MOLECULAR-CLONING OF THE GAMMA-GLUTAMYLCYSTEINE SYNTHETASE GENE OF SACCHAROMYCES-CEREVISIAE

被引:108
作者
OHTAKE, Y
YABUUCHI, S
机构
[1] Central Research Laboratories, Asahi Breweries Ltd., Tokyo, 143, Omori-kita 2-13-1, Ota-ku
关键词
YEAST; SACCHAROMYCES-CEREVISIAE; GLUTATHIONE; GLUTAMYLCYSTEINE SYNTHETASE;
D O I
10.1002/yea.320070907
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The 4.4 kb SphI DNA fragment (GSH1) that complements the gamma-glutamylcysteine synthetase-deficient mutation (gsh1) of Saccharomyces cerevisiae YH1 was cloned into vector plasmid YEp24. Gene disruption of the cloned fragment confirmed that this segment was the same gene as gsh1. Mutant strain YH1 with this plasmid not only restored gamma-glutamylcysteine synthetase (GSH-1) activity but the glutathione content and the growth rate. DNA sequence analysis of the SphI fragment showed that the GSH1 structural gene contained 2034 bp and predicted a polypeptide of 678 amino acids. The deduced amino acid sequence had about a 45% homology to that of rat kidney GSH-1, but a very low homology (about 26%) to that of Escherichia coli GSH-I. Northern analysis showed that GSHI had been transcribed into an approximately 2.7 kb mRNA fragment. Southern analysis showed that GSH1 mapped at chromosome X.
引用
收藏
页码:953 / 961
页数:9
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