IDIOPATHIC HEMOCHROMATOSIS DISEASE GENE

特发性血色沉着症(IH)是一种遗传性、隐性连锁疾病 人类主要组织相容性复合体内的人类白细胞抗原-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)，其特征是慢性和 全身性铁积累。我们最初的努力集中在 MHC基因多态性的分离及其在定位研究中的应用 通过基因分析定位缺陷。基于链接 不平衡研究和已发表的善意的描述 重组个体，现在可以明确地将 IH基因在250个定义明确且可接近的临界区内 Kb从人类白细胞抗原-H位点延伸至人类白细胞抗原F。整个地区都是 包含在重叠的粘粒和酵母人工染色体中 克隆人(YAC)帮助我们的实验室为 制作了这一第I类分地区的实物地图。 本提案的重点是候选基因的克隆和鉴定 位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(指示基因)的探针 作为利用整个YAC克隆的两项新技术，我们已经 鉴定了几个多基因家族，并克隆了一个截短的cdna 识别来自关键区域的9.5kb的转录，我们认为 几个强大的IH基因候选者之一。在筹资期间， 每一个新的克隆都将被测序，部分由外显子辅助 诱捕实验，并进一步表征。克隆人将作为 猪瘟病毒肠粘膜和肝组织RNA的Northern杂交研究 我们的实质性患者中包含正常和受影响的个人 人口。与Northern分析相一致，聚合酶链式反应探针也将 根据cdna序列制作，以测试池中的突变 从患者和正常人的单链或单链获得的c DNA 构象多态(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、使人衰弱的疾病 无序。

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.