胶原沉积抑制因子在大肠杆菌DH5α中的表达特性  

Expression features of human decorin in E.coli DH5α

作　　者：杨国嵘[1] 朱任之[2] 何晓霞[3] 薛永杰[1] 贺雪姣[1] 郁荣[1] 

机构地区：[1]解放军第一医院病理科,甘肃兰州730030 [2]兰州大学病理学研究所 [3]甘肃省人民医院脑系科,甘肃兰州730000

出　　处：《中国病理生理杂志》2007年第6期1219-1222,共4页Chinese Journal of Pathophysiology

基　　金：甘肃省科技攻关资助项目(No.GK972-2-81A);甘肃省自然科学基金资助项目(No.ZS001-A23-058-Y)

摘　　要：目的:研究人胶原沉积抑制因子(decorin)在大肠杆菌DH5α中的表达特性。方法:用SDS-PAGE法检测pGEX-4T-1-decorin融合克隆在大肠杆菌DH5α中表达,选择阳性表达克隆;测定1mmol/L异丙基-β-D-硫代半乳糖苷(IPTG)最佳诱导时间;用超声波裂解法分析GST-decorin融合蛋白的可溶性。用West-ern blotting进行定性分析。结果:用1mmol/LIPTG诱导时,融合蛋白可在大肠杆菌DH5α中表达;最佳诱导时间为4h;大部分融合蛋白以包涵体的形式存在;所表达蛋白为GST融合蛋白。结论:GST-decorin融合蛋白可在大肠杆菌DH5α中大量表达,但以包涵体的形式存在。AIM： To study the expression features of human decorin in E. coli DH5α. METHODS： The pGEX -4T - 1 - decorin fusion clone was expressed in E. coli DH5α. Positively expressed clone was selected by SDS - PAGE. The optimized inducing time by 1 mmol/L IPTG was determined. The solubility of GST - decorin fusion protein was analyzed by ultrasonic crush method, and its quality was examined by Western blotting. RESULTS： With the induction of 1 mmol/L isopropy - β - D - thiogalactoside （ IPTG）, the fusion protein was expressed in E. coli DHSα. The optimized inducing time by 1 mmol/L IPTG was 4 hours. Most of fusion protein existed in the form of inclusion body. The expressed protein was GST fusion protein. CONCLUSION： It is suggested that the fusion protein of GST - decorin may be expressed in E. coli DH5α in a large amount in the form of inclusion body.

关 键 词：DECORIN 融合蛋白质类 原核表达 

分 类 号：R363[医药卫生—病理学]