NEW APPROACH TO GENETIC MAPPING WITH DNA POLYMORPHORISMS

我们建议开发和应用一套技术，基于目标 用于DNA自动分析的寡核苷酸的依赖关系 序列多态性 这些技术将加快并增加 解决遗传连锁分析，扩大临床范围 可以映射到染色体位置并有助于染色体定位的条件。 疾病基因位点的鉴定。 这一建议的应用方面将允许比较 编码α基因的基因座中距离的遗传和物理测量 和T细胞抗原受体的β链， 关于人类DNA的流行、分布和核苷酸序列， 这些基因组区域的多态性。 T细胞分析 14号染色体上的受体α链基因座将被延伸以包括一个 一组跨越染色体长度的连锁标记， 测试自动化连锁分析的效率。 我们将开发一种能够快速分析大量 以自动化的方式从微小的组织样品中提取多态性序列。 这些序列信息用于设计等位基因特异性 将通过已建立的连锁的序列分析获得试剂 除了扫描基因组区域的多态性序列外， 使用变性梯度凝胶电泳。连锁图谱将 研究为T细胞受体位点和14号染色体建立了 从精子细胞中扩增的标记基因座中分离等位基因， 含有单倍体染色体互补体的。

We propose to develop and apply a set of techniques, based on the target dependent legation of oligonucleotides, for the automated analysis of DNA sequence polymorphisms. These techniques will speed up and increase the resolution of genetic linkage analysis, extend the range of clinical conditions that may be mapped to a chromosomal location and aid in the identification of the disease gene loci. The applied aspects of this proposal will permit a comparison between the genetic and physical measures of distance in the loci encoding the alpha and beta chains of the T-cell antigen receptor and will provide information regarding the prevalence, distribution and nucleotide sequence of human DNA polymorphisms in these genomic regions. The analysis of the T-cell receptor alpha chain locus on chromosome 14 will be extended to include a set of linkage markers spanning the length of the chromosome for realistic test of the efficiency of automated linkage analysis. We will develop an instrument capable of rapidly analyzing large numbers of polymorphic sequences from minute tissue samples in an automated fashion. These sequence information required for the design of allele-specific reagents will be obtained by sequence analysis of established linkage markers in addition to scanning genomic regions for polymorphic sequences using denaturing gradient gel electrophoresis. Linkage maps will be established for the T-cell receptor loci and for chromosome 14 by the study of segregation of alleles in amplified marker loci from sperm cells, containing haploid chromosomal complement.