Figure 6
From: Characterization of Norovirus RNA replicase for in vitro amplification of RNA
Effect of the terminal structure of dsRNA for the initiation of RNA replication. Two kinds of RNA template (10 pmol) ((i) Temp(GGG-UCCCC) and (ii) Temp(GGG-UCCC) (A)), of which secondary structures were predicted by Mfold (B), were incubated with NV3Dpol (4 pmol), respectively (reaction volume = 40 μL), and analyzed on a non-denaturing 10% PAGE. The PAGE image was visualized by SYBRgreenII staining (C). M; 10 bp DNA step ladder (Promega). Ref; both RNA templates were electrophoresed as references. Mfold predicted many other structures, but the stem-end (3’end and 5’end hybrid) structures were almost always conserved, namely, the probability of other 3’ structures (for (i), first appearance as #7 stable structure (ΔG of #1, #2,---,and -#7 structures were -12.83, -11.08, -10.49, -10.10, -9.39, -8.52, and -7.99 kcal/mol, respectively) was less than 0.03% .