Co-expression of genes encoding the capsid protein precusor P1-2A and protease 3C of foot-and-mouth disease virus Asia type 1 in insect cells

The capsid protein precursor P1-2A gene and protease 3C gene of foot-and-mouth disease virus(FMDV) Asia type 1 were amplified from the plasmid pTOP-P1-2A and pTOP-3C,respectively.P1-2A and 3C gene were inserted into Baculovirus transfer plasmid pFastBacTM Dual to construct recombinant transfer vector pDual-P1-2A-3C.The pDual-P1-2A-3C was then transformed into Escherichia coli DH10Bac competent cells,transposited with Baculovirus shuttle vector(Bacmid) and constructed recombinant transposition rBacmid-P1-3C.After the rBacmid-P1-3C transfected into Sf9 cells,the recombinant Baculovirus was harvested.The expressed proteins were analyzed by SDS-PAGE,Western blotting and Dot-ELISA.It was found that the expressed proteins could react with rabbit sera against FMDV Asia type 1.These results indicated that the expressed proteins were accurately expressed in Sf9 cells,and displayed nice specificity to FMDV Asia type 1 antisera and biologic activation.