WOODCHUCK VIRUS--MOLECULAR BIOLOGICAL STUDIES

土拨鼠病毒——分子生物学研究

• 批准号: 3803146
• 负责人: R H MILLER
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3803146
• 关键词: animal viral hepatitis; disease /disorder model; duck hepatitis B virus; genetic polymorphism; genetic promoter element; genetic strain; genome; hepatitis B virus group; marmots; molecular cloning; open reading frames; point mutation; site directed mutagenesis; transfection; virus DNA; virus assembly; virus genetics; virus replication

The woodchuck hepatitis virus (WHV) is taxonomically and serologically related to the hepatitis B virus (HBV). Infection with each of these viruses is associated with acute and chronic hepatitis and hepatic cell carcinoma in their respective hosts and these associations appear to be etiological in nature. Thus, WHV infection of woodchucks provides a relevant and convenient model for understanding HBV infections of humans. The following experiments were performed: (1) The variability of WHV genomes in an infected animal was determined. (2) Serum pools with and without defective virus were found to induce the same percentage of chronic carriers. This suggests that the presence of defective virus in a WHV inoculum is not a prerequisite for the establishment of persistent hepadnavirus infections. (3) Oligonucleotide directed mutagenesis was used to introduce point mutations in the precore and X genes as well as open reading frames 5 and 6 to determine the importance of these genes in virus replication using woodchuck transfection assays. (4) The WHV and HBV X gene promoters were cloned into the vector pSVOCAT to compare promoter strength in human and woodchuck cells. (5) A New York strain of WHV was cloned in preparation for DNA sequencing.

土拨鼠肝炎病毒（WHV）在分类学和血清学上是 与乙型肝炎病毒（HBV）有关。 感染其中每一种 病毒与急、慢性肝炎及肝细胞有关 各自宿主的癌症，这些关联似乎是 本质上是病因学。 因此，土拨鼠的 WHV 感染提供了 理解 HBV 感染的相关且方便的模型 人类。 进行了以下实验：（1）变异性 确定了受感染动物的 WHV 基因组。 (2) 血清池 发现有缺陷的病毒和没有缺陷的病毒都会诱导相同的情况 慢性携带者的百分比。 这表明存在 WHV 接种物中的缺陷病毒并不是 建立持续性肝炎病毒感染。 (3) 寡核苷酸 定向诱变用于在细胞中引入点突变 precore 和 X 基因以及开放阅读框 5 和 6 来确定 这些基因在土拨鼠病毒复制中的重要性 转染测定。 (4)克隆WHV和HBV X基因启动子 进入载体 pSVOCAT 以比较人类和 土拨鼠细胞。 (5) WHV纽约株克隆制备 用于 DNA 测序。