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A highly flexible tRNA acylation method for non-natural polypeptide synthesis

Nature Methods volume 3, pages 357359 (2006) | Download Citation

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  • A Corrigendum to this article was published on 01 August 2006

This article has been updated

Abstract

Here we describe a de novo tRNA acylation system, the flexizyme (Fx) system, for the preparation of acyl tRNAs with nearly unlimited selection of amino and hydroxy acids and tRNAs. The combination of the Fx system with an appropriate cell-free translation system allows us to readily perform mRNA-encoded synthesis of proteins and short polypeptides involving multiple non-natural amino acids.

*Note: In the version of this article initially published, the authors did not declare competing financial interests. They have filed a patent covering some of the information described in the paper and now declare competing financial interests.   This error has been corrected in the PDF version of the article.

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  • 19 July 2006

    In the version of this article initially published, the authors did not declare competing financial interests. They have filed a patent covering some of the information described in the paper and now declare competing financial interests. This error has been corrected in the PDF version of the article.

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Acknowledgements

We thank M. Komiyama for the use of MALDI instrumentation. This work was supported by grants from Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research (S) (16101007) and from the US National Institutes of Health (GM59159).

Author information

Affiliations

  1. Research Center for Advanced Science and Technology, The University of Tokyo, 153-8904 Tokyo, Japan.

    • Hiroshi Murakami
    • , Atsushi Ohta
    • , Hiroshi Ashigai
    •  & Hiroaki Suga
  2. Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 113-8656 Tokyo, Japan.

    • Atsushi Ohta
    • , Hiroshi Ashigai
    •  & Hiroaki Suga

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Hiroaki Suga.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Comparison of activity in flexizymes.

  2. 2.

    Supplementary Fig. 2

    In vitro selection using Hbi-DBE.

  3. 3.

    Supplementary Fig. 3

    In vitro selection using Phe-CME.

  4. 4.

    Supplementary Fig. 4

    Fx-independent acylation of tRNA.

  5. 5.

    Supplementary Fig. 5

    Acid substrates used in this study.

  6. 6.

    Supplementary Fig. 6

    Site-specific incorporation of various acids into GFP.

  7. 7.

    Supplementary Table 1

    Yields of acyl-tRNAs.

Word documents

  1. 1.

    Supplementary Methods

  2. 2.

    Supplementary Note

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DOI

https://doi.org/10.1038/nmeth877

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