Figure 5.
Effect of tRNA addition on uAUG recognition in vitro. Transcripts encoding different GFP forms, indicated above each panel, were translated in rabbit reticulocyte lysate complemented with 0–200 ng of tRNA per 5 µl assay. The resulting proteins were separated on a 15% 30:0.18 acrylamide/bisacrylamide gel. The gel was dried, and incorporated [35S]methionine was visualised by exposure to film (A). The bands were quantified. Percentage upstream initiation was calculated by dividing the amount of elongated GFP (#1 GFP or #23 GFP) by the total amount of GFP (B and C). The data were corrected for the number of methionines present in the different GFP forms. The open and closed circles represent two independent experiments.