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: 6162594
• 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/6162594
• 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 (NP-C) is an autosomal-recessive, neurovisceral disorder that is marked by neurological decline (progressive cognitive impairment, ataxia, dystonia, dysphagia, dysarthria, seizures, cataplexy and dementia) and hepatosplenomegaly resulting from cholesterol lipidosis. Our collaborators from NINDS have localized NP-C to 18q11-q12, more specifically in a 2 cM interval between markers D18S44 and D18S480. We used an integrative approach that involved the utilization of genomic and cDNA clones for complementation studies of NP-C cells, as well as the use of mouse backcross analysis of naturally occurring murine models to supplement the recombinant studies in reducing rapidly the candidate interval. Succesful contig assembly of the region in yeast artificial chromosome (YACs), (bacterial artificial chromosome) BACs and cosmids; sequence analyses; and mutation searches in candidate genes recently resulted in the identification of a 4.9 kb transcript that encodes a membrane protein of 142 kD that is mutated in NP-C patients. The role of this gene, NPC-1, in cholesterol trafficking and homeostasis will be examined using immunohistochemistry, cellular biology and modeling in other organisms.

尼曼-匹克C型（NP-C）是一种常染色体隐性遗传，神经内脏型， 一种以神经功能衰退为特征的疾病（进行性认知 损伤、共济失调、肌张力障碍、吞咽困难、构音障碍、癫痫发作、紧张症 和痴呆症）和胆固醇引起的肝脾肿大 前列腺增生症我们来自NINDS的合作者将NP-C定位于18 q11-q12， 更具体地，在标记D18 S44和D18 S480之间的2cM间隔中。 我们使用了一种综合方法，包括利用基因组 和cDNA克隆用于NP-C细胞的互补研究，以及 使用天然存在的鼠模型的小鼠回交分析， 补充重组研究，迅速减少候选 interval.酵母人工合成中该区域的重叠群成功组装 染色体（YACs）、（细菌人工染色体）BAC和cosmetic; 序列分析和突变搜索候选基因最近 结果鉴定了一个4.9kb的转录本， 在NP-C患者中突变的142 kD膜蛋白。的作用 这个基因，NPC-1，在胆固醇运输和体内平衡中， 使用免疫组织化学、细胞生物学和建模进行检查， 其他有机体。