绵羊肺炎支原体膜蛋白p56基因的克隆、表达及反应原性研究  

Cloning and Expression of Membrane Protein p56 Gene of Mycoplasma ovipneumoniae and Its Identification of Reactogenicity

作　　者：陈诚[1] 乔军[1] 孟庆玲[1] 刘田莉 胡政香 马玉[1] 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,甘肃兰州730046

出　　处：《西南农业学报》2016年第2期455-458,共4页Southwest China Journal of Agricultural Sciences

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：为了分析绵羊肺炎支原体(MO)膜蛋白p56的反应原性,本研究以MO新疆流行株为模板,设计特异性引物对p56抗原集中区段进行PCR扩增,将扩增产物克隆测序,进一步亚克隆至表达载体pET-28a(+)中,构建pET-28a(+)-p56原核表达载体。将重组载体转化至大肠杆菌BL21(DE3)感受态细胞中,用IPTG对重组菌进行诱导,SDS-PAGE和Western blot分析表达产物。结果表明,SDS-PAGE分析证实表达的重组融合蛋白相对分子量为44 kDa;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p56融合蛋白具有良好的反应原性,为进一步研发MO检测用抗原奠定了前期基础。In order to analyze the zeactogenicity of membrane protein p56 of Mycoplasma ovipneumoniae, p56 gene of MO Xinjiang isolate was amplified by PCR technique, then the amplified products were further subcloned in the expression vector pET-28a（ ＋ ） for construction of recombinant pET-28a（ ＋ ） -p56. The expression vector was transformed into E. coli competent cells of BL21 （ DE3 ） and induced by IPTG, and SDS-PAGE and western blot were conducted for analysis of expressed products. The results of SDS-PAGE showed that recombinant pro- tein was successfully expressed with a mass of molecule of 44 kDa. Western blot revealed that the recombinant protein could specifically react with anti-MO positive serum, which confirmed that the recombinant p56 protein had a good reactiongenicity. This study laid the foundation for the development of antigen to detect MO.

关 键 词：MO 膜蛋白 p56基因 克隆 原核表达 反应原性 

分 类 号：S852.62[农业科学—基础兽医学]