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. 2007 Mar 7;81(12):6690–6699. doi: 10.1128/JVI.02457-06

FIG. 1.

FIG. 1.

Two strategies to produce 2b-specific sRNAs (shsR2b and miR2b) in plant cells. (A) Locations of shsR2b- and miR2b-targeting sites in the 2b gene. The predicted base pairings between sRNAs and their target sequences are shown. (B) A schematic diagram of the shsR2b expression cassette. The cassette contains the Arabidopsis U6-29 promoter engineered with a BamHI site at positions −6 to −1, followed by the shRNA-coding sequence and a 7-thymidine termination signal. (C) A schematic diagram of the 35S promoter (35SPro)-driven miR2b expression cassette with a termination sequence from the nopaline synthase gene (NOS Ter). The miR171 (in red) and miR171* sequences in the stem of the Arabidopsis pre-miR171a (the upper hairpin) were replaced with 21-nt miR2b (in red and shadowed) and miR2b* sequences, resulting in miR2bprec (the lower hairpin). Note that two mismatches between miR171 and miR171* are maintained between miR2b and miR2b*. The free energy of pre-miR171a and that of miR2bprec are indicated.