绵羊肺炎支原体膜蛋白p56基因的克

发布时间：2018-02-11 00:17

本文关键词： MO 膜蛋白 p基因克隆原核表达反应原性　出处：《西南农业学报》2016年02期 　论文类型：期刊论文

【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。
[Abstract]:In order to analyze the reactivity of the membrane protein p56 of Mycoplasma pneumoniae, a specific primer was designed to amplify the concentrated region of p56 antigen using MO Xinjiang epidemic strain as template, and the amplified product was cloned and sequenced. The recombinant vector was subcloned into the expression vector pET-28a (pET-28a ()), and the expression vector pET-28a (pET-28a) (pET-28a) was constructed. The recombinant vector was transformed into E. coli BL21DDE3 cells. IPTG was used to induce SDS-PAGE and Western blot analysis of the expressed product. The results showed that the relative molecular weight of the expressed recombinant fusion protein was 44 kDa blot, which indicated that the recombinant protein could react specifically with MO positive serum. It was confirmed that the recombinant p56 fusion protein had good reactivity, which laid a foundation for the further development of antigen for MO detection.
【作者单位】：石河子大学动物科技学院;中国农业科学院兰州兽医研究所;
【基金】：国家国际科技合作专项(2014DFR31310) 兵团国际科技合作计划(2012BC006)
【分类号】：S852.62

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