猪流行性腹泻病毒S蛋白和M蛋白串联表达的研究  

Research on the co-expression of S protein and M protein of porcine epizootic diarrhea virus

作　　者：陈媛[1] 郑庆礼 李春燕[1] 袁晓琴[1] 王全溪[1] CHEN Yuan;ZHENG Qingli;LI Chunyan;YUAN Xiaoqin;WANG Quanxi(College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学动物科学院,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2018年第6期717-721,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：促进海峡两岸科技联合发展基金(U1405216)

摘　　要：猪流行性腹泻病毒(porcine epizootic diarrhea virus,PEDV)的M和S蛋白与中和抗体的形成有关,同时获得这两种蛋白质对预防和控制PED有重要的意义.本研究在实验室保存的M蛋白重组质粒的基础上构建了Pet-32a-M-S串联重组质粒.通过PCR鉴定、酶切鉴定和测序证实串联重组质粒构建成功.将Pet-32a-M-S质粒转染到BL21感受态细胞中,筛选诱导的最佳条件,最后进行Western blot鉴定.结果表明,串联重组蛋白最佳诱导条件为25℃、8 h、0.6 mmol·L-^(1)IPTG; Western blot鉴定结果表明,重组质粒能够同时诱导PEDV的S和M蛋白.因此,本试验成功获得PEDV S和M蛋白在体外的串联诱导表达,为PEDV亚单位疫苗的研究提供了基础.M and S protein of porcine epizootic diarrhea virus are related to the produce of neutralizing antibody. The simultaneousacquisition of the two proteins is greatly important to prevent and control of porcine epidemic diarrhea. We construct Pet-32a-M-S re-combinant plasmid on the basis of recombinant M protein plasmid preserved in our laboratory. Then, the recombinant plasmid wasverified by PCR, restriction enzyme digestion and secquencing. At last the recombinant plasmid was transfected into BL21 competentcells. The optimal conditions for induction were screened, and the antigenicity of the recombinant protein was identified by Westernblot. The results showed that the recombinant protein was induced at 25 ℃ for 8 h with 0.6 mmoL,L-1IPTG. It can be seen fromWestern blot identification that the M pretein and S protein were sucessfully expressed in vitro. Therefore the recombinant co-expres-sion of PEDV S protein and M protein is successfully induced in vitro, which provides a basis for the research of subunit vaccine ofPEDV.

关 键 词：猪流行性腹泻病 S基因 M基因 串联表达 

分 类 号：S852.65[农业科学—基础兽医学]