Simultaneous genomic overexpression of seven glycolytic enzymes in the yeast Saccharomyces cerevisiae

被引:115
作者
Hauf, J
Zimmermann, FK
Müller, S
机构
[1] Tech Univ Darmstadt, Inst Mikrobiol & Genet, D-64287 Darmstadt, Germany
[2] Sci Res & Dev Gmbh, D-61440 Oberursel, Germany
[3] Stratagene Europe, Gebouw Calif, NL-1101 CB Amsterdam, Netherlands
关键词
Saccharomyces cerevisiae; glycolysis; overexpression;
D O I
10.1016/S0141-0229(00)00160-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Fusions of the glycolytic genes TPI1, PGK1, ENO1, PYK1, PDC1, and ADH1 with the lacZ reporter gene of Escherichia coli and a lacZ fusion construct of a 390-bp fragment from the promoter of the HXT7 gene were assayed for beta-galactosidase activity. The glycolytic promoters were induced after addition of glucose to ethanol-grown cells, whereas the HXT7 promoter fragment showed a constitutive beta-galactosidase expression on both carbon sources. The genes coding for the seven enzymes of lower glycolysis Tdh, Pgk, Gpm, Eno, Pyk, Pdc, and Adh were simultaneously put under the control of the same strong promoter, a truncated HXT7 promoter that is constitutively active on ethanol as well as on glucose medium. Genomic expression of the glycolytic genes under the control of this promoter, resulted in an at least 2-fold overexpression. The gene MSG5 was isolated, coding for a protein phosphatase normally involved in cell cycle regulation, as a factor that possibly influences the expression of the HXT7 gene. However, overexpression of MSG5 had no effect on the expression of the HXT7/lacZ fusion, whereas a deletion of this gene resulted in a decreased expression of beta-galactosidase, (C) 2000 Elsevier science Inc. All rights reserved.
引用
收藏
页码:688 / 698
页数:11
相关论文
共 64 条
[1]   ISOLATION AND MOLECULAR ANALYSIS OF THE PHOSPHOGLUCOSE ISOMERASE STRUCTURAL GENE OF SACCHAROMYCES-CEREVISIAE [J].
AGUILERA, A ;
ZIMMERMANN, FK .
MOLECULAR & GENERAL GENETICS, 1986, 202 (01) :83-89
[2]  
ALBER T, 1982, Journal of Molecular and Applied Genetics, V1, P419
[3]   THE YDP PLASMIDS - A UNIFORM SET OF VECTORS BEARING VERSATILE GENE DISRUPTION CASSETTES FOR SACCHAROMYCES-CEREVISIAE [J].
BERBEN, G ;
DUMONT, J ;
GILLIQUET, V ;
BOLLE, PA ;
HILGER, F .
YEAST, 1991, 7 (05) :475-477
[4]  
Bergmeyer H.U., 1974, METHODEN ENZYMATISCH
[5]   SACCHAROMYCES-CEREVISIAE PHOSPHOGLUCOSE ISOMERASE AND FRUCTOSE BISPHOSPHATE ALDOLASE CAN BE REPLACED FUNCTIONALLY BY THE CORRESPONDING ENZYMES OF ESCHERICHIA-COLI AND DROSOPHILA-MELANOGASTER [J].
BOLES, E ;
ZIMMERMANN, FK .
CURRENT GENETICS, 1993, 23 (03) :187-191
[6]   Cloning of a second gene encoding 6-phosphofructo-2-kinase in yeast, and characterization of mutant strains without fructose-2,6-bisphosphate [J].
Boles, E ;
Gohlmann, HWH ;
Zimmerman, FK .
MOLECULAR MICROBIOLOGY, 1996, 20 (01) :65-76
[7]   The molecular genetics of hexose transport in yeasts [J].
Boles, E ;
Hollenberg, CP .
FEMS MICROBIOLOGY REVIEWS, 1997, 21 (01) :85-111
[8]   INDUCTION OF PYRUVATE DECARBOXYLASE IN GLYCOLYSIS MUTANTS OF SACCHAROMYCES-CEREVISIAE CORRELATES WITH THE CONCENTRATIONS OF 3-CARBON GLYCOLYTIC METABOLITES [J].
BOLES, E ;
ZIMMERMANN, FK .
ARCHIVES OF MICROBIOLOGY, 1993, 160 (04) :324-328
[9]   Vac7p, a novel vacuolar protein, is required for normal vacuole inheritance and morphology [J].
Bonangelino, CJ ;
Catlett, NL ;
Weisman, LS .
MOLECULAR AND CELLULAR BIOLOGY, 1997, 17 (12) :6847-6858
[10]   DNA-SEQUENCE ANALYSIS OF A 13 KBP FRAGMENT OF THE LEFT ARM OF YEAST CHROMOSOME-XV CONTAINING 7 NEW OPEN READING FRAMES [J].
CASAMAYOR, A ;
ALDEA, M ;
CASAS, C ;
HERRERO, E ;
GAMO, FJ ;
LAFUENTE, MJ ;
GANCEDO, C ;
ARINO, J .
YEAST, 1995, 11 (13) :1281-1288