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Characterization and regulation of the gamma-glutamyl transpeptidase gene from the fission yeast Schizosaccharomyces pombe

Authors
Park, HJLim, HWKim, KKim, IHPark, EHLim, CJ
Issue Date
Jan-2004
Publisher
CANADIAN SCIENCE PUBLISHING, NRC RESEARCH PRESS
Keywords
fission yeast; genomic DNA; gamma-glutamyl transpeptidase; Pap1; regulation; Schizosaccharomyces pombe
Citation
CANADIAN JOURNAL OF MICROBIOLOGY, v.50, no.1, pp 61 - 66
Pages
6
Journal Title
CANADIAN JOURNAL OF MICROBIOLOGY
Volume
50
Number
1
Start Page
61
End Page
66
URI
https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/15910
DOI
10.1139/W03-106
ISSN
0008-4166
1480-3275
Abstract
The structural gene for the putative gamma-glutamyl transpeptidase (GGT) was isolated from the chromosomal DNA of the fission yeast Schizosaccharomyces pombe. The determined sequence contained 3324 bp and encoded the predicted 630 amino acid sequence of GGT, which resembles counterparts in Homo sapiens, Rattus norvegicus, Saccharomyces cerevisiae, and Escherichia coli. The S. pombe cells harboring the cloned GGT gene showed about twofold higher GGT activity in the exponential phase than the cells harboring the vector only, indicating that the cloned GGT gene was functional. To monitor the expression of the S. pombe GGT gene, we fused the fragment 1085 bp upstream of the cloned GGT gene into the promoterless beta-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid pGT98. The synthesis of beta-galactosidase from the fusion plasmid in S. pombe cells was enhanced by treatments with NO-generating sodium nitroprusside (SN), L-buthionine-(S,R)-sulfoximine (BSO), and glycerol. The GGT mRNA level in the S. pombe cells was increased by SN and BSO. Involvement of Pap1 in the induction of the GGT gene by SN and BSO was observed.
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