Construction of glucose-repressible yeast expression vectors.
A set of two episomal yeast expression vectors, pYME1 and pYME2, were constructed. These Saccharomyces cerevisiae-Escherichia coli shuttle vectors each contain a modified yeast MAL6S (encoding maltase) promoter that is expressed constitutively, but is subject to carbon catabolite repression by glucose. Expression from this promoter is still dependent upon the presence of active MALR (regulatory) protein. These expression vectors are particularly useful because most S. cerevisiae strains are MAL+, thereby exhibiting a wider host range than GAL-based vector systems. These pYME1 and pYME2 vectors are capable of expression to levels comparable to GAL-based expression plasmids and much higher than a variety of other repressible promoter vectors. The vectors are identical, except that their multiple cloning sites (MCS) are in opposite orientations, making them convenient for inserting heterologous genes.
Yao
B
B
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
Marmur
J
J
Sollitti
P
P
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004274:DNA, Recombinant; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors; D005947:Glucose / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D000520:alpha-Glucosidases / Q000235:genetics
D004274:DNA, Recombinant; D000520:alpha-Glucosidases; D005947:Glucose