VIRAL PROTEINS IN ROTAVIRUS REPLICATION

由轮状病毒引起的急性胃肠炎是引起人类腹泻的主要原因， 婴儿死亡率。 由于人类的多种不同血清型 轮状病毒和缺乏保护不同血清型， 交叉攻毒试验，轮状病毒病的预防 可能会取得有限的成功。 疾病治疗方法的发展 了解每种轮状病毒的作用， 蛋白质在复制中起作用。 为此目的，建立一个 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 流程. 轮状病毒是分节的双链RNA病毒， 这取决于保护合成。 该项目的目的是 优化和表征轮状病毒SA11在一种 先前显示支持病毒dsRNA，mRNA， 和体外蛋白质合成。 该系统由病毒构建 指导RNA合成的模板和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。 的各种方法 将在系统中尝试制备模板和mRNA，以确定 支持最大双链RNA合成的那些。 后 优化，RNA复制，转录和核心颗粒组装 将在系统中详细描述轮状病毒的特征。 研究将 试图分离和表征不同种类的病毒 粒子存在于感染的细胞中，并将测定它们的能力， 体外模板RNA合成。 从该项目获得的结果将 提供轮状病毒分子生物学的更好理解 以及病毒蛋白在这些病毒复制中的作用 致病病毒

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.