Heterologous gene expression in continuous cultures of budding yeast |
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Authors: | Danilo Porro Enzo Martegani Bianca Marie Ranzi Lilia Alberghina |
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Institution: | (1) Dipartimento di Fisiologia e Biochimica generali, Sez. Biochimica comparata, Università di Milano, Via G. Celoria 26, 20133 Milan, Italy |
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Abstract: | Summary In a previous paper we have studied the expression of -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.
Offprint requests to: D. Porro |
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