Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli

Namthip Chumpolkulwong, Kensaku Sakamoto, Akiko Hayashi, Fumie Iraha, Naoko Shinya, Natsuko Matsuda, Daisuke Kiga, Akiko Urushibata, Mikako Shirouzu, Kenji Oki, Takanori Kigawa, Shigeyuki Yokoyama

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We analyzed the effect of nine 'rare' codons (AGA, AGG, AUA, CCC, CGA, CGG, CUA, GGA, and UUA) on gene expression in an Escherichia coli coupled transcription/translation cell-free system, in comparison with a cell-based expression system. Each reporter gene contained five consecutive repeats of a rare codon, or in some experiments, three consecutive repeats. The cell-free expression of the genes bearing the codons CGA, CUA, GGA, and UUA was not affected, although these codons, except for GGA, were inefficiently translated in E. coli cells. Translation of the remaining five codons (AGA, AGG, AUA, CCC, and CGG) was severely reduced in both systems, and was remarkably facilitated in the cell-free system based on an S30 extract from the E. coli cells overproducing 'minor' tRNAs for these codons.

Original languageEnglish
Pages (from-to)31-36
Number of pages6
JournalJournal of structural and functional genomics
Volume7
Issue number1
DOIs
Publication statusPublished - 2006 Mar 1

Keywords

  • Cell-free protein synthesis
  • Codon usage
  • Consecutive codons
  • Glutathione S-transferase
  • Minor tRNA

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Genetics

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  • Cite this

    Chumpolkulwong, N., Sakamoto, K., Hayashi, A., Iraha, F., Shinya, N., Matsuda, N., Kiga, D., Urushibata, A., Shirouzu, M., Oki, K., Kigawa, T., & Yokoyama, S. (2006). Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli. Journal of structural and functional genomics, 7(1), 31-36. https://doi.org/10.1007/s10969-006-9007-y