Optimization of programmed suppression in a cell-free protein synthesis system with unnatural amino acid S-(2-nitrobenzyl) cysteine

Taek Jin Kang, Hui Kyoung Song, Jin Ho Ahn, Cha Yong Choi, Hyun Joo

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

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 Mg2+ were crucial not only for the efficiency but also for the control of the exact suppression. In addition, sorne general optimization factors are reported in order to improve the efficiency in an unnatural amino acid mutagenesis.

Original languageEnglish
Pages (from-to)344-347
Number of pages4
JournalJournal of Microbiology and Biotechnology
Volume13
Issue number3
StatePublished - Jun 2003

Keywords

  • Cell-free protein synthesis
  • Erythropoietin
  • Salt concentration optimization
  • Suppressor tRNA
  • Unnatural amino acid mutagenesis

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