ENZYMATIC MECHANISM OF HERPES VIRUS DNA REPLICATION

这项研究计划的目的是了解分子 单纯疱疹病毒的复制机制 1（HSV-1）病毒DNA启动和持续。 我们建议 用纯化的酶在体外重建有序的，半- HSV-1基因组的保守复制。 我们预计 这些研究应该能让我们深入了解 真核基因组的复制由此启动。 他们 还应该产生关于一类复制的信息， 动物病毒对公共卫生具有重要意义。 的 调查将按照以下方针沿着组织： 1. HSV-I编码基因的鉴定、纯化和分析 复制蛋白 a. HSV-1 DNA聚合酶 B. HSV-1单链DNA结合蛋白（ICP 8） C. HSV-1诱导复制因子 D. HSV-1诱导的DNA解旋酶 e. HSV-1诱导引发酶 F. HSV-1诱导的oriS结合蛋白 G. 用纯化的蛋白质进行滚环DNA复制 2. 在HSV-1起点（oriS）启动DNA复制 3. HSV-1 L和S序列的体外倒位

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