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方文, 肖靓靓, 包怀恩, 牟荣. 亚洲带绦虫囊尾蚴特异性抗原筛选和鉴定[J]. 中国公共卫生, 2011, 27(11): 1431-1432. DOI: 10.11847/zgggws2011-27-11-29
引用本文: 方文, 肖靓靓, 包怀恩, 牟荣. 亚洲带绦虫囊尾蚴特异性抗原筛选和鉴定[J]. 中国公共卫生, 2011, 27(11): 1431-1432. DOI: 10.11847/zgggws2011-27-11-29
FANG Wen, XIAO Liang-liang, BAO Huai-en, . Screen and detection of specific antigens of Taenia asiatica cysticercus with two dimensional electrophoresis, western blotting and MS/MS spectrometry[J]. Chinese Journal of Public Health, 2011, 27(11): 1431-1432. DOI: 10.11847/zgggws2011-27-11-29
Citation: FANG Wen, XIAO Liang-liang, BAO Huai-en, . Screen and detection of specific antigens of Taenia asiatica cysticercus with two dimensional electrophoresis, western blotting and MS/MS spectrometry[J]. Chinese Journal of Public Health, 2011, 27(11): 1431-1432. DOI: 10.11847/zgggws2011-27-11-29

亚洲带绦虫囊尾蚴特异性抗原筛选和鉴定

Screen and detection of specific antigens of Taenia asiatica cysticercus with two dimensional electrophoresis, western blotting and MS/MS spectrometry

  • 摘要: 目的 对亚洲带绦虫囊尾蚴的特异性抗原进行筛选、鉴定和生物信息学分析.方法 6头20 d龄三元杂交乳猪随机分为实验组和对照组,实验组每头灌胃亚洲带绦虫虫卵悬液8.4万个;20 d后,分别制备实验组及对照组心脏血混合血清;收集、制备实验组亚洲带绦虫囊尾蚴蛋白并进行双向电泳分析,将凝胶蛋白斑点转移至聚偏氟乙烯膜,用实验组及对照组混合血清(均为1:10)为一抗,羊抗猪辣根过氧化物酶-IgG(1:200)为二抗,进行蛋白质印迹分析和质谱鉴定.结果 双向电泳凝胶共检测到(204±11)个蛋白质斑点,相对分子质量为14 400~94 000 kPa,等电点(pI)为3.0~10.0;筛选出6个特异性抗原,经质谱鉴定出其中2个蛋白,分别为ATP酶管家基因3-类蛋白2(AT-Pase family gene 3-like protein 2)和ATP合成酶β亚基(ATP synthase beta subunit),均与美国国家生物技术信息中心(NCBI)网上登录的亚洲带绦虫成虫蛋白一致,确定为亚洲带绦虫囊尾蚴特异性抗原.结论 获得2个亚洲带绦虫囊尾蚴特异性抗原,其免疫原性有待进一步研究,以便为研制抗囊尾蚴病疫苗提供参考.

     

    Abstract: Objective To screen and identify specific antigens of Taenia asiatica cysticercus and to predict the function of target proteins using bioinformatics method.Methods Six three-way cross hybrid pigs were randomly divided into experimental group and control group and each experimental pig was infected with 84 000 eggs.Serum samples were collected 40 days after the infection.The total protein of Taenia asiatica cysticercus was separated by two dimensional electrophoresis and western blotting was performed to find out distinct antigens.Proteins from the two groups were identified by electrospray ion trap mass spectrometry(ESI-Trap MS).Results Totally 204±11 spots were detected through Coomassie brilliant blue stained gels with relative molecular weight(Mr) of 14 400-94 000 kDa and isoelectric point(pI) of 3.0-10.0.Westernblotting showed 6 specific antigen spots with pool sera of infected pigs.Two of the 6 antigens with known functions were ascribed to ATPase family gene 3-like protein 2 and ATP synthase beta subunit.Conclusion Two specific antigens of Taenia asiatica cysticercus have been identified.

     

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