MAMMALIAN GENOME MAPPING OF TISSUE-SPECIFIC CDNAS

特定组织的cDNA将被快速映射到小鼠的染色体片段 为了便于对老鼠和人类基因组的定义 组织。一种种间杂交的小鼠，介于斯普雷特斯和一只 实验室自交系将允许定义cDNARFLV标记和 通过单倍型分析对大多数(如果不是全部)的cdna克隆进行定位。均衡 新生儿胸腺特异的c DNA文库将通过 一种新的固相体系的应用，其中的cDNA将结合和 从固定的基因组序列中洗脱出来。这项技术应该允许 杂交组织特异性DNA文库的构建 通过选择与人类结合的小鼠cDNA在哺乳动物物种之间 基因组序列。然后将绘制唯一的cDNA(约1000个) 经聚合酶链式反应扩增序列。由38个物种间组成的小组 回交的小鼠DNA已被鉴定为超过45个标记 整个小鼠基因组将被用于快速作图。此面板 将小鼠基因组分成470多个不同的片段。一个分层的 随后将利用作图方法将基因组划分为 1000个不同的细分市场。此外，间隔5 cm的参考标记物 在100到400次减数分裂事件中进行了分析，将进一步定义 新的cdna标记的定位。结果将提供一个 基于比较研究的人类区域定位假设 映射知识。选定的总共200个cDNA将用于 用原位方法精确地确定保守连锁群的边界 人类染色体上的杂交。这些cdna克隆也将部分 测序以提供已测序的标记位点，允许对YAC进行PCR筛选 文库，并使单链构象图谱成为可能 感兴趣的调查人员的多态现象。因为5‘和3’序列 将获得信息，这些成绩单将与 以前描述过的基因。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， 1000多个独特的新生儿胸腺转录本的图谱可以确定 基因组中是否存在转录活跃的区域 以协调的方式。作图的cdna克隆很容易制作。 可供其他调查人员使用，应极大地促进 关闭远程限制地图，并提供其他工具 用于构建小鼠和人类的基因富集区的重叠群 基因组。Cdna图谱也将与正在开发的 其他实验室的微卫星STS图谱。这将允许集成 努力定义小鼠基因组的重叠群。

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.