猪流行性腹泻病毒M基因的表达及鉴定  

IDENTIFICATION OF M PROTEIN EXPRESSION OF PORCINE EPIDEMIC DIARRHEA VIRUS

作　　者：吴玉璐[1] 虞凌雪[1] 程群[1] 王康[1] 于海[1] 刘光清[1] 童光志[1] 周艳君[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2012年第6期6-10,共5页Chinese Journal of Animal Infectious Diseases

基　　金：国家生猪现代产业技术体系建设项目(NYCYTX-009)

摘　　要：为了更好的研究近两年在我国爆发的猪流行性腹泻(porcine epidemic diarrhea,PED)疫情,本实验室收集不同发病地区的临床样品进行猪流行性腹泻病毒(Porcine epidemic diarrhea,PEDV)RT-PCR检测,并挑选8个阳性样品对其M基因进行扩增,克隆和测序。序列比对的结果显示,8个分离株与14个参考株之间的核苷酸以及氨基酸同源性分别是96.5%～99.9%和96.0%～100%,表明目前流行的PEDV其M基因是相对保守的。同时,我们将ZZ-1株的M基因克隆于原核载体pGEX-6p-1,转入菌株BL21中后获得了大量表达,重组蛋白大小约为50 kDa。利用PEDV阳性猪血清对纯化后的重组M蛋白进行Western blot分析,结果表明重组表达的M蛋白与临床阳性猪血清具有良好的免疫反应性。In order to understand recent outbreaks of porcine epidemic diarrhea（PED） in pig populations in China,we collected a few clinical samples from different farms with PED history.A total of 8 isolates were obtained and their M protein gene was sequenced and compared with other 14 reference strains.The nucleotide and amino acid identities among all these strains were 96.5%-99.9% and 96.0%-100%,respectively.The high identity of M gene sequences suggested genetic stability of the PED isolates recently prevalent in pig herds.Then,M gene of ZZ-1 isolate was inserted into pGEX-6p-1 vector that was transformed into BL21 cells.The expressed product was analyzed in SDS-PAGE and Western blotting.The results showed that M protein was expressed with molecular mass of 50 kDa and specifically reacted with PEDV positive swine serum.

关 键 词：猪流行性腹泻病毒 M蛋白 原核表达 

分 类 号：S852.659.6[农业科学—基础兽医学]