Cloning of P12X3C3D fragment of foot-and-mouth disease virus and construction of baculovirus expression vector

The multi-gene, P12X3C3D fragment, which contains the complete genes P1, 2A, 3C, 3D and part of 2B of foot-and-mouth disease virus, was amplified by the site mutation PCR. After being digested with SpeⅠand HindⅢ respectively, the gene was linked to the baculovirus transfer vector pFastBacHT. The results showed that the recombinant vector, pFB-P12X3C3D, was constructed successfully. Then the recombinant vector was transformed into DH10Bac E.coli, and the positive recombinant bacmid Bacmid-P12X3C3D was screened according to the resistant of Gmr and Kar. After the Sf9 insect cells were transfected with the DNA extracted from Bacmid-P12X3C3D, the recombinant baculovirus particles were obtained. The achieved baculovirus particles were identified by PCR. The results showed that the multi-gene, P12X3C3D, have been introduced into the recombinant baculovirus correctly.