Figure 1

Generation of a dual-reporter ASFV co-expressing Gluc and EGFP (rASFV-Gluc/EGFP). A Schematic representation of the strategy for constructing the transfer vector pOK12-Gluc/EGFP and the dual-reporter virus rASFV-Gluc/EGFP. B PCR confirmation of rASFV-Gluc/EGFP purity. PCR amplification of genomic DNA derived from the ASFV-WT and rASFV-Gluc/EGFP genomes was performed using the primer pair D-JD-F/R, and the amplified products were subjected to gel electrophoresis. C Observation of EGFP expression. PAMs were infected with ASFV-WT or rASFV-Gluc/EGFP (MOI = 0.5), and EGFP expression was directly visualized via fluorescence microscopy at 72 hpi, with representative fluorescence images captured. Scale bars = 400 μm. D Gluc activity assay. PAMs were infected with either ASFV-WT or rASFV-Gluc/EGFP (MOI = 0.5), and the supernatants were collected at 72 hours post infection (hpi) to measure Gluc activities, which were expressed as relative light units (RLUs). E Western blotting analysis. The expression levels of the p72 and A137R proteins in the PAMs infected with rASFV-Gluc/EGFP or ASFV-WT were analysed by western blotting.