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The Pseudozyma flocculosa actin promoter allows the strong expression of a recombinant protein in the Pseudozyma species

Abstract

Fungi belonging to the recently classified genus Pseudozyma possess some unique properties such as biocontrol activity, production of rare antimicrobial glycolipids and production of recombinant proteins. In this work, we report the first cloning of a promoter endogenous to the multi-faceted yeast-like Pseudozyma flocculosa, that of the actin gene. The promoter region lacked typical TATA or CAAT box but displayed three putative GC box and two CT-rich regions. As in other related basidiomycetes, only one copy of the actin gene was present in the genome of P. flocculosa. The activity of the actin promoter was compared to that of the HSP70 promoter from Ustilago maydis in two Pseudozyma species. In P. flocculosa, the actin promoter allowed the expression of a very high amount of GFP protein (27.8 mg g−1 total protein) compared to those obtained with the HSP70 promoter in liquid culture. By contrast, the levels of GFP expression obtained in liquid culture were similar with the actin or the HSP70 promoter in Pseudozyma antarctica. A similar pattern of GFP expression was observed in solid culture. The cloning of this new promoter offers a unique genetic tool to further exploit and study the unusual properties of fungi from the Pseudozyma genus.

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Acknowledgments

The authors would like to thank G. Marchand for the P. flocculosa genomic library and S. Laberge and D. Gagné of Agriculture and Agri-Food Canada for their help with the primer extension experiment. This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada (NSERC) and the Canada Research Chairs Program to R.R. Bélanger.

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Correspondence to Richard R. Bélanger.

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Neveu, B., Michaud, M., Belzile, F. et al. The Pseudozyma flocculosa actin promoter allows the strong expression of a recombinant protein in the Pseudozyma species. Appl Microbiol Biotechnol 74, 1300–1307 (2007). https://doi.org/10.1007/s00253-006-0786-7

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Keywords

  • Pseudozyma
  • Basidiomycetes
  • Actin
  • Promoter
  • Recombinant protein
  • Green fluorescent protein