Bu D F, Erlander M G, Hitz B C, Tillakaratne N J, Kaufman D L, Wagner-McPherson C B, Evans G A, Tobin A J
Department of Biology, University of California, Los Angeles 90024.
Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2115-9. doi: 10.1073/pnas.89.6.2115.
We report the isolation and sequencing of cDNAs encoding two human glutamate decarboxylases (GADs; L-glutamate 1-carboxy-lyase, EC 4.1.1.15), GAD65 and GAD67. Human GAD65 cDNA encodes a Mr 65,000 polypeptide, with 585 amino acid residues, whereas human GAD67 encodes a Mr 67,000 polypeptide, with 594 amino acid residues. Both cDNAs direct the synthesis of enzymatically active GADs in bacterial expression systems. Each cDNA hybridizes to a single species of brain mRNA and to a specific set of restriction fragments in human genomic DNA. In situ hybridization of fluorescently labeled GAD probes to human chromosomes localizes the human GAD65 gene to chromosome 10p11.23 and the human GAD67 gene to chromosome 2q31. We conclude that GAD65 and GAD67 each derive from a single separate gene. The cDNAs we describe should allow the bacterial production of test antigens for the diagnosis and prediction of insulin-dependent diabetes mellitus.
我们报道了编码两种人谷氨酸脱羧酶（GADs；L-谷氨酸1-羧基裂解酶，EC 4.1.1.15）即GAD65和GAD67的cDNA的分离和测序。人GAD65 cDNA编码一种分子量为65,000的多肽，含585个氨基酸残基，而人GAD67编码一种分子量为67,000的多肽，含594个氨基酸残基。两种cDNA均可在细菌表达系统中指导合成具有酶活性的GADs。每个cDNA与脑mRNA的单一物种以及人基因组DNA中的一组特定限制性片段杂交。用荧光标记的GAD探针与人染色体进行原位杂交，将人GAD65基因定位于10p11.23染色体，将人GAD67基因定位于2q31染色体。我们得出结论，GAD65和GAD67各自来源于一个单独的基因。我们描述的cDNA应能使细菌生产用于胰岛素依赖型糖尿病诊断和预测的检测抗原。
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