Fig. 3.

The biological characteristics analysis of PRV TK⁻/gE⁻/PrM-E⁺. a PRV TK⁻/gE⁻/PrM-E⁺ was grown on PK-15 cells sequentially for 20 passages and the total DNAs from virus-infected PK-15 cells were amplified by PCR with a specific primers for JEV E gene according to JEV SX09S-01 strain and analyzed by electrophoresis. M: marker DL2000; 1 ~ 11: cells infected with passages 11 to 20; 12: cells infected with PRV TK⁻/gE⁻/LacZ⁺; 13: H2O; b Genetic stability of protein identification for different generations by western blotting. PK-15 cells infected with different generations of PRV TK⁻/gE⁻/PrM-E⁺ were collected for JEV E protein expression detection by western blotting. 1 ~ 7: cells infected with passages 1, 2, 4, 8, 12, 16 and 20 of PRV TK⁻/gE⁻/PrM-E⁺. c Virus growth curves based on viral yield of supernatants harvested at different time points of PRV TK⁻/gE⁻/PrM-E⁺ or PRV TK⁻/gE⁻/LacZ⁺ infected PK-15 cells, and viral yield was measured by TCID₅₀