Design, synthesis and expression of a human interleukin-2 gene incorporating the codon usage bias found in highly expressed Escherichia coli genes.
A synthetic gene encoding human interleukin-2 (IL-2) was designed such that the codon usage bias resembled that found in highly expressed Escherichia coli genes. The percentage of preferred codons was increased from 43% in the native cDNA sequence to 85% in the synthetic sequence. The cDNA and synthetic IL-2 genes were placed under the control of the trc promoter and expressed in E. coli JM101. While Northern blot analysis of IL-2 mRNA from each genetic construct demonstrated equivalent message half-lives, immunoblot and bioactivity analyses showed the synthetic gene to direct the synthesis of up to 16 times more IL-2 than the native cDNA sequence.
Williams
D P
DP
Evans Department of Clinical Research, University Hospital, Boston, MA.
Regier
D
D
Akiyoshi
D
D
Genbauffe
F
F
Murphy
J R
JR
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003062:Codon; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005813:Genes, Synthetic; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic
D003062:Codon; D007376:Interleukin-2; D012333:RNA, Messenger