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. 2013 Jan;87(1):199–207. doi: 10.1128/JVI.06968-11

Fig 2.

Fig 2

Primer-dependent RNA synthesis. (a) Concentration dependence of primer-dependent RNA synthesis by wild-type BVDV NS5B. A 3′-recessed RNA primer-template duplex (duplex A) was used in this RdRp assay. The RdRp assay reaction mixtures contained between 0.5 and 5 μM wild-type BVDV NS5B, 0.1 μM 5′-end-labeled RNA, and 50 mM NaCl. (b) Primer extension by wild-type (WT) BVDV NS5B and its mutant derivatives in the presence of MgCl2 (white bars) or MnCl2 (gray bars) as the divalent metal ion. Reactions were carried out at 37°C for 2 h, using 0.250 μg of poly(C) template, 0.125 μg oligo(G)12 primer, 0.4 μCi/μl [α-32P]rGTP, and 50 nM NS5B in a total volume of 25 μl. [α-32P]rGMP incorporation was quantified by a DEAE filter binding scintillation assay.