MUTATIONAL SPECTRA OF PRODUCTS OF INCOMPLETE COMBUSTION AND PYROLYSIS

不完全燃烧和热解产物的突变谱

• 批准号: 3918445
• 负责人: WILLIAM G THILLY
• 金额: --
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3918445
• 关键词: B lymphocyte; DNA; benzopyrenes; blood proteins; carbopolycyclic compound; chemical addition; engine exhaust; environmental contamination; environmental toxicology; fossil fuel energy; gel electrophoresis; heat; human tissue; hypoxanthine phosphoribosyltransferase; mutagen testing; natural gene amplification; nucleic acid sequence; oxidation; petroleum oil; point mutation; pyrenes

Based on recent progress in our ability to amplify mutant DNA sequences by high fidelity polymerase chain reaction and to separate mutant and wild type DNA sequences using denaturing gradient electrophoresis, we propose three separate efforts, each necessary to the intended use of this new technology to study the rate of combustion effluents in mutation in humans. First, we need to understand the relationship between existing cellular modes of analysis of mutation in human cell samples and our proposed molecular mode. At present, mutants are enumerated as 6-thioguanine resistant cells, growing as colonies. These mutants have little or not activity for the enzyme hypoxanthine phosphoribosyl transferase (hpt). We have developed an approach which examines point mutations at the DNA sequence level in one or a few exons of hprt. These independent modes reveal partially overlapping sets of mutations, and their relationship requires careful definition. We propose to use the mutagens fluoranthene, cyclopenta(c,d)pyrene and benzo(alpha)pyrene in this characterization effort. Although not a major mutagen in incomplete combustion effluents, benzo(alpha)pyrene's mutational spectrum has been studied in bacteria, and our effort will permit an important human cell/bacterial comparison. Secondly and thirdly, we are combining with Projects C-1, C-2 and D-3 to study the quantitative dose and time dependence of mutation in human cells and mutation and tumor formation in mouse tissue. These studies will result in understanding of the relationships in time among protein adducts, DNA adducts and mutation after exposures to fluoranthene, cyclopenta(c,d)pyrene, benzo(alpha)pyrene and important pyrolysis products as shall be determined by research in Projects A-1,-2,-3,-4, B and D-1,-2, and -3.

基于我们最近在扩增突变DNA能力上的进展 通过高保真聚合酶链反应测定序列， 分离突变体和野生型DNA序列， 梯度电泳，我们提出了三个单独的努力，每一个 必要的预期用途，这项新技术的研究， 燃烧排出物在人类中的突变率。 首先，我们需要了解现有的 人类细胞样本突变分析的细胞模式， 我们提出的分子模式。 目前，突变体被列举 作为6-硫鸟嘌呤抗性细胞，作为菌落生长。 这些 突变体对次黄嘌呤酶几乎没有活性或没有活性 磷酸核糖转移酶（HPT）。 我们开发了一种方法 它在DNA序列水平上检查一个或多个基因组中的点突变， 一些HPRT的外显子。 这些独立的模式部分地揭示了 重叠的突变集合，它们的关系需要 仔细定义。 我们建议使用诱变剂荧蒽， 环戊二烯并（c，d）芘和苯并（α）芘 特征化的努力。 虽然不是主要的诱变剂， 不完全燃烧流出物，苯并（α）芘突变 我们已经在细菌中研究了光谱，我们的努力将使 一个重要的人类细胞/细菌比较。 第二和第三，我们正在结合项目C-1，C-2和C-3， D-3研究突变的定量剂量和时间依赖性 在人类细胞和突变和肿瘤形成在小鼠组织。 这些研究将有助于理解 蛋白质加合物、DNA加合物和突变之间的时间 接触荧蒽、环戊二烯并（c，d）芘、 苯并（α）芘和重要的热解产物， 通过项目A-1、A-2、A-3、A-4、A-B和D-1、A-2的研究确定， -3.