南方医科大学学报2012，Vol.32Issue(12)：1713-1717,5.DOI:10.3969/j.issn.1673-4254.2012.12.007
柯萨奇病毒A16型VP1～VP4基因克隆及其表达产物的抗原相关性分析
Cloning, expression and antigenic analysis of VP1-VP4 gene encoding the structural protein of Coxsackie virus A16
摘要
Abstract
Objective To clone and express VP1-VP4 genes encoding the structural proteins of Coxsackie virus A16 and analyze the antigenicity of the expressed recombinant proteins. Methods The VP1-VP4 cDNAs were amplified with RT-PCR from the extracted viral RNA and cloned into pMD19-T vectors. The VP1-VP4 genes were inserted to the multi-cloning sites of the plasmid pQE30a, and the protein expressions in E. coli M15 were induced by IPTG. After purification by washing with 8 mol/L urea under denaturing condition, the recombinant proteins were identified by Western blotting and ELISA for their immunogenicity against rabbit anrisera of Coxsackie virus A16 and enterovirus 71, respectively. Results The recombinant VP1-VP4 proteins were highly expressed in E. coli M15. The purified proteins could be recognized by rabbit antiserum of Coxsackie virus A16 and showed cross reactivity with the rabbit antiserum of Enterovirus 71. Conclusion The recombinant Coxsackie virus A16 VP1-VP4 proteins obtained possess good antigenicity.关键词
柯萨奇病毒A16型/VP1蛋白/VP2蛋白/VP3蛋白/VP4蛋白/克隆/原核表达/抗原反应性Key words
Coxsackie virus A16/VP1 protein/VP2 protein/VP3 protein/VP4 protein/clone/prokaryotic expression/antigenicity分类
医药卫生引用本文复制引用
宋远斌,何思杰,余楠,陈欣欣,王斌,车小燕,曾其毅..柯萨奇病毒A16型VP1～VP4基因克隆及其表达产物的抗原相关性分析[J].南方医科大学学报,2012,32(12):1713-1717,5.基金项目
国家重大科技专项课题(2009ZX10004-306) （2009ZX10004-306）
广州市医药卫生科技重大项目(201102A211007) （201102A211007）
广东省科技计划项目(2010B031600238) （2010B031600238）
广州市计划项目科技支撑计划(2010J-E421) （2010J-E421）
