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. Author manuscript; available in PMC: 2019 Aug 8.
Published in final edited form as: Bioconjug Chem. 2018 Feb 19;29(4):1209–1218. doi: 10.1021/acs.bioconjchem.8b00004

Figure 1.

Figure 1.

Illustration of FTL-X variant construction and production in E. coli. Human FTL was modified by PCR cloning method to incorporate amber stop codon (TAG) at desired position in the FTL cDNA. The FTL-X plasmid was cotransformed along with the pEVOL-AzF vector, which encodes the tRNA/aaRS pair for para-azido-L-phenylalanine (pAzF),33 into competent E. coli. Large scale production was performed by IPTG induction in the presence of pAzF. FTL-X was extracted by sonication and heat precipitation followed by purification by FPLC.