Construction Of Suicidal Eukaryotic Recombinant Expression Vector With FMDV(Type Asial) P12A3C Gene

Foot-and-mouth disease virus (FMDV) causes a highly contagious disease which is foot-and-mouth disease (FMD) in cloven-hoofed animals. Vaccine is one of major methods gainst FMD not only at present but also in the future. With the development of molecular biology and further understanding of structure and function of FMDV genome, DNA vaccine, especial suicidal DNA vaccine has become important direction of new-typed molecular vaccine research.The total RNA was extracted from FMDV Asial and the cDNA fragment of P12A or 3c was amplified with specific primers by RT-PCR. The purified cDNA fragment was inseted into respectively PMD-18T vector. The subclonging vector was identified by restriction end on unlease analysis and PCR. It was proved that the subclonging vector PMD-P12A or PMD-3c contains complete P12A or 3C gene by sequencing. PMD-P12A or PMD-3c digested by EcoRI and XhoI. then purified conjuncted transformed into E. coli JM109. The subclonging vector PMD-P12A3c was proved that contains complete P12A3C gene by same means. PMD-P12A3c and suicidal eucaryotic expression vector pshame-2a digested by Smal and BamHI respectively, then purified conjuncted transformed into E. coli JM109. The recombinant suicidal eucaryotic expression vector pshame-P12A3c was proved that contains complete P12A3C gene by same means. It can express speacial protein in BHK-21 cell by indirect fluorecsent antibody and ELISA.The suicidal recombinant expression vector with FMDV Asial P12A3C gene was constructed in the study, which was the base to research FMD suicidal DNA vaccine.