CANDIDATE GENE ANALYSIS--INTEGRATIVE EFFORT TO CLONE NIEMANN-PICK TYPE C DISEASE

候选基因分析--克隆尼曼-匹克C型疾病的综合努力

• 批准号: 2456801
• 负责人: D A TAGLE
• 金额: --
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2456801
• 关键词: Niemann Pick disease; animal genetic material tag; artificial chromosomes; genetic mapping; human genetic material tag; laboratory mouse; molecular cloning; molecular pathology; northern blottings; single strand conformation polymorphism; tissue /cell culture

Niemann-Pick type C (NPC) is an autosomal recessive lipid storage disorder that is characterized by psychomotor retardation, psychosis, cerebellar ataxia with extrapyramidal manifestations, supranuclear vertical opthalmoplegia, seizures, hepatosplenomegaly and foamy cells in the bone marrow. Genetic linkage analysis has linked the NPC gene to the pericentromeric region of human chromosome 18. We have further refined the location of the gene to the subchromosomal band 18q11-q12, between markers D18S44 and D18S480. This interval has been contigged in YACs and BACs clones. We are employing a concerted effort involving positional cloning strategies that takes advantage of the 1) rapid generation of a mouse genetic map that is easily integratable to the human physical mapping efforts, 2) the introduction of YACs into NPC rodent and human cells to assess for complementation by cholesterol esterification impairment and Filipin staining, and 3) the utilization of the information from the mouse genetic map and the YAC complementation to narrow down the candidate interval for the identification of candidate genes part of the positional cloning efforts. Evolutionarily conserved cDNA sequences will then be used to integrate the mouse map with the human map in an attempt to further narrow down the interval. Candidate genes have been isolated using both internal and 3' terminal exon trapping. These clones and additional ESTs mapped to this region will be tested for their ability to complement NPC cells and to search for the molecular lesion in NPC patient cell lines by SSCP and Northern blot analyses.

尼曼-匹克C型（NPC）是一种常染色体隐性遗传的脂质储存病 以精神发育迟滞，精神病， 小脑共济失调伴锥体外系表现，核上型 垂直眼肌麻痹，癫痫发作，肝脾肿大和泡沫细胞， 骨髓遗传连锁分析将NPC基因与 人类18号染色体的着丝粒周围区域。我们进一步完善了 该基因在亚染色体带18 q11-q12之间的位置 标记D18 S44和D18 S480。该区间已在YAC中重叠， BAC克隆。我们正在采取协调一致的努力， 克隆策略，利用1）快速生成的一个 小鼠基因图谱，很容易整合到人类的身体 作图工作，2）将YAC引入NPC啮齿动物和人类 细胞以评估胆固醇酯化的互补作用 损伤和Filipin染色，以及3）利用 从小鼠遗传图谱和YAC互补的信息， 缩小候选人识别的候选人间隔 基因是定位克隆工作的一部分。进化上保守 然后将cDNA序列用于将小鼠图谱与 人类地图，试图进一步缩小间隔。候选 已经使用内部和3'末端外显子分离基因 诱捕将这些克隆和映射到该区域的其他EST 测试它们补充NPC细胞的能力，并寻找 SSCP和北方印迹法检测鼻咽癌细胞系的分子损伤 分析。