THE MECHANISM OF SYNTHESIS OF SMALL VIRUSES

这项工作的目的是研究基因组的基本过程。 复制、病毒蛋白合成和病毒粒子组装及其相互作用 某些信使核糖核酸病毒之间的相互关系。我们尤其关注的是 这些过程是在分子水平上进行的，因此选择了 研究物理和遗传结构特别好的三种病毒 了解，并为此适当的合成系统和 方法论是可用的。我们将与)雀麦花叶病毒合作 (BMV)，一种分裂基因组的单链RNA植物病毒；b)黑甲虫 病毒(BBV)是一种分裂的基因组、单链RNA昆虫病毒；以及c) 南方菜豆花叶病毒(SBMV)是一种单基因组、单基因的植物病毒。 对于BMV和BBV，以及适当的SBMV，我们将确定 在运动学和结构上相关的中间产品和最终产品 基因组特别是亚基因组核糖核酸的合成。我们会 直接对合成的时间进程进行量化和测量 分馏放射性产品的测量和通过探测其 存在放射性的cDNA和cRNA。我们将研究病毒的结合 RNA和RNA片段与蛋白质和蛋白质片段一起分析 控制RNA复制和病毒粒子组装的相互作用。 我们将通过基因工程来实现病毒的大规模生产 用于酶重组研究的特定病毒蛋白， 抗体介导的抑制研究和RNA结合研究。 我们将设计病毒cDNA具有传染性的系统，并利用 这些用于调节和编码序列的功能研究， 遗传载体的构建及人工缺陷的研究进展 用于控制病毒感染的干扰颗粒。

The purpose of this work is to study the fundamental processes of genome replication, viral protein synthesis and virion assembly and their interrelationship for certain mRNA viruses. We are especially concerned with these processes at the molecular level and thus have selected for study three viruses whose physical and genetic structure is especially well understood, and for which appropriate synthesizing systems and methodologies are available. We will work with a) brome mosaic virus (BMV), a divided-genome, single-stranded RNA plant virus; b) black beetle virus (BBV) a divided-genome, single-stranded RNA insect virus; and c) southern bean mosaic virus (SBMV) a single genome, mRNA plant virus. For BMV and BBV, and as appropriate for SBMV, we will identify, and kinetically and structurally relate, the intermediate and final products of synthesis of the genomic and especially the subgenomic RNAs. We will quantitate and measure the time course of the syntheses by direct measurements of fractionated radioactive products and by probing their presence with radioactive cDNA and cRNA. We will study binding of viral RNA and RNA fragments with protein and protein fragments to analyze the interactions governing RNA replication and virion assembly. We will genetically engineer viral cDNA to give large-scale production of particular viral proteins for enzymatic reconstitution studies, antibody-mediated inhibition studies and RNA binding studies. We will devise systems in which viral cDNAs are infectious and utilize these for functional studies of regulatory and coding sequences, the construction of genetic vectors and the development of synthetic defective interfering particles for the control of viral infections.