FIGURE 5.

The expression of NbCA in Escherichia coli and in vitro replication assays with endogenous and exogenous templates. (A) Total proteins were extracted from E. coli that expressed vector only (vec) or NbCA with or without the induction of IPTG as indicated, separated on a 12% polyacrylamide gel/SDS, and stained with Coomassie blue. The eluents indicated as elute were the total proteins eluted from His-tag purification resin. In vitro replication assay involved use of the purified replicase complex from infected plants with the addition of E. coli-expressed proteins (A) to test the RdRp activity of the endogenous templates (B) and exogenous templates r138/40A (C) and Ba-77 (D). RdRp activity with the addition of E. coli-expressed eluent of vector only was set to 100%. Representative results are shown. The numbers shown above each bar are the mean relative RdRp activity with the standard errors derived from at least three independent experiments. ^∗∗∗p < 0.001 by Student’s t-test.