Construction of Recombinant Raculovirus Containing the P12A and 3C Genes of Foot-and-mouth Disease Virus Type O

The capsid protein precursor P12A gene of foot-and-mouth disease virus(FMDV) type O were amplified from the plasmid T-OP1,the P12A were inserted into the baculovirus transfer vector pFast Dual-3C to construct recombinant transfer vector pD-P12A3C,in which the P12A and 3C were under the control of PH promoter and P10 promoter respectively.The recombinant plasmids were transformed into Escherichia coli DH10Bac(Invitrogen) to construct the recombinant bacmid B-P12A3C,and then,transfected into Sf9 cells,the recombinant baculovirus was harvested.After amplied,the recombinant baculovirus were infected into Sf9 cells.The expressed proteins were analyzed by an indirect sandwich-ELISA and by immuno uorescent assay.These results indicated that the expressed proteins were accurately expressed in Sf9 cells,and displayed specificity to FMDV type O antisera and biologic activation.From this study,The recombinant baculovirus containing the the capsid(P1) and 3C protease coding regions of FMDV type O were successfully obtained,thus providing a basis for the research of FMDV type O empty capsid assembly in vitro and empty capsid vaccine.