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Orthogonal Protein Translation Using Pyrrolysyl-tRNA Synthetases for Single- and Multiple-Noncanonical Amino Acid Mutagenesis

Part of the Advances in Biochemical Engineering/Biotechnology book series (ABE,volume 162)

Abstract

To date, the two systems most extensively used for noncanonical amino acid (ncAA) incorporation via orthogonal translation are based on the Methanococcus jannaschii TyrRS/tRNA TyrCUA and the Methanosarcina barkeri/Methanosarcina mazei PylRS/tRNA PylCUA pairs. Here, we summarize the development and usage of the pyrrolysine-based system for orthogonal translation, a process that allows for the recombinant production of site-specifically labeled proteins and peptides. Via stop codon suppression in Escherichia coli and mammalian cells, genetically encoded biomolecules can be equipped with a great diversity of chemical functionalities including click chemistry handles, post-translational modifications, and photocaged sidechains.

Keywords

  • Expanded genetic code
  • Noncanonical amino acid
  • Orthogonal translation
  • Pyrrolysyl-tRNA synthetase
  • Stop codon suppression

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Correspondence to Nediljko Budisa .

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Baumann, T., Exner, M., Budisa, N. (2016). Orthogonal Protein Translation Using Pyrrolysyl-tRNA Synthetases for Single- and Multiple-Noncanonical Amino Acid Mutagenesis. In: Zhao, H., Zeng, AP. (eds) Synthetic Biology – Metabolic Engineering. Advances in Biochemical Engineering/Biotechnology, vol 162. Springer, Cham. https://doi.org/10.1007/10_2016_37

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