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Fluorescence-Activated Cell Sorting as a Tool for Recombinant Strain Screening

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Yeast Metabolic Engineering

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

Abstract

Metabolic engineering of microbial cells is the discipline of optimizing microbial metabolism to enable and improve the production of target molecules ranging from biofuels and chemical building blocks to high-value pharmaceuticals. The advances in genetic engineering have eased the construction of highly engineered microbial strains and the generation of genetic libraries. Intracellular metabolite-responsive biosensors facilitate high-throughput screening of these libraries by connecting the levels of a metabolite of interest to a fluorescence output. Fluorescent-activated cell sorting (FACS) enables the isolation of highly fluorescent single cells and thus genotypes that produce higher levels of the metabolite of interest. Here, we describe a high-throughput screening method for recombinant yeast strain screening based on intracellular biosensors and FACS.

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Author information

Authors and Affiliations

  1. Division of Systems and Synthetic Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden

    Christos Skrekas & Florian David

  2. The Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, Gothenburg, Sweden

    Christos Skrekas & Florian David

  3. Department of Genetics, Harvard Medical School, Boston, MA, USA

    Raphael Ferreira

Authors
  1. Christos Skrekas
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  2. Raphael Ferreira
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  3. Florian David
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Corresponding author

Correspondence to Florian David .

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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Skrekas, C., Ferreira, R., David, F. (2022). Fluorescence-Activated Cell Sorting as a Tool for Recombinant Strain Screening. 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_4

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  • DOI: https://doi.org/10.1007/978-1-0716-2399-2_4

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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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