羊种布鲁氏菌强毒株16M与疫苗株M5差异蛋白质组学分析
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摘要
目的从蛋白质整体水平分析羊种布鲁氏菌强毒株16M和疫苗株M5的差异蛋白,探讨羊种布鲁氏菌强毒株16M和疫苗株M5毒力机制。
方法利用丙酮干粉法分别提取布鲁氏菌强毒株16M和疫苗株M5的菌体全蛋白质,以双向聚丙烯酰胺凝胶电泳技术进行蛋白质分离,考马斯亮蓝染色,扫描获得双向电泳图谱,利用ImageMaster 2D Platinum4.0软件筛选出差异蛋白质,切点,制备质谱样,进行质谱分析,用MALDI-TOF-MS获得肽质量指纹图谱,数据库进行搜索,获得所鉴定蛋白质的生物学信息。
结果应用固相pH梯度双向凝胶电泳获得了重复性较好的2-DE凝胶图谱,观察布鲁氏菌强毒株16M和疫苗株M5的2-DE图谱,其蛋白表达模式非常相似,利用ImageMaster 2D Platinum 4.0软件自动识别蛋白点后,布鲁氏菌强毒株16M获得蛋白点856个,疫苗株M5蛋白点793个。从中共筛选出23个差异蛋白点进行质谱鉴定,其中20个蛋白点在数据库中匹配到了有意义的蛋白质。
通过质谱鉴定,得到的差异蛋白功能涉及糖的代谢、物质运输、蛋白的合成以及分子伴侣等。其中有13个蛋白在强毒株16M中表达上调,5个蛋白在疫苗株M5中表达上调。
Objective:To analyze different expression proteins of the brucella melitensis a virulent strain 16M with vaccine strain M5 by the technologies of proteomics, and explore the mechanism of virulence in B .melitensis and find out the diagnosis markers of the brucellosis.
Methods:Extracted total proteins of the brucella melitensis a virulent strain 16M and vaccine strain M5 by acetone cryochem,separated proteins by two-dimensional electrophoresis(2-DE),stained proteins by Coomassie brilliant blue, got the patterns,found out differential proteins, and obtained PMF(peptide mass fingerprinting) by MALDI-TOF-MS, gained related the information of proteins by database.
Results:In this study, 2-DE Patterns with high resolution and reproducibility from the proteins of the strains 16M and M5 was obtained. The total Protein distribution pattern of the strains16M and M5 2-DE is very similar.A total of 856 and 793 protein spots were detected for strains 16M and M5,respectively.20 out of 23 protein spots were identified by the technologies of mass spectrogram with bioinformatics.
20 protein spots identified by the technologies of mass spectrogram with bioinformatics seem to play important roles in a variety of pathways including sugar binding, protein production,lipid metabolism,and molecular chaperone.13 of 20 are upexpressed proteins in strain 16M,5 of 20 are upexpressed proteins in strain M5.
方法利用丙酮干粉法分别提取布鲁氏菌强毒株16M和疫苗株M5的菌体全蛋白质,以双向聚丙烯酰胺凝胶电泳技术进行蛋白质分离,考马斯亮蓝染色,扫描获得双向电泳图谱,利用ImageMaster 2D Platinum4.0软件筛选出差异蛋白质,切点,制备质谱样,进行质谱分析,用MALDI-TOF-MS获得肽质量指纹图谱,数据库进行搜索,获得所鉴定蛋白质的生物学信息。
结果应用固相pH梯度双向凝胶电泳获得了重复性较好的2-DE凝胶图谱,观察布鲁氏菌强毒株16M和疫苗株M5的2-DE图谱,其蛋白表达模式非常相似,利用ImageMaster 2D Platinum 4.0软件自动识别蛋白点后,布鲁氏菌强毒株16M获得蛋白点856个,疫苗株M5蛋白点793个。从中共筛选出23个差异蛋白点进行质谱鉴定,其中20个蛋白点在数据库中匹配到了有意义的蛋白质。
通过质谱鉴定,得到的差异蛋白功能涉及糖的代谢、物质运输、蛋白的合成以及分子伴侣等。其中有13个蛋白在强毒株16M中表达上调,5个蛋白在疫苗株M5中表达上调。
Objective:To analyze different expression proteins of the brucella melitensis a virulent strain 16M with vaccine strain M5 by the technologies of proteomics, and explore the mechanism of virulence in B .melitensis and find out the diagnosis markers of the brucellosis.
Methods:Extracted total proteins of the brucella melitensis a virulent strain 16M and vaccine strain M5 by acetone cryochem,separated proteins by two-dimensional electrophoresis(2-DE),stained proteins by Coomassie brilliant blue, got the patterns,found out differential proteins, and obtained PMF(peptide mass fingerprinting) by MALDI-TOF-MS, gained related the information of proteins by database.
Results:In this study, 2-DE Patterns with high resolution and reproducibility from the proteins of the strains 16M and M5 was obtained. The total Protein distribution pattern of the strains16M and M5 2-DE is very similar.A total of 856 and 793 protein spots were detected for strains 16M and M5,respectively.20 out of 23 protein spots were identified by the technologies of mass spectrogram with bioinformatics.
20 protein spots identified by the technologies of mass spectrogram with bioinformatics seem to play important roles in a variety of pathways including sugar binding, protein production,lipid metabolism,and molecular chaperone.13 of 20 are upexpressed proteins in strain 16M,5 of 20 are upexpressed proteins in strain M5.
引文
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