MOLECULAR BASIS OF CEREBRAL DEGENERATION

大脑退化的分子基础

• 批准号: 3393798
• 负责人: JOHN S O'BRIEN
• 金额: $ 17.13万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-09-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3393798
• 关键词: Sandhoff disease; Tay Sachs disease; affinity chromatography; beta N acetylhexosaminidase; beta galactosidase; complementary DNA; dogs; embryo /fetus; gangliosidosis GM1; gel electrophoresis; gene expression; gene therapy; genetic library; genetic manipulation; genetic mapping; genetic models; human subject; human tissue; immunogenetics; molecular cloning; molecular pathology; neurochemistry; nucleic acid sequence; prenatal diagnosis; radioimmunoassay; tissue /cell culture

Three human lysosomal genes, beta-galactosidase, hexosaminidase A and hexosaminidase B, will be cloned. cDNA's for each will be isolated from a human liver lambda gtll library by screening for antibody reactive plaques. Full-length cDNA's will be isolated and sequenced to establish colinearity with amino acid sequences for each protein. Comparisons with mutants with GM1 and GM2 gangliosidosis will be made after analysis of mRNA and genomic fragments to establish the nucleic acid defects in selected cell lines. Beta-galactosidase cDNA will be inserted into an expression vector and gene therapy will be attempted in beagles with GM1 gangliosidosis.

三个人溶酶体基因，β-半乳糖苷酶，氨基己糖苷酶A和 将克隆氨基己糖苷酶B。 每种基因的cDNA将从 通过筛选抗体反应性人肝λ GTII文库 斑块 全长cDNA将被分离和测序，以建立 与每种蛋白质的氨基酸序列共线性。 的比较 在mRNA分析后，将产生具有GM 1和GM 2神经节苷脂沉积症的突变体 和基因组片段，以确定所选细胞中的核酸缺陷， 细胞系 将β-半乳糖苷酶cDNA插入表达载体中， 载体和基因治疗将尝试在小猎犬与GM 1 神经节苷脂沉积症