Galactose-inducible expression systems in Candida maltosa using promoters of newly-isolated GAL1 and GAL10 genes.
The GAL1 and GAL10 gene cluster encoding the enzymes of galactose utilization was isolated from an asporogenic yeast, Candida maltosa. The structure of the gene cluster in which both genes were divergently transcribed from the central promoter region resembled those of some other yeasts. The expression of both genes was strongly induced by galactose and repressed by glucose in the medium. Galactose-inducible expression vectors in C. maltosa were constructed on low- and high-copy number plasmids using the promoter regions of both genes. With these vectors and the beta-galactosidase gene from Kluyveromyces lactis as a reporter, galactose-inducible expression was confirmed. Homologous overexpression of members of the cytochrome P-450 gene family in C. maltosa was also successful by using a high-copy-number vector under the control of these promoters.
Park
S M
SM
Department of Biotechnology, University of Tokyo, Japan.
Ohkuma
M
M
Masuda
Y
Y
Ohta
A
A
Takagi
M
M
D000595:Amino Acid Sequence; D002175:Candida / Q000235:genetics*; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D005690:Galactose / Q000378:metabolism / Q000502:physiology*; D005800:Genes, Fungal / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family; D011401:Promoter Regions, Genetic / Q000235:genetics; D001616:beta-Galactosidase
D003577:Cytochrome P-450 Enzyme System; D001616:beta-Galactosidase; D005690:Galactose