HIGH-RESOLUTION PHYSICAL MAP OF HUMAN CHROMOSOME 17

完整的高分辨率遗传（1-2 cM）和物理（1-2 Mb）地图 人类基因组正在构建中。 为此，我们建议 人类17号染色体高分辨率物理图谱的开发 细胞遗传学、体细胞遗传学和脉冲场凝胶的组合 映射策略。 这条染色体占人类基因组的3%， 代表大约100 Mb。 一个体细胞杂交图谱小组已经 将这条染色体分成大约20个区域， 基因和多态性标记已被定位在该染色体上。 的 位于17 q22 -23的选择性标记胸苷激酶使体细胞 对基因组分析特别有用的策略。 的具体目标 项目包括：1）通过以下方式扩大混合绘图小组： 在TK-啮齿动物中分离细胞遗传学明确定义的患者断点 细胞将染色体分成-40个区域。 这将作为一个 辐射诱导断点和脉冲场标测的参考图。 B）通过辐射a产生额外的随机断裂点（~ 100） 17、只有混血儿 我们将尝试快速筛选和鉴定克隆体 通过两种新的方法：RT-PCR检测人类的存在和数量 PCR产物或总杂交体的DNA和生物素原位杂交 DNA作为人中期染色体探针定位人染色体 片段 2)现有NotI连接克隆的定位和NotI连接克隆的构建 用作锚点的其它稀有切割酶连接克隆 地图上。 3)脉冲场凝胶法构建长距离限制性酶切图谱 使用克隆基因，匿名RFLP探针， 将先前映射到定义的物理 通过混合分析。 4)对于所有克隆的基因和保守的 序列（包括连接克隆），小鼠11号染色体上的同线性 将使用小鼠-大鼠体细胞杂交体进行测试， 通过合作安排绘制基因图谱。

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.