The glycoprotein gene of rabies virus and VP2 genes of canine parvovirus were cloned in pTargeT vector and the recombinant plasmids containing the genes in right orientation were selected. The sequencing of the insert genes were done to confirm the gene sequence and orientation. The recombinant plasmids thus selected were amplified and used to prepare recombinant plasmid DNA in large quantity which were used as DNA vaccines with ISA50 (seppic oil) as adjuvant. The DNA vaccines were found to be safe and gave full protection against rabies and canine parvovirus. Further trials of these vaccines showed that they can be used in the field for vaccination against these two diseases of dogs. The VP2 gene of infectious bursal disease virus (IBDV), 1367 bp, was synthesized in collaboration with Bangalore Genie, Bangalore. The VP2 gene insert was recloned in pVAX1 and the recombinant plasmid was designated as pVAX1.ibdvp2 and analysed by restriction enzyme analysis. 50 μg pVAX1.ibdvp2 plasmid DNA per chick was inocualted intramuscularly in 10 chicks and pVAX1 vector DNA alone was inocualted in 10 chicks, 10 vector alone and 10 healthy chicks were kept as control. All the chickens were challenged 21 days post vaccination. All the vaccinated chicks were fully protected while control birds died/suffered from disease, ex-pression studies showed the ex-pression of the recombinant plasmid in CEF cell cultures. The PCR amplification of full length 2916 bp FAV4 hexon gene was done successfully by PCR. The gene was cloned in pcDNA3.1/V5-His-TOPO vector (Invitrogen). Two clones containing the hexon gene in right orientation clone 9 and clone 6 were selected after restriction enzyme analysis. The sequencing of this recombinant plasmid confirmed the insert to be in right orientation and the sequence analysis revealed on ORF of 2814 bp coding for a 937 amino acid long poolypeptide with a molecular mass of 106.04 kDa. The recombinant plasmid was designated as pcDNA3.1-His-TOPO fav4hex. The sequencing of the insert gene revealed that it was in right orientation. The fav4hex recombinant plasmid expressed hexon protein in vero cells as determined by immunoperoxidase test.
