FIG. 5.

Primer extension analysis of RNA generated from a minigenome containing a minimal promoter at the 3′ terminus. (A) Schematic diagram illustrating the sequences at the 3′ termini of a wild-type minigenome and minigenome +13only, which contain 86 and 13 nucleotides, respectively, of wild-type RSV sequence at their 3′ termini. Le-specific sequence is shown as a thin black rectangle, the gene start sequence is illustrated with a hatched rectangle, the NS1 nontranslated region as a dotted rectangle, nonspecific sequence as a thin white rectangle, and CAT-specific sequence as a partial white rectangle. Minigenome wild-type 3C-A has the same 3′-terminal sequence as the wild-type minigenome except that it contains a C-to-A mutation at position 3 of the Le, indicated with a lightening symbol. (B) Primer extension analysis of total positive-sense RNA. Lane 1 represents RNA from cells transfected with a wild-type minigenome containing a C-to-A substitution at position 3 of the Le, lane 2 represents RNA produced from a wild-type minigenome, and lanes 3 and 4 represent RNA from cells transfected with the +13only minigenome, with lane 3 representing a negative control transfection in which the L plasmid was omitted.