【摘要】 目的构建IGF-1a,并将其表达及纯化。方法用多重PCR技术获得人IGF-1a基因,采用酶切与连接的方法,将其连接至pET28a载体,用IPTG诱导转化pET28a-Cpn10-IGF-1a表达载体的大肠杆菌,以镍金属鳌合亲和层析法纯化融合蛋白质。结果构建了IGF-1a的表达载体pET28a-Cpn10-IGF-1a,DNA序列测定结果表明构建正确;以镍金属鳌合亲和层析获得纯度为98%的蛋白质。结论用分子克隆技术正确克隆IGF-1a,并成功表达与纯化IGF-1a蛋白。更多还原

【Abstract】 Objective To construct the prokaryotic expression vector of IGF-1a,express and purify IGF-1a protein.Methods gene sequence were obtained by multiplex PCR method and the gene was cloned into pET28a plasmid successively to construet the prokaryotic expression vector(pET28a-Cpn10-IGF-1a),which recombined the gene of IGF-1a.The E.coli BL21(DE3) transformed with pET28a-Cpn10-IGF-1a were induced by IPTE for expression of IFG-1 protein.The protein was purified by magic Ni2+ affinity chromatography.Results The IGF-1a fusion protein was successfully expressed in BL21(DE3) and purified byNi affinity chromatography,the purity of IGF-1a fusion protein was over 98%.Conclusion The prokaryotic expression vector of pET28a-Cpn10-IGF-1a has been sucessfully cloned,and the protein has been successfully expressed and purifled.更多还原