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Efficient incorporation of unnatural amino acids into proteins in Escherichia coli

Nature Methods volume 3, pages 263–265 (2006)Cite this article

Abstract

We have developed a single-plasmid system for the efficient bacterial expression of mutant proteins containing unnatural amino acids at specific sites designated by amber nonsense codons. In this system, multiple copies of a gene encoding an amber suppressor tRNA derived from a Methanocaldococcus jannaschii tyrosyl-tRNA (MjtRNATyrCUA) are expressed under control of the proK promoter and terminator, and a gene encoding the desired mutant M. jannaschii tyrosyl-tRNA synthetase (MjTyrRS) is expressed under control of a mutant glnS (glnS') promoter.

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Figure 1: Improvement in suppression efficiencies.
Figure 2: Efficiency and fidelity of the new system.

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Acknowledgements

This work was supported by Department of Energy (ER46051). This is manuscript # 17854-CH of the Scripps Research Institute.

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  1. Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, 92037, California, USA

    Youngha Ryu & Peter G Schultz

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  1. Youngha Ryu
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  2. Peter G Schultz
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Correspondence to Peter G Schultz.

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The authors declare no competing financial interests.

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Ryu, Y., Schultz, P. Efficient incorporation of unnatural amino acids into proteins in Escherichia coli. Nat Methods 3, 263–265 (2006). https://doi.org/10.1038/nmeth864

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