小反刍兽疫病毒H蛋白的原核表达和免疫学活性鉴定  

Prokaryotic Expression and Immunoreactivity Identification of Peste des Petits Ruminants Virus Haemagglutinin Protein

作　　者：陶虹[1,2] 孙洁[1,2] 李健波 刘建利[1,2] 阮周曦[1] 张彩虹[1,2] 曹琛福[1] 吕建强[1] 花群义[1] 杨俊兴[1,2] 

机构地区：[1]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518045 [2]深圳市外来有害生物检测技术研发重点实验室,广东深圳518045

出　　处：《动物医学进展》2014年第11期6-9,共4页Progress In Veterinary Medicine

基　　金：国家863计划项目(2012AA101301);国家科技支撑计划项目(2013BAD12B03)

摘　　要：研究小反刍兽疫病毒(Peste des petits ruminants virus,PPRV)H蛋白生物学活性,为建立PPRV抗体检测方法提供材料。根据GenBank已发表的PPRV H蛋白质序列,设计上、下游引物,以PPRV核酸为模板进行PCR扩增,扩增产物克隆至pET52/LIC载体中,构建了pET52/LIC-PPRV H表达载体。将pET52/LIC-PPRV H重组质粒转化大肠埃希菌BL21(DE3)进行融合表达,经SDS-PAGE电泳分析,可见PPRV H蛋白成功得到了表达,融合蛋白的分子质量约为75ku,Western blot分析表明所表达的重组蛋白可与PPRV阳性血清发生特异性反应,说明表达的PPRV H蛋白可以作为建立PPRV抗体检测的蛋白。In order to study the biological activity of hemagglutinin protein of peste des petits ruminants vi-rus （PPRV）and supply materials for establishing immunological assay for detection of antibodies against PPRV,the PPRV H gene was amplified by RT-PCR using the specific primers designed according to the H gene sequence of PPRV published in GenBank.The amplified gene was cloned into a prokaryotic expres-sion vector pET52/LIC to obtain a prokaryotic expression vector pET52/LIC-PPRV H.Then the recombi-nant pET52/LIC-PPRV H was transformed into E.coli BL21 （DE3）for protein expression induced by IPTG.The expressed protein was tested by SDS-PAGE and Western blot,and result showed that the ex-pressed PPRV H protein was approximately 75 ku and could reacted with the positive sera against PPRV. The expressed PPRV H protein could be used as an antigen for PPRV antibody detection.

关 键 词：小反刍兽疫病毒 H蛋白 原核表达 

分 类 号：S852.65[农业科学—基础兽医学]