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Synthetic Gene Networks pp 211–225Cite as

Expressed Protein Modifications: Making Synthetic Proteins

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 813))

Abstract

Techniques to manipulate cellular gene expression such that amino acid analogs not encoded by the genetic code are incorporated into a polypeptide chain have recently gained increasing interest. The so-called noncanonical amino acids often have unusual properties that can be translated into target proteins by reprogrammed ribosomal protein synthesis. Residue-specific substitution of a specific canonical amino acid by its analogs provokes global effects in the resulting protein congeners that include improved stability or catalytic activity, reduced redox sensitivity, as well as altered spectral properties. Thus, the approach holds great promise for the engineering of synthetic proteins.

This contribution describes a protocol for the incorporation of a noncanonical amino acid into a target protein expressed in an appropriate amino acid auxotrophic E. coli strain.

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Acknowledgments

The author would like to thank Katrin Biermann for recording the growth curve and the determination of the limiting tyrosine concentration shown in Fig. 2. This work was supported by a grant from the Ministry of Science, Research and the Arts of Baden-Württemberg (Az: 33-720.830-6-04) to B. Wiltschi.

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  1. Faculty of Biology, University of Freiburg, Freiburg, Germany

    Birgit Wiltschi

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  1. Birgit Wiltschi
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Correspondence to Birgit Wiltschi .

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  1. Inst. Biologie II, Zentrum für Biologische Signalstudien, Universität Freiburg, Stübeweg 51, Freiburg, 79108, Germany

    Wilfried Weber

  2. Dept. Ingenieurwissenschaften, Biosysteme (BSSE), ETH Zürich, Mattenstr. 26, Basel, 4058, Switzerland

    Martin Fussenegger

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Wiltschi, B. (2012). Expressed Protein Modifications: Making Synthetic Proteins. In: Weber, W., Fussenegger, M. (eds) Synthetic Gene Networks. Methods in Molecular Biology, vol 813. Humana Press. https://doi.org/10.1007/978-1-61779-412-4_13

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  • DOI: https://doi.org/10.1007/978-1-61779-412-4_13

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