键 词：猪流行性乙型脑炎病毒 Ea基因 原核表达 抗原性鉴定 

分 类 号：S858.28[农业科学—临床兽医学]