本文選題：乙型肝炎病毒 + RNA　； 參考：《福建醫(yī)科大學(xué)》2012年碩士論文

【摘要】：乙型肝炎病毒（hepatitis B virus, HBV）是一種部分雙鏈環(huán)狀嗜肝DNA病毒，在醫(yī)學(xué)和公共衛(wèi)生上具有重要的意義。從世界范圍來看，大約有350000000的人感染乙肝病毒，其中亞洲地區(qū)占了75%。HBV是慢性肝炎、肝硬化和肝癌的主要致病原因，每年導(dǎo)致1000000人死亡。 HBV基因組剪接變異體(spliced variants of hepatitis B virus genomes)是由3.5Kb前基因組RNA (pregenomic RNA, pgRNA)經(jīng)剪接后逆轉(zhuǎn)錄而成的一類亞基因組DNA。其中長度為2.2kb的HBV剪接變異體占了80%以上，，其剪接類型分為單剪接型和雙剪接型，對2.2Kb HBV剪接變異體的功能研究顯示，此類變異體可能導(dǎo)致肝細胞凋亡并與HBV的持續(xù)性感染密切相關(guān)，但其確切致病機制迄今未明。本研究旨在利用免疫沉淀技術(shù)探索能夠與雙剪接型2.2Kb剪接變異體編碼的剪接特異性新蛋白（Hepatitis B Doubly Spliced Protein, HBDSP）相互作用的肝細胞蛋白，以此深入了解其致病性。 本課題第一部分，我們運用免疫沉淀（Immunoprecipitation）及基質(zhì)輔助激光解吸電離飛行時間質(zhì)譜鑒定（MALDI-TOF-MS）的方法獲得了四個與HBDSP相互作用的肝細胞蛋白，分別為：通用轉(zhuǎn)錄因子II,i (General transcription factor III, GTF2I或TFII-I)、葡萄糖調(diào)節(jié)蛋白78(Glucose-regulated protein, GRP78)、熱休克蛋白70(Heat shock protein70kDa, HSP70)和鉀離子通道(Potassiumvoltage-gated channel, KVβ.2)。本課題第二部分，首先通過免疫共沉淀（Co-Immunoprecipitation, Co-IP）實驗來作驗證，結(jié)果表明在肝細胞中HBDSP與TFII-I蛋白存在相互作用，而HBDSP與GRP78和KVβ.2沒有相互作用。進一步通過GST pull-down實驗證實HBDSP與TFII-I蛋白在體外具有相互作用。TFII-I蛋白是一個多功能轉(zhuǎn)錄因子，不僅能夠促進染色質(zhì)重塑，同時在細胞有絲分裂周期中具有重要作用。HBDSP與TFII-I的相互作用，可能影響肝癌細胞的分裂和增殖。本研究為深入探討HBDSP的致病性奠定基礎(chǔ)。
[Abstract]:Hepatitis B virus (HBV) is a partial double stranded circular hepatitis virus (HBV), which is of great significance in medicine and public health. Worldwide, about 350000000 people are infected with hepatitis B virus, of which 75. HBV is the leading cause of chronic hepatitis, cirrhosis and liver cancer in Asia. The splicing variant of variants of hepatitis B virus genomeses is a subgenome DNA, which is composed of 3.5kb pregenomic RNAs (pgRNAs) spliced and reverse transcribed. Among them, the length of 2.2kb splicing variants accounted for more than 80%, and the splicing types were divided into single splicing type and double splicing type. The function of 2.2 Kb HBV splicing variant was studied. Such variants may lead to hepatocyte apoptosis and are closely related to HBV persistent infection, but its exact pathogenesis has not been clarified. The aim of this study was to explore the hepatocyte proteins which can interact with the splicing specific protein (HBDSPN) encoded by double splicing variant 2.2Kb splicing variants by immunoprecipitation technique in order to understand the pathogenicity of Hepatitis B Doubly splicing protein (HBDSP). In the first part of this study, we obtained four hepatocyte proteins interacting with HBDSP by using immunoprecipitation and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). GTF2I, GTF2I, GTF2I, Glucose-regulated Glucose-regulated protein, GRP78A, heat shock protein 70heat shock 70kDa, HSP70) and Potassiumvoltage-gated channel, KV 尾. In the second part of this thesis, the co-immunoprecipitation (Co-IPT) experiment was carried out to verify the interaction between HBDSP and TFII-I protein in hepatocytes, but no interaction between HBDSP and GRP78 and KV 尾 .2 in hepatocytes. Furthermore, it was proved by GST pull-down that HBDSPs interact with TFII-I protein in vitro. TFII-I protein is a multifunctional transcription factor, which can not only promote chromatin remodeling, but also promote chromatin remodeling. The interaction between HBDSP and TFII-I may affect the division and proliferation of hepatoma cells. This study lays a foundation for further discussing the pathogenicity of HBDSP.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R373

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