口蹄疫病毒O型病毒VP1蛋白表达及鉴定

Expression and Identification of Foot and Mouth Disease Virus VP1 Protein in Escherichia coli Expression System

利用大肠杆菌p ET-30a表达系统表达口蹄疫病毒(FMD)O型野毒株完整的VP1蛋白,并以Western blot试验进行鉴定。结果表明:VP1蛋白以可溶性表达,大小约为25 ku,可与标准O型口蹄疫病毒阳性血清特异结合。由此说明,口蹄疫病毒VP1原核表达蛋白有良好的反应原性,为构建口蹄疫病毒病毒样颗粒(Virus-like particles,VLPs)和快速特异诊断试剂研制奠定基础。

In this study,the Escherichia coli p ET-30 a expression system was used to express the VP1 protein of foot and mouth disease（ FMD） serotype O. A wild stain of（ FMD） serotype O was chosen and a recombinant plasmid which contained complete VP1 protein（ p ET-30a-O-VP1） was constructed. The combinant VP1 was expressed in BL21,then the expressed products were purified and identified through Ni affinity chromatography and Western blot test. Western blot test indicated that the VP1 can be specifically recognized by the standard positive serum of FMD virus serotype O.The results suggested that the recombination protein of deer FMD VP1 possess a good antigenicity,which will be the basis for production of the particle like vaccine and development of a specifically diagnostic reagent of FMD.