LINKAGE MAPPING HUMAN TELOMERES AND CENTROMERES

这项研究计划的目标是1)确定 个体端粒附近DNA的多态基因座 人类染色体，并将其用于连锁研究，以尝试 关闭遗传图谱，以及2)识别并放置在 对单个人类着丝粒的多态基因座进行连锁分析， 提供有用的染色体参考点，用于遗传和 物理映射。 人端粒特异性和着丝粒特异性人工酵母 染色体(YAC)克隆以及着丝粒的其他探针 并且候选端粒相邻序列将被提供为 由合作者开始编写材料。人的染色体定位 端粒和着丝粒探针将在4 端粒特异和着丝粒特异的杂交 序列，并进行遗传连锁分析。 我们计划用着丝粒和端粒构建图书馆 YAC克隆，筛选出揭示多态的探针， 对40个CEPH参考家系进行基因分型，并整合 使用连锁分析程序将标记添加到我们的遗传图谱 打包CRI-MAP。此外，我们将对现有的基因进行分型 着丝粒特异探针和现有的端粒相邻探针 关于参考家系和着丝粒基因数据的利用 由我们的合作者提供，使我们能够将这些 先前识别的RFLP添加到映射中。对于任何端粒RFLP 这不会链接到遗传图谱(由所有其他人绘制的图谱 标准应跨越染色体)，YAC步行实验 将进行努力，以连接物理结束的 染色体和遗传图谱。 这项拟议的研究最近才变得可行， 将定义遗传图谱的边界并促进合并 物理和基因图谱。我们还将开发高效的 用于跟踪内部资源和数据的数据管理系统 我们的实验室，用于与其他实验室的交流，以及 为已建立的国家资源贡献大型数据集 数据库。

The goals of this research proposal are 1) to identify polymorphic loci from DNA adjacent to telomeres of individual human chromosomes, and to use them in linkage studies to attempt closure of the genetic maps, and 2) to identify and place on the linkage maps polymorphic loci for individual human centromeres, providing useful chromosome reference points for both genetic and physical mapping. Human telomere-specific and centromere-specific Yeast Artificial Chromosome (YAC) clones as well as other probes for centromeres and candidate telomere adjacent sequences will be provided as starting materials by collaborators. Chromosomal localization of the telomere and centromere probes will be determined by 4 hybridization to telomere-specific and centromere-specific sequences, and by genetic linkage analysis. We plan to construct libraries from the centromere and telomere YAC clones, screen them for probes that reveal polymorphism, genotype the 40 CEPH reference pedigrees, and integrate the markers into our genetic maps using the linkage analysis program package CRI-MAP. In addition, we will genotype existing centromere-specific probes and existing telomere adjacent probes on reference pedigrees and make use of centromeric genotypic data provided by our collaborators, enabling us to incorporate these previously identified RFLPs into the maps. For any telomere RFLP that does not link to the genetic map (a map that by all other criteria should span the chromosome), YAC walking experiments will be conducted in an effort to link the physical end of chromosome with the genetic map. The proposed research, which has only recently become feasible, will define the boundaries of genetic maps and facilitate merging of physical and genetic maps. We will also develop efficient data management systems for tracking of resources and data within our laboratory, for communication with other laboratories, and for contributing large data sets to established national resource databases.