Cell-free translation reconstituted with purified components

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

doi:10.1038/90802

Cell-free translation reconstituted with purified components

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

^*この研究の対応する著者

研究成果: Article › 査読

1268 被引用数 (Scopus)

抄録

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).

本文言語	English
ページ（範囲）	751-755
ページ数	5
ジャーナル	Nature biotechnology
巻	19
号	8
DOI	https://doi.org/10.1038/90802
出版ステータス	Published - 2001
外部発表	はい

ASJC Scopus subject areas

バイオテクノロジー
バイオエンジニアリング
応用微生物学とバイオテクノロジー
分子医療
生体医工学

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10.1038/90802

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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).",

author = "Yoshihiro Shimizu and Akio Inoue and Yukihide Tomari and Tsutomu Suzuki and Takashi Yokogawa and Kazuya Nishikawa and Takuya Ueda",

note = "Funding Information: Note: Supplementary information can be found on the Nature Biotechnology website in Web Extras (http://biotech.nature.com/web_extras) Acknowledgments We thank S. Yokoyama, The University of Tokyo, for providing the expression plasmid encoding the ArgRS gene. This work was supported by a grant (JSPS-RFTF 96100306) from The Japan Society for the Promotion of Science (to T.U.).",

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AU - Shimizu, Yoshihiro

AU - Inoue, Akio

AU - Tomari, Yukihide

AU - Suzuki, Tsutomu

AU - Yokogawa, Takashi

AU - Nishikawa, Kazuya

AU - Ueda, Takuya

N1 - Funding Information: Note: Supplementary information can be found on the Nature Biotechnology website in Web Extras (http://biotech.nature.com/web_extras) Acknowledgments We thank S. Yokoyama, The University of Tokyo, for providing the expression plasmid encoding the ArgRS gene. This work was supported by a grant (JSPS-RFTF 96100306) from The Japan Society for the Promotion of Science (to T.U.).

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Cell-free translation reconstituted with purified components

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