ADDUCT AND REPAIR MAPPING IN ONCOGENES

特异性诱变剂可重复激活特定细胞中的特定癌基因， 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。 可再现 突变可能是由于该核苷酸位置被修饰 经常或经常误修。 我们开发了一种新技术， 连接介导的聚合酶链反应，以绘制烷基化加合物 和紫外光诱导的加合物在体内的核苷酸水平， 在测序凝胶上的分辨率;甚至在 核苷酸分辨率。 有了这一发现，我们将涉及可重复的 致癌基因激活的热点转变为加合物形成的热点，或者 加合物修复的慢点。 加合频率测绘技术 将适合于其他烷基化剂，和苯并芘， 二甲基苯并蒽和黄曲霉毒素，致癌基因的诱变剂 activation. p53与几种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.

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.