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Characterisation of the Mucor circinelloides regulated promoter gpd1P

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Abstract

The promoter of the Mucor circinelloides gpd1 gene encoding glyceraldehyde-3-phosphate dehydrogenase (gpd1P) was recently cloned and used for the production of recombinant proteins, such as the Aspergillus niger glucose oxidase 1 (GOX). This represents the first example of the application of a strong and regulated promoter from this fungus for recombinant protein production. The original 741-bp gpd1P promoter fragment conferred hexose-dependent expression of GOX in M. circinelloides. To understand the regulatory mechanisms involved in gpd1P-driven expression and to develop improved promoter fragments, deletion derivatives of gpd1P were constructed. These derivatives were fused to the A. niger gox1 gene and used to construct strains containing a single copy of the expression cassette. GOX activity was detected in strains containing the full-length gpd1P and also in strains containing a 713-bp or a 361-bp derivative. Expression levels for the 361-bp derivative were high and comparable, regardless of the carbon source used. This promoter represents a useful derivative for constitutive heterologous gene expression in M. circinelloides.

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Acknowledgements

This work was partly supported by the Danish Ministry of Science, Technology and Development, via the EUKA-Center. Ulla Poulsen is acknowledged for excellent technical assistance and the rest of the Department of Fungal Biotechnology and participants in the EUKA-Center for fruitful suggestions.

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Author notes

  1. Gitte G. Larsen

    Present address: Plant Research Department, Risø National Laboratory, Frederiksborgvej 399, P.O. 49, 4000, Roskilde, Denmark

Authors and Affiliations

  1. Department of Fungal Biotechnology, Biotechnological Institute, Kogle Allé 2, 2970, Hørsholm, Denmark

    Gitte G. Larsen, Karen F. Appel, Anne-Mette Wolff & José Arnau

  2. Center for Process Biotechnology, Biocentrum–DTU, 2800, Lyngby, Denmark

    Jens Nielsen

Authors
  1. Gitte G. Larsen
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  2. Karen F. Appel
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  3. Anne-Mette Wolff
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  4. Jens Nielsen
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  5. José Arnau
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Corresponding author

Correspondence to José Arnau.

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Communicated by U. Kück

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Larsen, G.G., Appel, K.F., Wolff, AM. et al. Characterisation of the Mucor circinelloides regulated promoter gpd1P. Curr Genet 45, 225–234 (2004). https://doi.org/10.1007/s00294-003-0484-2

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  • DOI: https://doi.org/10.1007/s00294-003-0484-2

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