1Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly-243122
2Department of Botany & Microbiology, Gurukula Kangri University, Hardwar-249404
* Author for correspondence; E-mail: raia1@rediffmail.com
The bacterial culture, E. coli DH5α containing recombinant plasmid pTargeT.rabgp which contained rabies glycoprotein gene insert was amplified in LB medium containing ampicillin 100 μg/ml. The overnight grown culture was used for plasmid DNA isolation. Expression of recombinant plasmid DNA was done in Vero cells using effectene transfection reagent. Transfected vero cell monolayer were examined by immunoperoxidase test and positive staining was observed showing expression of rabies glycoprotein. ELISA test also revealed glycoprotein antigen in the transfected cells. The SDS-PAGE analysis of transfected vero cells as well as sera from vaccinated dogs after 21 days of vaccination revealed rabies glycoprotein at 70kDa level, thus confirming the expression of the rabies glycoprotein by the recombinant plasmid.
Rabies virus, glycoprotein gene, expression
