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.
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Abbreviations
- GST:
-
glutathione S-transferase
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Acknowledgements
We thank Dr. Takashi Yabuki and Mr. Takayoshi Matsuda for technical advice and valuable discussions. This work was supported by the RIKEN Structural Genomics/Proteomics Initiative (RSGI), the National Project on Protein Structural and Functional Analyses and the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Chumpolkulwong, N., Sakamoto, K., Hayashi, A. et al. Translation of ‘rare’ Codons in a Cell-free Protein Synthesis System from Escherichia coli . J Struct Funct Genomics 7, 31–36 (2006). https://doi.org/10.1007/s10969-006-9007-y
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DOI: https://doi.org/10.1007/s10969-006-9007-y