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Preparation of Pichia pastoris Expression Plasmids

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Recombinant Protein Production in Yeast

Part of the book series: Methods in Molecular Biology ((MIMB,volume 866))

Abstract

When planning any heterologous expression experiment, the very first critical step is related to the design of the overall strategy, hence to the selection of the most adapted expression vector. The very flexible Pichia pastoris system offers a broad range of possibilities for the production of secreted, endogenous or membrane proteins thanks to a combination of various plasmid backbones, selection markers, promoters and fusion sequences introduced into dedicated host strains. The present chapter provides some guidelines on the choice of expression vectors and expression strategies. It also brings the reader a complete toolbox from which plasmids and fusion sequences can be picked and assembled to set up appropriate expression vectors. Finally, it provides standard starting protocols for the preparation of the selected plasmids and their use for host strain transformation.

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Author information

Authors and Affiliations

  1. Département Récepteurs et Protéines Membranaires, Centre National de la Recherche Scientifique, Ecole Supérieure de Biotechnologie de Strasbourg, Illkirch, France

    Christel Logez, Fatima Alkhalfioui & Renaud Wagner

  2. Division of Molecular Biosciences, Imperial College London, London, UK

    Bernadette Byrne

Authors
  1. Christel Logez
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  2. Fatima Alkhalfioui
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  3. Bernadette Byrne
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  4. Renaud Wagner
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Corresponding author

Correspondence to Renaud Wagner .

Editor information

Editors and Affiliations

  1. School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, B4 7ET, United Kingdom

    Roslyn M. Bill

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Logez, C., Alkhalfioui, F., Byrne, B., Wagner, R. (2012). Preparation of Pichia pastoris Expression Plasmids. In: Bill, R. (eds) Recombinant Protein Production in Yeast. Methods in Molecular Biology, vol 866. Humana Press. https://doi.org/10.1007/978-1-61779-770-5_3

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  • DOI: https://doi.org/10.1007/978-1-61779-770-5_3

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-769-9

  • Online ISBN: 978-1-61779-770-5

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