Fig. 4. Site-specific incorporation of BocK and OMeY into EGFP-39-TAG-151-TGA. (A) The scheme of dual suppression. (B) Expression of the full-length EGFP reporter, measured by its characteristic fluorescence in cell-free extract, upon transfecting HEK293T cells with pAcBac3-EcTyrTGA-EGFP** and pAcBac1-PylTAG (Fig. S5†), in the presence or absence of the two ncAAs. (C) Fluorescence microscopy images of the cells for the same experiment. (D) ESI-MS of the IMAC-purified protein confirms a mass consistent with the incorporation of the desired ncAAs.
