摘要
本文利用兼并引物成功克隆到土元类胰蛋白酶样丝氨酸蛋白酶基因,通过构建该基因的原核重组表达载体及IPTG体外诱导,包涵体蛋白复性,Ni 2+柱亲和层析纯化,最终获得了重组表达目的蛋白.经体外检测,该蛋白具有较好的溶栓活性,这为揭示土元溶栓作用的分子基础及其进一步的开发利用奠定了基础.
In order to reveal the molecular basis of E.sinensis thrombolytic effect,trypsin-like serine protease gene was cloned through degenerate primers.The prokaryotic expression plasmid was constructed and recombinant protein was expressed by inducing with isopropyl-β-D-thiogalactopyranoside（IPTG）and purified by Ni 2＋affinity chromatography column after inclusion body refolding.Recombinant protein was confirmed to have a fibrinolytic activity in vitro.This study will lay the foundation for the future utilization.
出处
《河南大学学报（自然科学版）》
CAS
2015年第2期187-191,共5页
Journal of Henan University:Natural Science
基金
河南省科技厅基础研究项目(122300410127)
河南省教育厅自然科学研究计划项目(2009A180002)
河南省重大科技攻关项目(132102110066)
河南省科技攻关项目(132102110112)
关键词
土元
类胰蛋白酶样丝氨酸蛋白酶
原核表达
溶栓活性
Eupolyphaga sinensis
trypsin-like serine protease
prokaryotic expression
fibrinolytic activity
