Evolution of Amber Suppressor tRNAs for Efficient Bacterial Production of Proteins Containing Nonnatural Amino Acids

Authors

  • Jiantao Guo,

    1. Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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    • These authors contributed equally to this work.

  • Charles E. Melançon III,

    1. Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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    • These authors contributed equally to this work.

  • Hyun Soo Lee,

    1. Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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  • Dan Groff,

    1. Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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  • Peter G. Schultz Prof.

    1. Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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  • This work is supported by NIH grants R01 GM062159 (P.G.S.) and F32 GM080067 (C.E.M.), and the Skaggs Institute for Chemical Biology.

Abstract

original image

Applied evolution: Regions of the M. jannaschii tyrosyl tRNACUA thought to interact with elongation factor Tu were randomized, and the resulting tRNA libraries were subjected to in vitro evolution. The tRNAs identified resulted in significantly improved yields of proteins containing nonnatural amino acids. In some cases, the degree of improvement varied in an amino acid dependent manner.

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