Analysis of disease associated genes in polycystic kidney disease

多囊肾疾病相关基因分析

• 批准号: 08671302
• 负责人: ANDO Asako
• 金额: $ 1.41万
• 依托单位: Tokai University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671302/
• 关键词: polycystic kidney disease; HLA; HKE6; Ke6; alcohol dehydrogenase; disease associated gene; gene expression; アルコール脱水酸化酵素

In order to identify a disease associated gene which is involved in the development of human polycystic kidney disease (PKD), we have constructed a physical map of the 250-kb segment and searched new genes within the segment on the centromeric side of the human major histocompatibility complex (MHC) including a human homologue of the mouse Ke6 gene, HKE6.. The mouse Ke6 gene is a candidate gene responsible for the development of PKD.From the analysis of a YAC clone and cosmid clones which span the centromeric side of the HLA class II region, 8 new genes were identified. The HKE6 gene (alcohol dehydrogenase-like gene) was mapped on the 100 kb centromeric side of the HLA-DP subregion. The genomic sequence of a 42,801 bp long region encoded by one cosmid clone in the RING1, HKE6 and HKE4 subregions was determined by the shotgun method. The HKE6 gene is composed of eight exons-and seven introns with the same exon-intron organization as observed in the mouse Ke6 gene. The 25 kb region proximal to the RING1 gene includes an extensive dense cluster of Alu repeats (about 1.2 Alu per kb) and no gene has been identified in this so far. A 1.0-kb faint message from the HKE6 gene was detected in the kidney of PKD patients. In these patients, no large-scale deletion or translocation, was detected using genomic Southern hybridization analysis by the HKE6 cDNA insert as a probe. Additionally, no nucleotide deletion, insertion, or base replacement was detected by the direct sequencing of PCR products from 640-bp of the 5' upstream region and exon 1-4 regions in the HKE6 gene.

为了鉴定与人类多囊肾病（PKD）发展相关的疾病相关基因，我们构建了250kb片段的物理图谱，并在人类主要组织相容性复合物（MHC）着丝粒侧的片段内搜索了新基因，包括小鼠Ke6基因的人类同源物HKE6。小鼠Ke6基因是负责发展的候选基因。 PKD。通过对 YAC 克隆和跨越 HLA II 类区域着丝粒侧的粘粒克隆的分析，鉴定出 8 个新基因。 HKE6 基因（乙醇脱氢酶样基因）定位于 HLA-DP 亚区的 100 kb 着丝粒侧。通过鸟枪法测定了 RING1、HKE6 和 HKE4 亚区中一个粘粒克隆编码的 42,801 bp 长区域的基因组序列。 HKE6基因由八个外显子和七个内含子组成，其外显子-内含子组织与在小鼠Ke6基因中观察到的相同。靠近 RING1 基因的 25 kb 区域包括大量密集的 Alu 重复序列（每 kb 约 1.2 Alu），但迄今为止尚未在其中鉴定出任何基因。在 PKD 患者的肾脏中检测到来自 HKE6 基因的 1.0 kb 微弱信息。在这些患者中，使用 HKE6 cDNA 插入片段作为探针进行基因组 Southern 杂交分析，未检测到大规模缺失或易位。此外，通过对 HKE6 基因 5' 上游区域和外显子 1-4 区域的 640 bp 的 PCR 产物进行直接测序，未检测到核苷酸缺失、插入或碱基替换。

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