MOLECULAR CHARACTERIZATION OF THE MURINE PCY MUTATION

鼠 PCY 突变的分子特征

• 批准号: 3247171
• 负责人: DAVID D WOO
• 金额: $ 18.63万
• 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/3247171
• 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（改编自申请人的摘要）：多囊性 肾脏疾病（PKD）是一组以以下特征为特征的疾病： 严重增大的肾脏中存在大量囊肿。 在人类中，这些疾病可以通过常染色体显性获得或遗传 （ADPKD）或常染色体隐性遗传（ARPKD）形式。 ADPKD 是最常见的， 男性显性遗传性肾脏疾病，而 ARPKD 的发生相对 很少。临床上，ADPKD 是慢性肾病的一个主要原因。 人类衰竭，占所有需要慢性治疗的患者的 10% 透析或肾移植。 ADPKD 的确切分子损伤 未知，除了透析和移植之外 姑息治疗，不存在根治性治疗。 DBA/2 小鼠纯合子 pcy 突变导致一种具有病理性的多囊肾病 表型与常染色体最常见的形式非常相似 人类显性多囊肾病。 本次活动的总体目标 建议结合使用小鼠遗传学和定位克隆 分离小鼠 pcy 基因并阐明分子机制的策略 该基因中导致多囊的突变的基础 表型。 随附的实验计划建议：1）产生 来自 pcy/pcy x Mus m 中 600 只多囊 F2 动物的 DNA 组合。板栗 用于谱系分析的种间杂交。 该面板将有一个 分辨率0.1厘米； 2) 为当前 31 个中的每一个定义 RFLP 位于小鼠 9 号染色体区域的可用遗传标记 含有 pcy 基因座； 3）使用定义的进行谱系分析 RFLP 用于查找与 pcy 连锁小于 0.1 cM 的标记； 4） 从 DBA 和 pcy/pcy 生成 P1 基因组和单向 cDNA 文库 肾脏； 5) 分离并定位对应于 200 kb 的 8-10 个 P1 克隆 通过染色体行走寻找包含 pcy 基因的区域 每个克隆内有新的 RFLP； 6) 在这200 kb区域中搜索基因；和 7) 表征并筛选每个候选基因的特征 与pcy多囊肾表型一致，鉴别pcy 基因。