Fig. 2.

Identification of PRV TK⁻/gE⁻/PrM-E⁺ and expression of JEV PrM-E in PK-15 cells. a The total DNAs from virus-infected PK-15 cells were amplified by PCR with a pair of specific primers for JEV E gene according to JEV SX09S-01 strain and analyzed by electrophoresis. M: marker DL2000; 1 ~ 13: cells infected with different recombinant PRV clones; 14: cells infected with PRV TK⁻/gE⁻/LacZ⁺; 15: H₂O; 16: positive control, plasmid pIE-CAG-PrM-E-BGH. b & c PK-15 cells infected with PRV TK⁻/gE⁻/PrM-E⁺ or parental virus PRV TK⁻/gE⁻/LacZ⁺ strain were collected for JEV E (B) and PrM (C) protein expression detection by western blotting. The proteins were detected in cells at 72 h post infection with the recombinant virus. Primary antibody is mAbs for E or for PrM and secondary antibody is goat anti-mouse IgG-HRP. Lane 1: cell lysates of PRV TK⁻/gE⁻/PrM-E⁺; Lane 3: cell lysates of PRV TK⁻/gE⁻/LacZ⁺