發(fā)布時間：2018-04-13 12:59

  本文選題：HCV + 全長基因�。� 參考：《中國科學(xué)院研究生院（武漢病毒研究所）》2005年碩士論文

【摘要】：丙型肝炎病毒(Hepatitis C Virus, HCV)是輸血后及散在性的非甲非乙型肝炎(Non-A, Non-B Hepatitis, NANBH) 主要病因。缺乏可以在實驗室內(nèi)穩(wěn)定擴增病毒的細(xì)胞系統(tǒng)和動物模型嚴(yán)重阻礙了HCV 病毒本身的研究發(fā)展和由其感染引起的疾病的治療方法的研究進步。各國科學(xué)家一方面致力于擴增病毒的細(xì)胞系和動物模型的發(fā)展,另一方面利用分子生物學(xué)的手段對HCV 開展了基礎(chǔ)性的研究。在如今分子生物學(xué)研究發(fā)展迅猛的背景下,HCV 的研究水平也很快得到了很大提高。 本研究通過分段RT-PCR 的方法分段得到了HCV 的全基因組克隆,分析得到所獲得的克隆為1b亞型并且與HCV-Shanghai最為接近;通過對所采集的HCV陽性血清UTR 進行測序分析,得到目前在武漢地區(qū)流行的主要HCV 基因型為1 型。另外采用桿狀病毒系統(tǒng)在昆蟲細(xì)胞中表達HCV 結(jié)構(gòu)蛋白E1,并采用Confocal 顯微鏡觀察E1 在昆蟲細(xì)胞中的表達分布。 本論文包括以下三個章節(jié): 第一章對HCV 的發(fā)現(xiàn)、其基因組結(jié)構(gòu)和基因型的特點和分布、檢測手段、治療方法和E1 蛋白做了簡要綜述,對利用桿狀病毒表達系統(tǒng)表達外源蛋白的廣泛應(yīng)用作了簡要介紹。 第二章描述了采用分段RT-PCR的方法從湖北省HCV 感染陽性病人血清中克隆得到了HCV 的全基因組,通過軟件對其進行基因型和進化樹的分析。最終認(rèn)為所克隆的HCV 基因組為1b 亞型,與目前在NCBI 上發(fā)布的HCV-Shanghai株親緣關(guān)系最為接近。但是在比較中發(fā)現(xiàn)在湖北株內(nèi)有363 nt 的缺失,而同時又替換了285nt。在基因組內(nèi)這285nt 替換了NS5b 的C 端和3’UTR 的前端。而替換的285nt 中有153nt 位與NS5b 內(nèi)而剩余132nt 位于3’UTR 內(nèi)。通過分析發(fā)現(xiàn)替換序列在結(jié)構(gòu)上與原序列相似,但是功能分析有待進一步的實驗證實。 第三章通過克隆得到HCV UTR 和core 的部分基因,測序分析和軟件對比
[Abstract]:Hepatitis C virus (HCV) is the main cause of non-A, Non-B hepatitis B after blood transfusion.The lack of cell systems and animal models that can stably amplify the virus in the laboratory seriously impedes the development of HCV itself and advances in the treatment of diseases caused by its infection.On the one hand, scientists from all over the world are committed to the development of cell lines and animal models for the amplification of viruses; on the other hand, molecular biology is used to carry out basic research on HCV.With the rapid development of molecular biology, the research level of HCV has been greatly improved.In this study, the whole genome clone of HCV was obtained by segmenting RT-PCR, the clone obtained was 1b subtype and closest to HCV-Shanghai, and the HCV positive serum UTR was sequenced.The main genotype of HCV in Wuhan is genotype 1.In addition, baculovirus system was used to express HCV structural protein E1 in insect cells, and Confocal microscope was used to observe the expression distribution of E1 in insect cells.This thesis includes the following three chapters:In the first chapter, the discovery of HCV, the characteristics and distribution of its genome structure and genotypes, the methods of detection, treatment and E1 protein were briefly reviewed, and the wide application of the expression of exogenous proteins by baculovirus expression system was briefly introduced.In the second chapter, the whole genome of HCV was cloned from the sera of HCV positive patients in Hubei province by using the method of segmented RT-PCR. The genotypes and evolutionary trees of HCV were analyzed by software.It was concluded that the cloned HCV genome was subtype 1b, which was most closely related to the HCV-Shanghai strain published on NCBI.However, 363 NT deletion was found in Hubei strain, and 285 NT was replaced at the same time.Within the genome, the 285nt replaces the C end of NS5b and the front end of 3'UTR.In the alternative 285nt, there are 153nt bits and NS5b bits, while the remaining 132nt is located in 3'UTR.It is found that the substitution sequence is similar to the original sequence in structure, but the functional analysis needs further experimental verification.In chapter 3, some genes of HCV UTR and core were cloned, sequenced and compared with software.
【學(xué)位授予單位】：中國科學(xué)院研究生院（武漢病毒研究所）
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R346

【引證文獻】

相關(guān)博士學(xué)位論文 前1條

1 石立瑩;副粘病毒Tianjin株全基因組測序及生物信息學(xué)分析[D];天津醫(yī)科大學(xué);2007年

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本文編號：1744595