HIGH-RESOLUTION PHYSICAL MAP OF HUMAN CHROMOSOME 17

完整的高分辨率遗传(1-2厘米)和物理(1-2 Mb)图谱 人类基因组目前正在构建中。为此，我们建议 人类17号染色体高分辨率物理图谱的建立 细胞遗传学、体细胞遗传学和脉冲场凝胶的组合 绘制战略图。这种染色体占人类基因组的3%， 代表大约100Mb。体细胞杂交作图小组已经成立 将这个染色体分成大约20个区域，并克隆了100多个 基因和多态标记已被定位在该染色体上。这个 17q22-23上的可选择标记胸苷激酶使体细胞 对基因组分析特别有用的策略。《行动纲领》的具体目标 项目包括：1)扩大混合测绘小组：a) 分离TK-啮齿动物细胞遗传学明确的患者断裂点 细胞将染色体分成-40个区域。这将作为一种 辐射诱发断点和脉冲场映射的参考图。 B)通过辐照a产生额外的随机断点(-100 17只是混血儿。我们将尝试快速筛选克隆人并确定其特征 通过两种新的方法：Alu-PCR检测人类的存在和数量 Alu-PCR产物或全杂交种的DNA和生物素原位杂交 DNA作为针对人类中期染色体的探针定位人类 细分市场。2)现有Noti连锁克隆的作图和构建 额外的稀有切割酶连接克隆将用作锚点 地图上的。3)远距离限制图的脉冲场凝胶构建 使用克隆的基因、匿名的RFLP探针和 链接先前已映射到定义的物理的克隆 时间间隔由混合分析确定。4)对于所有克隆的基因和保守的 小鼠11号染色体上的序列(包括连锁克隆)、同步性 将使用小鼠和大鼠的体细胞杂交进行测试，并放置在小鼠身上 通过协作安排的遗传图谱。

Complete high-resolution genetic (1-2 cM) and physical (1-2 Mb) maps of the human genome are now being constructed. Toward that end, we propose development of a high-resolution physical map for human chromosome 17 by a combination of cytogenetic, somatic cell genetic, and pulsed-field gel mapping strategies. This chromosome comprises 3% of the human genome, representing about 100 Mb. A somatic cell hybrid mapping panel already divides this chromosome into approximately 20 regions, and over 100 cloned genes and polymorphic markers have been mapped on this chromosome. The selectable marker thymidine kinase at 17q22-23 makes somatic cell strategies especially useful for genome analysis. Specific goals of the project include: 1) Expansion of the hybrid mapping panel by: a) isolation of cytogenetically well-defined patient breakpoints in TK- rodent cells to divide the chromosome into -40 regions. This will serve as a reference map for radiation induced breakpoints and pulsed-filed mapping. b) Production of additional random breakpoints (-100) by irradiation of a 17 only hybrid. WE will attempt to rapidly screen and characterize clones by two novel approaches: Alu-PCR to detect presence and amount of human DNA and biotin in situ hybridization of Alu-PCR products or total hybrid DNA as probe against human metaphase chromosomes to localize human segments. 2) Mapping of existing NotI linking clones and construction of additional rare-cutting enzyme linking clones to be used as anchor points of the map. 3) Pulsed-field gel construction of long-range restriction map of the entire chromosome using cloned genes, anonymous RFLP probes, and linking clones which have been previously mapped to defined physical intervals by hybrid analysis. 4) For all cloned genes and conserved sequences (including linking clones), synteny in the mouse on chromosome 11 will be tested using mouse-rat somatic cell hybrids and placed on the mouse genetic map through collaborative arrangements.