大肠埃希氏菌表达FMDV3AB非结构蛋白的纯化复性与活性检测  

Purification,renaturation and activity assay of FMDV non-struct ural protein 3AB expressed in Escherichia coli

作　　者：曹轶梅[1] 卢曾军[1] 刘在新[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病毒学重点实验室,甘肃兰州730046

出　　处：《中国兽医科技》2003年第8期22-25,共4页Chinese Journal of Veterinary Science and Technology

基　　金："十五"国家科技攻关项目 (2 0 0 2BA5 14A 181);国家"973"资助项目 (G19990 1190 1)

摘　　要：采用超声波裂解细菌 ,高浓度尿素裂解包涵体和金属螯合亲合层析的方法对大肠埃希氏菌中表达的FMDV 3AB非结构蛋白进行纯化 ,经SDS PAGE分析 ,3AB融合蛋白纯度达 90 %以上。采用稀释法和氧化型、还原型谷胱甘肽系统相结合方法对融合蛋白进行复性 ,通过Westernblotting分析 ,表明复性后的 3AB融合蛋白能与抗FMDV抗体发生特异性结合。ELISA检测表明 ,复性后的 3AB融合蛋白有很好的抗原活性。The non-structural 3AB fusion protein of foo t -and-mouth disease virus expressed in E.coli Rosetta(DE3), wa s obtained with the purity up to 90% by Ni-NTA His Bind Resin affinity chromato graphy. Western blotting showed specific Ag-Ab binding band between the antiser um and the 3AB fusion protein after the protein was purified and renatured.By E LISA detection,all serum of experimentally infected animals had a positive resul t,but animals that neither vaccinated nor infected had a negative result.All thi s made a good ground for the further studies on purification and renaturation of inclusion bodies.

关 键 词：口蹄疫病毒 3AB非结构蛋白 纯化 复性 抗原活性 大肠埃希氏菌表达 

分 类 号：S852.659.6[农业科学—基础兽医学]