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An artificially split class 3 intein

MicroPubl Biol .. 2023-09; 
Tia M Ariagno , John S Smetana , Christopher W Lennon
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Gene Synthesis lasmids used in this study (pET-Duet1 MBP-DnaB1 IntN / DnaB1IntC-sfGFP; pET-Duet1 MBP-DnaB1 IntN / DnaB1IntCsfGFP C118A) were synthesized (Genscript) and were transformed into Escherichia coliBL21(DE3) for protein expression. Get A Quote
PCR Cloning and Subcloning Get A Quote


Inteins excise themselves from precursor polypeptides through protein splicing, joining N- and C-exteins with a peptide bond. Split inteins are expressed as separate polypeptides that undergo protein trans splicing (PTS). Here, we demonstrate PTS can be achieved using an artificially split class 3 intein. Because class 3 inteins use an internal initiating nucleophile near the C-extein junction, rather than the first residue of the intein, both catalytic nucleophiles are present on a single polypeptide. This results in a compact arrangement of catalytic nucleophiles for PTS compared to the standard arrangement for split class 1 inteins.