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Competent Cell Preparation and Transformation of Pichia pastoris

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Yeast Metabolic Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2513))

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Abstract

During the past three decades, the methylotrophic yeast Pichia pastoris (recently reclassified as Komagataella phaffii) has gained widespread acceptance as a system of choice for heterologous protein expression. One of the reasons that this yeast is used so frequently is the simplicity of techniques required for its molecular genetic manipulation. There are several different protocols available for introducing DNA into P. pastoris using electroporation or heat shock. We describe here a shortened protocol for cell preparation and transformation that works reliably with either prototrophic markers or antibiotic selection in this host. This procedure utilizes the most efficient portions of the electroporation and heat-shock transformation protocols to yield a method that is both time-saving and effective.

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Acknowledgments

This work was funded by undergraduate research funds from the University of the Pacific and NIH-AREA grants GM65882-01, GM65882-02, GM65882-03, and GM129758-01 to J.L.-C. and G.P.L.-C.

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Authors and Affiliations

  1. Department of Biological Sciences, University of the Pacific, Stockton, CA, USA

    Joan Lin-Cereghino, Christopher A. Naranjo & Geoff P. Lin-Cereghino

Authors
  1. Joan Lin-Cereghino
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  2. Christopher A. Naranjo
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  3. Geoff P. Lin-Cereghino
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Corresponding author

Correspondence to Geoff P. Lin-Cereghino .

Editor information

Editors and Affiliations

  1. Division of Industrial Biotechnology, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden

    Valeria Mapelli

  2. Division of Industrial Biotechnology, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden

    Maurizio Bettiga

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Lin-Cereghino, J., Naranjo, C.A., Lin-Cereghino, G.P. (2022). Competent Cell Preparation and Transformation of Pichia pastoris. In: Mapelli, V., Bettiga, M. (eds) Yeast Metabolic Engineering. Methods in Molecular Biology, vol 2513. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2399-2_7

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  • DOI: https://doi.org/10.1007/978-1-0716-2399-2_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2398-5

  • Online ISBN: 978-1-0716-2399-2

  • eBook Packages: Springer Protocols

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