HIGH-RESOLUTION GENE MAPPING BY IN SITU HYBRIDIZATION

通过原位杂交进行高分辨率基因图谱

• 批准号: 2208685
• 负责人: DAVID C WARD
• 金额: $ 37.45万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-05-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2208685
• 关键词: animal genetic material tag; artificial chromosomes; biomedical automation; chromosomes; digital imaging; fluorescent dye /probe; genetic library; genetic mapping; genetic markers; genome; human genetic material tag; human subject; in situ hybridization; laboratory mouse; nucleic acid repetitive sequence; sequence tagged sites

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.

该项目的总体目标是促进建设 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 小组合作，完成一项低 使用 FISH 技术解析每个人类染色体的 YAC 重叠群图 分析 800 个 YAC 重叠群（约 85% 基因组覆盖率） 验证染色体位置、确定嵌合克隆含量的日期 并识别错误的重叠群。 孤儿 YACS（没有重叠群分配且 无 STS 标记）也将进行 FISH 定位，数百个重叠群也将进行 FISH 定位 与基因定义的 STS 进行鉴定，以便直接相关 可以制作遗传图谱和细胞遗传学图谱之间的图谱。 当，以及如果 必要的，来自表现出嵌合状态的重叠群的单个YAC克隆 假关节将单独进行 FISH 定位以识别和剔除 有问题的克隆。 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。 我们将 还准备、排列和分发染色体特异性 YAC 子库 将通过筛选高复杂度的基因组YAC文库来制作 正常小鼠和罗伯逊融合小鼠产生的 PCR 产物 染色体。 将为小鼠构建高分辨率地图 1 号染色体，人类和小鼠染色体中含有锌指的区域 基因簇以及包含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 绘图服务 基因组界中任何有新发现的研究人员，但是 未定位的基因，或一种新的转基因小鼠，其染色体信息 转基因的位点将提供丰富的信息。 最后，我们将充分 自动化我们的数字成像显微镜，既提高了吞吐量 FISH 实验的数量增加了 10 倍，并允许多重分析 筛选高密度、网格化的 YAC 克隆阵列 组合标记的荧光探针。