IDENTIFICATION OF TANGIER DISEASE AND HSN 1 GENES

目标：这个项目的主要目标是识别导致 遗传性神经病。该项目将调查丹吉尔病 遗传性感觉神经病1型(HSN1)家系 利用连锁分析和位置克隆技术。这个 潜在的假设是，这些技术将识别 TD和HSN1的疾病相关基因。《公约》的具体目标 项目是：1)在一种大的丹吉尔病中建立基因连锁 谱系并绘制相关联区域的物理地图；2)使用 进化的9号染色体物理图谱以完善定位 HSN1区；3)启动TD和HSN1基因的搜索 在它们各自绘制的基因座内，通过识别 两个疾病基因座内的候选基因。方法：在目标1中，我们将 进行全基因组筛查以确定遗传标记(简称 简单序列标记的串联重复序列多态(SSR) 用TD分离，然后用重组分析来提纯 连锁区。然后我们将使用克隆的基因组DNA进入人工 染色体来构建连接区域的物理图谱。在AIM 2，我们将初步确认HSN1基因座与标记的连锁 D9S318和D9S176，然后选择其 克隆的基因组DNA横跨该区域，以实现进一步的精细物理 映射。在目标3中，我们将使用现有的cdna文库和 人工染色体，由项目的初始部分定义，在 克隆基因的多种杂交和扩增技术 映射到这两种疾病的基因座上。此外，我们将使用 人类基因组数据库以识别任何先前特征的 映射到疾病基因的基因。在确定了这个基因之后 方式，我们将在疾病家系中筛查他们，使用单一 链构象多态和直接测序 检测任何突变。意义：这些疾病的识别-- 相关基因将：1)阐明潜在的分子缺陷 这两种神经病；2)增加了我们对正常的认识 感觉神经功能；3)在TD的情况下，定义新的基因或 高密度脂蛋白和细胞内关键基因家族 胆固醇代谢。

Aims: The broad goal of this project is to identify genes which cause hereditary neuropathies. The project will investigate Tangier disease (TD) and Hereditary sensory neuropathy type-1 (HSN1) pedigrees using linkage analysis and positional cloning techniques. The underlying hypothesis is that these techniques will identify the disease-associated genes for TD and HSN1. The specific aims of the project are to: 1) establish gene linkage in a large Tangier Disease pedigree and develop a physical map of the linked region; 2) use the evolving physical map of chromosome 9 to refine the localization of the HSN1 region; 3) initiate a search for the TD and HSN1 genes within their respective mapped loci, through the identification of candidate genes within the two disease loci. Methods: In aim 1, we will carry out a genome wide screen to identify genetic markers (short tandem repeat polymorphisms of simple sequence markers (SSRs)) that segregate with TD and then use recombination analysis to refine the region of linkage. We will then use genomic DNA cloned into artificial chromosomes to construct a physical map of the linked region. In aim 2, we will initially confirm linkage of the HSN1 locus to markers D9S318 and D9S176 and then select artificial chromosomes whose cloned genomic DNA spans the region for further fine physical mapping. In aim 3, we will use available cDNA libraries and the artificial chromosomes, defined by the initial parts of the project, in various hybridization and amplification techniques to clone genes mapped to the loci of the two diseases. Furthermore, we will, using the Human Genome Database to identify any previously characterized genes mapping to the disease loci. Having identified the genes in this manner, we will screen them in the disease pedigrees, using single strand conformational polymorphism and straight sequencing and to detect any mutations. Significance: The identification of these disease- associated genes will: 1) elucidate the molecular defects underlying these two neuropathies; 2) increase our understanding of normal sensory nerve function; 3) in the case of TD, define a new gene or gene family crucial to high density lipoprotein and intracellular cholesterol metabolism.