A novel heterologous gene expression system in Saccharomyces cerevisiae using the isocitrate lyase gene promoter from Candida tropicalis.
We have found that the upstream region of the isocitrate lyase gene (UPR-ICL) from the n-alkane-utilizing yeast Candida tropicalis was functional in Saccharomyces cerevisiae as a novel promoter with nonfermentable carbon sources, such as oleic acid, acetate, ethanol, and glycerol/lactate. The expression of two foreign genes coding for beta-galactosidase from Escherichia coli (LacZ) and glutamate decarboxylase from rat brain was carried out under the control of UPR-ICL. Expression of LacZ was repressed by glucose and enhanced over 300-fold by acetate. When an expression vector pWI3 containing multicloning sites between UPR-ICL and the transcriptional terminator of the isocitrate lyase gene (TERM-ICL) was used, the smaller isoform of glutamate decarboxylase (GAD65) was highly produced in a soluble and active form. These results demonstrate that the novel expression system using UPR-ICL and TERM-ICL from C. tropicalis is useful for the production of heterologous proteins in S. cerevisiae.
Kanai
T
T
Department of Synthetic Chemistry and Biological Chemistry, Kyoto University, Japan.
Atomi
H
H
Umemura
K
K
Ueno
H
H
Teranishi
Y
Y
Ueda
M
M
Tanaka
A
A
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology; D002175:Candida / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005796:Genes / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics; D005947:Glucose / Q000494:pharmacology; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000378:metabolism; D007522:Isocitrate Lyase / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012331:RNA, Fungal / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D019346:Sodium Acetate / Q000494:pharmacology; D013728:Terminator Regions, Genetic / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D012331:RNA, Fungal; D012333:RNA, Messenger; D019346:Sodium Acetate; D001616:beta-Galactosidase; D005968:Glutamate Decarboxylase; D007522:Isocitrate Lyase; D005947:Glucose