PACKAGING OF THE SEGMENTED GENOME OF BACTERIOPHAGE PHI 6

噬菌体 PHI 6 分段基因组的包装

• 批准号: 3285204
• 负责人: LEONARD E MINDICH
• 金额: $ 24.01万
• 依托单位: PUBLIC HEALTH RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-01-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3285204
• 关键词: DNA replication; Escherichia coli; Pseudomonas; bacterial virus; binding proteins; complementary DNA; double stranded RNA; enzyme mechanism; gene expression; genetic transcription; genetic translation; genome; host organism interaction; hydrolase; molecular cloning; mutant; nucleic acid sequence; nucleocapsid; plasmids; replicase; tissue /cell culture; virus genetics

Bacteriophage phi6 contains three pieces of double-stranded RNA in its nucleocapsid. The pieces possess different genetic information but have similar termini. The packaging of the genomic segments is precise. The aim of this proposal is to determine how the packaging system is able to select one of each of the chromosomes. Specifically, the regions of the three chromosomes necessary for packaging will be identified. The order of packaging and the requirements for packaging in terms of amount of RNA will be determined in an in vitro packaging system that we have developed. The entire genome of phi6 has been cloned as cDNA and completely sequenced. Procapsids are produced from cDNA copies of segment L in both Escherichia coli and Pseudomonas phaseolicola, the natural host of phi6. These procapsids are capable of packaging single-stranded viral RNA, synthesizing the complementary minus strand to form double-stranded RNA and then transcribing this dsRNA to form plus ssRNA. Procapsids will be isolated and used to study both the replicase and packaging reactions. The composition of procapsids, will be varied so as to identify the proteins involved in general packaging, segment specific packaging, replication and transcription. RNA transcripts made from cDNA constructions of the genomic segments can be packaged by the procapsids. The regions of the RNA involved with packaging will be identified and manipulated so as to clarify the interaction between the RNA and the packaging and replication machinery. The nature of the mechanisms of selective packaging in viruses of segmented genomes is currently unknown. The elucidation of the mechanism for phi6 has relevance to the packaging of other segmented-genome viruses. Several members of the reoviridae, e.g., rotavirus and blue tongue virus, are the etiologic agents of important human and animal diseases.

噬菌体 phi6 的内部含有三段双链 RNA 核衣壳。 这些碎片拥有不同的遗传信息，但 类似的终点站。 基因组片段的包装是精确的。 这 该提案的目的是确定包装系统如何能够 选择每条染色体之一。 具体而言，各地区 将鉴定包装所需的三个染色体。 订单 包装的种类和包装数量的要求 RNA 将在我们拥有的体外包装系统中测定 发达。 phi6的整个基因组已被克隆为cDNA并完全 已测序。 原衣壳是由片段 L 的 cDNA 拷贝产生的 大肠杆菌和绿豆假单胞菌，它们是细菌的天然宿主 phi6。 这些衣壳能够包装单链病毒 RNA，合成互补负链形成双链 RNA，然后转录该 dsRNA 形成 plus ssRNA。 原衣壳 将被分离并用于研究复制酶和包装 反应。 原衣壳的组成会有所不同，以便 识别一般包装、特定细分市场中涉及的蛋白质 包装、复制和转录。 RNA 转录本由 基因组片段的 cDNA 构建体可以通过以下方式包装： 原衣壳。 涉及包装的 RNA 区域将是 识别和操纵，以澄清之间的相互作用 RNA 以及包装和复制机器。 病毒选择性包装机制的本质 分段基因组目前未知。 的阐明 phi6 的机制与其他包装有关 分段基因组病毒。 呼肠孤病毒科的几个成员， 例如，轮状病毒和蓝舌病毒是下列疾病的病原体： 重要的人类和动物疾病。