Fig. 5: Construction of UAA-dependent synthetic auxotroph. | Nature Communications

Fig. 5: Construction of UAA-dependent synthetic auxotroph.

From: Directed-evolution of translation system for efficient unnatural amino acids incorporation and generalizable synthetic auxotroph construction

Fig. 5

a Amber suppression efficiency of the AzF system and the BTA system is tested in the presence of indicated concentration of AzF or BTA. Statistical significance is quantified with one-way ANOVA (***p < 0.001, p = 5.74E–7 and 4.24E–6, respectively). b Schematic illustration of the procedure of engineering UAA-dependent synthetic auxotroph. The pTKRed vector carries the lambda Red recombinase system, and the pChira vector carries AzFRS-2 (under the control of oxb20 promoter), 3C11-chPheT (under the control of lpp promoter), and the dnaN-TAG gene (under the control of tac promoter). c Escape frequencies of E. coli strains at day 14 post-plating with indicated numbers of TAG in the dnaN gene. The black line represents escape frequency below the detection limit. d Growth curve of various UAA-dependent synthetic auxotrophs in the presence or absence of AzF. e Escape frequencies of E. coli strains at day 14 post-plating with three TAGs in the indicated genes. The black line represents escape frequency below the detection limit. Error bars represents ±standard error of the mean from three biologically independent experiments. The box plots showing the minima, maxima, center, bounds of box and whiskers and percentiles. Source data are provided as a Source Data file.

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