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

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.