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、病毒蛋白质合成和病毒粒子组装及其 某些mRNA病毒的相互关系。 我们尤其关注 在分子水平上进行这些过程，因此选择了 研究三种病毒，它们的物理和遗传结构特别好， 理解，并为适当的合成系统和 方法可用。 我们将研究a）雀麦花叶病毒 (BMV)，一种基因组分裂的单链RNA植物病毒; B）黑甲虫 病毒（BBV），一种分裂的基因组、单链RNA昆虫病毒;和 南方菜豆花叶病毒（SBMV）是一种单基因组、mRNA植物病毒。 对于BMV和BBV以及SBMV，我们将确定， 在动力学和结构上相关的中间产物和最终产物， 基因组尤其是亚基因组RNA的合成。 我们将 通过直接定量和测量合成的时间过程， 分馏放射性产品的测量，并通过探测它们的 存在放射性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进行分析， 控制RNA复制和病毒体组装的相互作用。 我们将对病毒cDNA进行基因工程改造， 用于酶促重建研究的特定病毒蛋白， 抗体介导的抑制研究和RNA结合研究。 我们将设计出病毒cDNA具有感染性的系统， 这些用于调控和编码序列的功能研究， 基因载体的构建和合成缺陷型 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.