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. Author manuscript; available in PMC: 2022 Jun 17.
Published in final edited form as: Methods Mol Biol. 2017;1657:331–345. doi: 10.1007/978-1-4939-7240-1_26

Fig. 1.

Fig. 1

UV photo-cross-linking using genetic code expansion. (a) Structures and mechanism of photoactivation of pAzF and Bpa noncanonical amino acids. (b) Expression system and plasmids for the incorporation of noncanonical amino acids into a protein of interest. This sytem requires two plasmids: the expression plasmid (pBrew) in which the protein of interest containing the TAG amber stop codon is C-terminally fused to superfolder GFP, and the machinery plasmid (pDule2) that expresses the orthogonal tRNA synthetase that recognizes and charges its cognate amber-suppressing tRNA (also expressed on this plasmid) with the noncanonical amino acid. Only full-length protein containing the noncanonical amino acid is fluorescent