Heterologous gene expression in continuous cultures of budding yeast.
In a previous paper we have studied the expression of beta-galactosidase from Escherichia coli, driven from the inducible GAL1-10/CYC1 hybrid promoter, in batch cultures of budding Saccharomyces cerevisiae and have described operating conditions for maximal productivity. In this paper we show that the plasmid instability in continuous cultures can be overcome by utilizing appropriate selection markers and a high copy number vector. The maximal level of expression is influenced by the dilution rate. Moreover, enzyme accumulation appears to depend also upon the degree of oxygenation. A possible explanation of these modulations is discussed, taking into account the interactions of the UAS-GAL and TATA-CYC1 elements.
Dipartimento di Fisiologia e Biochimica generali, Università di Milano, Italy.
D
D
Porro
E
E
Martegani
B M
BM
Ranzi
L
L
Alberghina
D004926:Escherichia coli / Q000235:genetics; D005784:Gene Amplification; D015870:Gene Expression; D005819:Genetic Markers; D014644:Genetic Variation; D010100:Oxygen / Q000494:pharmacology; D010957:Plasmids; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D005819:Genetic Markers; D001616:beta-Galactosidase; D010100:Oxygen