FIGURE 1.

Schematic diagram of CHv-BAC-ΔLORF5 construction. (A) DPV genome structure. (B) Detection of protein expression of LORF5 gene in eukaryotic plasmid-transfected cells by western blotting. 293T and DEF cells were transfected and harvested at 24 hpi (293T) and 36 hpi (DEF cells). Proteins were detected using a mouse anti-FLAG MAb. (C) The principle of knocking out the DPV LORF5 gene. In the first step of homologous recombination, the LORF5 gene was replaced by the Kan resistance gene through 40-bp homology arms (sequences a and b). In the second step, L-arabinose induced recombinase to recognize the I-SecI cleavage site and delete the Kan fragment. Then, the LORF5 gene ORF was deleted in the DPV genome without any reservation.