Division of Virology, Defence R&D Establishment (DRDE), Jhansi Road, Gwalior-474002, India.
Abstracts of the papers presented at the International Conference of Indian Virological Society on “Emerging and Re-emerging viral Diseases of the Tropics and Subtropics” at Indian Agricultural Research Institute, New Delhi, India, December 11–14, 2007.
Japanese encephalitis (JE) virus is one the most important arbovirus causing encephalitis among humans. JE is endemic in most parts of India and outbreaks are now reported at regular intervals. JE virus is a member of genus flavivirus of the family Flaviviridae and transmitted by Culex mosquitoes. Being a viral disease, there is no appropriate therapeutics available. Therefore, rapid and reliable diagnosis helps in effective patient management and to undertake suitable control measures. Domain III of envelope protein is reported to be primarily responsible for receptor binding and elicits neutralizing antibodies. Hence the development of recombinant protein against domain III might be useful towards an effective diagnostics reagents. In the present study, the molecular characterization of an Indian isolate of Japanese encephalitis virus was carried out by cloning and sequencing the domain III of envelope gene. A set of suitable primers targeting domain III was used to amplify a 345 bp amplicon. The PCR product was gel purified and cloned in to pET vector. The cloning was confirmed by RE analysis. The presence of insert was further confirmed by nucleotide sequencing. The BLAST search revealed that this isolate is most closely related to another JE virus isolated in 1956 at Vellore. This will be further sub cloned into an expression vector for the development of indigenous recombinant protein to be used in as a suitable diagnostic reagent.
