GENETICS OF HERPESVIRUS-HOST CELL INTERACTIONS

协同控制的规制现象分析 单纯疱疹病毒1型（HSV-1）的顺序合成 在感染培养物中的细胞期间纯化蛋白质是 这项研究。 我们的目的是确定病毒的调控序列， 基因，它们响应的调节信号以及 它们相互作用以控制病毒基因表达。 病毒蛋白 被分为三个时间组，称为阿尔法，贝塔和 γ的 α蛋白作为正调控元件， β和γ蛋白的表达。 主要的实验工具，在 我们的调控研究，是细胞转化为限制性区域的 病毒基因组 这些转化的细胞是由 感染病毒突变体的表达信息， 转换序列。 将转化细胞以获得携带 调节蛋白ICP 4的α基因和糖蛋白的β基因 B直接观察α基因对β基因的诱导作用 产品 这种构建将提供仅能够表达 一种免疫原性活性糖蛋白， 病毒感染 伽玛基因调控将探讨使用这一点， 系统 细胞将针对γ gC基因和β gC基因进行转化。 TK基因用于诱导表达的直接比较。 Alpha的影响 和β基因产物以及病毒DNA合成对这些诱导的影响 将研究两类基因。 转化HSV-1的细胞 节将作为允许细胞的隔离宿主范围 变种人 琥珀无意义突变体也将被寻找。 纯化的ICP 4将 在体外检查与gB调节序列的特异性结合 基因 结合位点将通过DNA足迹检测 法 另外的病毒和/或细胞因子的参与， 将探索特异性结合。 突变体的影响 ICP 4蛋白和gB调节区的序列改变将 被衡量。 HSV-1 Eco RI F片段跨越的区域将被 重新检查形态学和致癌转化能力。 Rat 2 用含有HSV-1的质粒将tk-细胞转化为tk+ TK基因和Eco RI F片段，以确保每只果蝇具有 收到了这些病毒序列 ICP 4可能发挥作用的可能性 将探索形态转化。

The analysis of the regulatory phenomena controlling the coordinately and sequentially ordered synthesis of herpes simplex virus type 1 (HSV-1) sepcified proteins during infection of cells in culture is the objective of this research. Our aim is to identify the regulatory sequences on viral genes, the regulatory signals to which they respond and the mechanisms by which these interact to control viral gene expression. The viral proteins have been classified into three temporal groups termed Alpha, Beta and Gamma. Alpha proteins act as positive regulatory elements for the expression of Beta and Gamma proteins. The primary experimental tool, in our regulatory studies, is cells transformed for restricted regions of the viral genome. These transformed cells are genetically manipulated by infection with viral mutants for information on expression of the transforming sequences. Cells will be transformed for plasmids carrying an Alpha gene for the regulatory protein ICP4 and a Beta gene for glycoprotein B to look directly at the induction of the Beta gene by the Alpha gene product. This construction will provide a cell capable of expressing only one of the immunogenically active glycoproteins induced in the course of viral infections. Gamma gene regulation will be explored using this system. Cells will be transformed for both the Gamma gC gene and the Beta TK gene for direct comparison of induced expression. The effects of Alpha and Beta gene products and of viral DNA synthesis on the induction of these two classes of genes will be explored. The cells transformed for HSV-1 segments will serve as permissive cells for the isolation of host range mutants. Amber nonsense mutants will also be sought. Purified ICP4 will be examined in vitro for specific binding to regulatory sequences of the gB gene. Sites of binding will be detected by the DNA footprinting technique. The involvement of additional viral and/or cellular factors in specific binding will be explored. The effects of mutant alterations in the ICP4 protein and sequence alteration in the gB regulatory region will be measured. The region spanned by the HSV-1 Eco RI F fragment will be reexamined for morphological and oncogenic transformation capacity. Rat 2 tk- cells have been transformed to tk+ with a plasmid containing the HSV-1 TK gene and the Eco RI F fragment to insure that each transformant has received these viral sequences. The possibility that ICP4 may play a role in morphological transformation will be explored.