IDIOPATHIC HEMOCHROMATOSIS DISEASE GENE

特发性血色病（IH）是一种遗传性、隐性遗传性疾病， 人类主要组织相容性复合体内的HLA-A位点 (MHC)位于染色体6p21.3上。 这种疾病可能是最常见的 遗传性疾病携带者频率为0.10），其标志是慢性的， 全身铁积累。 我们最初的努力集中在 MHC多态性分离及其在定位中的应用 通过基因分析定位缺陷。 基于连锁 不平衡研究和公开描述的善意 重组个体，现在可以明确地分配 IH基因在250的明确定义和可接近的关键区域内 kb从HLA-H基因座延伸至HLA-F。 整个地区都是 包含在重叠粘粒和酵母人工染色体中 克隆（YACs），这有助于我们的实验室， 为这一第一类次区域绘制实物地图。 该提案的重点是克隆和表征候选人 存在于上述基因组区间内的疾病基因。 使用dna 富含CpG二核苷酸序列的探针（指示基因）以及 作为两项利用YAC克隆的新技术，我们已经 鉴定了几个多基因家族，并克隆了一个截短的cDNA 从关键区域识别9.5kb转录本，我们认为 几个强IH基因候选者之一。 在资助期间， 每一个新的克隆都将被测序， 捕获实验，并进一步表征。 克隆人将作为 肠粘膜和肝RNA的北方印迹探针， 正常人和受影响的个人包含在我们的大量病人 人口 与北方分析相一致，PCR探针也将被 基于cDNA序列进行，以测试池内的突变 从患者和正常人中获得的cDNA， 构象多态性（SSCP）或变性梯度凝胶 电泳（DGGE）分析。 这项研究的结果应该有助于 早期发现和预防这种常见的和使人衰弱的 disorder.

Idiopathic Hemochromatosis (IH) is a heritable, recessive disorder linked to the HLA-A locus within the human Major Histocompatibility Complex (MHC) on chromosome 6p21.3. The disease is perhaps the most common genetic disorder carrier frequency of 0.10) whose hallmark is chronic and systemic iron accumulation. Our initial efforts have centered on the isolation of MHC polymorphisms and their utilization in positionally localizing the defect through genetic analysis. Based on linkage disequilibrium studies and the published description of a bona fide recombinant individual, it is now possible to unequivocally assign the IH gene to within a well-defined and approachable critical region of 250 kb extending from the HLA-H locus through HLA-F. This entire region is contained within overlapping cosmid and Yeast Artificial Chromosome clones (YACs) which have aided our laboratory in contributing to the creation of a physical map of this class I subregion. This proposal focuses on the cloning and characterization of candidate disease genes residing within the above genomic interval. using DNA probes rich in CpG dinucleotide sequences (indicative of genes) as well as two novel technologies utilizing whole YAC clones, we have already identified several multigene families and have cloned a truncated cDNA recognizing a 9.5kb transcript from the critical region which we believe to one of several strong IH gene candidates. During the funding period, each of the novel clones will be sequenced, aided in part by exon trapping experiments, and further characterized. Clones will serve as probes on Northern blots of intestinal mucosa and liver RNA derived from normals and affected individuals contained within our substantial patient population. In concert with Northern analyses, PCR probes will also be made based on cDNA sequences in order to test for mutations within pools of cDNA derived from patients and normals by either Single-Strand Conformation Polymorphism (SSCP) or Denaturing Gradient Gel Electrophoresis (DGGE) analyses. Results from this study should aid in the early detection and prevention of this common and debilitating disorder.