| 牛霉形体免疫相关性蛋白P51的筛选与鉴定 |
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| 引用本文： | 李媛,陈维,呼太飞,刘洋,宋志强,孙文静,辛九庆.牛霉形体免疫相关性蛋白P51的筛选与鉴定[J].中国兽医科学,2011(5). |
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| 作者姓名： | 李媛 陈维 呼太飞 刘洋 宋志强 孙文静 辛九庆 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室动物细菌病研究室国家牛传染性胸膜肺炎指定检测实验室;东北农业大学动物医学学院;黑龙江省勃利县动物卫生监督所; |
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| 基金项目： | 国家自然科学基金项目(31072131); 哈尔滨市科技攻关项目(2010AA6AN019) |
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| 摘 要： | 为了筛选免疫原性蛋白,建立牛霉形体的快速诊断方法,利用二维凝胶(2-D)电泳、Western-blot分析及蛋白质谱分析,筛选出一个免疫相关性蛋白P51。以牛霉形体中国分离株Hubei-1为模板,扩增了p51基因,并克隆到原核表达载体pET32a上。结果显示,重组蛋白pET32a-p51与牛霉形体阳性血清的Dot-blotting试验结果为阳性,而Western-blotting试验结果为阴性。以pET32a-p51重组蛋白为包被抗原的ELISA试验表明其能与自然感染和人工感染牛霉形体的阳性血清发生特异性反应,而与灭活疫苗免疫的阳性血清不发生特异性反应,与牛传染性胸膜肺炎的阳性血清没有交叉反应。表明P51可能是牛霉形体特有的一种构象依赖性免疫相关蛋白,是一种有前景的诊断用抗原。
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| 关 键 词： | 牛霉形体 牛传染性胸膜肺炎 二维凝胶电泳 质谱 |
Screening and identification of immuno-related protein P51 of Mycoplasma bovis |
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| Affiliation: | LI Yuan1,CHEN Wei2,HU Tai-fei3,LIU Yang1,SONG Zhi-qiang1,SUN Wen-jing2,XIN Jiu-qing1(1.National Contagious Bovine Pleuropneumonia Designate Detection Laboratory/State Key Laboratory of Veterinary Biotechnology/Division of Bacterial Diseases/Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China,2.College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,3.Heilongjiang Province Boli County Animal Sanitary Supervision Institute,Boli 154500,... |
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| Abstract: | To select the immuno-related proteins of Mycoplasma bovis and establish the quickly diagnostic method of M.bovis,2-DE,Western-blot and mass spectrometric were used to filter out an immunoprotein P51.A p51 gene was amplified by PCR from a M.bovis strain isolated from Hubei-1,and was cloned into pET32a vector to express.The highly purified protein was obtained by Ni-NTA system.Dot-blot showed that recombinant protein pET32a-p51 could be probed with M.bovis positive bovine antiserum and could be considered to ... |
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| Keywords: | Mycoplasma bovis contagious bovine pleuropneumonia (2-D)two-dimensional gel electrophoresis mass spectrometric |
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