Dependence of JEV RdRp activity on Mn 2+ ion . (A) RdRp assays were performed with the poly(A)/(U) 20 template (A) in the absence (lane 1) or in the presence of increasing concentrations of MgCl 2 or MnCl 2 (lanes 2–6 and 7–11; 0.5, 1.0, 2.5, 5.0, and 10 mM of MgCl 2 and MnCl 2 , respectively). (B) RdRp assays were performed with the 83-nt RNA representing the 3' end of the plus-strand JEV genome, in the absence of metal ions (lane 1) or in the presence of 2.5 mM of the divalent metal ion indicated above the autoradiogram (lanes 2 and 3). RdRp products were denatured and resolved on a medium size (20 × 20 cm) denaturing 5% polyacrylamide gel, and subjected to autoradiography.
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