绵羊肺炎支原体核糖体蛋白30S RPS11基因的克

发布时间：2018-01-23 21:16

本文关键词： MO SRPS 克隆原核表达免疫原性反应原性　出处：《家畜生态学报》2017年05期 　论文类型：期刊论文

【摘要】：本研究以绵羊肺炎支原体(Mycoplasma ovipneumoniae,MO)新疆流行株基因组DNA为模板,通过PCR扩增30S RPS11基因,对其进行分子特征性分析,并将其抗原表位集中区进行原核表达,分析重组蛋白免疫原性。结果表明,30S RPS11基因全长405bp,编码134个氨基酸,该蛋白是一种受磷酸化调控的蛋白,含有一个核糖体蛋白S11信号。SDSPAGE结果显示,筛选的抗原表位集中区重组蛋白分子质量为29kDa;Western blot分析显示,该重组蛋白可被MO阳性血清特异性识别,说明其具有良好的反应原性;小鼠免疫试验表明,重组蛋白诱导机体产生的间接血凝抗体效价可达1∶32~1∶64,表明其具有较强的免疫原性。本试验首次证实MO核糖体蛋白(30SRPS11)具有良好的免疫原性和反应原性,为MO血清学诊断及亚单位疫苗研发提供了科学依据。
[Abstract]:In this study, Mycoplasma pneumoniae (Mycoplasma ovipneumoniae MO) Xinjiang strains 30S genomic DNA as template, RPS11 gene was amplified by PCR and analyzed the molecular characteristics of it, and the epitope concentration area for prokaryotic expression analysis of recombinant protein immunogenicity. The results showed that the full-length 405bp gene of 30S RPS11. Encoding 134 amino acids. The protein is a protein by phosphorylation regulation, containing a ribosomal protein S11 signal.SDSPAGE showed that screening epitope recombinant protein molecular mass concentration area is 29kDa; Western blot analysis showed that the recombinant protein was MO positive serum shows its immunogenicity good response; mice immunological tests showed that the recombinant protein induced indirect hemagglutination antibody titer of 1: 32~1: 64, show that it has strong immunogenicity. This experiment confirmed that the first MO Ribosomal protein (30SRPS11) has good immunogenicity and reactivity, which provides a scientific basis for MO serological diagnosis and subunit vaccine development.

【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) 国家自然科学基金(31360596)
【分类号】：S852.62
【正文快照】： 绵羊肺炎支原体(Mycoplasma ovipneumoni-ae,MO)是绵羊、山羊呼吸道感染的重要致病菌之一,病羊是主要的传染源,传播方式主要为接触感染,如吸入病羊的呼吸道飞沫而发病。MO可粘附在绵羊呼吸道粘膜表面引起感染,发病羊主要表现为干咳、消瘦和发育不良。该病在全球范围内广泛分布

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