To Compare The Immunological Results Of Foot-and-Mouth Disease DNA Vaccine With Triple Codon AAA Of Kozak Sequences

Objective: To compare the expression and immunizing efficiency of Foot-and-Mouth Disease DNA vaccine with six different Kozak sequences. Methods: Six different Kozak sequences (KZ1—KZ6) were inserted into the initiation site of the antigenic gene coding VP1 within the FMD DNA vaccine and connected with the vector pVAX1, FMD DNA vaccine were constructed to immunize female Balb/c mice aged 6—8 weeks. The anti-VPl antibodies values in peripheral blood of the experimental animals were analyzed using indirect enzyme_linked immunosorbent assay. after the antigenic gene coding VP1 had been rebuilt, And then the sequences were inserted into the fusion expression vector pEGFP-N1 of Green Fluorescent protein, And then the sequences were inserted into the fusion expression vector pEGFP-Nl, The in vitro expression plasmids containing GFP were constructed and transfected Hela cells for transient expression. The in vitro expression level was analysized. Result: There was a significant difference of expression efficiency in vitro in Hela cells between the pKG6 and other groups (p<0.05). The mice produced significantly higher anti-VPl antibody level after DNA immunization with pKV6 than that of other recombinant plasmids (p<0.05). Conclusion: FMD DNA vaccine was constructed by Kozak sequence KZ6 with triple codon AAA have much higher expression efficiency and can induce stronger immune response than other recombinant DNA vaccine because of its high expression efficiency.