ASSEMBLY AND APPLICATION OF RECOMBINANT RHABDOVIRUSES

重组弹状病毒的组装和应用

• 批准号: 2633456
• 负责人: John K. Rose
• 金额: $ 40.19万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2633456
• 关键词: CD4 molecule; G protein; Vesiculovirus; glycoprotein biosynthesis; glycoproteins; laboratory mouse; membrane activity; membrane proteins; polymerase chain reaction; recombinant virus; tissue /cell culture; virus assembly; virus envelope; virus protein; virus replication; viruslike particle

DESCRIPTION (adapted from applicant s abstract): The long term objective of the project is to understand basic aspects of enveloped virus assembly and replication. Two model viral systems, vesicular stomatitis virus (VSV), and a new minimal virus system based partially on VSV will be used. VSV is the prototype of the rhabdovirus group and is one of the simplest and best characterized enveloped viruses. Recently Dr. Rose reported development of a system in which recombinant VSVs can be recovered from a complete DNA copy of the genome, and they have also reported a stable, highly efficient gene expression vector based on VSV. Additional studies will be performed to define RNA packaging limits in VSV and to generate new vectors suitable for expression of multiple genes. Such vectors are likely to have important application in vaccine development. The recombinant VSV system will be exploited in studies on the protein domains and modifications that are critical to efficient assembly of the glycoprotein into infectious particles. Basic studies will also be performed on incorporation of foreign membrane proteins into the VSV envelope. These studies will define the signals required for protein incorporation in the presence or absence of the wild-type VSV glycoprotein (G). An understanding of the rules governing efficient glycoprotein incorporation into virus particles could permit the design of novel vaccines derived by incorporating foreign glycoproteins into VSV particles. Other studies will be directed toward deriving VSVs with new targeting specificity based on incorporation of CD4 into the viral envelope. VSV is ideal for such studies because it grows rapidly to very high titers and is easily isolated in much larger quantities than most other enveloped viruses. Cell lines expressing the L and/or P genes of VSV will also be developed to permit the growth of defective VSVs lacking these genes and expressing very large amounts of foreign genetic material. Expression of the VSV G protein from a self-replicating RNA derived from Semliki Forest virus results in production of enveloped, infectious, self-propagating virus particles that contain G protein as their single structural protein. Dr. Rose s lab will continue basic studies on these minimal virus particles to determine the role of VSV G protein in their formation, to use them in a genetic selection of revertants of G protein fusion mutants, and to determine if they display any pathogenicity in animals.

描述（改编自申请人的摘要）：长期目标 该项目旨在了解包膜病毒组装的基本方面和 复制。 两种模型病毒系统，水泡性口炎病毒（VSV）和 将使用部分基于 VSV 的新的最小病毒系统。 VSV 是 弹状病毒组的原型，是最简单和最好的之一 具有包膜病毒的特征。 最近，Rose 博士报告了 可以从完整 DNA 拷贝中恢复重组 VSV 的系统 他们还报道了一种稳定、高效的基因 基于VSV的表达载体。 将进行额外的研究 定义 VSV 中的 RNA 包装限制并生成适合的新载体 多个基因的表达。 此类载体可能具有重要作用 在疫苗研发中的应用。 重组VSV系统将是 在蛋白质结构域和修饰的研究中被利用 对于将糖蛋白有效组装成感染性物质至关重要 颗粒。 还将对外资企业的设立进行基础研究 膜蛋白进入 VSV 包膜。 这些研究将定义 在存在或不存在的情况下蛋白质掺入所需的信号 野生型 VSV 糖蛋白 (G)。 对管理规则的理解 有效地将糖蛋白掺入病毒颗粒中可以允许 通过将外来糖蛋白掺入而衍生的新型疫苗的设计 VSV 颗粒。 其他研究将针对使用新的 VSV 进行衍生 基于 CD4 掺入病毒包膜的靶向特异性。 VSV 非常适合此类研究，因为它可以快速生长到非常高的滴度 并且比大多数其他包封的产品更容易隔离更多的数量 病毒。 表达VSV的L和/或P基因的细胞系也将是 开发用于允许缺乏这些基因的有缺陷的 VSV 生长，并且 表达大量外源遗传物质。 源自自复制 RNA 的 VSV G 蛋白表达 塞姆利基森林病毒会产生有包膜的、传染性的、 含有G蛋白作为单一病毒颗粒的自繁殖病毒颗粒 结构蛋白。 Rose博士的实验室将继续对这些进行基础研究 最小病毒颗粒以确定 VSV G 蛋白在病毒中的作用 形成，将它们用于 G 蛋白回复体的遗传选择 融合突变体，并确定它们是否表现出任何致病性 动物。