大肠埃希氏菌表达猪瘟病毒E_2蛋白的纯化  

Expression and purification of E_2 protein of classical swine fever virus in Escherichia coli

作　　者：魏旭文[1] 尚佑军[1] 孙世琪[1] 靳野[1] 刘湘涛[1] 马军武[1] 韩雪清[1] 谢庆阁[1] 

机构地区：[1]中国农业科学院兰州兽医研究所农业部畜禽病毒学重点开放实验室,甘肃兰州730046

出　　处：《中国兽医科技》2003年第10期55-57,共3页Chinese Journal of Veterinary Science and Technology

基　　金：国家"973"项目 (G19990 1190 5 )

摘　　要：采用 pPROEX HTB融合蛋白表达系统 ,将猪瘟病毒E2 基因与 6×His的编码序列在大肠埃希氏菌BL2 1(DE3)中进行融合表达 ,表达蛋白质主要以包涵体形式存在。用 8mol/L尿素溶解包涵体后 ,利用镍离子金属螯合亲和层析法纯化蛋白。结果 ,在SDS PAGE上显示出一 17ku的目的蛋白条带 ,纯度达到 90 %以上。纯化后的蛋白变性。With the pPROEX HTB fusion expression system, the 6×His E 2 protein was expressed in Escherichia coli. The expressed protein was in the form of inclusion bodies.Inclusion bodies were solubilized in 8 mol/L urea,and purified directly by immobilized metal ion affinity chromatography (IMAC), purification product was target protein by SDS PAGE analysis. The product appears as a single band on SDS PAGE ,with a purity of 90%. Denaturation and Renaturation in vitro was also carried out. The result showed the renaturation protein can be recognized by the positive serum of classical swine fever virus.

关 键 词：大肠埃希氏菌 猪瘟病毒 E2蛋白 纯化 融合表达 包涵体 金属螯合亲和层析 

分 类 号：S858.28[农业科学—临床兽医学]