Improved heterologous gene expression in Escherichia coli by optimization of the AT-content of codons immediately downstream of the initiation codon.
Escherichia coli is the most frequently used host for heterologous gene expression. This study focuses on the effect of AT-rich codons immediately downstream of the initiation codon of the target gene. The third to sixth codons of ndx3, a Nudix hydrolase gene from Thermus thermophilus HB8, were engineered by introducing several silent mutations. As a result, the expression level of ndx3 increased in proportion to the AT-content in the third to sixth codons. This result suggests that incorporation of AT-rich codons can be utilized as a general strategy for improving the expression efficiency of a recombinant protein.
Nishikubo
Takashi
T
Department of Biology, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.
Nakagawa
Noriko
N
Kuramitsu
Seiki
S
Masui
Ryoji
R
D020856:AT Rich Sequence / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D018387:Codon, Initiator / Q000235:genetics*; D004926:Escherichia coli; D015870:Gene Expression; D011755:Pyrophosphatases / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics; D000096964:Nudix Hydrolases
D001426:Bacterial Proteins; D018387:Codon, Initiator; D011755:Pyrophosphatases; D011994:Recombinant Proteins; D000096964:Nudix Hydrolases