Figure 1.

Effect of increasing concentrations of 2A on PRF efficiency. (A) Mutations introduced into the TMEV PRF signal in reporter plasmid pDluc. Nucleotide coordinates refer to the TMEV genome. All constructs (including WT) contain a U to C mutation in the loop to remove the −1 frame stop codon UAA (red). (B–E) RNAs containing the TMEV PRF signal were translated in WG (B) or RRL (D) in the presence of increasing concentrations of 2A, or 2A dialysis buffer (DB). Products generated by ribosomes that do not frameshift (stop) or that enter the −1 reading frame (−1 FS) are indicated. Frameshifting efficiencies estimated from densitometry are indicated above lanes. Markers are in lane 1. The PRF efficiencies are shown in (C) and (E).