MECHANISMS OF CHEMICAL MUTAGENESIS IN MAMMALIAN CELLS

我们计划在中国开展一项突变研究计划。 哺乳动物细胞。作为突变的目标，这些研究将 使用导入哺乳动物细胞的小细菌基因(Gpt) 作为逆转录病毒穿梭载体的一部分并整合到 哺乳动物前病毒形式的染色体DNA。我们有 最近开发了这个系统，并展示了它的潜力 提供有关哺乳动物细胞突变的信息 特定基数变化的级别。研究的目标是 基因突变的分子机制研究进展 哺乳动物细胞。 拟议的研究将利用穿梭载体转换的GPT+ 小鼠细胞和我们拥有的大量GPT+突变体 已经分离的，以特定的碱基为特征的 以及其他待分离的转化系和突变体。 主要的具体目标是：(1)确定基数变化 自发发生的突变或作为突变的结果 通过化学或物理因素，(2)分析它们之间的关系 在核苷酸池扰动和诱变剂基础上 DNA的变化，(3)通过以下方式确定分子机制 哺乳动物细胞中哪些突变基因可以逆转；(4)分离 哺乳动物细胞抑制突变体，(5)决定是否 Gpt基因的突变谱因个体不同而不同。 接受物种或航天飞机的整合地点 载体，(6)研究哺乳动物细胞的错配修复，(7) 将穿梭载体中的gpt基因导入小鼠体内并进行分析 GPT+小鼠突变的分子机制。

We propose to carry out a program of research on mutation in mammalian cells. As the target for mutations, these studies will use a small bacterial gene (gpt) introduced into mammalian cells as part of a retroviral shuttle vector and integrated into mammalian chromosomal DNA in a proviral form. We have recently developed this system and demonstrated its potential for providing information on mutations in mammalian cells at the level of specific base changes. The goal of the research is the elucidation of the molecular mechanisms of mutation in mammalian cells. The proposed studies will utilize shuttle vector transformed Gpt+ mouse cells and the large number of Gpt+ mutants that we have already isolated and characterized in terms of specific base changes, and other transformed lines and mutants to be isolated. The major specific aims are: (1) determine the base changes in mutations occuring spontaneously or as the result of mutagenesis by chemical or physical agents, (2) analyze the relationshp between nucleotide pool perturbation and mutagen-induced based changes in DNA, (3) determine the molecular mechanisms by which mutant genes in mammalian cells can revert; (4) isolate mammalian cell suppressor mutants, (5) determine if the mutational spectra for the gpt gene are different depending upon the recipient species or the site of integration of the shuttle vector, (6) examine mismatch repair in mammalian cells, (7) introduce the gpt gene in a shuttle vector into mice and analyze the molecular mechanisms of mutation in Gpt+ mice.