The Expression of the γ-Glutamylcysteine Synthetase Gene of Escherichia coli B in Saccharomyces cerevisiae(Biological Chemistry)

概要

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The complete coding region of the gene (gshI) for γ-glutamylcysteine synthetase (GSH-I) of Escherichia coli B was fused with a promoter fragment of Saccharomyces cerevisiae, P8, which was newly cloned in this study. Since the P8 fragment functioned not only as a transcriptional promoter but also the code for the amino terminal residue, the fused gshI was translated into the fused GSH-I in S. cerevisiae. The GSH-I activity and glutathione (GSH) content of the transformant ubcreased nire than 1,000-fold and about 3-fold, respectively. DNA sequencing of the fused gshI revealed that there were two TATA boxes at positions -25 and -165, two CAAT boxes at -248 and -278, and two possible mRNA start sites at -17 and -7 from the putative initiation codon ATG. The fused gshI. Though the calculated molecular weight was 112,668, the purified fused GSH-I had a molecular weight of 55,000 and was immunologically identical to the GSH-I of E. coli B. These results suggested that the fused GSH-I was posttranslationally processed to an active, small protein.
社団法人日本農芸化学会の論文
1988-11-23