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High-Throughput Protein Production and Purification pp 321–333Cite as

High-Throughput Isolation of Soluble Protein Domains Using a Bipartite Split-GFP Complementation System

Part of the Methods in Molecular Biology book series (MIMB,volume 2025)

Abstract

The identification of soluble, folded domains of proteins is a recurring task in modern molecular biology. We detail a protocol for identifying compact soluble protein domains using a self-assembling two-part split-GFP comprised of a detector fragment (GFP β-strands 1 through 10, or GFP1–10) and a tagging fragment (GFP β-strand 11, or GFP11). The assay is performed in E. coli cells and in cell extracts. A selection step insures the protein fragments are in frame and contain no stop codons, while an inverse PCR is used to enrich protein fragment libraries containing a specific target sequence.

Key words

  • Split-GFP
  • Domain trapping
  • Protein solubility
  • Protein tagging
  • Protein fragment complementation

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  • DOI: 10.1007/978-1-4939-9624-7_15
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References

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Author information

Authors and Affiliations

  1. Institut de Pharmacologie et de Biologie Structurale, IPBS, Université de Toulouse, CNRS, UPS, Toulouse, France

    Amélie Massemin & Jean-Denis Pedelacq

  2. Centre de Recherche en Cancérologie de Toulouse (CRCT), Inserm, Université Paul Sabatier-Toulouse III, CNRS, Toulouse, France

    Stéphanie Cabantous

  3. Bioscience Division, MS-M888, Los Alamos National Laboratory, Los Alamos, NM, USA

    Geoffrey S. Waldo

Authors
  1. Amélie Massemin
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  2. Stéphanie Cabantous
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  3. Geoffrey S. Waldo
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  4. Jean-Denis Pedelacq
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Corresponding author

Correspondence to Jean-Denis Pedelacq .

Editor information

Editors and Affiliations

  1. Architecture et Fonction des Macromolécules Biologiques (AFMB), Unité Mixte de Recherche (UMR) 7257, Centre National de la Recherche Scientifique (CNRS), Aix-Marseille Université, Marseille cedex 9, France

    Dr. Renaud Vincentelli

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Massemin, A., Cabantous, S., Waldo, G.S., Pedelacq, JD. (2019). High-Throughput Isolation of Soluble Protein Domains Using a Bipartite Split-GFP Complementation System. In: Vincentelli, R. (eds) High-Throughput Protein Production and Purification. Methods in Molecular Biology, vol 2025. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9624-7_15

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  • DOI: https://doi.org/10.1007/978-1-4939-9624-7_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9623-0

  • Online ISBN: 978-1-4939-9624-7

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