CLONING OF THE AICARDI AND GOLTZ SYNDROME GENES

AIcardi 和 Goltz 综合征基因的克隆

• 批准号: 2269292
• 负责人: HUDA Y ZOGHBI
• 金额: $ 24.74万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-02-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2269292
• 关键词: SDS polyacrylamide gel electrophoresis; anencephalus; animal tissue; artificial chromosomes; chromosome deletion; chromosome translocation; complementary DNA; computer assisted sequence analysis; cytogenetics; developmental genetics; gene expression; gene rearrangement; genetic mapping; genetic markers; human genetic material tag; human subject; hybrid cells; in situ hybridization; molecular cloning; northern blottings; nucleic acid sequence; phenotype; polymerase chain reaction; pulsed field gel electrophoresis; radiation genetics; sequence tagged sites; sex linked trait; skin hyperplasia; southern blotting

Aicardi and Goltz syndromes are both X-linked dominant disorders with lethality in males. We have collected cell lines from a number of patients with chromosomal rearrangements showing clinical features of Aicardi syndrome, Goltz syndrome, or both. These patents suffer from a combination of clinical features and developmental defects which include agenesis of the corpus callosum, seizures, developmental delay, retinal lesions, microphthalmia and skin defects. The phenotypes of these patients show significant overlap, suggesting that the same gene, or two or more contiguous genes, may be involved. The variability of the phenotype may be due to the effects of X-inactivation. In all these cases the chromosomal rearrangement results in a terminal deletion involving the Xp22.2-Xp22.3 region, strongly suggesting that the loci for Aicardi and Goltz syndromes map to this region. Therefore: these cell lines represent ideal tools to perform positional cloning of the genes which are mutated in Aicardi and Goltz syndromes. Somatic cell hybrids from these cell lines will be established in order to separate the abnormal X chromosome from the normal one. Precise mapping of the Aicardi and Goltz syndrome critical regions will be obtained by testing the hybrid cell lines with numerous existing and newly generated markers from Xp22.2Xp22.3. We will then perform overlap cloning using yeast artificial chromosome (YAC) clones obtained from several libraries, including a library specifically made from a hybrid retaining the human Xp2l-pter region. YAC subclones will be used for the identification of genes from the region. The expression pattern and sequence analysis of the genes will be examined in normal individuals and in patients affected by Aicardi and Goltz syndromes to determine their involvement in the pathogenesis of these diseases. Further studies will include the characterization of the structure and function of the disease genes and of their products.

AICARDI和GOLTZ综合征都是X连锁的主要疾病 男性的致命性。 我们已经从多个的细胞系收集了 染色体重排的患者显示 AICARDI综合征，Goltz综合征或两者兼而有之。 这些专利遭受了 临床特征和发育缺陷的结合，包括 call体的发育不全，癫痫发作，发育延迟，视网膜 病变，微观心脏和皮肤缺陷。 这些表型 患者表现出明显的重叠，表明相同的基因或两个 或可能涉及更多连续的基因。 可变性 表型可能是由于X灭活的影响。 在所有这些 病例染色体重排会导致末端缺失 涉及XP22.2-XP22.3区域，强烈表明 AICARDI和GOLTZ综合征映射到该区域。 因此：这些细胞 线代表执行基因位置克隆的理想工具 在AICARDI和GOLTZ综合征中突变。 体细胞杂种 从这些细胞系中将建立以分离 正常染色体异常X染色体。 精确的映射 AICARDI和GOLTZ综合征关键区域将通过测试获得 具有许多现有和新生成的标记的混合细胞系 来自xp22.2xp22.3。然后，我们将使用酵母进行重叠克隆 从几个库中获得的人造染色体（YAC）克隆， 包括专门由混合动力保留人类的图书馆 XP2L-pter区域。 YAC亚克隆将用于识别 来自该地区的基因。 表达模式和序列分析 这些基因将在正常个体和受影响的患者中检查 AICARDI和GOLTZ综合征确定它们参与 这些疾病的发病机理。 进一步的研究将包括 疾病基因结构和功能的表征和功能 他们的产品。