Fig. 2

Analysis of plasmids transfection (magnification 100 x). (A) The minireplicon was produced by transfection of the pCMV-CDVmini plasmid into Vero cells, and RdRP was generated by virus incubation before plasmid transfection. The production and replication of viral microreplicons depend on the viral polymerase supplied by CDV-infected cells. (B), (C), (D), and (E) Analysis of different rescue systems based on minigenome expression. (B) BSR cells were transfected with 2.0 µg of pCMV-CDVmini plasmid and 3 µg of pCMV-NPL plasmid. (C) BSR cells were transfected with 2.0 µg of pCMV-CDVmini plasmid, 1 µg of pCMV-NP and 1 µg of pCMV-L. (D) BSR cells were transfected with 2.0 µg of pCMV-CDVmini plasmid, 0.5 µg of pCMV-N, 0.5 µg of pCMV-P, and 1 µg of pCMV-L. (E) BSR cells were transfected with 2.0 µg of pCMV-CDVmini plasmid. (F) Cytopathic effect (CPE) induced in Vero cells co-transfected with plasmids pCMV-NPL and puCMVZJ. Arrows indicate syncytia in the cells that were observed at 5 days after transfection. The arrow indicates the formation of syncytia in the cells, which was observed 5 days after transfection. (G) Control cells. (H) Sequencing results of the mutation sites in the recombinant virus genome. Asterisks demonstrate that the A → G and A → T substitutions at viral nucleotides 8975 and 8976 to create a Pme I restriction site in the recombinant viral genome