Construction and Identification of Eukaryotic Expression Vector for P1-2A-3C Gene of Foot and Mouth Disease Virus Type Asia 1

Objective To construct a eukaryotic expression vector for the P1-2A-3C gene of foot and mouth disease virus (FMDV) type Asia 1.Methods Amplify viral structural protein precursor P1-2A gene and non-structural protein 3C (proteinase) gene by RT-PCR from the vesiculae of bovine in epidemic area of foot and mouth disease and clone into vector pMD18-T.Digest the constructed recombinant plasmid pMD18-T-P1-2A-3C with restriction endonuclease,and insert the obtained P1-2A-3C gene fragment downstream to the pCMV promoter of pVAXⅠ.Transfect HeLa cells with the constructed eukaryotic expression vector pVAXⅠ-P1-2A-3C in mediation of liposome,and identify the expressed product by IFA.Results Restriction analysis proved that eukaryotic expression vector pVAXⅠ-P1-2A-3C was correctly constructed.The HeLa cells transfected with the constructed recombinant plasmid showed obvious yellowish-green fluorescence under fluorescent microscope,which indicated the expression of P1-2A-3C gene.Conclusion pVAXⅠ-PI-2A-3C might be used as a candidate FMDV DNA vaccine.