人类乙型肝炎病毒聚合酶反式调节新基因HBVDNAPTP1的克隆表达与部分功能研究

发布时间：2019-01-10 08:51

【摘要】：乙型肝炎病毒的持续感染至今仍然是一个全球性的健康问题，不仅引起急慢性病毒性肝炎，而且与肝纤维化、肝细胞癌的发生发展密切相关。然而对于HBV感染目前尚无特效的治疗技术和方法，这主要是由于HBV感染引起的病毒性肝炎发病机制还有诸多方面没有研究清楚。目前认为，，HBV进入肝细胞后，病毒基因组及其编码的蛋白与肝细胞基因及蛋白之间的相互作用是决定HBV复制、表达、免疫逃逸、慢性感染、肝纤维化及致恶性转化的关键因素。特别是近年来随着对HBV致病机制研究的深入，发现在HBV与肝细胞相互作用的过程中涉及到复杂的反式调节机制，肝炎病毒蛋白在肝细胞中的表达对于肝细胞的基因表达谱产生影响，这一作用可能与其致病机制有关。寻找肝细胞中与HBV各抗原成分反式调节相关的基因，并进一步弄清具体作用机制、作用效应及连带影响，对明确HBV的致病机制，寻找有效防治方法具有重要意义。对于筛选得到的未知功能基因的克隆表达与功能研究是病毒性肝炎发病机制研究领域中创新知识的重要源泉，同时也是人类基因组计划和后基因组计划的重要内容。利用转染了HBV DNA聚合酶三个功能基因片段表达载体的肝细胞和仅转染空载体的肝细胞为材料，提取其mRNA应用SSH技术研究了HBV DNAP的反式调节靶基因，筛选得到一未知功能基因，经斑点杂交技术验证后，确定其基因编码序列，命名为乙型肝炎病毒DNA聚合酶反式调节蛋白1(GenBank Accession：AY450389)。发现一种新基因，将其编码的新蛋白结构与生物学功能，与生物学和临床医
[Abstract]:The persistent infection of hepatitis B virus is still a global health problem, not only causing acute and chronic viral hepatitis, but also closely related to the occurrence and development of liver fibrosis and hepatocellular carcinoma. However, there is no effective treatment for HBV infection, which is mainly due to the pathogenesis of viral hepatitis caused by HBV infection is still not clear. At present, it is believed that the interaction between the viral genome and its encoded proteins and the genes and proteins of hepatocytes after HBV enters into hepatocytes is the key factor that determines the replication, expression, immune escape, chronic infection, liver fibrosis and malignant transformation of HBV. Especially in recent years, with the further study of the pathogenesis of HBV, it has been found that the complex trans-regulation mechanism is involved in the interaction between HBV and hepatocytes. The expression of hepatitis virus protein in hepatocytes has an effect on the gene expression profile of hepatocytes, which may be related to its pathogenesis. It is important to find out the genes related to the transregulation of HBV antigens in hepatocytes, and to find out the specific mechanism, effects and associated effects of HBV. It is of great significance to clarify the pathogenetic mechanism of HBV and to find effective methods for its prevention and treatment. Cloning, expression and functional study of unknown genes obtained from screening are important sources of innovative knowledge in the field of viral hepatitis pathogenesis research, and are also important contents of the human genome project and post-genome project. Using hepatocytes transfected with three functional gene fragments of HBV DNA polymerase and hepatocytes transfected with empty vector as materials, mRNA was extracted to study the trans-regulatory target gene of HBV DNAP by SSH technique, and an unknown functional gene was screened out. The gene coding sequence was confirmed by dot blot, and named DNA polymerase trans regulatory protein 1 (GenBank Accession:AY450389) of hepatitis B virus. Discovery of a new gene that encodes new protein structures and biological functions, with biological and clinical medicine
【学位授予单位】：大连医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R373

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