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

  • Jin-Ho Ahn
  • Mi-Yeon Hwang
  • In-Seok Oh
  • Kyung-Moon Park
  • Geun-Hee Hahn
  • Cha-Yong Choi
  • Dong-Myung Kim
Article

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.

Keywords

cell-free protein synthesis unnatural amino acid in vitro suppression tRNA chloramphenicol acetyltransferase erythropoietin 

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Copyright information

© The Korean Society for Biotechnology and Bioengineering 2006

Authors and Affiliations

  • Jin-Ho Ahn
    • 1
  • Mi-Yeon Hwang
    • 2
  • In-Seok Oh
    • 2
  • Kyung-Moon Park
    • 3
  • Geun-Hee Hahn
    • 4
  • Cha-Yong Choi
    • 1
    • 2
  • Dong-Myung Kim
    • 4
  1. 1.Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of EngineeringSeoul National UniversitySeoulKorea
  2. 2.School of Chemical and Biological Engineering, College of EngineeringSeoul National UniversitySeoulKorea
  3. 3.Department of Chemical System EngineeringHongik UniversityChungnamKorea
  4. 4.Department of Fine Chemical Engineering and ChemistryChungnam National UniversityDaejeonKorea

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