重组人胰岛素原基因高效表达和纯化的研究  

A Study on Efficient Expression and Purification for Recombinant Human Proinsulin Gene

作　　者：黄秀梅[1] 叶子坚[1] 李鹤宾[1] 

机构地区：[1]厦门医学高等专科学校药学系,福建厦门361008

出　　处：《江西农业大学学报》2014年第2期418-421,共4页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(31201969);厦门市科技计划项目(3502Z20093041)

摘　　要：克隆人胰岛素原基因,通过密码子优化和融合蛋白化学水解位点和纯化标签优化,构建人胰岛素原高效原核表达系统,为重组人胰岛素原的高效表达纯化制备奠定基础。根据大肠杆菌密码子偏好性优化设计人胰岛素原基因,通过PCR技术获得重组基因,构建原核表达载体pET-32a-proinsulin,转化大肠杆菌表达株E.coli BL21,筛选人胰岛素原高效表达菌株,建立诱导表达和纯化融合蛋白技术方法。克隆获得优化的人胰岛素原重组基因,构建原核高效表达载体,获得优化的诱导表达方法和纯化的人胰岛素原。该研究为建立胰岛素高效制备的优化条件奠定基础。To optimize human proinsulin codon , fusion protein chemical hydrolysis sites and purification tags and construct human proinsulin prokaryotic expression system for high efficient expression and purifica -tion.Methods：Designing proinsulin gene according to the Escherichia coli password preference .Getting the pro-insulin gene by PCR technology and construct the cloning prokaryotic expression vector pET -32a-proinsulin. The pET-32a-proinsulin vector was imported into E.coli BL21.Screening human proinsulin high expression strain,establishing the method for proinsulin induced expression and fusion protein purification .The optimiza-tion proinsulin gene was obtained , a high effective prokaryotic expression vector was constructed , the better method for proinsulin expression and purified human proinsulin were obtained .The study provides the founda-tion for increasing insulin output .

关 键 词：人胰岛素原 蛋白表达 基因重组 pET-32a 

分 类 号：Q578[生物学—生物化学]