THE GENE FOR HUMAN BETA GALACTOSIDASE-A

这项研究的第一个目标是分析该基因的启动者 为了研究影响α-半乳糖苷酶A基因表达的因素 特别是控制这种基因的表达，以及这类基因的表达 基因大体来说。溶酶体酶的水平是已知的不同的。 广泛地，至少在啮齿动物中，在器官发育和变化期间 在给定细胞内的溶酶体数。抗原刺激 免疫细胞的变化导致溶酶体酶显著增加 级别。目前尚不清楚这些变化是否发生在 转录或转录后。此外，还存在一些机制 来协调调节这类酶的水平，因为 一种酶对另一种酶的相对比例保持相对恒定， 尽管酶的绝对水平可能会有很大的差异。我们有 最近分离出一个含有人类启动子的基因组克隆 α-半乳糖苷酶A，这是迄今为止分离到的第一个基因组克隆 对于任何溶体水解酶，它应该是有价值的建议 基因调控的研究。这项研究的第二个目标是 继续开发一种可能的治疗策略 法布里病，一种遗传性代谢紊乱的原型。上一首 研究表明，外源α-半乳糖苷酶具有 A以纠正Fabry成纤维细胞的代谢缺陷，并临床 试验表明，酶替代疗法具有潜在的效果。 细菌、酵母和杆状病毒产生的人α-半乳糖苷酶A 将使用用于酶替代的细胞培养系统进行评估 法布里病。克隆的α-半乳糖苷酶A的cDNA将是 被引入培养的Fabry细胞中，以监测其整合和 表情。

The first objective of this research is to analyze the promoter of the alpha-galactosidase A gene in order to investigate the factors that control the expression of this gene in particular, and of this class of genes in general. The levels of lysosomal enzymes are known to vary widely, at least in rodents, during organ development and during changes in the number of lysosomes within a given cell. Antigenic stimulation of immune cells results in a dramatic increase in lysosomal enzyme levels. It is not known if these changes occur at the level of transcription or posttranscriptionally. In addition, mechanisms exist to coordinately regulate the levels of this class of enzymes, since the relative ration of one enzyme to another remains relatively constant, although the absolute levels of the enzymes may vary widely. We have recently isolated a genomic clone containing the promoter for the human alpha-galactosidase A, which is the first genomic clone isolated to date for any lysomal hydrolase, and it should be valuable for the proposed studies of gene regulation. The second objective of this research is to pursue the development of one possible strategy for the treatment of Fabry disease, a prototype inherited metabolic disorder. Previous studies have demonstrated the ability of exogenous alpha-galactosidase A to correct the metabolic defect in Fabry fibroblasts, and clinical trials have suggested the potential effectiveness of enzyme replacement. Human alpha-galactosidase A produced in bacteria, yeasts, and baculovirus will be evaluated using a cell culture system for enzyme replacement in Fabry disease. The cloned cDNA for alpha-galactosidase A will be introduced into Fabry cells in culture to monitor its integration and expression.