VIRAL PROTEINS IN ROTAVIRUS REPLICATION

轮状病毒复制中的病毒蛋白

• 批准号: 3444720
• 负责人: JOHN T PATTON
• 金额: $ 9.54万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-01 至 1989-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3444720
• 关键词: Rotavirus; acute infectious nonbacterial gastroenteritis; cell free system; communicable disease control; double stranded RNA; genetic transcription; molecular cloning; nucleocapsid; radiotracer; virulence; virus genetics; virus replication; viruslike particle

Acute gastroenteritis caused by rotaviruses is a leading cause of human infant mortality. Because of the multiple distinct serotypes of human rotaviruses and the lack of protection by different serotypes in cross-challenge experiment, prevention of rotavirus disease by vaccination may meet with oly limited success. Development of methods for disease prevention would be helped by information on the role that each rotavirus protein plays in replication. Towards this end, the establishment of an in vitro system that supports rotavirus RNA replication and particle assembly would provide a method for examining the role of viral proteins in these processes. Rotaviruses are segmented, double-stranded RNA virsuses whose replication is dependent upon protection synthesis. The purpose of this project is to optimize and characterize rotavirus SA11 replication and morphgensis in a cell-free system that was previously shown to support viral dsRNA, mRNA, and protein synthesis in vitro. The system is constructed from viral templates to direct RNA synthesis and a micrococcal nuclease-treated rabbit reticulocyte lysate to translate mRNAs into proteins. Various methods for preparing templates and mRNAs will be tried in the system to determine those which support maximum double-strand RNA synthesis. After optimization, the RNA replication, transcription and core particle assembly of rotavirus will be characterized in detail in the system. Studies will attempt to separate and characterize the different species of viral particles present in infected cells and will assay their ability to template RNA synthesis in vitro. RResults obtained from this project will provide a better understanding of the molecular biology of the rotaviruses as well as the role of viral proteins in the replication of these pathogenic viruses.

轮状病毒引起的急性胃肠炎是人类胃肠道疾病的主要原因 婴儿死亡率。 由于人类有多种不同的血清型 轮状病毒和不同血清型缺乏保护 交叉攻击实验，通过疫苗接种预防轮状病毒病 可能只会取得有限的成功。 疾病方法的开发 有关每种轮状病毒的作用的信息将有助于预防 蛋白质参与复制。 为此，建立了一个 支持轮状病毒RNA复制和颗粒组装的体外系统 将提供一种方法来检查病毒蛋白在这些中的作用 流程。 轮状病毒是分段的双链 RNA 病毒，其复制 取决于保护合成。 该项目的目的是 优化和表征轮状病毒 SA11 的复制和形态发生 先前显示支持病毒 dsRNA、mRNA、 和体外蛋白质合成。 该系统是由病毒构建的 指导 RNA 合成的模板和经过微球菌核酸酶处理的兔子 网织红细胞裂解物将 mRNA 翻译成蛋白质。 各种方法用于 准备模板和mRNA将在系统中尝试以确定 那些支持最大双链RNA合成的。 后 优化、RNA 复制、转录和核心颗粒组装 轮状病毒的特征将在系统中进行详细表征。 研究将 尝试分离和表征不同种类的病毒 颗粒存在于受感染的细胞中，并将检测它们的能力 体外模板RNA合成。 R从该项目获得的结果将 更好地了解轮状病毒的分子生物学 以及病毒蛋白在这些病毒复制中的作用 致病病毒。