ADDUCT AND REPAIR MAPPING IN ONCOGENES

癌基因中的加合物和修复图谱

• 批准号: 3199311
• 负责人: GERALD P HOLMQUIST
• 金额: $ 17.2万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3199311
• 关键词: DNA repair; RNA splicing; aflatoxins; alkylating agents; benzanthracenes; benzopyrenes; chemical synthesis; disease /disorder model; epoxides; fibroblasts; fluorine; gel electrophoresis; gene frequency; gene mutation; genetic mapping; genetic promoter element; laboratory mouse; mutagens; nitrosourea; nonvisual photosensitivity; nucleic acid chemical synthesis; nucleic acid sequence; oncogenes; polymerase chain reaction; site directed mutagenesis; sulfonate; transcription factor; ultraviolet radiation

Specific mutagens reproducibly activate specific oncogenes in a specific tissue. Resulting tumors often show the oncogene to have a specific mutation at one particular nucleotide position. The reproducible mutation may be due to this nucleotide position being either modified frequently or misrepaired frequently. We developed a novel technique, the Ligation Mediated-Polymerase Chain Reaction, to map alkylated adducts and ultraviolet light induced adducts in vivo at the nucleotide level of resolution on sequencing gels; even repair rates were quantified at nucleotide resolution. With this discovery, we will relate reproducible hot spots for oncogene activation to hot spots for adduct formation or to slow spots for adduct repair. Technology for mapping adduct frequency will be tailored to other alkylating agents, and to benzopyrene, dimethyl-benzanthracene, and aflatoxin, mutagens responsible for oncogene activation. Results from adduct mapping of p53 and several ras and myc oncogenes in murine tissue culture systems will be applied to mouse model systems.

特定的诱变剂可重复激活特定的特定癌基因 组织。 由此产生的肿瘤通常表明癌基因具有特定的 在一个特定的核苷酸位置的突变。 可再现 突变可能是由于该核苷酸位置被修饰 经常或经常误导。 我们开发了一种新颖的技术， 连接介导的 - 聚合酶链反应，以绘制烷基化性合并 和紫外线在体内诱导的核苷酸水平的加合物 测序凝胶的分辨率；甚至在 核苷酸分辨率。 通过这一发现，我们将联系可重现 热点癌基因激活加入的热点形成或 缓慢的斑点维修。 映射加合物频率的技术 将针对其他烷基剂量身定制 二甲基苯乙烯和黄曲霉毒素，负责癌基因的诱变剂 激活。 p53和几个RAS和MYC的加合物映射的结果 鼠组织培养系统中的癌基因将应用于小鼠模型 系统。