ASSEMBLY AND APPLICATION OF RECOMBINANT RHABDOVIRUSES

描述(改编自申请者S摘要)：远景目标 该项目是为了了解包膜病毒组装的基本方面，并 复制。两种模型病毒系统，水疱性口炎病毒(VSV)和 将使用部分基于VSV的新的最小病毒系统。VSV是 是横纹病毒组的原型，是最简单和最好的 描述了包膜病毒的特征。最近，罗斯博士报告了 一种可从完整DNA拷贝中恢复重组VSV的系统 他们还报告了一种稳定、高效的基因 基于VSV的表达载体。将进行更多研究，以 在VSV中定义RNA包装限制，并生成适用于 多基因的表达。这样的载体很可能具有重要的 在疫苗开发中的应用。重组VSV系统将是 在对蛋白质结构域和修饰的研究中被利用 有效地将糖蛋白组装成有感染性的 粒子。还将进行关于外国公司注册的基础研究 膜蛋白进入VSV包膜。这些研究将定义 存在或不存在时蛋白质掺入所需的信号 野生型VSV糖蛋白(G)。对规则的理解 有效地将糖蛋白整合到病毒颗粒中可以使 外源糖蛋白介导的新型疫苗的设计 VSV颗粒。其他研究将针对衍生具有新的VSV 靶向特异性基于将CD4结合到病毒包膜中。 VSV是此类研究的理想选择，因为它可以迅速生长到非常高的滴度 与大多数其他包膜相比，很容易分离出更多的量 病毒。表达疱疹病毒L和/或P基因的细胞株也将 发展成允许缺乏这些基因的有缺陷的VSV生长和 表达了大量的外来遗传物质。 VSV G蛋白在自复制核糖核酸中的表达 塞姆利基森林病毒导致产生包膜的，传染性的， 含有G蛋白的自我繁殖病毒颗粒 结构蛋白。罗斯·S博士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蛋白在其 形成，用于G蛋白回复突变体的遗传选择 融合突变体，并确定它们是否在 动物。

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.