Regulated secretion of MuGM-CSF in Saccharomyces cerevisiae via GAL1:MF alpha 1 prepro sequences.
Murine granulocyte-macrophage colony-stimulating factor (GM-CSF) was expressed in Saccharomyces cerevisiae using a novel regulated secretion system. This system involves the fusion of the GAL1 upstream regulatory region to the signal sequence of the alpha mating pheromone, and the integration of this GAL1:MF alpha 1 prepro:MuGM-CSF construct into the yeast chromosome. These constructs were very stable under both selective and nonselective conditions: after 30 generations of growth no plasmid loss was observed. The expression and secretion of MuGM-CSF were analyzed by biological assays and Western blots of yeast culture medium and yeast cell extracts. Expression of MuGM-CSF was regulated by galactose induction. In addition, expression levels were proportional to the number of tandem copies of the gene inserted into the yeast chromosome.
Shaw
K J
KJ
Schering Corporation, Bloomfield, NJ 07003.
Frommer
B R
BR
Anagnost
J A
JA
Narula
S
S
Leibowitz
P J
PJ
D000818:Animals; D003115:Colony-Stimulating Factors / Q000235:genetics*; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000235:genetics*; D007120:Immunochemistry; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013728:Terminator Regions, Genetic
D003115:Colony-Stimulating Factors; D006133:Growth Substances; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA