title: Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli.
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.
authors.affiliation: Genomic Sciences Center, RIKEN, Tsurumi, Yokohama 230-0045, Japan.
authors.forename: Namthip
authors.initials: N
authors.lastname: Chumpolkulwong
authors.forename: Kensaku
authors.initials: K
authors.lastname: Sakamoto
authors.forename: Akiko
authors.initials: A
authors.lastname: Hayashi
authors.forename: Fumie
authors.initials: F
authors.lastname: Iraha
authors.forename: Naoko
authors.initials: N
authors.lastname: Shinya
authors.forename: Natsuko
authors.initials: N
authors.lastname: Matsuda
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kiga
authors.forename: Akiko
authors.initials: A
authors.lastname: Urushibata
authors.forename: Mikako
authors.initials: M
authors.lastname: Shirouzu
authors.forename: Kenji
authors.initials: K
authors.lastname: Oki
authors.forename: Takanori
authors.initials: T
authors.lastname: Kigawa
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
mesh_terms: 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
chemical_list: D003062:Codon; D015801:Helminth Proteins; D011994:Recombinant Proteins; D005982:Glutathione Transferase