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. Author manuscript; available in PMC: 2020 Dec 19.
Published in final edited form as: Cell Chem Biol. 2019 Nov 6;26(12):1743–1754.e9. doi: 10.1016/j.chembiol.2019.10.008

Figure 6. One-Pot ncAA Incorporation into sfGFP Using 759.T7.Opt Lysates.

Figure 6.

(A) sfGFP produced in vitro from cell extracts derived from induced BL21 Star (DE3) cells as well as 759.T7.Opt cells, without supplementation with purified T7RNAP. The indicated sfGFP amber mutant variants were synthesized in the presence of the complete pAcF OTS. At least three independent reactions were performed per condition, and one standard deviation is shown.

(B) Mass spectra of the +28 charge states of the indicated sfGFP variants obtained by top-down mass spectrometry illustrating site-specific incorporation of one to several pAcF residues. Deconvoluted experimental (Exper) and theoretical (Theor) monoisotopic mass values for each variant are listed. Major peaks are highlighted in each spectrum with color.

See also Figure S2.