Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Optimization of Programmed Suppression in a Cell-Free Protein Synthesis System with Unnatural Amino Acid S-(2-Nitrobenzyl)cysteine

  • HYUN JOO (Department of Molecular Science and Technology/Life Sciences, Ajou University) ;
  • KANG, TAEK JIN (School of Chemical Engineering, College of Engineering, Seoul National University) ;
  • HUI KYOUNG SONG (Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University) ;
  • JIN HO AHN (Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University) ;
  • CHA YONG CHOI (School of Chemical Engineering, College of Engineering, Seoul National University, Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University)
  • Published : 2003.06.01
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Abstract

Unnatural amino acid S-(2-nitrobenzyl)cysteine was incorporated into human erythropoietin by using a programmed suppression of nonsense codon in a cell-free protein synthesis system. Several controlling factors affecting the operational efficiency of the suppression were investigated and optimized. The amount of suppressor tRNA and the concentration of $Mg^2+$ were crucial not only for the efficiency but also for the control of the exact suppression. In addition, some general optimization factor are reported in order to improve the efficiency in an unnatural amino acid mutagenesis.

Keywords

References

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