广东药学院学报2012，Vol.28Issue(5)：556-559,4.DOI:10.3969/j.issn.1006-8783.2012.05.023
肠道病毒71型外壳蛋白VP1在大肠杆菌的表达和纯化
Expression and purification of recombinant capsid protein VP1 of enterovirus 71 in Escherichia coli
摘要
Abstract
Objective To express and purify of recombinant capsid protein VP1 of enterovirus 71 in E. coli for preparation of anti-VPl monoclonal antibody. Methods Kecombinant VP1 was induced to produce in transformed E. coli BL21 with IPTG and identified by SDS-PAGE. VP1 was purified by IMAC using a Ni-NTA agarose column. The antigenicity of VP1 was analyzed by Western blot. The optimal expression condition of VP1 was also identified. Results VP1 was mainly expressed in the form of inclusion bodies with molecular size of about 36 000 and migrated as single band after purification by SDS-PAGE. The protein was detected with anti-His tag and anti-EV71 polyclonal antibodies respectively by Western blot. The optimal expression condition for VP1 was defined as 37℃, 1.25 mmol/L IPTG and 4 h. Conclusion EV71 VP1 could be expressed in E. coli BL21 and possesses antigen activity after purification and renaturation. This work provides a basis for further study.关键词
肠道病毒71型/VP1/原核表达/纯化Key words
enterovirus 71 / VP1 / prokaryotic expression/ purification分类
医药卫生引用本文复制引用
吴亚安,邓丽芬,龚雨婷,陈端萍,谢端,肖承荣,蔡陈珣,赵晓蓉..肠道病毒71型外壳蛋白VP1在大肠杆菌的表达和纯化[J].广东药学院学报,2012,28(5):556-559,4.基金项目
广东省大学生创新实验项目(1057310005) （1057310005）
