绵羊肺炎支原体p60蛋白N端与C端的表达及其免疫学特性研究

发布时间：2018-02-25 20:29

本文关键词： 绵羊肺炎支原体 p蛋白表达　出处：《中国畜牧兽医》2017年02期 　论文类型：期刊论文

【摘要】：为分析绵羊肺炎支原体(Mycoplasma ovipneumoniae,MO)膜表面脂蛋白p60的免疫学特性,本研究分别扩增p60蛋白N端与C端的基因,并通过重叠延伸PCR将C端基因序列中的TGA(1 459-1 461bp)定点突变为同义密码子TGG。将扩增的基因片段分别插入到pET-32a(+)表达载体上,重组质粒分别转化至大肠杆菌BL21(DE3)感受态细胞中进行诱导表达,得到分子质量约57ku的p60-N蛋白和分子质量约为30.5ku的p60-C蛋白;再对纯化后的重组蛋白进行Western blotting分析,结果表明重组蛋白均可与MO阳性血清发生特异性免疫学反应,表明表达的重组蛋白p60-N和p60-C均有良好的反应原性。本研究为筛选MO主要的免疫原性蛋白和建立特异的血清学诊断方法提供了依据。
[Abstract]:In order to analyze the immunological characteristics of membrane surface lipoprotein p60 of Mycoplasma ovis pneumoniae MOA, the N-terminal and C-terminal genes of p60 protein were amplified, respectively. The TGA(1 459-1 461bp in C terminal gene sequence was mutated into synonymous codon TGG by overlapping extension PCR. The amplified gene fragments were inserted into pET-32a () expression vector, respectively. The recombinant plasmids were transformed into Escherichia coli BL21DE3) and induced to express the p60-N protein with molecular weight of 57ku and p60-C protein with molecular weight of 30.5ku. The purified recombinant protein was analyzed by Western blotting. The results showed that the recombinant protein could react specifically with MO positive serum. The results showed that the recombinant proteins p60-N and p60-C had good reactivity. This study provided a basis for screening the major immunogenic proteins of MO and establishing a specific serological diagnostic method.
【作者单位】：内蒙古农业大学兽医学院微生物与免疫实验室;
【基金】：内蒙古自治区科技计划“舍饲肉羊疫病动态防控关键技术研究”(201502070) 国家科技支撑计划“重点牧区”生产生态生活“保障技术集成与示范”(2012BAD13B00)
【分类号】：S852.62

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