MOLECULAR GENETIC STUDIES OF POLYCYSTIC KIDNEY DISEASE

多囊肾病的分子遗传学研究

基本信息

批准号：
3235772
负责人：
Clair A. Francomano
金额：
$ 8.94万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-07-01 至 1989-11-30
项目状态：
已结题

项目摘要

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.

几条证据表明地下膜的改变结构可能是观察到的临床和形态变化的基础成人多囊性肾脏疾病（APKD）。拟议的研究是设计的回答问题：在一个或多个编码基因中进行突变地下膜蛋白[IV型胶原蛋白层蛋白，Nidogen，Entactin或硫酸乙酰肝素蛋白聚糖（HSPG）核心蛋白]原因（APKD）？这提出了以下方法：（1）编码APKD中主要基底膜蛋白的基因人口将通过Southern印迹分析确定。不寻常的模式将与在对照组中看到的那些进行比较，以确定是否是否前者是基因改变的结果。人类cDNA探针的pro探针目前可用于这些研究。层粘连蛋白A和B2，Nidogen，Entactin和HSPG的鼠基因核心蛋白将用于分离同源的人cDNA探针。（2）这些基因结构的更微妙的变化将由链接分析。有多个受影响个人的大家庭将检查APKD表型和特定基因的cosegregation 通过多态性限制位点检测其继承的等位基因。三个大型多代家族适合这些研究；其他家庭将在早期的早期进行确定和表征拟议的项目。（3）在（1）或（2）表明其中一个基因感兴趣的突变，对mRNA的研究将执行由该基因编码的，以查明涉及的区域突变。（4）（1）或（2）确定的突变基因将为克隆并测序以证明突变的性质。（5）寡核苷酸探针将用于突变和正常等位基因并用于研究多个家庭，以确定 APKD中的遗传异质性。