TY - JOUR
T1 - Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli
AU - Chumpolkulwong, Namthip
AU - Sakamoto, Kensaku
AU - Hayashi, Akiko
AU - Iraha, Fumie
AU - Shinya, Naoko
AU - Matsuda, Natsuko
AU - Kiga, Daisuke
AU - Urushibata, Akiko
AU - Shirouzu, Mikako
AU - Oki, Kenji
AU - Kigawa, Takanori
AU - Yokoyama, Shigeyuki
N1 - Funding Information:
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.
PY - 2006/3
Y1 - 2006/3
N2 - 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.
AB - 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.
KW - Cell-free protein synthesis
KW - Codon usage
KW - Consecutive codons
KW - Glutathione S-transferase
KW - Minor tRNA
UR - http://www.scopus.com/inward/record.url?scp=33747682140&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33747682140&partnerID=8YFLogxK
U2 - 10.1007/s10969-006-9007-y
DO - 10.1007/s10969-006-9007-y
M3 - Article
C2 - 16703415
AN - SCOPUS:33747682140
SN - 1345-711X
VL - 7
SP - 31
EP - 36
JO - Journal of Structural and Functional Genomics
JF - Journal of Structural and Functional Genomics
IS - 1
ER -