| Foot-and-mouth disease (FMD) is an extremely contagious animal disease caused by foot-and-mouth disease virus (FMDV), which mainly infects cloven-hoofed animals such as swine, cattle and sheep. FMD seriously impair healthy development of animal husbandry and result in considerable economic losses. Therefore, all the countries of the world pay much attention to its research and prevention. Because the traditional inactivated vaccine has insecure factor, so it is very necessary to develop efficient, safe and economical FMDV Genetic Engineering Vaccine.In this paper, the P12A and 3C genes were obtained from FMDV OMâ…¢strain of type O using special primers by RT-PCR and cloned into pMD18-T plasmid respectively. After sequencing and analyzing with DNAstar, it was indicated that the p1-2A and 3C genes were derived from OMâ…¢strain. Then the two genes were cleaved by double digestion and subcloned orientationally into shuttle vector pAdTrack-CMV, being selected with kanamycin and screened by digestion, a recombinant shuttle plasmid was obtained and naned pAdTrack-CMV/P12A3C; Using the pAdEasy system, the recombinant adenovirus DNA bearing exogenous FMDV genes by employing homologous recombination was efficiently constructed, the resultant construction was linearized with PmeI and transformed together with a supercoiled adenoviral vector pAdEasy-1 into E. coli strain BJ5183; In the end, the recombinant adenoviral construct was cleaved with PacI to expose its inverted terminal repeats, and transfected into HEK293 packaging cell line. The strain of recombinant adenovirus containing P1-2A gene and 3C gene from OMâ…¢strain was successfully obtained. P1-2A,3C have hereditary stability in recombinant adenovirus. Using double-antibody sandwich ELISA, immunization test proved the adenovirus vaccine can produce special antibody to P1.This research provided the basis for research replication-deficient adenovirus vector vaccine containing P1-2A,3C gene of FMDV.
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