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A Glucose-Derepressed Promoter for Expression of Heterologous Products in the Filamentous Fungus Aspergillus nidulans

Bio/Technology volume 11, pages 815–818 (1993)Cite this article

Abstract

We describe a putative binding sequence (GCGGGGC) for the glucose-responsive represser protein CreA at two positions upstream of the transcription start site of the alcohol dehydrogenase I (alcA) gene of Aspergillus nidulans. To positively identify the putative binding sites as CreA-specifie, the GCGGGGC blocks were mutated at five internal nucleotide positions to GTACTAC and reintroduced into the wild type alcA promoter driving expression of the endogenous alcohol dehydrogenase I gene. This CreA-binding site variant was then transformed into an AlcR constitutive A. nidulans host strain (T2625) and growth was monitored in the presence of the non-metabolized glucose analogue, 2-deoxyglucose. Positive transformants were selected by their ability to grow using ethanol as a carbon source hi the presence of 2-deoxyglucose. Similar CreA binding site variant alcA promoters should permit the alcA-driven expression of heterologous genes in A. nidulans in the presence of glucose, the preferred carbon source for biomass accumulation and provides a model for controlling carbon-catabolite regulated expression in other expression systems.

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Author notes

  1. William E. Hintz: Corresponding author at Dept. of Biology, University of Victoria, P.O. Box 1700, Victoria, B.C. V8W 2Y2.

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  1. Allelix Biopharmaceuticals Inc., 6850 Goreway Drive, Mississauga, Ontario, LAV 1P1, Canada

    William E. Hintz & Peter A. Lagosky

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  1. William E. Hintz
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  2. Peter A. Lagosky
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Hintz, W., Lagosky, P. A Glucose-Derepressed Promoter for Expression of Heterologous Products in the Filamentous Fungus Aspergillus nidulans. Nat Biotechnol 11, 815–818 (1993). https://doi.org/10.1038/nbt0793-815

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