Fig. 8.

Enhancement of viral replication through mutation of the 2K peptide. (A) Resistance and replication analyses using WNV replicon. The effect of the G6871A mutation in the 2K peptide on resistance to lycorine was quantified using a transient replicon (Rluc2A-Rep) assay. Equal amounts (10 μg) of WT and G6871A mutant replicon RNAs were electroporated into BHK-21 cells. The transfected cells were immediately treated with lycorine (1.5 μM) or without lycorine (1% DMSO as controls). Luciferase activities were measured at the indicated time points post-transfection. Average results from three independent experiments are presented. (B) Growth kinetics of WT and 2K peptide G6871A mutant viruses. Vero and C6/36 cells were infected with the WT and the 2K peptide mutant viruses at an MOI of 0.05. After 1-h incubation, the cells were washed three times with PBS and the medium was replenished. Viral titers in culture fluids were quantified at the indicated time points using plaque assays.