ADDUCT AND REPAIR MAPPING IN ONCOGENES

特定的诱变剂可重复激活特定的癌基因 组织。由此产生的肿瘤通常显示癌基因具有特定的 在一个特定的核苷酸位置发生突变。可复制的 突变可能是由于该核苷酸位置被修改 频繁地或者经常地误修。我们开发了一种新技术， 配基介导的聚合酶链式反应定位烷基化加合物 和紫外光在体内诱导的核苷酸水平的加合物 对测序凝胶的分辨率；甚至修复率在 核苷酸解析。有了这一发现，我们将把可复制的 癌基因激活的热点到加合物形成的热点或 内收物修复缓慢的部位。一种测绘加合物频率的技术 将被量身定做用于其他烷基化试剂，以及苯并(A)芘， 二甲基苯并菲和黄曲霉毒素是致癌基因的诱变剂 激活。P53和几种ras和myc加合物图谱的结果 小鼠组织培养系统中的癌基因将应用于小鼠模型 系统。

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.