Mapping of sequences in the 5’ region and 3’ UTR of tomato ringspot virus RNA2 that facilitate cap-independent translation of reporter transcripts in vitro
Fig 5
Cap analog inhibits translation of uncapped VRV transcripts.
(A) Cap analog was added at increasing concentrations ranging from 0 to 150 picomoles per 50 μl translation reaction volume that also contained one picomole of reporter transcripts (uncapped VRV or capped ARA) and the luminescence activity of the translation reactions was measured. Experiments were repeated four times using varying concentrations of cap and a representative result is shown. Error bars represent the standard deviation of three technical repeats. (B) Translation inhibition of uncapped VRV or capped ARA (cARA) transcripts in the presence of a 125-molar excess of cap analog. Reactions were conducted as above. Translation of 1 picomole of either transcript (VRV or cARA) was reduced significantly in the presence of 125 picomoles of cap analog (**, P<0.001, Student’s t-test, two-tailed, n = 4). Results are presented as an average of four independent experiments, each with three technical repeats. Error bars represent the standard deviation.