Construction Of A Bi-functional Vaccine Vector Of Asia Ⅰ Foot-and-mouth Disease Virus

At present, slaughter, isolation and vaccination are major methods to control FMD, and vaccination before epidemic of FMD is the specific efficient method to protect animals against virus entry. But a challenge to the development of vaccines against microbiology is the exist of immunization black and the latent period, FMD would outbreak when animals challenged with virus during the immunization black as well as animals inoculate the vaccine during the latent period. In order to solve this problem, we introduced RNAi into the bi-functional vaccine research, using Asia I/JS strain as the model to confirm the feasibility of the conception.1. Replaced the CMV promoter of vector pBudCE4.1 by U6 promoter, developed a dual-promoter vector pBudCE4.1-U6.2. Tested the functions of the promoters of vector pBudCE4.1-U6 by expressing EGFP and Interfering EGFP expression. Results demonstrated that both CMV promoter and U6 promoter could play its role.3. We successfully constructed three bi-functional plasmids expressing shRNAs targeting 3D gene of Asia I/JS strain FMDV and expressing immunization protein P12A+3C simultaneously.4. The antiviral potential induced by bi-functional plasmid was evident in the face of challenge of with a homologous FMDV and the inhibition was inconspicuousness.5. Detection of indirect immunofluorescence showed that immunization protein was expressed in BHK-21 cells.