Fig. 2. Incorporation efficiency and structural tolerance of NBAK in mNG. (A) Schematic of the 192 selected candidate modification sites out of the 236 amino acid residues in the mNG β-barrel. (B) GCE-mediated NBAK incorporation and efficiency assay via fluorescence ratio (+/−UAA). (C) and (D) Incorporation efficiency and expression levels of 1-NBAK (C) and 2-NBAK (D) across 192 sites, yielding 105 and 87 bright variants, respectively, for subsequent photostability screening (see Fig. S1 and S3 for detailed residue positions). Schematic diagrams were created with BioRender.com. mNG, mNeonGreen; GCE, genetic code expansion.
