中国工程科学Issue(1)：67-73,7.
星突江鲽胰岛素样生长因子I的体外重组表达及生物活性分析
Prokaryotic expression and bioactivity analysis of growth hormone and insulin-like growth factor-I from Platichthys stellatus
摘要
Abstract
The mature peptide domain of insulin-like growth factor-I from Platichthys stella-tus were amplified with specific primers based on its cDNA sequence. Then the matured peptide fragment was subcloned into the prokaryotic expression vector pET-28a to successfully con-struct IGF-I/pET-28a recombinant plasmid which were highly expressed in E.coli BL21(DE3) after being induced by IPTG with special fusion polypeptides containing His6 at their N-termi-nus. The obtained IGF-I polypeptide expressed in form of inclusion bodies with molecular weight of 12.1 kD and maximally accounted for 39.8%of the whole bacterial protein post 3 h induction with 0.5 mmol/L IPTG at 37℃. Western blotting analysis indicated fusion polypep-tides had the antigenicity to His6 antibody. The inclusion bodies were denaturalized using 6 mol/L guanidine HCl,purified using Ni-NTA affinity chromatography and annealed by gradient dialy-sis in urea,then purified proteins with molecular weight of 12.1 kD which was obtained from IGF-I recombinant bacterium. The proliferation experiment showed recombinant IGF-I protein could significantly promote the proliferation of human embryonic kidney cells HEK293T at 0.6 μg/mL and inhibit the proliferation with 1.8 μg/mL which verified its biological activity. Therefore,the IGF-I prokaryotic expression system was successfully constructed in the present study and biologically active IGF-I fusion protein was obtained. The present results would be helpful for better understanding the roles of IGF-I in growth regulation and development of high effective additive for growth promotion of Platichthys stellatus.关键词
星突江鲽/胰岛素样生长因子I/原核表达/生物活性Key words
Platichthys stellatus/insulin-like growth factor-I/prokaryotic expression/bio-activity分类
农业科技引用本文复制引用
徐永江,臧坤,柳学周,史宝,陈圣毅..星突江鲽胰岛素样生长因子I的体外重组表达及生物活性分析[J].中国工程科学,2015,(1):67-73,7.基金项目
国家863计划项目（2012AA10A413）；鲆鲽类现代产业技术体系项目（CARS-50）；山东省自然科学基金项目 （）
