ASSEMBLY AND APPLICATION OF RECOMBINANT RHABDOVIRUSES

描述（改编自申请人的摘要）： 该项目旨在了解包膜病毒组装的基本方面， 复制的 两个模型病毒系统，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，并且是最简单和最好的病毒之一。 以包膜病毒为特征。 罗斯博士最近报告了 可以从完整DNA拷贝回收重组VSV的系统 他们还报道了一种稳定、高效的基因 表达载体的构建。 将进行其他研究， 定义VSV中的RNA包装限度，并产生适合于 多个基因的表达。 这些载体可能具有重要的 应用于疫苗开发。 重组VSV系统将 在蛋白质结构域和修饰的研究中， 关键是将糖蛋白有效组装成感染性的 粒子 还将对外国公司的合并进行基础研究。 膜蛋白进入VSV包膜。 这些研究将确定 蛋白质掺入所需的信号，在存在或不存在 野生型VSV糖蛋白（G）。 对管理规则的理解 有效的糖蛋白掺入到病毒颗粒中可以允许 设计通过将外源糖蛋白掺入 VSV粒子。 其他研究将针对用新的 靶向特异性基于CD 4掺入病毒包膜。 VSV是这类研究的理想选择，因为它生长迅速，滴度非常高 并且比大多数其他被包裹的物质更容易被隔离， 病毒。 表达VSV的L和/或P基因的细胞系也将在本发明的方法中进行。 开发以允许缺乏这些基因的缺陷VSV的生长， 表达大量的外来遗传物质。 VSV G蛋白从来自于以下的自我复制RNA的表达： 塞姆利基森林病毒导致产生有包膜的，传染性的， 含有G蛋白作为其单个的自繁殖病毒颗粒 结构蛋白 罗斯博士的实验室将继续对这些基础研究 最小的病毒颗粒，以确定VSV G蛋白在其 形成，将它们用于G蛋白回复突变体的遗传选择 融合突变体，并确定它们是否显示任何致病性， 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.