HIGH-RESOLUTION GENE MAPPING BY IN SITU HYBRIDIZATION

该项目的总体目标是促进 人类和小鼠染色体的详细物理和遗传图谱。 我们 将继续与CEPH/Genethon集团正在进行的工作， 用FISH技术解析人类每条染色体的YAC重叠群图 为了分析800个YAC重叠群（约85%基因组覆盖率）组装成 验证染色体位置，确定嵌合克隆含量的日期 并鉴定假重叠群。 孤儿YACS（没有重叠群分配， 没有STS标记）也将被FISH定位，数百个重叠群也将被FISH定位 与基因定义的STS相关联， 基因图谱和细胞遗传图谱之间的联系。 何时以及是否 来自显示嵌合性的重叠群的必需的单个YAC克隆， 假关节将分别进行FISH映射，以识别和清除 克隆问题 与麻省理工学院基因组中心合作，我们将共同 构建小鼠基因组的低分辨率YAC重叠群图谱。 我们将 还制备、排列和分发染色体特异性YAC亚文库 这将通过筛选具有高复杂性的基因组YAC文库来实现 正常和罗伯逊融合小鼠产生的PCR产物 染色体 将构建小鼠的高分辨率图谱 1号染色体，人类和小鼠染色体中含有锌指的区域 基因簇，以及含有已知疾病的其他染色体位点 基因或有趣的基因家族。 我们将提供FISH映射服务 基因组学界的任何一位研究者， 未定位的基因，或新的转基因小鼠，其中染色体上的信息 转基因的基因座将提供信息。 最后，我们将全面 自动化我们的数字成像显微镜，既增加了吞吐量， FISH实验的10倍，并允许多重 筛选YAC克隆的高密度网格阵列， 组合标记的荧光探针。

The overall objective of this project is to facilitate the construction of detailed physical and genetic maps of human and murine chromosomes. We will continue ongoing work with the CEPH/Genethon group to complete a low resolution YAC contig map of each human chromosome by using FISH techniques to analyze the 800 YAC contigs (approx. 85% genome coverage) assembled to date to verify chromosomal location, to determine chimeric clone content and to identify false contigs. Orphan YACS (with no contig assignment and no STS marker) also will be FISH mapped as will several hundred contigs identified with genetically defined STSs so that a direct correlation between the genetic and cytogenetic maps can be made. When, and if necessary, individual YAC clones from contigs exhibiting chimerism and false joints will be FISH mapped separately to identify and weed out problem clones. In collaboration with MIT Genome Center, we will jointly construct a low resolution YAC contig map of the mouse genome. We will also prepare, arrange and distribute chromosome-specific YAC sublibraries that will be made by screening the genome YAC library with high complexity PCR products generated from normal and Robertsonian fusion mouse chromosomes. High resolution maps will be constructed for murine chromosome 1, regions of human and mouse chromosomes containing zinc finger gene clusters, as well as other chromosomal loci containing known disease genes or interesting gene families. We will provide a FISH mapping service to any investigator in the genome community who has a newly identified, but unmapped gene, or a novel transgenic mouse where information on chromosomal locus of the transgene would be informative. Finally, we will fully automate our digital imaging microscopes, both to increase the throughput of FISH experiments by a factor of 10 fold and to permit the multiplex screening of high density, gridded arrays of YAC clones with combinatorially labeled fluorescence probes.