不同截短片段基因1b型丙型肝炎病毒核心蛋白功能研究

发布时间：2018-02-04 12:38

本文关键词： 丙型肝炎病毒核心蛋白蛋白激酶R 信号转导及转录激活子3 丙型肝炎病毒核心蛋白凋亡基因芯片基因表达谱　出处：《浙江大学》2005年博士论文　论文类型：学位论文

【摘要】：丙型肝炎病毒(HCV)核心蛋白(CORE)是构成病毒的核壳蛋白,在HCV持续感染及肝癌(HCC)的发病机制中起重要作用。不同基因型HCV编码CORE结构和功能存在一定的差异,且同一病毒株编码的不同功能区域的功能不同。我国HCV感染主要流行株基因型为基因1b型。因此,研究基因1b型CORE及其不同截短片段在HCV持续感染及肝癌发病机制中的作用,对防治我国丙型肝炎具有重要的实际意义。蛋白激酶R(PKR)是干扰素(IFN)发挥抗病毒主要工具,在抵抗HCV的持续感染及HCC的发病机制中起重要作用,近年来受到特别的重视。信号转导及转录激活子(STAT)家族在介导细胞信号传导中起关键作用,其中STAT3被认为是癌性转录因子,可以介导多种不同的生理和病理过程,包括细胞生长,分化凋亡,胚胎的发育,转化,炎症和免疫反应。基于对CORE研究发现来自HCC癌组织中HCV的CORE能与PKR发生直接作用,CORE能激活STAT3而诱发肝细胞的恶性转化,PKR又可作为接头蛋白。本课题进一步对CORE与PKR、STAT3相互作用进行了研究,并对相互作用所依赖的功能区域进行了定位,提出了三者相互作用的模式,探讨三者在HCV致病机制,为抗HCV治疗提供新的靶位。目的 (1) 探讨基因1b型HCV不同截短片段CORE在HCV致病机制中的作用; (2) HCV病毒蛋白或CORE对PKR及STAT3表达水平的影响; (3) CORE与PKR及STAT3的相互作用在HCV致病机制中的作用,定位相互作用所依赖的各自功能区域; (4) 建立CORE与PKR及STAT3相互作用可能模式。方法 (1) 构建7个基因1b型HCV不同截短片段的谷胱苷肽S转移酶(GST)CORE融合蛋白的原核表达质粒pGEX 4T-1/CORE。表达纯化获得不同截短片段的GST-CORE及
[Abstract]:Hepatitis C virus (HCV) core protein (Corel) is the core-shell protein that makes up the virus. It plays an important role in the pathogenesis of HCV persistent infection and hepatocellular carcinoma (HCC). The structure and function of CORE encoded by different genotypes of HCV are different. Different functional regions of the same virus strain encode different functions. The main genotype of HCV infection in China is genotype 1b. To study the role of 1b type CORE and its different truncated fragments in the pathogenesis of HCV persistent infection and hepatocellular carcinoma. Protein kinase protein kinase (PKR) is the main antiviral tool of interferon interferon (IFN). It plays an important role in resisting the persistent infection of HCV and the pathogenesis of HCC. In recent years, special attention has been paid to the family of signal transduction and activator of transcription (STAT), which plays a key role in mediating cell signal transduction. Among them, STAT3 is considered to be a cancerous transcription factor. It mediates a variety of physiological and pathological processes, including cell growth, differentiation and apoptosis, embryonic development, transformation, inflammation and immune response. Based on the study of CORE, it was found that CORE from HCC carcinomas can directly interact with PKR. Core can activate STAT3 and induce malignant transformation of hepatocytes. PKR can also be used as a joint protein. In this study, the interaction between CORE and PKR- STAT3 was further studied, and the functional regions dependent on the interaction were located. A model of interaction among the three was proposed to explore the pathogenesis of HCV and to provide a new target for anti-HCV therapy. Purpose 1) to explore the role of different truncated CORE fragments of 1b type HCV in the pathogenesis of HCV. (2) the effect of HCV virus protein or CORE on the expression of PKR and STAT3; (3) the interaction of CORE with PKR and STAT3 in the pathogenesis of HCV, and the location of the respective functional regions on which the interaction depends; (4) to establish a possible model for the interaction of CORE with PKR and STAT3. Method Construction of glutathione S-transferase (GST) with different truncated fragments of 1b type HCV. The prokaryotic expression plasmid pGEX 4T-1 / Core of CORE fusion protein was expressed and purified to obtain different truncated GST-CORE and
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R373

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