Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli.
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.
Chumpolkulwong
Namthip
N
Genomic Sciences Center, RIKEN, Tsurumi, Yokohama 230-0045, Japan.
Sakamoto
Kensaku
K
Hayashi
Akiko
A
Iraha
Fumie
F
Shinya
Naoko
N
Matsuda
Natsuko
N
Kiga
Daisuke
D
Urushibata
Akiko
A
Shirouzu
Mikako
M
Oki
Kenji
K
Kigawa
Takanori
T
Yokoyama
Shigeyuki
S
D000818:Animals; D002474:Cell-Free System; D003062:Codon / Q000235:genetics; D004926:Escherichia coli; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D015801:Helminth Proteins / Q000096:biosynthesis* / Q000235:genetics; D014176:Protein Biosynthesis / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics; D013045:Species Specificity
D003062:Codon; D015801:Helminth Proteins; D011994:Recombinant Proteins; D005982:Glutathione Transferase