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Marker and promoter effects on heterologous expression in Aspergillus nidulans

  • Applied Genetics and Molecular Biotechnology
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Abstract

To study the effects of selection marker, promoter type, and copy number on heterologous expression in Aspergillus nidulans, strains were constructed with single- and multicopy plasmid integrations bearing a reporter gene (lacZ) under the control of either an inducible (alcA) or constitutive (gpdA) promoter and one of three Aspergillus nutritional marker genes (argB, trpC, or niaD). β-Galactosidase activity in the transformants varied over three orders of magnitude, with the majority of levels in the range of 5×103–1×104 U/mg. Significant differences in mean expression levels were found when comparing single-copy transformants with the same promoter but a different marker. Transformants with the argB marker had the highest average expression, ∼threefold over the trpC or niaD clones. For each promoter, maximal expression for the set was seen in the range of the single-copy clones, implying that increasing the copy number does not reliably increase expression in Aspergillus.

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Acknowledgement

This research was funded by a grant from Merck.

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Authors and Affiliations

  1. Department of Chemical Engineering,, University of California, Berkeley, CA, 94720, USA

    David Lubertozzi & Jay D. Keasling

  2. Department of Bioengineering,, University of California, Berkeley, CA, 94720, USA

    Jay D. Keasling

  3. Synthetic Biology Department,, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA

    Jay D. Keasling

  4. Berkeley Center for Synthetic Biology,, University of California, 717 Potter Street, Building 977, Mail code 3224, Berkeley, CA, 94720-3224, USA

    Jay D. Keasling

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  1. David Lubertozzi
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Correspondence to Jay D. Keasling.

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Lubertozzi, D., Keasling, J.D. Marker and promoter effects on heterologous expression in Aspergillus nidulans . Appl Microbiol Biotechnol 72, 1014–1023 (2006). https://doi.org/10.1007/s00253-006-0368-8

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  • DOI: https://doi.org/10.1007/s00253-006-0368-8

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