Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar-243122, Uttar Pradesh.
Abstracts of the papers presented at the 16th Annual Convention and International Symposium of Indian Virological Society on “Management of Vector-Borne Viruses” at International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru-502324, Hyderabad, India, February 7–10, 2006.
Canine distemper virus (CDv) is a worldwide, highly contagious disease of young dogs of particularly 3 to 6 months of age, having high mortality. The glycoproteins hemagglutinin (H) and fusion (F) encoded by H and F gene of the virus genome, respectively, that are present on virus envelope are major immunogenic proteins which elicit protective immunity in the host. In the present study, complete 1810 bp coding region of H gene of CDV was amplified by RT-PCR. The RNA extracted from the commercially available vaccine by TRIZOL reagent was reverse transcribed using random hexamer primers and MMLV-RT. From cDNA, the complete H gene was amplified using Taq DNA polymerase and specific primers. The authenticity of PCR product was verified by its size in 1% agarose gel and nested PCR using internal primers. After confirmation, PCR product was cloned in pTARGET mammalian expression vector. The recombinant pTARGET containing H gene was confirmed by digestion with restriction enzyme Acc I, which resulted in fragments of 7387 bp and 133 bp confirming the clone to have right orientation of H gene. The expression of this recombinant construct in vitro in Vero cells and its use as DNA vaccine is under way.
