Fig. 5. Site-specific incorporation of two ncAAs into EGFP-39-TAG-151-TGA using EcLeuTAG + PylTGA dual suppression system. (A) Scheme of the 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-EcLeuTAG-EGFP** and pAcBac1-PylTGA (Fig. S5†), in the presence or absence of the two ncAAs. (C) ESI-MS of the IMAC-purified protein confirms a mass consistent with the incorporation of the desired ncAAs.
