MOLECULAR GENETIC STUDIES OF POLYCYSTIC KIDNEY DISEASE

多囊肾病的分子遗传学研究

• 批准号: 3235769
• 负责人: Clair A. Francomano
• 金额: $ 9.53万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1989-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3235769
• 关键词: alleles; basement membrane; complementary DNA; electron microscopy; endonuclease; fibroblasts; gene expression; gene mutation; genes; genetic manipulation; heparan sulfate; human population genetics; human subject; leukocytes; messenger RNA; molecular genetics; molecular pathology; polycystic kidney

Several lines of evidence suggest that alterations of basement membrane structure may underlie the clinical and morphologic changes observed in adult polycystic kidney disease (APKD). The proposed studies are designed to answer the question: Do mutation in one or more of the genes encoding basement membrane proteins [Type IV collagen laminin, nidogen, entactin, or heparan sulfate proteoglycan (HSPG) core protein] cause (APKD)? The following approaches are proposed: (1) The structure and organization of the genes encoding the major basement membrane proteins in an APKD population will be determined by Southern blot analysis. Unusual patterns will be compared to those seen in a panel of controls to determine if the former are the result of gross gene alterations. Human cDNA probes for pro Alpha1(IV) collagen and laminin chain B1 are currently available for these studies. Murine genes for laminin A and B2, nidogen, entactin, and HSPG core protein will be used to isolate the homologous human cDNA probes. (2) More subtle alterations in the structure of these genes will be detected by linkage analysis. Large families with multiple affected individuals will be examined for cosegregation of a the APKD phenotype and specific gene alleles whose inheritance is detected by polymorphic restriction sites. Three large, multi-generation families are suitable for these studies; other families will be ascertained and characterized in the early phase of the proposed project. (3) In individual or families in whom either (1) or (2) indicates mutation in one of the genes interest, studies of mRNA encoded by this gene will be performed to pinpoint the region involved in the mutation. (4) Mutant genes identified by either (1) or (2) will be cloned and sequenced to demonstrate the nature of the mutation. (5) Oligonucleotide probes will be synthesized for mutant and normal alleles and used to study multiple families in an effort to determine the extent of genetic heterogeneity in APKD.

几条证据表明，基底膜的改变 结构可能是临床和形态学变化的基础， 成人多囊肾病（APKD）。 拟议的研究旨在 来回答这个问题：一个或多个编码 基底膜蛋白[IV型胶原层粘连蛋白，巢蛋白，巢蛋白，或 硫酸乙酰肝素蛋白聚糖（HSPG）核心蛋白]（APKD）原因？ 的 提出了以下方法：（1）结构和组织 编码APKD中主要基底膜蛋白的基因 群体将通过Southern印迹分析来确定。 不寻常的模式 将与对照组中的结果进行比较，以确定 前者是基因突变的结果。 人原癌基因cDNA探针 α 1（IV）胶原蛋白和层粘连蛋白链B1目前可用于这些 问题研究 层粘连蛋白A和B2、巢蛋白、巢蛋白和HSPG的小鼠基因 核心蛋白将用于分离同源的人cDNA探针。 （二） 这些基因结构的更微妙的变化将被发现， 连锁分析 有多个受影响者的大家庭将 检查APKD表型和特定基因的共分离 通过多态性限制位点检测遗传的等位基因。 三个大的、多代的家庭适合于这些研究; 其他家庭将在早期阶段确定和表征， 拟议的项目。 (3)在个人或家庭中，（1）或 (2)表明其中一个感兴趣的基因发生了突变， 将执行该基因编码，以确定参与的区域， 突变 (4)通过（1）或（2）鉴定的突变基因将被 克隆并测序以证明突变的性质。 （五） 将合成突变和正常等位基因的寡核苷酸探针 并用于研究多个家庭，以确定 APKD的遗传异质性