本文選題：型豬鏈球菌 + 毒力調(diào)控因子CovR��； 參考：《中國人獸共患病學(xué)報》2016年05期

【摘要】：目的通過比較2型豬鏈球菌中國強毒株05ZYH33及其covR基因突變株△covR蛋白表達(dá)譜差異,質(zhì)譜鑒定差異表達(dá)蛋白,分析CovR在蛋白表達(dá)調(diào)控中的作用。方法首先將05ZYH33和△covR在THB培養(yǎng)基中培養(yǎng)至對數(shù)期,然后裂解細(xì)菌制備蛋白樣品。第一向等電聚焦電泳在pH3~10的IPG膠條上完成后進行SDS-PAGE電泳,電泳膠經(jīng)掃描分析后選取蛋白點進行質(zhì)譜鑒定。結(jié)果 05ZYH33和△covR全菌蛋白裂解液經(jīng)二維電泳分別得到559和491個蛋白點,經(jīng)比對發(fā)現(xiàn)兩菌株蛋白表達(dá)量差異3倍以上蛋白點40個,經(jīng)質(zhì)譜鑒定出15個蛋白,主要涉及細(xì)胞代謝的酶類如谷氨酸脫氫酶、腺苷酸激酶、PTS系統(tǒng)成分等,以及分子伴侶蛋白如GroEL和Dnak等;電泳分離得到△covR特異蛋白點124個,質(zhì)譜鑒定出15個,主要為參與細(xì)胞糖代謝過程中的酶,如磷酸甘油酸激酶、甘油醛-3-磷酸脫氫酶、丙酮酸激酶等;質(zhì)譜鑒定05ZYH33特異表達(dá)蛋白5個。結(jié)論鑒定05ZYH33和△covR差異表達(dá)蛋白35個,部分蛋白涉及細(xì)菌毒力、宿主細(xì)胞粘附、細(xì)胞分裂等生命過程,同時蛋白分子伴侶在△covR中的表達(dá)變化說明CovR的調(diào)控可能發(fā)生在蛋白修飾水平,為研究CovR在調(diào)控細(xì)菌毒力中的作用和分子機制奠定基礎(chǔ)。
[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.
【作者單位】： 南京師范大學(xué)生命科學(xué)學(xué)院;南京軍區(qū)軍事醫(yī)學(xué)研究所;
【基金】：國家自然科學(xué)基金(Nos.81571965;81471920) 江蘇省自然科學(xué)基金(No.BK20151091) 江蘇省333工程科研資助項目(BRA2014363)聯(lián)合資助~~
【分類號】：R378.12

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