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A Strategy for Production of Correctly Folded Disulfide-Rich Peptides in the Periplasm of E. coli

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Heterologous Gene Expression in E.coli

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1586))

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Abstract

Recombinant expression of disulfide-reticulated peptides and proteins is often challenging. We describe a method that exploits the periplasmic disulfide-bond forming machinery of Escherichia coli and combines this with a cleavable, solubility-enhancing fusion tag to obtain higher yields of correctly folded target protein than is achievable via cytoplasmic expression. The protocols provided herein cover all aspects of this approach, from vector construction and transformation to purification of the cleaved target protein and subsequent quality control.

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Acknowledgments

This work was supported by the Australian Research Council (DP130103813) and the Australian National Health and Medical Research Council (Principal Research Fellowship and Project Grant APP1063798 to G.F.K.). B.C.-A. is supported by an Australian Postgraduate Award from the Australian Government.

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Authors and Affiliations

  1. Institute for Molecular Bioscience, The University of Queensland, 306 Carmody Road, St. Lucia, QLD, 4067, Australia

    Natalie J. Saez, Ben Cristofori-Armstrong, Raveendra Anangi & Glenn F. King

Authors
  1. Natalie J. Saez
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  2. Ben Cristofori-Armstrong
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  3. Raveendra Anangi
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  4. Glenn F. King
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Corresponding authors

Correspondence to Natalie J. Saez or Glenn F. King .

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Editors and Affiliations

  1. SGC, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United Kingdom

    Nicola A. Burgess-Brown

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Saez, N.J., Cristofori-Armstrong, B., Anangi, R., King, G.F. (2017). A Strategy for Production of Correctly Folded Disulfide-Rich Peptides in the Periplasm of E. coli . In: Burgess-Brown, N. (eds) Heterologous Gene Expression in E.coli. Methods in Molecular Biology, vol 1586. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6887-9_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6887-9_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6885-5

  • Online ISBN: 978-1-4939-6887-9

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