绵羊肺炎支原体HSP70基因的克隆、表达及免疫原性分析  

Cloning,Expression of HSP70 Gene of Mycoplasma ovipneumoniae and Its Immunogenicity of the Recombinant Protein

作　　者：胡政香 乔军[1] 孟庆玲[1] 赵海龙[1] 陈诚[1] 赞哈尔.波拉提 马玉[1] 刘田莉 才学鹏[2] 陈创夫[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]中国农业科学院兰州兽医研究所,兰州730046

出　　处：《西北农业学报》2014年第12期8-12,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家国际科技合作专项(2014DFR31310);兵团国际科技合作计划(2012BC006)

摘　　要：根据GenBank登录的MO HSP70基因序列,设计特异性引物对MO新疆分离株HSP70基因进行扩增,克隆入T载体中测序。将HSP70基因亚克隆入表达载体pGEX-4T-1,构建重组表达载体pGEX4T-1-HSP70,并转化入大肠埃希氏菌BL21(DE3),用IPTG进行诱导重组菌,SDS-PAGE和Western blot分析表达产物。SDS-PAGE分析证实表达的重组融合蛋白分子质量为53ku;Western blot分析表明该重组蛋白可与MO阳性血清发生特异性免疫学反应,证实表达的重组HSP70具有良好的反应原性。重组HSP70蛋白免疫小鼠后可诱导机体产生特异性抗体,证实该重组蛋白具有良好的免疫原性。According to MO HSP70 gene sequences accessed in GenBank,apair of specific primers was designed to amplify HSP70 gene,and the amplified product was cloned into T vector and sequenced.Then HSP70 gene was subcloned into pGEX4T-1vector for expression in E.coli BL21（DE3）.Recombinant plasmid pGEX4T-1-HSP70 was transformed into E.coli,and SDS-PAGE and western blot were conducted for analysis of expressed products after IPTG induction.SDS-PAGE showed that recombinant protein was successfully expressed with a mass of molecule of 53 ku.Western blot revealed that the recombinant protein can specifically react with anti-MO polyclonal antibody,which confirmed that the recombinant HSP70 protein was of good reactiongenicity.The recombinant HSP70 protein can induce the body to produce specific antibodies in mice after immunization,which confirms that the recombinant protein has good immunogenicity.

关 键 词：MO HSP70基因 载体构建 原核表达 

分 类 号：S852.62[农业科学—基础兽医学]