猪塞尼卡谷病毒VP1蛋白的原核表达及其多克隆抗体制备

Prokaryotic expression of VP1 protein of swine Seneca Valley virus and preparation of polyclonal antibodies against the protein

为获得猪塞尼卡谷病毒(Seneca Valley virus,SVV)衣壳蛋白VP1及其多克隆抗体,本文通过大肠杆菌原核表达系统表达VP1基因,并纯化VP1蛋白作为抗原免疫大鼠,采用ELISA和Western blot检测高免血清的特异性和抗体效价。结果:VP1主要以包涵体形式得到高效表达,并纯化得到单一条带的VP1蛋白。表达的VP1蛋白能与SVV阳性血清发生特异性反应。纯化的融合蛋白VP1免疫大鼠获得的高免血清效价高于1∶32000,且与真核表达的VP1蛋白能够发生特异性结合反应。本研究为SVV的VP1蛋白功能研究奠定了物质基础。

To obtain the antigen of capsid protein VP1 of the Seneca Valley virus(SVV)and the corresponding polyclonal antibodies,the VP1 gene was cloned and expressed in the E.coli prokaryotic expression system and was purified.Then,the purified protein was used to immunize rats in order to prepare the polyclonal antibodies against the VP1 protein.The titer of the polyclonal antibodies was determined by Elisa.Specificity of the polyclonal antibodies was analyzed by Western blotting.The results showed that the VP1 gene was mainly expressed in the inclusion body of E.coli.SDS-PAGE analysis proved that there was a clear single target band for the purified VP1 protein.The expressed VP1 protein could be specifically identified by the SVV-positive serum.The titer of the polyclonal antibodies from the rats was determined to be higher than 1:32000.The polyclonal antibody could specifically recognize the expressed VP1 protein in eukaryotic cells.