Tunable translational control using site-specific unnatural amino acid incorporation in Escherichia coli

Translation of target gene transcripts in Escherichia coli harboring UAG amber stop codons can be switched on by the amber-codon-specific incorporation of an exogenously supplied unnatural amino acid, 3-iodo- L -tyrosine. Here, we report that this translational switch can control the translational efficiency at any intermediate magnitude by adjustment of the 3-iodo- L -tyrosine concentration in the medium, as a tunable translational controller. The translational efficiency of a target gene reached maximum levels with 10 -5 M 3-iodo- L -tyrosine, and intermediate levels were observed with suboptimal concentrations (approximately spanning a 2-log 10 concentration range, 10 -7 to 10 -5 M). Such intermediate-level expression was also confirmed in individual bacteria.