Cell-free translation reconstituted with purified components

Yoshihiro Shimizu, Akio Inoue, Yukihide Tomari, Tsutomu Suzuki, Takashi Yokogawa, Kazuya Nishikawa, Takuya Ueda

Research output: Contribution to journalArticle

1014 Citations (Scopus)

Abstract

We have developed a protein-synthesizing system reconstituted from recombinant tagged protein factors purified to homogeneity. The system was able to produce protein at a rate of about 160 μg/ml/h in a batch mode without the need for any supplementary apparatus. The protein products were easily purified within 1 h using affinity chromatography to remove the tagged protein factors. Moreover, omission of a release factor allowed efficient incorporation of an unnatural amino acid using suppressor transfer RNA (tRNA).

Original languageEnglish
Pages (from-to)751-755
Number of pages5
JournalNature Biotechnology
Volume19
Issue number8
DOIs
Publication statusPublished - 2001 Aug 20
Externally publishedYes

Fingerprint

Proteins
Affinity chromatography
Recombinant proteins
Transfer RNA
Affinity Chromatography
Recombinant Proteins
Amino acids
Amino Acids

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Cite this

Shimizu, Y., Inoue, A., Tomari, Y., Suzuki, T., Yokogawa, T., Nishikawa, K., & Ueda, T. (2001). Cell-free translation reconstituted with purified components. Nature Biotechnology, 19(8), 751-755. https://doi.org/10.1038/90802

Cell-free translation reconstituted with purified components. / Shimizu, Yoshihiro; Inoue, Akio; Tomari, Yukihide; Suzuki, Tsutomu; Yokogawa, Takashi; Nishikawa, Kazuya; Ueda, Takuya.

In: Nature Biotechnology, Vol. 19, No. 8, 20.08.2001, p. 751-755.

Research output: Contribution to journalArticle

Shimizu, Y, Inoue, A, Tomari, Y, Suzuki, T, Yokogawa, T, Nishikawa, K & Ueda, T 2001, 'Cell-free translation reconstituted with purified components', Nature Biotechnology, vol. 19, no. 8, pp. 751-755. https://doi.org/10.1038/90802
Shimizu Y, Inoue A, Tomari Y, Suzuki T, Yokogawa T, Nishikawa K et al. Cell-free translation reconstituted with purified components. Nature Biotechnology. 2001 Aug 20;19(8):751-755. https://doi.org/10.1038/90802
Shimizu, Yoshihiro ; Inoue, Akio ; Tomari, Yukihide ; Suzuki, Tsutomu ; Yokogawa, Takashi ; Nishikawa, Kazuya ; Ueda, Takuya. / Cell-free translation reconstituted with purified components. In: Nature Biotechnology. 2001 ; Vol. 19, No. 8. pp. 751-755.
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