Expression of Recombinant VP2 Protein of Canine Parvovirus in Escherichia coli

บทคัดย่อ

Canine parvovirus (CPV) appears to be endemic in almost all populations of wild and domesticated dogs. It causes serious contagious enteric disease. The VP2 protein of CPV is a major capsid protein and plays an important role in the host immune response. In the present study, the
recombinant VP2 was expressed in Escherichia coli (E. coli) using the pBAD expression system. Virus DNA from the infected feces was extracted and used to amplify the whole VP2 gene by using specific primers. Subsequently, the whole VP2 gene was ligated with plasmid pBAD202/D-TOPO and used to transform into the E. coli strain TOP10. The SDS-PAGE analysis revealed a specific band approximately 80 kDa and was found mainly in the pellet of the bacterial lysate. The optimum time and concentration of arabinose for expression of recombinant VP2 protein was 8 h and 0.002%, respectively. By dot blot and Western blot analysis, the recombinant VP2 protein showed specific interaction with mouse antihistidine monoclonal antibody and rabbit anti-CPV hyperimmune serum. The recombinant protein VP2 might be a useful tool for the development of a diagnostic test for the detection of CPV and a vaccine against CPV.