Enhanced expression of heterologous proteins by the use of a superinducible vector in budding yeast.
We report the effects of a strong overexpression of the GAL4 activator protein on the expression of UASGAL regulated genes, obtained by cloning the GAL4 gene and the GAL1-10 upstream activating sequence (UASGAL)-lacZ fusion in the same high copy number plasmid. Comparable amounts of active enzyme were obtained by host strains usually producing different levels of cloned proteins due to their different genetic background. The transformed cells constitutively produced low levels of beta-galactosidase (1-2% of total proteins) both in glucose and in raffinose minimal media. Nevertheless, expression was still inducible and a tenfold induction could be rapidly obtained by the addition of 0.5% (w/v) galactose to the culture, even when glucose was still present in the medium.
Porro
D
D
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, Italy.
Lotti
M
M
Martegani
E
E
Ranzi
B M
BM
Alberghina
L
L
D004268:DNA-Binding Proteins; D004790:Enzyme Induction / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005809:Genes, Regulator / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D007763:Lac Operon; D011994:Recombinant Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics*
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D001616:beta-Galactosidase