MOLECULAR GENETIC ANALYSIS OF THE PRADER-WILLI SYNDROME LOCUS

普拉德-威利综合征基因座的分子遗传学分析

• 批准号: 5212586
• 负责人: MARC E LALANDE
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5212586
• 关键词: Prader Willi syndrome; biochemical evolution; brain; chromosome deletion; chromosome disorders; chromosome translocation; clone cells; cytogenetics; genomic imprinting; human genetic material tag; human tissue; laboratory mouse; library; mental retardation; molecular cloning; molecular genetics; nucleic acid sequence

Prader-Willi syndrome (PWS) is a genetic disease characterized by mental retardation, obesity and hypogonadism. The majority of patients display a characteristic deletion of chromosome 15q11q13. This deletion is always of paternal origin. In cases where there is no detectable deletion, uniparental maternal disomy of chromosome 15q is observed. These findings indicate that the PWS disease locus is genetically imprinted. We propose to more narrowly delineate the PWS critical region of deletion overlap by identifying and cloning the breakpoint junction of an unbalanced translocation associated with PWS. Cosmid and lambda phage libraries constructed from flow sorted chromosome 15q11q13 material as well as yeast artificial chromosome clones will be used to clone the breakpoint junction. These reagents will also be used to construct a contig of the PWS critical region. Potential transcription units in the PWS critical region will then be identified. We will determine whether any of the transcribed sequences which map to the PWS critical region display allele-specific expression. This will be accomplished using a PCR-based assay for short tandem repeats which occur in the 3' untranslated regions of the cDNAs from the PWS critical region. A gene from the PWS critical region which is expressed exclusively from the paternal chromosome 15 will be classified as a candidate gene for PWS.

Prader-Willi综合征（PWS）是一种以精神为特征的遗传疾病 迟钝，肥胖和性能不足。 大多数患者显示 15Q11Q11Q11Q11Q11Q11Q11的特征缺失。 此删除是 永远是父亲的起源。 如果没有可检测到的情况 观察到缺失，染色体15q染色体的单亲孕产妇疾病。 这些发现表明，PWS疾病基因座是遗传的 印记。 我们建议更狭窄地描绘PWS关键区域 通过识别和克隆断点连接的删除重叠 与PW相关的不平衡易位。 Cosmid和Lambda 噬菌体库由流量排序的染色体15q11q13材料构建 以及酵母人造染色体克隆将用于克隆 断点连接。 这些试剂也将用于构建 PWS关键区域的重叠群。 潜在的转录单元 然后将确定PWS关键区域。 我们将确定是否 映射到PWS临界区域的任何转录序列 显示特定于等位基因的表达式。 这将使用 基于PCR的分析，用于在3'中发生的短串联重复序列 来自PWS关键区域的CDNA的未翻译区域。 一个基因 来自PWS关键区域，该区域仅从 父亲15号染色体将被归类为PWS的候选基因。