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.
出处
《中国兽医科技》
CSCD
北大核心
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 )