Figure 2

Identification of the recombinant virus rPRV HN1201-EGFP-Luc. A PK-15 cells were infected with purified recombinant rPRV HN1201-EGFP-Luc virus and visualized by fluorescence and bright field microscopy. B Sequence chromatogram showing the DNA sequences of PRV HN1201 (top) and rPRV HN1201-EGFP-Luc (bottom) at the genetic homologous recombination sites. C Cells were infected with rPRV HN1201-EGFP-Luc (MOI = 0.1) for 12 h. The mRNA of firefly luciferase in the cells was detected with RT-PCR. D PK-15 cells (left) and ST cells (right) were either mock infected or infected with PRV HN1201 or rPRV HN1201-EGFP-Luc (MOI = 0.1) for 24 h and then lysed for immunoblotting analysis with antibodies against firefly luciferase and the viral proteins gB and gE. β-Actin was used as the loading control. E PK-15 cells were infected with rPRV HN1201-EGFP-Luc (MOI = 0.1) for 24 h and then lysed for luciferase assays. PK-15 cell lysate was used as a negative control, and PK-15 cells transfected with donor plasmid lysate were used as a positive control. This experiment was performed three times, and the results are shown as the mean ± SD.