THE GENE FOR HUMAN BETA GALACTOSIDASE-A

本研究的第一个目的是分析启动子的启动子， α-半乳糖苷酶A基因，以研究 特别是控制这种基因的表达， 基因一般 已知溶酶体酶的水平 广泛地，至少在啮齿动物中，在器官发育和变化期间， 在一个给定的细胞内溶酶体的数量。 抗原刺激 免疫细胞的大量增殖导致溶酶体酶 程度. 目前尚不清楚这些变化是否发生在 转录或转录后。 此外，还有一些机制 协调调节这类酶的水平，因为 一种酶与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变化很大。 我们有 最近分离了一个基因组克隆，其中含有人类 α-半乳糖苷酶A，这是迄今为止分离的第一个基因组克隆 对于任何溶酶体水解酶，它应该是有价值的建议 基因调控的研究。 本研究的第二个目的是 寻求制定一项可能的战略， 法布里病，一种遗传性代谢紊乱的原型。 先前 研究表明外源α-半乳糖苷酶 A纠正法布里成纤维细胞中的代谢缺陷， 试验表明酶替代物的潜在有效性。 在细菌、酵母和杆状病毒中产生的人α-半乳糖苷酶A 将使用细胞培养系统进行酶替代评价， 法布里病。 α-半乳糖苷酶A的克隆cDNA将被 引入培养的法布里病细胞中以监测其整合， 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.