MOLECULAR GENETIC STUDIES OF POLYCYSTIC KIDNEY DISEASE

多囊肾病的分子遗传学研究

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/3235772
• 关键词: alleles; basement membrane; collagen; complementary DNA; electron microscopy; endonuclease; entactin; fibroblasts; gene expression; gene mutation; genes; genetic manipulation; heparan sulfate; human population genetics; human subject; laminin; leukocytes; membrane proteins; messenger RNA; molecular genetics; molecular pathology; nucleic acid hybridization; nucleic acid probes; 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.

多项证据表明基底膜的改变 结构可能是观察到的临床和形态学变化的基础 成人多囊肾病（APKD）。 拟议的研究设计 回答这个问题：编码一个或多个基因是否发生突变 基底膜蛋白[​​IV 型胶原层粘连蛋白、nidogen、entactin 或 硫酸乙酰肝素蛋白多糖（HSPG）核心蛋白]导致（APKD）？ 这 建议采用以下方法： (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。 不寻常的图案 将与控制面板中看到的进行比较，以确定是否 前者是基因总体改变的结果。 人类 cDNA 探针 Alpha1(IV) 胶原蛋白和层粘连蛋白链 B1 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类 cDNA 探针。 (2) 这些基因结构的更微妙的改变将被检测到 连锁分析。 有多名受影响人员的大家庭将 检查 APKD 表型和特定基因的共分离 其遗传通过多态性限制位点检测的等位基因。 三个大的多代家庭适合这些研究； 其他家庭将在早期阶段得到确定和定性 拟议的项目。 (3) 在个人或家庭中，如果 (1) 或 (2) 表示感兴趣的基因之一发生突变，mRNA 研究 由该基因编码的基因将被执行以查明所涉及的区域 突变。 (4) 由(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。 (5) 将为突变和正常等位基因合成寡核苷酸探针 并用于研究多个家庭以确定 APKD 中的遗传异质性。

项目成果

Achondroplasia is not caused by mutation in the gene for type II collagen.

软骨发育不全不是由 II 型胶原蛋白基因突变引起的。

• DOI: 10.1002/ajmg.1320290433
• 发表时间: 1988
• 期刊: American journal of medical genetics
• 作者: Francomano,CA;Pyeritz,RE
• 通讯作者: Pyeritz,RE