FIGURE 3.

Impact of deleting nucleotides 1 and 2 of the minigenome template on RSV RNA replication in BSR-T7 cells. (A,B) The panels are arranged as in Figure 2. (C) 5′-RACE sequence analysis of the RNA products generated from the Δ1 and Δ2 mutant templates: sequence analysis of the input minigenome plasmid DNA isolated from Δ1 (i) and Δ2 (iv) transfected cells. 5′-RACE sequence trace of the corresponding Δ1 (ii) and Δ2 (v) replication products (tailed with dCTP). All sequence traces are shown as template sense DNA. The Le sequence (underlined); sites of mutation in the templates (arrows); and the reintroduced nucleotides in the replication products. Note that the minor sequence that can be detected (beneath the 5′-RACE sequence trace) corresponds to the sequence of input plasmid DNA. Agarose gel electrophoresis of the 5′-RACE products generated from Δ1 and Δ2 mutant RNA samples from transfections that were lacking or containing the L polymerase plasmid, as indicated (iii and vi).