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Heterologous Expression of Membrane Proteins pp 67–85Cite as

Membrane Protein Expression in Lactococcus lactis

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 601))

Abstract

Membrane proteins play key roles in cellular physiology, and they are important drug targets. Approximately 25% of all genes identified in sequenced genomes are known to encode membrane proteins; however, the majority have no assigned function. Although the resolution of soluble protein structure has entered the high-throughput stage, only 100 high-resolution structures of membrane proteins have been described until now. Lactococcus lactis is a gram-positive lactic bacterium that has been used traditionally in food fermentations, but it is now used widely in biotechnology for large-scale overproduction of heterologously expressed proteins. Various expression vectors based on either constitutive or inducible promoters exist. The nisin-inducible controlled gene expression (NICE) system is the most suitable for recombinant membrane protein expression allowing for fine control of gene expression based on the autoregulation mechanism of the bacteriocin nisin. Recombinant membrane proteins can be produced with affinity tags for efficient detection and purification from crude membrane protein extracts. The purpose of this chapter is to provide a detailed protocol for the expression of membrane proteins and their detection using the Strep-tag II affinity tag in L. lactis.

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Acknowledgments

This work has been supported by the Commissariat à l’Energie Atomique (CEA-PM project, A.F., S.B., and N.R.). We thank Shaun Peters and Daphné Seigneurin-Berny for critical reading of the manuscript. We thank Igor Mierau (NIZO, The Netherlands) for kindly allowing us to illustrate this chapter with the figure on the NICE system.

Author information

Authors and Affiliations

  1. Laboratoire de Physiologie Cellulaire Végétale, CNRS (UMR-5168)/CEA/INRA (UMR-1200), Université Joseph Fourier, iRTSV, CEA-Grenoble, France

    Annie Frelet-Barrand

  2. Laboratoire de Physiologie Cellulaire Végétale, Grenoble, France

    Sylvain Boutigny

  3. CEA, DSV, iRTSV, Grenoble, France

    Sylvain Boutigny & Norbert Rolland

  4. INRA, Grenoble, France

    Sylvain Boutigny & Norbert Rolland

  5. Université Joseph Fourier, Grenoble, France

    Sylvain Boutigny & Norbert Rolland

  6. The Medical Research Council, Dunn Human Nutrition Unit, Cambridge, UK

    Edmund R. S. Kunji

  7. Laboratoire de Physiologie Cellulaire Végétale, CNRS, Grenoble, France

    Norbert Rolland

Authors
  1. Annie Frelet-Barrand
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  2. Sylvain Boutigny
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  3. Edmund R. S. Kunji
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  4. Norbert Rolland
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Corresponding author

Correspondence to Annie Frelet-Barrand .

Editor information

Editors and Affiliations

  1. CNRS - UMR 6543, Université Nice, parc Valrose, Nice CX 02, 06108, France

    Isabelle Mus-Veteau

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Frelet-Barrand, A., Boutigny, S., Kunji, E.R.S., Rolland, N. (2010). Membrane Protein Expression in Lactococcus lactis . In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology™, vol 601. Humana Press. https://doi.org/10.1007/978-1-60761-344-2_5

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  • DOI: https://doi.org/10.1007/978-1-60761-344-2_5

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-343-5

  • Online ISBN: 978-1-60761-344-2

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