2型猪链球菌中国强毒株及其covR基因突变株的蛋白质组学研究

发布时间：2018-06-03 05:53

本文选题：型猪链球菌 + 毒力调控因子CovR　；参考：《中国人兽共患病学报》2016年05期

【摘要】：目的通过比较2型猪链球菌中国强毒株05ZYH33及其covR基因突变株△covR蛋白表达谱差异,质谱鉴定差异表达蛋白,分析CovR在蛋白表达调控中的作用。方法首先将05ZYH33和△covR在THB培养基中培养至对数期,然后裂解细菌制备蛋白样品。第一向等电聚焦电泳在pH3~10的IPG胶条上完成后进行SDS-PAGE电泳,电泳胶经扫描分析后选取蛋白点进行质谱鉴定。结果 05ZYH33和△covR全菌蛋白裂解液经二维电泳分别得到559和491个蛋白点,经比对发现两菌株蛋白表达量差异3倍以上蛋白点40个,经质谱鉴定出15个蛋白,主要涉及细胞代谢的酶类如谷氨酸脱氢酶、腺苷酸激酶、PTS系统成分等,以及分子伴侣蛋白如GroEL和Dnak等;电泳分离得到△covR特异蛋白点124个,质谱鉴定出15个,主要为参与细胞糖代谢过程中的酶,如磷酸甘油酸激酶、甘油醛-3-磷酸脱氢酶、丙酮酸激酶等;质谱鉴定05ZYH33特异表达蛋白5个。结论鉴定05ZYH33和△covR差异表达蛋白35个,部分蛋白涉及细菌毒力、宿主细胞粘附、细胞分裂等生命过程,同时蛋白分子伴侣在△covR中的表达变化说明CovR的调控可能发生在蛋白修饰水平,为研究CovR在调控细菌毒力中的作用和分子机制奠定基础。
[Abstract]:Objective to compare the differentially expressed protein profiles of 05ZYH33 and covR gene mutants of Streptococcus suis type 2, identify the differentially expressed proteins by mass spectrometry, and analyze the role of CovR in the regulation of protein expression. Methods 05ZYH33 and covR were cultured in THB medium for a logarithmic period, and then the bacteria were used to prepare protein samples. First, the isoelectric focusing electrophoresis was carried out on the IPG strip of pH3~10 and then SDS-PAGE electrophoresis was carried out. After scanning analysis, the protein spots were selected for mass spectrometry identification. Results 559 and 491 protein spots were obtained by two-dimensional electrophoresis in 05ZYH33 and covR whole bacterial protein cleavage solution. The protein expression difference between the two strains was found to be more than 3 times of 40, and 15 proteins were identified by mass spectrometry. The enzymes involved in cell metabolism, such as glutamate dehydrogenase, adenylate kinase, PTS system, and molecular chaperones such as GroEL and Dnak, were identified by mass spectrometry, and 124 specific protein sites of covR were isolated by electrophoretic electrophoresis, and 15 of them were identified by mass spectrometry. The enzymes involved in cell glucose metabolism, such as phosphoglycerate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase, etc., were identified by mass spectrometry for 5 proteins specifically expressed by 05ZYH33. Conclusion 35 differentially expressed proteins of 05ZYH33 and covR were identified. Some proteins were involved in bacterial virulence, host cell adhesion and cell division. At the same time, the expression of protein chaperones in covR suggests that the regulation of CovR may occur at the level of protein modification, which lays a foundation for the study of the role and molecular mechanism of CovR in the regulation of bacterial virulence.
【作者单位】：南京师范大学生命科学学院;南京军区军事医学研究所;
【基金】：国家自然科学基金(Nos.81571965;81471920) 江苏省自然科学基金(No.BK20151091) 江苏省333工程科研资助项目(BRA2014363)联合资助~~
【分类号】：R378.12

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