ENZYMATIC MECHANISM OF HERPES VIRUS DNA REPLICATION

这个研究项目的目的是理解分子中的 详细说明单纯疱疹病毒复制的机制- 1(HSV-1)病毒DNA被启动并持续。我们建议 用纯化的酶体外重组有序、半重组 单纯疱疹病毒1型基因组的保守复制。我们期待着 这些研究应该能让我们对这种机制有一个深入的了解。 启动真核生物基因组复制的基因。他们 还应生成有关复制类的信息 对公共卫生具有重大意义的动物病毒。这个 调查工作将按以下思路进行： 1.单纯疱疹病毒1型编码蛋白的鉴定、纯化和分析 复制蛋白 A.单纯疱疹病毒1型DNA聚合酶 单纯疱疹病毒1型单链DNA结合蛋白(ICP8) C.单纯疱疹病毒1型诱导复制因子(S) D.单纯疱疹病毒1型诱导的DNA解旋酶 E.HSV-1诱导的启动子酶 F.HSV-1诱导的Oris结合蛋白 G.用纯化的蛋白质进行滚动循环DNA复制 2.在HSV-1起始处启动DNA复制(ORIS) 3.单纯疱疹病毒1型L和S序列的体外倒置

The aim of this research program is to understand in molecular detail the mechanism by which the replication of Herpes simplex- 1 (HSV-1) virus DNA is initiated and sustained. We propose to reconstitute in vitro with purified enzymes the orderly, semi- conservative replication of the HSV-1 genome. We expect that these studies should provide us with an insight into the mechanism by which replication of a eukaryotic genome is initiated. They should also yield information about the replication of a class of animal viruses that are of great public health importance. The investigation will be organized along the following lines: 1. Identification, purification and analysis of HSV-1 encoded replication proteins a. HSV-1 DNA polymerase b. HSV-1 single-stranded DNA binding protein (ICP8) c. HSV-1 induced replication factor(s) d. HSV-1 induced DNA helicase e. HSV-1 induced primase f. HSV-1 induced oriS binding protein g. Rolling circle DNA replication with purified proteins 2. Initiation of DNA replication at an HSV-1 origin (oriS) 3. In vitro inversion of HSV-1 L and S sequences