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作 者:张双翔[1] 程振涛[2,3] 唐宇[1] 冉隆仲[1] 李涛[1] 周碧君[2,3] 文明[2,3] 王开功[2,3] 王慧[2]
机构地区:[1]贵州省动物疫病预防控制中心,贵州贵阳550008 [2]贵州大学动物科学学院,贵州贵阳550025 [3]贵州省动物疫病研究室,贵州贵阳550025
出 处:《中国畜牧兽医》2014年第7期74-80,共7页China Animal Husbandry & Veterinary Medicine
基 金:黔西南州种草养羊产业发展省州科技合作专项项目(黔西南科合[2012]5号);贵州省自然科学技术基金(黔科合J字[2011]2332号);贵州省农业科技攻关项目(黔科合NY字[2011-3105]号);贵州省动物疫病与兽医公共卫生重点实验室培育(黔科合计Z字[2011]4008号);贵州省重大科技专项计划资助项目(黔科合重大专项字[2011]6009号)
摘 要:本试验旨在了解绵羊肺炎支原体(Mo)标准Y98株Hsp70基因生物学特性,以期利用该基因表达蛋白建立ELISA方法。本研究应用DNAStar、Bioedit 7.0、ClustalX 3.0、Mega 4.0软件与Protparam、TMpred、IEDB在线工具对Hsp70蛋白的理化参数、跨膜结构、信号肽、二级结构、B细胞表位及与其他支原体Hsp70蛋白进化关系进行了比对分析。结果显示,Mo Y98株Hsp70蛋白理论分子质量为66.14ku,pI值为5.08,为稳定的可溶性蛋白。该蛋白无跨膜结构,不分泌信号肽,存在多处具有免疫原性的肽段,具有形成抗原表位的优势结构,是建立免疫学方法较为理想的靶蛋白。与多种支原体Hsp70基因进化关系分析结果表明,该蛋白与Mo进化关系最近,与丝状支原体簇成员亲缘关系较远,为进一步分析Mo致病机理提供了理论依据。The assay was aimed to establish the ELISA by Mycoplasma ovipneumoniae (Mo) strain Y98 Hsp70 proteinwhich biological characteristics should be analyzed. The physical and chemical parameters, transmembrane, signal peptide, sec-ondary strucure,B cell epitopes were predicted and analyzed by DNAStar, Bioedit 7.0, ClustalX 3.0, Mega 4.0 softwares andProtparam,TMpred,IEDB online tools in this study. The results showed that the Hsp70 protein of Mo strain Y98 was stableand soluble which predicted molecular weight was 66.14 ku and pI was 5.08. The protein had no transmembrane and did notsecrete signal peptide. It was also proved that the protein was ideal target protein to establish immune method due to the advan-tage structure of epitope existed. It was indicted that the evolutionary relationships between Mo and Mycoplasma hyopneumon-iae was close which was far compared with Mycoplasma mycoides cluster by constructed phylogenetic trees of Hsp70 gene. Itwould provide theoretical basis to analyze pathogenesis of Mo further.
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