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: 3759186
• 负责人: MARCIA BUDARF
• 金额: --
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 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/3759186
• 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.

研究研究的一部分孔子先天性心脏病（CHD）患者 已显示其微缺失为22q11.2。 该地区删除了 很大，很可能为几个连续基因编码。 开始 了解该特定染色体片段的半合子如何 引起所见心脏缺陷，必须谨慎 分析和鉴定的基因。 为了实现这一目标，我们建议 该区域的详细物理地图，并使用此地图隔离 重叠的克隆基因组DNA片段来自YAC和宇宙。 脱氧核糖核酸 来自冠心病患者选定子集的样品将用于缩小 关键区域。 特别是，患者的断点 将研究用较小的删除来进行研究以剖析 该区域内的基因可能负责构造 这些患者观察到畸形。 将使用多种方法 识别和分离从该区域的cDNA。 这些方法将 包括新分离的染色体22特异性基因的映射，直接 选择使用YAC和映射到关键区域的CDNA和 从大型DNA序列的宇宙和计算机分析 关键区域的比例测序。 在此确定的成绩单 通过PCR的杂交来确认和分离方式 基于策略。 因此，重要的发育调节基因将 被识别和表征。 它们在冠心病的发病机理中的作用 将被检查。