MAPPING, GENE IDENTIFICATION AND DETECTION OF MUTATIONS IN CHROMOSOME 22

22 号染色体突变的绘图、基因鉴定和检测

• 批准号: 6110277
• 负责人: MARCIA L BUDARF
• 金额: --
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-26 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6110277
• 关键词: animal tissue; complementary DNA; computer assisted sequence analysis; congenital heart disorder; developmental genetics; gene deletion mutation; genetic mapping; genetic markers; genetic transcription; human genetic material tag; nucleic acid sequence; polymerase chain reaction; pulsed field gel electrophoresis

A subset of conotruncal congenital heart disease (CHD) patients studied have been shown to have microdeletions of 22q11.2. The region deleted is large and is likely to code for several contiguous genes. To begin to understand how hemizygosity of this particular chromosomal segment gives rise to the cardiac defects seen, this region must be carefully analyzed and the genes identified. To accomplish this we propose to make a detailed physical map of the region and use this map to isolate overlapping cloned genomic DNA fragments from YACs and cosmids. DNA samples from a selected subset of CHD patients will be used to narrow the critical region. In particular, the breakpoints of patients who represent with smaller deletions will be studied to dissect out the gene(s) within the region likely to be responsible for the conotruncal malformations observed in these patients. Multiple methods will be used to identify and isolate cDNAs from the region. These methods will include mapping of newly isolated chromosome 22-specific genes, directly selecting for cDNAs which map to the critical region using YACs and cosmids and computer analysis of the DNA sequence obtained from large scale sequencing of the critical region. Transcripts identified in this manner will be confirmed and isolated by either hybridization of PCR- based strategies. Thus, important developmentally regulated genes will be identified and characterized. Their role in the pathogenesis of CHD will be examined.

研究了圆锥动脉干先天性心脏病 (CHD) 患者的子集 已被证明具有 22q11.2 的微缺失。 该地区已删除 很大并且可能编码几个连续的基因。 开始 了解这个特定染色体片段的半合性 引起可见的心脏缺陷，必须仔细检查该区域 分析并鉴定基因。 为了实现这一目标，我们建议 该地区的详细物理地图，并使用该地图来隔离 来自 YAC 和粘粒的重叠克隆基因组 DNA 片段。 脱氧核糖核酸 来自选定的先心病患者子集的样本将用于缩小范围 临界区。 特别是，患者的断点 将研究具有较小删除的代表以剖析 该区域内的基因可能负责圆锥干 在这些患者中观察到的畸形。 将使用多种方法 从该区域识别并分离 cDNA。 这些方法将 包括新分离的 22 号染色体特异性基因的映射，直接 使用 YAC 选择映射到关键区域的 cDNA，并 粘粒和从大获得的DNA序列的计算机分析 关键区域的尺度测序。 在此确定的转录本 方式将通过PCR-杂交来确认和分离 为基础的策略。 因此，重要的发育调控基因将 被识别和表征。 它们在 CHD 发病机制中的作用 将接受检查。