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.
出处
《西南农业学报》
CSCD
北大核心
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)
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
Swine Japanese encephalitis virus
Ea gene
Prokaryotic expression
Identification of antigenic
