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地激活特定的癌基因 组织。 由此产生的肿瘤通常显示癌基因具有特定的 某一特定核苷酸位置的突变。 可重现的 突变可能是由于该核苷酸位置被修改 经常或经常误修。 我们开发了一项新技术， 连接介导的聚合酶链反应，以绘制烷基化加合物 和紫外线在体内核苷酸水平诱导加合物 测序凝胶上的分辨率；甚至维修率也被量化为 核苷酸分辨率。 有了这个发现，我们将把可重复的 癌基因激活的热点到加合物形成的热点或 加合物修复的慢点。 绘制加合物频率的技术 将针对其他烷化剂和苯并芘进行定制， 二甲基苯并蒽和黄曲霉毒素是致癌基因的诱变剂 激活。 p53 与几个 ras 和 myc 的加合物作图结果 小鼠组织培养系统中的癌基因将应用于小鼠模型 系统。