多型FMDV VP1表位融合蛋白SLP-MultiVP1的纯化及鉴定

Purification and characterization of VP1 epitope based fusion protein of multitype FMDV

口蹄疫(foot-and-mouth disease,FMD)是由口蹄疫病毒(foot-and-mouth disease virus,FMDV)主要感染偶蹄兽引起的烈性传染病。本研究对已构建的含有VP1表位肽基因的原核表达载体pGEX-SLP-MultiVP1进行IPTG诱导表达,经SDS-PAGE试验在菌体裂解物中证明了80 000融合蛋白的存在。通过Pierce?GST Spin Purification Kit亲和层析和切胶相结合的方法,成功纯化出融合蛋白GST-SLP-MultiVP1,并用Prescission Protease将该融合蛋白中的GST标签切掉,获得大小为54 000的目的蛋白SLP-MultiVP1,并用Western Blot试验证明了该蛋白与牛A、O、AsiaⅠ型疫苗株阳性血清均有特异性的结合。本试验进一步为口蹄疫VP1表位肽的免疫原性研究奠定了基础。

Foot-and-mouth disease（FMD）was a highly contagious disease,which is mainly caused by FMDV infection of cloven-hoofed animals.In this study,constructed pGEX-SLP-MultiVP1 prokaryotic expression genetic vector which contains VP1 epitope by IPTG induction expression,and the result of SDS-PAGE indicated that fusion protein existed in cell lysate and relative molecular mass were about 80 000.The fusion protein was purified successfully by method of combining Pierce？GST Spin Purification Kit with gel slices and obtained the target protein which has relative molecular mass of 54 000 with GST tag excised by Prescission Protease.Each-type FMDV vaccine positive serum with A,O and AsiaⅠwere able to bind the target protein GST-SLP-MultiVP1 specifically through the method of Western blot.This experiment laid a foundation for further research on immunogenicity of FMD VP1 epitope.