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Cloning and characterisation of the glyceraldehyde 3-phosphate dehydrogenase gene of Candida bombicola and use of its promoter

  • Original Paper
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Journal of Industrial Microbiology & Biotechnology

Abstract

The glyceraldehyde-3-phosphate dehydrogenase gene (GPD) of the sophorolipid producing yeast Candida bombicola was isolated using degenerated PCR and genome walking. The obtained 3,740 bp contain the 1,008 bases of the coding sequence and 1,613 and 783 bp of the upstream and downstream regions, respectively. The corresponding protein shows high homology to the other known GPD genes and is 74% identical to the gyceraldehyde-3-phosphate dehydrogenase of Yarrowia lipolytica. The particular interest in the C. bombicola GPD gene sequence originates from the potential use of its promoter for high and constitutive expression of homologous and heterologous genes. Southern blot analysis did not give any indication for the presence of multiple GPD genes and it can therefore be expected that the promoter can be used for efficient and high expression. This hypothesis was further confirmed by the biased codon usage in the GPD gene. GDP promoter fragments of different lengths were used to construct hygromycin resistance cassettes. The constructs were used for the transformation of C. bombicola and all of them, even the ones with only 190 bp of the GPD promoter, were able to render the cells resistant to hygromycin. The efficacy of a short GPD promoter can be a convenient characteristic for the construction of compact expression cassettes or vectors for C. bombicola. The GenBank accession number of the sequence described in this article is EU315245.

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Acknowledgments

The authors wish to thank Ecover Belgium NV and the Bijzonder Onderzoekfonds of Ghent University for financial support (grants A05/003 and 01D18604).

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Authors and Affiliations

  1. Laboratory of Industrial Microbiology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Inge N. A. Van Bogaert, Sofie L. De Maeseneire, Wim Soetaert & Erick J. Vandamme

  2. Ecover Belgium NV, Industrieweg 3, 2390, Malle, Belgium

    Dirk Develter

Authors
  1. Inge N. A. Van Bogaert
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  2. Sofie L. De Maeseneire
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  3. Dirk Develter
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  4. Wim Soetaert
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  5. Erick J. Vandamme
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Correspondence to Inge N. A. Van Bogaert.

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Van Bogaert, I.N.A., De Maeseneire, S.L., Develter, D. et al. Cloning and characterisation of the glyceraldehyde 3-phosphate dehydrogenase gene of Candida bombicola and use of its promoter. J Ind Microbiol Biotechnol 35, 1085–1092 (2008). https://doi.org/10.1007/s10295-008-0386-x

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  • DOI: https://doi.org/10.1007/s10295-008-0386-x

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