大肠杆菌中可溶性表达重组人成纤维细胞生长因子21及制备  

Soluble Expression and Preparation of Recombinant Human Fibroblast Growth Factor 21 in E.coli

作　　者：范开[1] 谭克海[1] 张淳[1] 张益[1] 

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400054

出　　处：《重庆理工大学学报（自然科学）》2011年第5期18-22,50,共6页Journal of Chongqing University of Technology：Natural Science

基　　金：国家科技支撑计划资助项目(2008DAI63D05)

摘　　要：为在大肠杆菌中高效、非融合、可溶性表达重组人成纤维细胞生长因子21(rhFGF-21),研究了纯化方法,以获得具有较高生物活性的rhFGF-21。全基因合成人成纤维细胞生长因子21cDNA序列,克隆进原核表达载体pET-3c,在转化大肠杆菌BL21(DE3)PlysS中进行表达。表达产物经盐析、层析等方法纯化后,利用3T3-L1细胞鉴定其促葡萄糖摄取的生物学活性。结果表明:rhFGF-21在BL21(DE3)PlysS实现了高效、非融合、可溶性表达,目的蛋白占菌体总蛋白的20%左右;表达产物经纯化后,纯度可达95%以上;表达产物能够明显促进3T3-L1细胞对葡萄糖的吸收。该方法成功实现了rhFGF-21在大肠杆菌中高效可溶性表达。To effectively express human Fibroblast Growth Factor 21 in the form of solubility without fusion and purify it from E. coli, the gene of human FGF-21 was integrated and inserted into pET-3c, and itconstructed the recombinant vector pET-3c-FGF21 which was subsequently transformed into BL21 （DE3）PlysS. The engineering bacteria was induced by IPTG to express recombinant human FGF-21 . The recombinant protein＇s biological function of uptake glucose on 3T3-L1 was tested after purifying with centrifugation, salting and chromatography. The recombinant vector pET-3c-FGF21 was successfully constructed,and the human FGF-21 was expressed by soluble pattern in E. coli ,and took more than 20% of total protein. The FGF-21 ＇ s purity reached 95% after the process of purifying. The result on 3T3-L1 showed that the rhFGF-21 had an apparent up-regulation of glucose uptake. The rhF-GF-21 with good biological activity can be effectively expressed in E. coli in the form of solubility, which lays a good foundation for subsequent research and drug development of human FGF-21.

关 键 词：人成纤维细胞生长因子21 可溶性表达 葡萄糖摄取 

分 类 号：Q812[生物学—生物工程]