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Preparation method forEscherichia coli S30 extracts completely dependent upon tRNA addition to catalyze cell-free protein synthesis

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Abstract

A simple method for depletingE. coli S30 extracts of endogenous tRNA has been developed. An ethanolamine-Sepharose® column equilibrated with water selectively captured the tRNA molecules inE. coli S30 extracts. As a result, S30 extracts filtered through this column became completely dependent upon the addition of exogenous tRNA to mediate cell-free protein synthesis reactions. We anticipate that the procedures developed and described will be particularly useful forin vitro suppression reaction studies designed to introduce unnatural amino acids into protein molecules.

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Authors and Affiliations

  1. Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University, 151-742, Seoul, Korea

    Jin-Ho Ahn & Cha-Yong Choi

  2. School of Chemical and Biological Engineering, College of Engineering, Seoul National University, 151-742, Seoul, Korea

    Mi-Yeon Hwang, In-Seok Oh & Cha-Yong Choi

  3. Department of Chemical System Engineering, Hongik University, Jochiwon, 339-701, Chungnam, Korea

    Kyung-Moon Park

  4. Department of Fine Chemical Engineering and Chemistry, Chungnam National University, 305-764, Daejeon, Korea

    Geun-Hee Hahn & Dong-Myung Kim

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  1. Jin-Ho Ahn
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  2. Mi-Yeon Hwang
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  4. Kyung-Moon Park
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  5. Geun-Hee Hahn
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  7. Dong-Myung Kim
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Correspondence to Dong-Myung Kim.

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Ahn, JH., Hwang, MY., Oh, IS. et al. Preparation method forEscherichia coli S30 extracts completely dependent upon tRNA addition to catalyze cell-free protein synthesis. Biotechnol. Bioprocess Eng. 11, 420–424 (2006). https://doi.org/10.1007/BF02932309

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  • DOI: https://doi.org/10.1007/BF02932309

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