FIGURE 5.

Biosynthesis and incorporation of OMeY in HEK293T protein. (a) pLenti‐MfnG plasmid encodes a CMV promoter‐driven MfnG. pAcBac2.tR4‐OMeYRS/EGFP* encodes a CAG‐promoter‐driven EGFP mutant, CMV‐promoter‐driven EcTyrRS, and two copies of E. coli tRNA_Tyr ^CUA and B. stearothermophilus tRNA_Tyr ^CUA. (b) Fluorescence imaging of EGFP signal of HEK293T cells with fed OMeY and HEK293T‐MfnG transfected with pAcBac2.tR4‐OMeYRS/EGFP*. (c) SDS‐PAGE gel analysis of purified EGFP mutants with OMeY from cells with exogenously fed OMeY (+OMeY) or biosynthesized OMeY (bio). (d) Mass spectra of EGFP mutants purified from HEK293T (left) or HEK293T‐MfnG (right)