IDENTIFICATION OF TANGIER DISEASE AND HSN 1 GENES

目的：该项目的主要目标是确定导致 遗传性神经病该项目将调查丹吉尔病 (TD)和遗传性感觉神经病1型（HSN 1）家系 使用连锁分析和定位克隆技术。的 潜在的假设是，这些技术将识别 TD和HSN 1的疾病相关基因。的具体目标 项目是：1）在一个大型丹吉尔病中建立基因连锁 系谱，并制定一个物理地图的链接区域; 2）使用 不断发展的9号染色体物理图谱， HSN 1区域; 3）启动对TD和HSN 1基因的搜索 在它们各自的定位位点内，通过鉴定 两个疾病位点内的候选基因。方法：在目标1中，我们将 进行全基因组筛选，以确定遗传标记（简称 简单序列标记（SSR）串联重复多态性）， 分离与TD，然后使用重组分析，以改善 联系区域。然后我们将使用基因组DNA克隆到人工 染色体来构建连锁区域的物理图谱。在aim中 2，我们将初步确认HSN 1位点与标记的连锁 D9 S318和D9 S176，然后选择人工染色体， 克隆的基因组DNA跨越该区域， 映射.在目标3中，我们将使用可用的cDNA文库和 人工染色体，由该项目的初始部分定义， 各种杂交和扩增技术来克隆基因 与这两种疾病的基因位点对应。此外，我们将使用 人类基因组数据库，以确定任何先前表征 定位到疾病位点的基因。在确定了这些基因之后， 以这种方式，我们将在疾病谱系中筛选它们，使用单一的 链构象多态性和直链测序， 检测任何突变意义：这些疾病的鉴别- 相关基因将：1）阐明潜在的分子缺陷， 这两种神经病变; 2）增加我们对正常 感觉神经功能; 3）在TD的情况下，定义一个新的基因或 对高密度脂蛋白和细胞内 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.