| 1. The glycoprotein (G) of Rabies virus (RV) is a major antigen for protective immune responses against RV infection. In this study, G gene of RV was optimized according to mammalian favored codons and cloned into eukaryotic expression vector pCAGGS to contruct recombinant plasmid pCAGG-RVG, and the G protein was expressed in transected BHK-21cells identified by indirect immunofluorescence assay and western bolt. In addintion, Beagle dogs were immunized with500μg of pCAGG-RVG via intramuscular inoculation for primary and booster vaccination at4weeks interval. The results of virus neutralization demonstrated that the dogs developed the neutralization antibody (NA) at average of3.81IU/mL to RV post the initial vaccination, and the NA titers reached a pike level at average of8.21IU/mL in2weeks post the booster. Thereafter, the NA in the dogs maintained at high titre (2.88IU/mL) during54weeks of the experimental period.0.5IU/mL of the minimum NA titer required for protecting animals from challenge with street RV. Therefore, the recombinant plasmid pCAGG-RVG has the potential to serve as a DNA vaccine against rabies in dogs.2. To evaluation the immunogenicity of Fusion protein(F), Hemagglutinin protein(H), Matrix protein(M) and Nucleocapsid protein(N) in dogs against canine distemper virus(CDV). F, H, M, N gene of CDV was optimized according to mammalian favored codons and cloned into eukaryotic expression vector pCAGGS to contruct recombinant plasmid pCAGG-CDVF, pCAGG-CDVH, pCAGG-CDVM, pCAGG-CDVN, and the F, H, M, N protein was expressed in transected BHK-21cells identified by indirect immunofluorescence assay. In addintion, we desigened A(admix F+H+M) and B(admix F+H+M+N) two test groups, six Beagles of each groups were immunized with500μg of each plasmid via intramuscular inoculation for primary and booster vaccination at4weeks interval. The results of virus neutralization demonstrated that both DNA vaccines of A and B test groups showed sensitivity and specificity immunogenicity. After first inmunication, The group which contain pCAGG-CDVN express Nucleoprotein was relatively slow, but remarkable enhancement effect were showed after booster. Supposedly of immune memory cells were induced because of nucleoprotein. Thereafter, the NA (A and B) in the dogs maintained at high titre (25and26) during54weeks of the experimental period. Therefore, both two groups of recombinant plasmid has the potential to serve as a DNA vaccine against canine distemper in dogs. |
