牛布鲁氏菌不同种强毒株膜蛋白质组学差异的比较

Comparative Analysis of Brucella Virulent Strains Membrane Proteomic Differences

[目的]比较牛的不同种类布鲁氏菌的菌株膜蛋白质组学的差异。[方法]采用丙酮干粉方法,提取布鲁氏菌强毒株16M和疫苗株M5的全蛋白,具有双向聚丙烯酰胺凝胶电泳分离蛋白质,用考马斯亮蓝染色、扫描获取二维电泳图谱,用ImageMaster TM2D Platinum6.0-TOF软件筛选出差异蛋白质酶切,准备样品进行质谱分析,通过MALDI.0软件在不同的肽质量指纹质谱搜索数据库,通过生物信息技术进行蛋白质识别。[结果]应用固相pH梯度图获得良好的重复性2-DE凝胶,观察布鲁氏菌强毒株16M和疫苗株M5 2-DE图谱,其蛋白质的表达模式非常相似,使用ImageMaster TM2D4.0软件自动识别蛋白质点,检测布鲁氏菌16 M蛋白的蛋白质点856,疫苗株M5的蛋白质点793。在数据库中选择有意义的蛋白质23种进行选择质谱鉴定。蛋白质功能的差异是由质谱鉴定得到的,涉及糖代谢、物质运输、蛋白质合成和分子伴侣等方面,有13种在16M和疫苗株M5的蛋白高表达。[结论]通过质谱鉴定,获得的蛋白质功能涉及糖代谢、物质运输、蛋白质合成和分子伴侣等,有13种在16M和疫苗株M5的蛋白高表达。

[ Objective ] To compare membrane proteomic differences of different Brucella virulent strains of cattle. [ Method ] Use acetone dry powder method to extract whole proteins of Brucella virulent strain 16 M and vaccine strain M5, two-dimenslonal polyacrylamide gel electro- phoresis to separate proteins, Kaumas Coomassie brilliant blue staining and scanning to obtain two-dimensional electrophoretogram, Im- ageMaster TM2D Platinum 6.0-TOF to screen different protein digestion, prepare samples for mass spectral analysis, use MALDI . 0 to search and identify proteins in databases on the basis of bio-information technology. [ Result] Two-dimensional gel electrophoresis shows that repeat- able 2-DE gel can be obtained by using solidoid pH graduated symbols; according to 2-DE spectrum of Brueella virulent strain 16 M and vac- cine strain M5, expression patterns of their proteins are similar; [mageMaster TM21M. 0 is used to identify the protein spots, that of Brueella virulent strain 16 M is 856, that of vaccine strain M5 is 793. Twenty-three significant proteins were chosen from the database for mass spec- trum identification. [ Conclusion ] Differences of protein functions are obtained from the mass spectrum identification, 13 of them are included in 16M and M5, involving glycometabolism, material transport, protein synthesis and molecular chaperones.