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. Author manuscript; available in PMC: 2015 Jan 3.
Published in final edited form as: Angew Chem Int Ed Engl. 2014 Jan 3;53(1):319–323. doi: 10.1002/anie.201308584

Scheme 1.

Scheme 1

Codon recoding with Sec. A) Synthesis of the E. coli Sec-containing formate dehydrogenase (FDH) from the wild type fdhF gene requires recoding the UGA stop codon at position 140 to Sec by Sec-tRNASec, elongation factor SelB, and the SECIS mRNA hairpin. Replacing the UGA with any of the 64 NNN triplets at codon 140 in fdhF and co-expressing a cognate Sec-tRNASecNNN is expected to yield SelB and SECIS-dependent sense codon-recoded Sec-containing FDHH (B) that may compete with canonical AA insertion (C) directed by native aminoacyl-tRNAs and EF-Tu dependent protein synthesis. In this case (C), SECIS is still present in the fdhF mRNA, but SECIS is not expected to interact with EF-Tu directed decoding.