Figure 4. VSV L in vitro RNA synthesis assay.
(A) De novo RNA synthesis by VSV L, in the absence and presence of P, on a synthetic template corresponding to the first 19 nt of the leader (Le19). Activity assays were set using 0.2 μM of Le19, 0.2 μM of protein and [α32P]-GTP as the radio-labeled nucleotide. Reactions were quenched by the addition EDTA/formamide and analyzed on a 20% polyacrylamide/7 M urea gel. De novo initiation product sizes are indicated on the left. (B) RNA synthesis by VSV L on a naked 50 (Le50) nucleotides long RNAs, and the encapsidated RNA (N-RNA) templates. (C) RNA synthesis by MACV L on a naked RNA (left), or a naked mutated RNA (right).