Construction And Immune Effection Of DNA Vaccine Against Classical Swine Fever E2 And Foot-and-Mouth Disease VP1 Linker

The major antigen region E2 gene of Chinese C-strain virus,amplified by RT-PCR and nPCR,were ligated into eukaryotic expression plasmids pcDNA3.1(+) with foot and mouth diseases virus (FMDV)VP1,and the recombinant plasmid pcDNA3.1/VP1E2 were successfully constructed.The insert position,size and reading frame were right using PCR,restriction digestion and the sequence analysis. The PK-15 cells were transfected by the recombinant plasmids pcDNA3.1/VP1E2。The E2 and VP1 antigens had been detected with RT-PCR and direct fluoroscopic technology.Immunological effects of the recombinants plasmids pcDNA3/VP1E2.The rabbits were injected intramuscularly with 100ug each of the pcDNA3/VP1E2,And administered again in the same way after the fourth,sixth and eighth week,respectively.The VP1 and E2 antibody were detected with ELISA and the VP1 antibody dilution was detected using direct haemoaglutination.The rabbits were challenged with C-strain virus.The results shows that the eukaryotic expression plasmids pcDNA3/VP1E2 were constructed successfully,and the VP1 and E2 protein were expressed after transfected the PK-15 cells.The E2 specific antibody was detected after the rabbits were immunited,but the VP1 specific antibody was not found.