吉林大学学报（医学版）2016，Vol.42Issue(4)：642-647,6.DOI:10.13481/j.1671-587x.20160402
融合表达载体 pET22b-SUMO-FGFR4的构建及其在大肠杆菌中表达条件的优化
Construction of fusion expression vector pET22b-SUMO-FGFR4 and optimization of expression conditions in E.coli
摘要
Abstract
Objective:To design the small ubiquitin modification-fibroblast growth factor receptor 4 (SUMO-FGFR4) fusion gene and construct the expression vector pET22b-SUMO-FGFR4, to optimize the expression conditions. Methods:The SUMO-FGFR4 fusion gene was obtained by Overlap PCR and was connected to pET22b;the recombinant expression vector pET22b-SUMO-FGFR4 was obtained. The influence of lactose concentration, induction time,induction temperature,induction point and adding mode of lactose in the expression levels was observed,and the best induction condition was determined; then the solubility of recombinant protein was analyzed.Results:The SUMO-FGFR4 fusion protein was highly expressed,the molecular weight of the fusion protein was about 40 000 and it could bind with FGFR4 specific antibody.When the lactose concentration was 1.0 g·L-1 ,the induction time was 3 h,the induction temperature was 37℃,the value of A (600)was 0.8,the expression level was highest;but adding mode of lactose had no remarkable effect on the protein expression.The expression level of recombinant protein induced by lactose was higher than IPTG.SUMO-FGFR4 protein existed in a form of inclusion body.Conclusion:The SUMO-FGFR4 fusion protein is expressed successfully in this study while lactose is used as inducer and the best expression conditions are confirmed.关键词
小泛素相关修饰物/成纤维细胞生长因子受体 4/重组融合蛋白类Key words
small ubiquitin-like modifier/fibroblast growth factor receptor 4/recombinant fusion proteins分类
生物科学引用本文复制引用
刘微,姚杨,马萧萧,邓裕宣,梅迪,刘磊,王会岩..融合表达载体 pET22b-SUMO-FGFR4的构建及其在大肠杆菌中表达条件的优化[J].吉林大学学报（医学版）,2016,42(4):642-647,6.基金项目
国家自然科学基金资助课题（81273421）；吉林省卫计委青年项目资助课题（2015Q042）；吉林省吉林市科技局科技发展计划项目资助课题（20156427）；吉林省教育厅大学生创新创业训练项目资助课题 （）
