盖他病毒E2蛋白的表达纯化以及抗原性鉴定  

EXPRESSION AND CHARACTERIZATION OF RECOMBINANT E2 PROTEIN OF GETAH VIRUS

作　　者：陆莹梅 魏建超[2] 夏鹏[2] 吴亚玲[2] 黎倩倩[2] 武专昌[2] 齐鹏飞[2] 石坤[2] 李玉明[2] 邱亚峰[2] 李蓓蓓[2] 刘珂[2] 邵东华[2] 周望平[1,3] 马志永[2] 

机构地区：[1]湖南农业大学动物医学院,长沙410128 [2]中国农业科学院上海兽医研究所,上海200241 [3]湖南省畜牧兽医研究所,长沙410000

出　　处：《中国动物传染病学报》2015年第2期15-20,共6页Chinese Journal of Animal Infectious Diseases

基　　金：上海市科技兴农重点攻关项目(沪农科攻字(2013)第5-6号);公益性行业(农业)科研专项(201303045);国家自然科学青年基金(31302116)

摘　　要：参考Gen Bank(EU015066)盖他病毒(Getah virus,GETV)SH05-6中结构蛋白E2的全基因序列并对其进行密码子优化。将优化后的基因片段克隆至原核表达载体p ColdⅠ中,构建重组表达载体p Cold-E2。重组质粒转化E.coli BL21(DE3)感受态细胞,经IPTG低温诱导后,SDS-PAGE和Western blot进行分析。结果显示,在相对分子质量46 k Da处出现目的条带,与预期相符,且占大肠杆菌表达蛋白总量的80%。结果证明E2基因在大肠杆菌中获得了高效表达,并且能与抗血清发生特异性反应。本研究为GETV快速检测方法的建立和GETV流行病学调查奠定了基础。The E2 gene of Getah virus strain SH05-6 was optimized and synthesized according to the complete genome sequence published in GenBank （EU015066）. The synthesized E2 gene was cloned into the expression vector pColdlfor generation of a recombinant plasmid pCold I -E2. The plasmid pCold I -E2 was then transformed into Eocoli BL21 （DE3） competent cells following induction with IPTG. The expression of the target E2 protein was detected in SDS-PAGE and Westem blot. The results showed that the recombinant E2 was a 46 kDa protein corresponding to the molecular mass of the expected E2 and accounted for approximately 80% of total protein expressed in E.coli. The recombinant E2 protein reacted with antiserum against Getah virus. The availability of the recombinant E2 protein has laid a foundation for development of rapid detection and epidemiological investigation methods for Getah virus.

关 键 词：盖他病毒 原核表达 E2基因 

分 类 号：S852.659.6[农业科学—基础兽医学]