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A Cassette Approach for the Identification of Intein Insertion Sites

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Split Inteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1495))

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Abstract

Over the past decade split inteins have established themselves as powerful tools for protein engineering, protein semisynthesis, and protein functional control approaches. Their key advantage lies in the protein trans-splicing (PTS) reaction that enables posttranslational protein assembly from two independent, even synthetic, peptide precursors. However, since most split intein applications deal with fragmentation and modification of proteins, various issues can arise, ranging from reduced stability to impairment of protein folding. In this chapter I address how the usage of DNA encoded intein cassettes can streamline and speed up the identification of functional split intein insertion sites in novel proteins of interest (POI).

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Acknowledgements

I thank the Mootz lab for continuous support and suggestions, in particular Joachim Zettler for his contribution to the PTS project. The author acknowledges funding from the Fonds der Chemischen Industrie, the National Institutes of Health (R01 GM074868 and R01 DK083834), and the Kieckhefer Foundation.

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Authors and Affiliations

  1. Clayton Foundation Laboratories for Peptide Biology, The Salk Institute for Biological Studies, La Jolla, California, 92037, USA

    Tim Sonntag

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  1. Tim Sonntag
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Correspondence to Tim Sonntag .

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Editors and Affiliations

  1. Institute of Biochemistry Department of Chemistry and Pharmacy, University of Muenster, Münster, Germany

    Henning D. Mootz

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Sonntag, T. (2017). A Cassette Approach for the Identification of Intein Insertion Sites. In: Mootz, H. (eds) Split Inteins. Methods in Molecular Biology, vol 1495. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6451-2_16

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  • DOI: https://doi.org/10.1007/978-1-4939-6451-2_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6449-9

  • Online ISBN: 978-1-4939-6451-2

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