MAPPING AND CLONING THE 17Q-LINKED BREAST CANCER LOCUS

乳腺癌易感等位基因的定位 17号染色体q是第一个成功绘制的常见癌症图谱。 的 这项建议的目标是克隆和遗传特性，这17 q 乳腺癌易感基因 就像定位常见疾病一样 与特定染色体的配对被证明是困难和容易出错的， 随后的精细结构作图和基因分离将是更多的 比罕见的孟德尔特征更难。 的独特特征 犹他州人口的遗传分析将是一个重要的资产， 这个项目 我们的项目将是许多联系在一起的一个 联合提供必要资源， 17 q易感基因座的分析。 成功的关键是 一个项目是能够识别大的信息kinestrams映射 问题研究 信息量很大的运动， 将研究乳腺癌易感性的分离。 联动 分析将确定亲属的易感性是否与 17 q. 将围绕这些区域绘制一幅信息量很大的标记图。 易感基因座 我们已经鉴定出了位于 乳腺癌易感性基因座和特征的混合面板 由K创建。Fournier基因周围有断点。 粘粒 文库将从一个Fournier杂交体构建，该杂交体包含10到 15兆碱基的人类DNA，包括含有该基因的区域。 在候选基因内进行遗传和物理定位的克隆 区域用作成核点以形成该区域的重叠群。 最终的产物将是YAC重叠群，这是一个罕见的YAC重叠群的脉冲场图。 切割机，以及具有遗传边界的区域的粘粒重叠群 定义易感性位点必须位于的候选区域。 我们已经排除了七个潜在的候选基因（EDBH 17、ERB B2、HOX 2、 NM 23、WNT 3、RARA和Prohibitin）;因此， 几乎可以肯定易感基因座是一个新基因。 克隆 将通过消减杂交选择来自cDNA文库的 杂交、cDNA与亲和柱杂交和直接 粘粒克隆与cDNA克隆杂交。 所有的cDNA都将 筛查病例和对照组之间的DNA序列差异， 定义了内含子-外显子边界。 我们将描述 17 q连锁易感基因位点的作用， 年龄特异性染色体畸变 我们将研究偏侧性的问题， 疾病，并试图确定任何组织病理学特征， 与17号染色体相关的乳腺癌 生殖史数据将 收集并评估与易感基因座的相互作用。 如果在研究过程中发现了17 q基因，我们将 筛选每个家族的突变，并对所述分析进行分层 在特定变异的基础上。

The localization of a susceptibility allele for breast cancer to chromosome 17q is the first successful mapping of a common cancer. The goal of this proposal is to clone and genetically characterize this 17q breast cancer susceptibility locus. Just as localizing common diseases to a specific chromosome has proven to be difficult and error-prone, the subsequent fine structure mapping and gene isolation will be more difficult than for rare mendelian traits. The unique characteristics of the Utah population for genetic analysis will be an essential asset for this project. Our project will be one of many tied together in a scientific consortium jointly providing the resources necessary for the analysis of the 17q susceptibility locus. A key to the success of this project is the ability to identify large informative kindreds for mapping studies. Highly informative kindreds with a high likelihood of segregating a breast cancer susceptibility will be studied. Linkage analysis will determine whether a kindred's susceptibility is linked to 17q. A map of highly informative markers will be developed around the susceptibility locus. We have identified recombinants which flank the breast cancer susceptibility locus and characterized a hybrid panel created by K. Fournier with breakpoints surrounding the gene. A cosmid library will be constructed from a Fournier hybrid which contains 10 to 15 megabases of human DNA including the region containing the gene. Clones which map genetically and physically within the candidate gene region are used as nucleation points to develop a contig of the region. The final product will be a YAC contig, a pulsed field map of rare cutters, and a cosmid contig of the region with genetic boundaries defining the candidate region where the susceptibility locus must lie. We have eliminated seven potential candidate genes (EDBH17, ERB B2, HOX2, NM23, WNT3, RARA, and Prohibitin) from the region; therefore, it is virtually certain that the susceptibility locus is a novel gene. Clones derived from cDNA libraries will be selected by subtractive hybridization, hybridization of cDNAs to an affinity column, and direct hybridization of cosmid clones to cDNA clones. All cDNAs will be screened for DNA sequence differences between cases and controls once intron-exon boundaries are defined. We will characterize the phenotypic effect of the 17q-linked susceptibility locus in terms of its site and age-specific penetrance. We will examine questions of laterality of disease and try to identify any histopathologic features characteristic of 17q-linked breast cancer. Data on reproductive history will be gathered and interaction with the susceptibility locus will be assessed. If, during the course of this study the 17q gene is identified, we will screen for mutations in each family and stratify the analyses described on a mutation-specific basis.