Fig. 5.

TBK incorporation into proteins, using genetically expanded cell-free protein synthesis. a) Cell-free kinetics of GFP Y35X production in the presence of different TBK concentrations. b) Cell-free kinetics of RFP K15X production in the presence and absence of TBK. c) ESI mass spectra of purified RFP K15X with TBK, before and after boc-deprotection. d) Anti-ubiquitin Western blot of genetically expanded cell-free protein synthesis of the 36.4 kDa Ub-Intein-CBD construct. In the left panel: Cell-free reactions of incorporation of PrK and TBK in different concentrations. The right panel presents a comparison between TBK incorporated into two different branching sites, K48X and K63X, in the ubiquitin construct in the presence or absence of TBK, the bend in site 63 in the absence of TBK could be a result of stop codon readthrough.