重组人胰岛素原在大肠杆菌中的可溶性表达  

Soluble Expression of Recombinant Human Proinsulin in E. coli

作　　者：吕丹 闫亚丽 景丽芳 张秋荣 常莉 刁爱坡[1] 李玉银[1] Lü Dan;YAN Yali;JING Lifang;ZHANG Qiurong;CHANG Li;DIAO Aipo;LI Yuyin(Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science ＆ Technology, Tianjin 300457, Chin)

机构地区：[1]工业发酵微生物教育部重点实验室,天津科技大学生物工程学院,天津300457

出　　处：《天津科技大学学报》2017年第6期21-25,共5页Journal of Tianjin University of Science & Technology

基　　金：大学生创新创业训练计划资助项目(201510057058);天津科技大学大学生实验室创新基金资助项目(1504A302X)

摘　　要：目前重组胰岛素主要用于糖尿病的治疗.通过大肠杆菌(E.coli)密码子优化,设计人胰岛素原基因,建立利用大肠杆菌表达可溶性重组胰岛素原的技术方法.聚丙烯酰胺凝胶电泳(SDS-PAGE)结果显示,重组质粒p His-Nus Ainsulin诱导表达的大肠杆菌可溶性表达融合蛋白His-Nus A-insulin.重组蛋白His-Nus A-insulin可溶性表达的最适条件为0.1,mmol/L IPTG作用下37,℃诱导4,h,重组蛋白产物经过Ni柱纯化、500,mmol/L咪唑洗脱,得到纯度较高的1.059,μg/μL的重组蛋白His-Nus A-insulin,重组蛋白His-Nus A-insulin经烟草蚀纹病毒蛋白酶(TEV)酶切作用后,获得可溶的人胰岛素原.Recombinant insulin is used to treat diabetes at present. A human proinsulin gene was designed using optimizedE. coli codons. A method of expressing soluble recombinant proinsulin in E. coli was established. The SDS-PAGE results indicated that the recombinant fusion protein His-NusA-insulin was solubly expressed in E. coli, which was inducibly ex-pressed by recombinant plasmid pHis-NusA-insulin. The optimum protein expression conditions are 0.1,mmol/L IPTG in-duction at 37,℃ for 4,h. The recombinant protein product was purified with Ni column, and eluted by 500,mmol/L imida-zole. The concentration of high purity His-NusA-insulin is 1.059,μg/μL. Tobacco etch virus protease（TEV）was used to cleave His-NusA-insulin to produce the soluble recombinant proinsulin.

关 键 词：人胰岛素原 重组蛋白 NUSA 

分 类 号：Q786[生物学—分子生物学]