ISOLATION, CHARACTERIZATION AND OVEREXPRESSION OF THE YEAST GENE, GLR1, ENCODING GLUTATHIONE-REDUCTASE

Using degenerate oligodeoxyribonucleotides based on the N-terminal amino acid (aa) sequence of a yeast glutathione reductase (GR) CNBr-generated peptide fragment and a conserved C-terminal region of known GR aa sequences, the yeast gene encoding GR, GLR1, was isolated using PCR followed by screening of a yeast genomic DNA plasmid library. GLR1 encodes a 467-aa protein with a deduced M(r) of 51545. Comparison with Escherichia coli and human GR sequences reveals 49.8% aa identity. Yeast cells transformed with a multicopy plasmid containing the genomic clone overproduced GR activity sixfold. GLR1 was found not to be an essential gene.