MOLECULAR CHARACTERIZATION OF MURINE PCY MUTATION

鼠 PCY 突变的分子特征

• 批准号: 2144863
• 负责人: DAVID D WOO
• 金额: $ 21.74万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1997-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2144863
• 关键词: disease /disorder model; genetic disorder; genetic library; genetic mapping; laboratory mouse; molecular cloning; polycystic kidney; restriction fragment length polymorphism

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.

描述(改编自申请者摘要)：多囊 肾脏疾病(PKD)是一组以 巨大的肾脏内存在大量的囊性肿块。 在人类中，这些疾病可以在常染色体显性遗传中获得或遗传 (ADPKD)或常染色体隐性遗传(ARPKD)。ADPKD是最常见的， 男性以遗传性肾病为主，而ARPKD相对较多 很少见。临床上，ADPKD是慢性肾脏疾病的主要原因之一。 人类的失败，占所有需要慢性疾病的患者的10% 透析或肾移植。ADPKD的确切分子病变(S) 都是未知的，除了透析和移植，这是 姑息治疗，目前还没有根治的方法。DBA/2小鼠为纯合子 Pcy突变发展为一种具有病理性的多囊肾病 表型与最常见的常染色体非常相似 人类显性多囊肾病。这个项目的总体目标是 建议使用老鼠遗传学和位置克隆相结合的方法 小鼠pcy基因的分离及其分子机制的研究 该基因突变(S)导致多囊变的基础 表型。随附的实验计划建议：1)生产一种 600只多囊F2代小鼠的DNA图谱 用于系谱分析的种间杂交。此面板将有一个 分辨率0.1厘米；2)定义目前31个中的每一个的RFLP 位于小鼠9号染色体区域的可用遗传标记 包含pcy基因座；3)使用定义的 寻找小于0.1 cM的与pcy连锁的标记；4) 利用DBA和pcy/pcy构建P1基因组和单向cDNA文库 肾脏；5)分离和定位对应于200kb的8-10个P1克隆 在搜索的同时通过染色体行走包含pcy基因的区域 在每个克隆中添加新的RFLP；6)在该200kb区域中搜索基因；以及 7)对每个候选基因进行特征描述和筛选 与Pcy多囊肾表型一致以确定Pcy 吉恩。