天津科技大学学报2017，Vol.32Issue(6)：21-25,5.DOI:10.13364/j.issn.1672-6510.20160103
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
摘要
Abstract
Recombinant insulin is used to treat diabetes at present.A human proinsulin gene was designed using optimized E.colicodons.A method of expressing soluble recombinant proinsulin inE.coli was established.The SDS-PAGE results indicated that the recombinant fusion protein His-NusA-insulin was solubly expressed inE.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/NusAKey words
human proinsulin/recombinant protein/NusA分类
生物科学引用本文复制引用
吕丹,闫亚丽,景丽芳,张秋荣,常莉,刁爱坡,李玉银..重组人胰岛素原在大肠杆菌中的可溶性表达[J].天津科技大学学报,2017,32(6):21-25,5.基金项目
大学生创新创业训练计划资助项目(201510057058) （201510057058）
天津科技大学大学生实验室创新基金资助项目(1504A302X) （1504A302X）
