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: 3840626
• 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/3840626
• 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; nucleic acid sequence; oxidation; petroleum oil; point mutation; polymerase chain reaction; 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.

基于我们扩增突变 DNA 能力的最新进展 通过高保真聚合酶链式反应序列 使用变性分离突变型和野生型 DNA 序列 梯度电泳，我们提出了三个单独的努力，每个 必要的这项新技术的预期用途来研究 燃烧废气对人类的突变率。 首先，我们需要了解现有的关系 人类细胞样本突变分析的细胞模式和 我们提出的分子模式。 目前已枚举突变体 作为 6-硫鸟嘌呤抗性细胞，以集落形式生长。 这些 突变体对次黄嘌呤酶具有很少或没有活性 磷酸核糖基转移酶（hpt）。 我们开发了一种方法 它检查 DNA 序列水平上的点突变之一或 HPRT 的一些外显子。 这些独立模式部分揭示了 重叠的突变组，以及它们的关系需要 仔细的定义。 我们建议使用诱变剂荧蒽， 环戊(c,d)芘和苯并(α)芘 表征努力。 虽然不是主要诱变剂 不完全燃烧流出物、苯并(α)芘的突变 光谱已经在细菌中进行了研究，我们的努力将允许 重要的人类细胞/细菌比较。 第二和第三，我们正在与项目C-1、C-2和 D-3 研究突变的定量剂量和时间依赖性 人类细胞中的突变和小鼠组织中的肿瘤形成。 这些研究将导致对以下关系的理解： 蛋白质加合物、DNA加合物之间的时间以及突变后 暴露于荧蒽、环戊(c,d)芘、 苯并（α）芘和重要的热解产物 通过项目 A-1、-2、-3、-4、B 和 D-1、-2 的研究确定，以及 -3。