CHARACTERIZATION OF PKD1 GENE

常染色体显性遗传性多囊肾病（ADPKD）是一种常见的遗传性疾病， 影响1/1000人的疾病，通常导致严重的 肾衰竭或中风的发病率。先前的定位克隆 努力确定了小于500个碱基的遗传间隔 含有PKD 1基因的cDNA，并从该区域鉴定了23个cDNA; 这项提案的长期目标是查明和研究基因， 导致PKD 1的突变。最初，三核苷酸重复从 将分析间期的疾病相关扩张， 临床上与ADPKD相似。所有新多态 标记将用于连锁研究。聚合酶扩增的基因座 将在囊肿来源的组织中检查链反应， 杂合性如果通过研究不稳定的基因 该战略将采用系统的方法，如单一的， 链构象多态性分析（SSCP）扫描区域 基因突变的转录本PKD 1基因序列分析，基因组 结构和表达模式。表型 将研究突变的相关性。第二阶段 项目将取决于基因的性质和预测的基因产物。 考虑实验以开发抗体来研究空间分布。 和基因产物的时间分布。该项目还 预期开发或确认体外和体内 多囊肾病的病因有哪些

Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder that affects 1/1000 persons, often leading to significant morbidity from renal failure or stroke. Previous positional cloning efforts have defined a genetic interval of less than 500 kilobases containing the PKD1 gene and identified 23 cDNAs from the region; the long-term aim of this proposal is to pinpoint and study the gene and mutations that cause PKD1. Initially, trinucleotide repeats from the interval will be analyzed for disease-related expansions, which have been shown to cause disorders clinically similar to ADPKD. All new polymorphic markers will be used in linkage studies. Loci amplified by the polymerase chain reaction will be examined in cyst-derived tissues for loss of heterozygozity. If the gene is not identified by studying unstable elements, the strategy will employ systematic methods such as single- stranded conformation polymorphism analysis (SSCP) to scan regional transcripts for mutations. Analysis of the PKD1 gene sequence, genomic structure, and expression pattern will be performed. Phenotypic correlations of mutations will be investigated. The second phase of the project will depend on the nature of the gene and predicted gene product. Experiments are contemplated to develop antibodies to study the spatial and temporal distribution of the gene product. In addition, the project contemplates the development or confirmation of in vitro and in vivo models of polycystic kidney disease abnormalities.