Figure 1.
Expression of functionally active PPRV L protein. A VDS cells were transfected with the indicated plasmid (2 μg) in 6-well dishes and cultured for 48 h. After lysis of the transfected cells, expressed proteins were analysed by SDS-PAGE and Western blot using mouse anti-V5 tag. B Cells (VDS or VDS-L) in 6-well dishes were transfected with the indicated plasmids and incubated for 8 days. The transfected cells were subjected to 1 freeze-thaw cycle to release infectious virus and the clarified supernatant used to infect fresh VDS or VDS-L cells. After 7 days, the cells were harvested and the expression of virally-encoded GFP analysed by SDS-PAGE and Western blot. Actin was used as a loading control. C VDS or VDS-L cells (~4 × 105) were lysed directly in SDS-PAGE sample buffer and analysed by SDS-PAGE and Western blot using mouse anti-V5 tag. D As B, except that no plasmid encoding the L protein was included.