Biotechnology and Bioprocess Engineering:BBE

  • 제11권5호
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  • Pages.420-424
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  • 2006
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  • 1226-8372(pISSN)
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  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Preparation Method for Escherichia coliS30 Extracts Completely Dependent upon tRNA Addition to Catalyze Cell-free Protein Synthesis

  • Ahn, Jin-Ho (Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University) ;
  • Hwang, Mi-Yeon (School of Chemical and Biological Engineering, College of Engineering, Seoul National University) ;
  • Oh, In-Seok (School of Chemical and Biological Engineering, College of Engineering, Seoul National University) ;
  • Park, Kyung-Moon (Department of Chemical System Engineering, Hongik University) ;
  • Hahn, Geun-Hee (Department of Fine Chemical Engineering and Chemistry, Chungnam National University) ;
  • Choi, Cha-Yong (Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University) ;
  • Kim, Dong-Myung (Department of Fine Chemical Engineering and Chemistry, Chungnam National University)
  • 발행 : 2006.10.30
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초록

A simple method for depleting E. coliS30 extracts of endogenous tRNA has been developed. An $ethanolamine-Sepharose^{(R)}$ column equilibrated with water selectively captured the tRNA molecules in E. 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 for in vitro suppression reaction studies designed to introduce unnatural amino acids into protein molecules.

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