河南农业大学学报2018，Vol.52Issue(3)：350-355,6.
分子伴侣对A型FMDV结构蛋白VP1在大肠杆菌中可溶性表达的促进作用
Facilitaion of molecular chaperones on soluble protein expression of recombinant A-FMDV VP1 in E.coli
摘要
Abstract
In order to obtain soluble recombinant A-VP1 protein,A type of FMDV VP1 gene fragment was amplified by PCR using pUC57-MM-VP1 as a template.The fragment was inserted into expression vector pET-28a to construct the plasmid of pET-28a-MM-VP1,which was later transformed into bacterial expression host E.coli BL21(DE3).The expression of the protein was induced by IPTG to obtain soluble A type VP1 protein.However,SDS-PAGE showed that VP1 protein was highly expressed in the form of inclusion bodies under inducing temperature of 18 ℃.In order to improve the solubility of VP1 protein,the pET-28a-MM-VP1 was co-transfected with the pTF16 into E.coli BL21(DE3).The results showed that TF16 significantly increased the expression level of A-VP1,and optimal induction conditions are 18 ℃,0.1 mol·L-1 of IPTG,2 g·L-1 of arabinose with inducing time of 20 h.The results of Western-Blot showed that the obtained soluble protein of A type VP1 could be identified by standard positive serum of FMDV,which confirmed that the fusion protein had good a responsiveness.关键词
分子伴侣/VP1蛋白/原核表达/可溶性表达Key words
Molecular chaperone/VP1 protein/prokaryotic expression/solubility分类
农业科技引用本文复制引用
崔颖磊,刘运超,冯华,刘畅,王彦伟,杨棋,王冬雨,张改平..分子伴侣对A型FMDV结构蛋白VP1在大肠杆菌中可溶性表达的促进作用[J].河南农业大学学报,2018,52(3):350-355,6.基金项目
国家重点研发计划项目(2016YFD0500704 （）
2017YFD0501103) （）
