MOLECULAR CHARACTERIZATION OF MURINE PCY MUTATION

描述（改编自申请人的摘要）： 肾脏疾病（PKD）是一组特征在于 存在大量的囊肿在整个巨大的肾脏。 在人类中，这些疾病可以以常染色体显性遗传方式获得或遗传。 常染色体隐性遗传（ARPKD）。 ADPKD是最常见的， 男性显性遗传性肾病，而ARPKD相对 很少。临床上，ADPKD是慢性肾功能衰竭的主要原因。 它占所有需要长期治疗的患者的10%。 透析或肾移植。 ADPKD的确切分子病变 是未知的，除了透析和移植， 姑息性的，不存在治愈性治疗。 DBA/2纯合子小鼠 pcy突变发展为一种病理性多囊肾病 表型与最常见的常染色体 多囊肾是一种常见的人类疾病。 一项建议是将小鼠遗传学和定位克隆相结合， 分离小鼠pcy基因并阐明其分子机制的策略 该基因突变的基础负责多囊性 表型。 所附的实验计划建议：1）生产一种 来自600只多囊F2动物（pcy/pcy x Mus m）的DNA组。栗色 种间杂交用于系谱分析。 这个小组将有一个 分辨率为0.1 cM; 2）为31个目前 位于小鼠9号染色体区域的可用遗传标记 3）使用定义的pcy基因座进行谱系分析; RFLPs以发现与小于0.1cM的pcy连锁的标记; 4） 从DBA和pcy/pcy中产生P1基因组和单向cDNA文库 5）分离和定位对应于200 kb的P1克隆， 在搜索pcy基因的同时， 在每个克隆内新的RFLP; 6）在该200 kb区域中搜索基因;和 7)表征和筛选每个候选基因的特征 与PCY多囊肾表型一致，以鉴定PCY 基因

DESCRIPTION (Adapted from the Applicant's Abstract): The polycystic kidney diseases (PKD) are a group of disorders characterized by the presence of a large number of cysts throughout grossly enlarged kidneys. In humans, the diseases can be acquired or inherited in autosomal dominant (ADPKD) or autosomal recessive (ARPKD) forms. ADPKD is the most common, dominantly inherited kidney disease of man while ARPKD occurs relatively rarely. Clinically, ADPKD represents a major cause of chronic renal failure in man and it accounts for 10% of all patients requiring chronic dialysis or renal transplantation. The exact molecular lesion(s) of ADPKD are unknown and except for dialysis and transplantation, which are palliative, no curative treatment exists. DBA/2 mice homozygous for the pcy mutation develop a form of polycystic kidney disease with pathological phenotypes remarkably similar to the most prevalent form of autosomal dominant human polycystic kidney disease in man. The overall goal of this proposal is to use a combination of mouse genetics and positional cloning strategies to isolate the murine pcy gene and to elucidate the molecular basis of the mutation(s) in this gene responsible for the polycystic phenotype. The accompanying experimental planproposes to: 1) produce a DNA panel from 600 polycystic F2 animals in a pcy/pcy x Mus m. castaneus interspecific cross for pedigree analysis. This panel will have a resolution of 0.1 cM; 2) define RFLP for each of the 31 currently available genetic markers positioned in the region of mouse chromosome 9 containing the pcy locus; 3) perform pedigree analysis using the defined RFLPs to find markers that are linked to pcy at less than 0.1 cM; 4) produce P1 genomic and unidirectional cDNA libraries from DBA and pcy/pcy kidneys; 5) isolate and map 8-10 P1 clones corresponding to a 200 kb region encompassing the pcy gene by chromosome walking while searching for new RFLP within each clone; 6) search for genes in this 200 kb region; and 7) characterize and screen each candidate gene for characteristics consistent with the pcy polycystic kidney phenotype to identify the pcy gene.