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. Author manuscript; available in PMC: 2016 Dec 1.
Published in final edited form as: Nat Biotechnol. 2015 Nov 16;33(12):1272–1279. doi: 10.1038/nbt.3372

Figure 4.

Figure 4

Evaluation of multi-site nsAA incorporation by evolved aaRS variants expressed on multi-copy plasmids. Production of GFP(3UAG) (a), ELP(10UAG)-GFP (b) and ELP(30UAG)-GFP (c) by progenitor and evolved orthogonal translation systems expressed on multi-copy plasmids in the genomically recoded organism compared with WT (no UAG) proteins. Production of ELP(30UAG)-GFP by progenitor and evolved orthogonal translation systems expressed on multi-copy plasmids in the genomically recoded organism in the presence of variable pAcF (d) or pAzF (e) concentrations (n = 3, error bars, s.d. *P < 0.05 indicates comparison of evolved aaRS with pAcFRS in the +pAcF or +pAzF condition). (f) Efficiency and specificity of progenitor and evolved pAcFRS and pAzFRS variants for 14 phenylalanine derivatives, as assayed by ELP(30UAG)-GFP fluorescence. Data shown are the average of two independent experiments each with n = 3.