ISOLATION OF THE HUNTINGTON'S DISEASE GENE

亨廷顿病基因的分离

• 批准号: 2265610
• 负责人: JOHN J WASMUTH
• 金额: $ 26.7万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-02-01 至 1998-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2265610
• 关键词: Huntington's disease; alleles; complementary DNA; gene expression; gene mutation; genetic crossing over; genetic mapping; genetic markers; genetic recombination; heterozygote; homozygote; human genetic material tag; human tissue; hybrid cells; linkage mapping; molecular cloning; molecular genetics; molecular pathology; nucleic acid probes; nucleic acid sequence; nucleic acid structure; tissue /cell culture; yeasts

The long term goals of this project are to isolate the Huntington's disease (HD) gene, identify the most common mutation in the gene which causes the disease and, ultimately, understand at the molecular level how a mutant allele at this locus produces the neuropathology of the disease in heterozygotes. A combination of physical mapping strategies will be employed to compile a very high resolution physical map of a region of about 2.5 Mb of DNA which now appears to be the most narrowly defined location of the disease gene. This will include several approaches to rapidly isolate and map many new DNA probes throughout this segment of chromosomal band 4p16.3. Somatic cell hybrids which retain chromosomes 4 from several important recombinants with HD will be isolated and extensively characterized. This provides a means for unequivocal haplotyping of a large number of polymorphic loci which will allow a more precise locilization for the disease gene to be made. All of the DNA from the minimal physical region shown to contain the HD gene will be isolated in overlapping cosmid and YAC clones. The cloned DNA will be thoroughly examined to identify genomic sequences likely to represent transcribed regions. Full lengih cDNA clones and genomic clones representing candidates for the HD gene will be isolated and used to compare the structure, sequence and expression of these genes in normal individuals, HD heterozygotes and HD homozygotes in order to identify a specific alteration which represents the most common HD mutation. The achievement of this goal will eventually lead to an understanding of the function of the normal HD gene produce and provide insight into how the presence of a mutant gene product disorder. This information will hopefully point to possible completely prevent the onset of symptoms.

该项目的长期目标是隔离亨廷顿氏病 （HD）基因，确定基因中最常见的突变，导致该基因 疾病，最终在分子水平上了解突变体如何 该基因座的等位基因在 杂合子。 物理映射策略的结合将是 用于编译一个非常高的分辨率的物理图 大约2.5 MB的DNA现在似乎是最狭义的DNA 疾病基因的位置。 这将包括几种方法 迅速分离并绘制整个部分的许多新DNA探针 染色体带4P16.3。 保留染色体4的体细胞杂种4 从几种具有HD的重要重组者中，将分离出来，并 广泛的表征。 这提供了明确的手段 大量多态基因座的单倍型，这将允许更多 为了制定疾病基因的精确量差。 来自 显示包含HD基因的最小物理区域将分离 在重叠的宇宙和YAC克隆中。 克隆的DNA将彻底 检查以识别可能代表转录的基因组序列 地区。 代表的完整Lengih cDNA克隆和基因组克隆 HD基因的候选者将被隔离并用于比较 这些基因在正常个体中的结构，序列和表达， 高清杂合子和高清纯合子，以识别特定 代表最常见的HD突变的改变。 成就 这个目标最终将导致对 正常的HD基因产生并提供有关A的存在的见解 突变基因产物障碍。 希望这些信息将指向 可能会完全防止症状发作。