Cloning of whole ORF coding genes of FMDV and construction of recombinant adenoviral pAdTrack-CMV

In order to develop a recombinant adenoviral pAdTrack-CMV vaccine,genes of a whole ORF coding genes of foot-and-mouth disease virus(FMDV) were amplified by RT-PCR and inserted subsequently into a pMD18T vector and sequenced.The gene was cloned into the vector pAdTrack-CMV to construct the recombinant adenoviral pAdTract-CMV/ORF.The recombinant pAdTrack/ORF was identified by PCR,digestion with restriction endonucleases and sequencing.The sequencing results showed that the amplified L,P1,P2 and P3 gene's nucleotide sequences shared 98.8%,97.9%,99.0% and 97.6% homology with that of Akesu/58,respectively.PCR amplification and digestion with XbaⅠ+NotⅠ confirmed that the recombinant adenoviral pAdTrack-CMV contained the target genes of 6.9kb.The results showed that the recombinant pAdTrack/ORF containing the whole ORF gene of O type FMDV was constructed successfully.