VIRAL PROTEINS IN ROTAVIRUS REPLICATION

轮状病毒引起的急性胃肠炎是人类 婴儿死亡率。因为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作用 交叉挑战实验，接种疫苗预防轮状病毒病 可能只会取得有限的成功。疾病治疗方法的发展 有关每种轮状病毒的作用的信息将有助于预防 蛋白质在复制中起作用。为此目的，建立一个国际劳工组织 支持轮状病毒RNA复制和颗粒组装的体外系统 将提供一种方法来研究病毒蛋白在这些 流程。 轮状病毒是分段的双链RNA病毒，其复制 依赖于保护合成。这个项目的目的是 轮状病毒SA11复制和形态发生的优化和特征 以前被证明支持病毒dsRNA，mRNA， 和蛋白质的体外合成。该系统是由病毒构建的 指导RNA合成的模板和微球菌核酸酶处理的兔 网织红细胞裂解物将mRNAs翻译成蛋白质。不同的方法来实现 将在系统中尝试准备模板和mRNA，以确定 那些支持最大双链RNA合成的基因。之后 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详细描述。研究将会 尝试分离和鉴定不同种类的病毒 粒子存在于受感染的细胞中，并将测试它们的能力 模板RNA的体外合成。从这个项目中获得的结果将 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。

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.