血管内皮生长因子D基因的原核表达及纯化  

Prokaryotic Expression and Purification of Vascular Endothelial Growth Factor D

作　　者：耿明伟[1] 张春[2] 王春雨[1] 曾范丽[1] 王全凯[1] 

机构地区：[1]吉林农业大学中药材学院,长春130118 [2]吉林农业大学发展学院,长春130600

出　　处：《经济动物学报》2008年第4期245-248,共4页Journal of Economic Animal

基　　金：农业部948项目

摘　　要：利用GST融合基因表达系统原核表达血管内皮生长因子D,并进行纯化。诱导重组质粒pGEX-VEGF-D在大肠杆菌BL21(DE3)中表达,经超声破菌后,采用GST蛋白纯化系统进行纯化,并用3C-Protease酶切去标签GST,所得产物进行15%SDS-PAGE电泳。结果表明:大肠杆菌经诱导,高效表达出分子质量约56 ku的GST-VEGF-D融合蛋白;纯化后的蛋白经酶切后电泳显示只有一条带,纯度相对较高,可基本满足生物学活性测定,为研究VEGF-D蛋白的结构和功能提供了有利的条件。To express, purify and identify VEGF- D fusion protein, E· coli BL21（DE3） containing recombinant plasmid pGEX-6p-1-VEGF-D was induced by IPTG. The bacteria was lyzed with lysozymc. Expressed product was purified by GST＇protein purification system. GST was cut by 3C-protease. And then the expressed fusion protein was identified by SDS-PAGE. The result showed that the relative molec- ular mass of expressed product was 56 000, which was consistent with that of GST-VEGF-D fusion protein. The VEGF-D protein without GST was successfully purified. The purified protein could be used to study biological activity of VEGF-D.

关 键 词：GST-VEGF-D 融合蛋白 原核表达 纯化 

分 类 号：Q786[生物学—分子生物学]