Construction Of Bi-functional Expression Vector Of AsiaⅠ FMDV And Its Expression

Foot-and-mouth disease (FMD) is an extremely contagious animal disease caused by Foot-and-mouth disease virus (FMDV), which dose serious harm to Animal Husbandry Production and causes great economic loss to those countries that spreads FMD. At present , vaccination is still one of an important methods to control FMD , But a challenge to the deveopment of vaccines against microbiology is the exist of immunization black and the latent period, diseases would out break when animals challenged with virus during the immunization black as well as animals inoculate the vaccine during the latent period. In this study, to solve this problem, a bi-funcational vaccine was studied with AsiaI/JQ strain.1.A DNA fragment from the 5'UTR and structural protein P1+2A and nonstructural 3C of Foot-and-mouth Virus was amplified by RT-PCR, they were inserted into pMD18-T vector for sequencing.2. Then the fragment AS gene were subcloned into the MCSâ… and the P1+2A+3C gene of FMDV was subcloned into the MCSâ…¡of the vector pQCXIX. The recombinant plasmid named pQC-AS-P1+2A+3C was identified by enzyme digestion, PCR and sequencing. The results showed the recombinant expression vector had Antisense nucleotide and P1+2A+3C that could be constructed.3.Recombinant plasmids pQC-AS-P1+2A+3C was constructed , and then pseudovirus was obtained by transfacting recombinant retroviral vector pQC-AS-P1+2A+3C into the Ampho-pack293 package cells. BHK-21 cells were infected by pseudovirus mediated in polybrene. RT-PCR showed the Antisense nucleotide has been transcripted in BHK-21 cells.4. Detection of indirect immunofluorescence showed that P1+2A structural protein was expressed in BHK-21 cellsThe bi-functional expression vector, which constructed in this study, would lay a foundation for studying on bi-functional vaccine of AsiaI FMD.