摘要
目的利用蛋白质组学技术研究肠道病毒71型(EV71)编码3C蛋白对宿主细胞蛋白表达的影响。方法在293T细胞中瞬时过表达EV71 3C蛋白,应用双向荧光差异凝胶电泳(2D-DIGE)检测3C表达后293T细胞中蛋白的差异。选取表达水平有明显变化的蛋白用Western blot进行验证。结果在293T细胞中过表达3C蛋白后,可引起5个蛋白表达水平发生明显改变,其中4个蛋白表达上调,1个蛋白(HSPA4)表达下调。Western blot验证表明过表达3C可降低HSPA4的表达,HeLa细胞中HSPA4表达为对照的46.8%±10.1%(P<0.05)。结论过表达EV713C蛋白可引起细胞内蛋白表达差异。
Objective To explore the effects of 3C protease on host cell proteins by proteomics techniques.Methods 293T cells were transfected with plasmid encoding EV71 3C,cell lysates were analyzed by using two-dimensional fluorescence difference gel electrophoresis（2D-DIGE） and mass spectrometry（MS） analysis.Down-regulated protein was confirmed by Western blot analysis.Results The expression of EV71 3C in 293T cells was confirmed.A total of 5 altered proteins were identified after 3C overexpression.Among them,expression of 4 proteins increased,while expression of 1 protein decreased.Down-regulated protein HSPA4 was confirmed by Western blot.HSPA4 expression was determined by performing densitometric analyses in HeLa cells.The results showed that HSPA4 expression was 46.8%±10.1%（P0.05） compared to normal cells.Conclusion Overexpression of EV71 3C regulates the protein expression in 293T cells.
出处
《基础医学与临床》
CSCD
北大核心
2011年第6期652-655,共4页
Basic and Clinical Medicine
基金
国家973计划(2011CB504903)
