ENVIRONMENTAL MUTAGENESIS AND DNA REPAIR

我们研究的长期目标是分析烷基化损伤。 以果蝇为模型多细胞动物系统的诱变 哪种遗传学、分子遗传学和生物化学可以系统地 用于DNA修复和突变诱导的研究。通过隔离 突变敏感(MUS)突变体，我们的目标是识别DNA修复基因和 并确定这些MUS基因在构成中的作用 以及可诱导的DNA修复途径和突变。我们的方法包括 包括体内和体外实验。体外实验被用来 分离MUS突变体并检测这些突变体对突变的影响 诱导，使用与性连锁的隐性致死试验或玫瑰 单基因系统。性连锁隐性致死试验提供了一种 DNA修复缺陷突变体的客观快速检测技术 用于突变诱导频率的改变。玫瑰色的单曲 基因系统，与强大的重组DNA技术相结合 提供了一种详细的分子分析方法，用于分析在 不存在已知的DNA修复能力。初步研究 结合后一种系统，论证了该系统的可行性 用于对这种高等真核生物的诱变进行分子分析。在……里面 利用从mus突变体制备的细胞培养进行的体外实验可以 确定烷基化修复缺陷。非程序性DNA的测量 合成为修复用小鼠串蛋白的普查提供了一种方法 缺陷，目前的研究已经确定了一些基因座 几种烷基化剂引起的UDS缺乏。这些和其他 间接研究表明，需要更灵敏的方法来检测 烷基化引起的NDA损伤分析。初步研究表明 论证了直接层析分析的可行性 烷基化引起的果蝇DNA碱基损伤。因为高 高效液相色谱提供了最灵敏的直接方法 为了详细分析烷基化修复缺陷的性质，我们 提出了基于这项技术的进一步实验。最直接的 作为一种动物模型继续研究该系统的重要性 对人类健康的影响已被充分证明：DNA缺陷 修复能力直接与许多主要的人类 疾病。

The long term objective of our research is an analysis of alkylation damage and mutagenesis in Drosophila as a model multicellular animal system in which genetics, molecular genetics and biochemistry can be systematically exploited for studies of DNA repair and mutation induction. By isolating mutagen-sensitive (mus) mutants, we aim to identify DNA repair genes and their products and determine the roles of these mus genes in constitutive and inducible DNA repair pathways and mutagenesis. Our methods involve both in vivo and in vitro experiments. In vitro experiments are used to isolate mus mutants and examine the effects of these mutants on mutation induction, using either the sex-linked recessive lethal test or the rosy single gene system. The sex-linked recessive lethal test provides an objective and rapid technique for examining DNA repair-deficient mutants for alterations in the frequency of mutation induction. The rosy single gene system, in combination with powerful recombinant DNA techniques provides a method for detailed molecular analysis of mutants induced in the presence of absence of known DNA repair deviciencies. Preliminary studies with this latter system have demonstrated the feasibility of this system for the molecular analysis of mutagenesis in this higher eukaryote. In vitro experiments utilizing cell cultures prepared from the mus mutants can identify alkylation repair defects. Measurements of unscheduled DNA synthesis provide a method for the general survey of mus strins for repair deficiencies, and current studies have identified a number of loci deficient in UDS induced by several alkylating agents. These and other indirect studies have indicated the need for more sensitive methods for the analysis of alkylation-induced NDA damage. Preliminary studies have demonstrated the feasibility of direct chromatographic analysis for alkylation-induced base lesions in Drosophila DNA. Because high performance liquid chromatography provides the most sensitive direct method for detailed analysis of the nature of alkylation repair defects, we propose further experiments whichrely on this technology. The immediate importance of continued study of this system as an animal model with implications for human health has been amply demonstrated: defects in DNA repair ability have been directly associated with a number of major human diseases.