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Noncanonical Amino Acids pp 147–154Cite as

Using Amber and Ochre Nonsense Codons to Code Two Different Noncanonical Amino Acids in One Protein Gene

Part of the Methods in Molecular Biology book series (MIMB,volume 1728)

Abstract

In Escherichia coli, conventional amber and ochre stop codons can be separately targeted by engineered amber-suppressing Methanocaldococcus jannaschii tyrosyl-tRNA synthetase-tRNAPyl and ochre-suppressing Methanosarcina maezi pyrrolysyl-tRNA synthetase-tRNAPyl pairs for coding two different noncanonical amino acids in one protein gene. Here, we describe the application of this approach to produce a protein with two distinct chemical functionalites which can be selectively labeled with two fluorescent dyes.

Key words

  • Double incorporation
  • Protein labeling
  • Amber codon
  • Ochre codon
  • MjTyrRS
  • PylRS
  • Noncanonical amino acid

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Author information

Authors and Affiliations

  1. Department of Chemistry, Texas A&M University, College Station, TX, USA

    Jeffery M. Tharp & Wenshe R. Liu

Authors
  1. Jeffery M. Tharp
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  2. Wenshe R. Liu
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Corresponding author

Correspondence to Wenshe R. Liu .

Editor information

Editors and Affiliations

  1. Structural and Computational Biology Unit & Cell Biology and Biophysics Unit, EMBL, Heidelberg, Germany

    Edward A. Lemke

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Tharp, J.M., Liu, W.R. (2018). Using Amber and Ochre Nonsense Codons to Code Two Different Noncanonical Amino Acids in One Protein Gene. In: Lemke, E. (eds) Noncanonical Amino Acids. Methods in Molecular Biology, vol 1728. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7574-7_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7574-7_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7573-0

  • Online ISBN: 978-1-4939-7574-7

  • eBook Packages: Springer Protocols

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