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Selection of Heterologous Protein-Producing Strains in Yarrowia lipolytica

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Recombinant Protein Production in Yeast

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1923))

Abstract

Yarrowia lipolytica has emerged as an alternative expression system for heterologous protein production and enzyme evolution. Several different expression systems dedicated for this species have been developed, ranging from the simple cloning of expression vectors to recently developed high-throughput methodologies using efficient cloning and assembly such as Gateway and Golden Gate strategies. The latter strategies, due to their modular character, enable multiple vector construction and the construction of expression cassettes containing different genes or a gene under different promoters of various strengths.

Here, we present the Golden Gate cloning strategy for the construction of multiple expression cassettes, the transformation into Y. lipolytica, and the selection of efficient enzyme-producing strains using an insect alpha-amylase as a reporter detected via a thermal cycler-based microassay.

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Acknowledgment

The experiments conducted by EC were financially supported by the Polish Ministry of Sciences and Higher Education (Project No IP2015 011074).

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Authors and Affiliations

  1. Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, France

    Paul Soudier, Macarena Larroude, Tristan Rossignol & Jean-Marc Nicaud

  2. Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Poznan, Poland

    Ewelina Celińska

Authors
  1. Paul Soudier
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  2. Macarena Larroude
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  3. Ewelina Celińska
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  4. Tristan Rossignol
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  5. Jean-Marc Nicaud
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Corresponding author

Correspondence to Jean-Marc Nicaud .

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Editors and Affiliations

  1. Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) and Austrian Centre of Industrial Biotechnology (acib), Vienna, Austria

    Brigitte Gasser

  2. Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) and Austrian Centre of Industrial Biotechnology (acib), Vienna, Austria

    Diethard Mattanovich

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Soudier, P., Larroude, M., Celińska, E., Rossignol, T., Nicaud, JM. (2019). Selection of Heterologous Protein-Producing Strains in Yarrowia lipolytica. In: Gasser, B., Mattanovich, D. (eds) Recombinant Protein Production in Yeast. Methods in Molecular Biology, vol 1923. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9024-5_6

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  • DOI: https://doi.org/10.1007/978-1-4939-9024-5_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9023-8

  • Online ISBN: 978-1-4939-9024-5

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