Fig. 2.

Effect of mutations in the TCV 5′ UTR on translation of a single-luciferase construct in protoplasts. (A) Positions of point mutations in the 5′ UTR. Names of mutations are bracketed. The location of the 5′ TE after completion of these experiments is shown. (B) Translational activity in protoplasts of single-luciferase constructs containing the 5′ UTR (wt and mutant) and 3′ UTR+. Luciferase was assayed at 18 h after transfection. None, no 5′ or 3′ sequences; 5′ UTR, 5′ UTR only; 3′ UTR+, 3′ region (positions 3661 to 4054) only; 5′+3′, 5′ UTR and 3′ UTR+. Data from at least three replicate experiments are shown. Standard deviation bars are given. (C) TCV gRNA containing mutations denoted above the lanes were assayed for accumulation in protoplasts at 40 h postinoculation (hpi). Levels of accumulation and standard deviations in three independent experiments are given below the lanes.