MAMMALIAN GENOME MAPPING OF TISSUE-SPECIFIC CDNAS

组织特异性cDNA将快速定位到小鼠染色体片段 为了便于定义小鼠和人类基因组 organization.小家鼠与一只小家鼠的种间杂交 实验室近交系将允许定义cDNA RFLV标记， 通过单倍型分析对大多数（如果不是全部）cDNA克隆作图。均衡 新生儿胸腺特异性cDNA文库将通过 一种新的固相系统的应用，其中cDNA将被结合， 从固定的基因组序列洗脱。这项技术应该允许 产生交叉杂交的组织特异性cDNA文库 通过选择与人类结合的小鼠cDNA， 基因组序列然后将独特的cDNA（约1000个）定位 PCR扩增后的序列。一组38个种间 回交小鼠DNA已被表征为超过450个标记 整个小鼠基因组将用于快速作图。此面板 将老鼠的基因组分成470多个不同的片段。分层 随后将利用作图方法将基因组分为 1000个不同的片段。此外，以5 cM间隔的参考标记物 在100到400次减数分裂事件中进行分析， 新的cDNA标记的定位。结果将提供一个 应用比较法进行人类区域定位 地图知识。选择的cDNA，总共200个，将用于 精确地定义保守连锁群的边界， 人类染色体上的杂交cDNA克隆也将部分地 测序以提供测序标记位点，允许PCR筛选YAC 文库，并使单链构象的映射 感兴趣的研究人员的多态性。由于5'和3'序列 将获得信息，这些成绩单将与 以前的特征基因。新的成绩单将提供一个资源， 用于胸腺细胞成熟的分子遗传学研究。此外该 绘制超过1000个独特的新生儿胸腺转录本可以确定 基因组中是否有转录活跃的区域 以协调的方式。定位的cDNA克隆将很容易制备 提供给其他研究人员，并应大大促进 关闭长距离限制性酶切图谱，并提供额外的工具 用于小鼠和人类基因丰富区域的重叠群构建 基因组cDNA图谱也将与开发中的 微卫星STS地图在其他实验室。这将允许整合 定义小鼠基因组重叠群的努力。

Tissue specific cDNAs will be rapidly mapped to mouse chromosomal segments in order to facilitate the definition of both mouse and human genomic organization. An interspecific mouse cross between Mus spretus and a laboratory inbred strain will allow definition of cDNA RFLV markers and mapping by haplotype analysis of most if not all cDNA clones. Equalized neonatal thymus specific cDNA libraries will be obtained by the application of a novel solid phase system in which cDNAs will be bound and eluted from immobilized genomic sequences. This technique should allow the generation of a library of tissue specific cDNAs that cross hybridize between mammalian species by the selection mouse cDNAs that bind to human genomic sequences. Unique cDNAs (approximately 1000) will then be mapped after PCR amplification of sequences. A panel of 38 interspecific backcross mouse DNAs that has been characterized for over 45O markers throughout the mouse genome will be used for rapid mapping. This panel divides the mouse genome into over 470 different segments. A stratified mapping approach will be subsequently utilized to divide the genome into 1000 different segments. In addition, reference markers at 5 cM intervals that have been analyzed in 100 to 400 meiotic events will further define the mapped location of new cDNA markers. The results will provide a putative human regional localization by the application of comparative mapping knowledge. Selected cDNAs, a total of 200, will be used to precisely define the borders of conserved linkage groups by in situ hybridization on human chromosomes. The cDNA clones will also be partially sequenced to provide sequenced tagged sites, allow PCR screening of YAC libraries, and enable the mapping of single stranded conformational polymorphisms by interested investigators. Since 5' as well as 3' sequence information will be obtained, these transcripts will be compared with previously characterized genes. Novel transcripts will provide a resource for molecular genetic studies of thymocyte maturation. In addition, the mapping of over 1000 unique neonatal thymus transcripts may determine whether there are regions of the genome that are transcriptionally active in a coordinate fashion. The mapped cDNA clones will be made easily available to other investigators and should dramatically facilitate closure of long range restriction maps as well as provide additional tools for contig construction of gene rich regions of both the mouse and human genome. The cDNA map will also be integrated with the developing microsatellite STS maps in other laboratories. This will allow integration of efforts to define a contig of the mouse genome.