FIG. 4.

Transient replication of wild-type and mutant forms of the GFP-11E1 protein. (A) Activities of the wild-type GFP-11E1, the phosphorylation mutations, and the corresponding NES mutations (NEm) in transient replication assays. Linearized ori (arrow) in DpnI⁻ lanes reflects the total amount of both the residual input ori and newly replicated ori plasmids, while DpnI⁺ lanes represent the newly replicated ori DNA. The signals from the pMT2-E2 plasmid served as a control for the complete DpnI digestion, equal ori plasmid input, transfection efficiency, and plasmid recovery. (B) Signals of replicated origin DNA (ori band from DpnI⁺ lane) were quantified, and the ratios to that obtained with wild-type E1 (100%) were presented as relative replication activities. WT, wild type.