Figure 11.
Effect of AMP-PNP, a non-hydrolyzable analog of ATP, on mRNA translation in a cell-free system. The wheat germ cell-free system programmed with 5′UTRObelin-GFP-3′UTRTMV mRNA (500 nM) was incubated in batch as described in ‘Materials and Methods’ section at the ATP concentration of 0.1 mM. AMP-PNP was added to the reaction mixture to the final concentration of 2 mM in 10 (filled diamond) or 60 (filled square) minutes after translation start. In control reactions the same amount of ATP was added after 10 min (open square), or the reaction was conducted without any addition (open circle). Arrows indicate the time of AMP-PNP or ATP addition. The synthesis of GFP was recorded by the measurement of fluorescence in 2 µl aliquots taken at the indicated time points.