Construction of a recombinant baculovirus expressing the major capsid protein [VP6] of bovine rotavirus

ABSTRACT

The present study reports the expression of VP6, the major inner capsid protein of bovine rotavirus Nebraska calf diarrhea virus [NCDV] strain in a baculovirus expression system. The full-length DNA copies of RNA segment 6 [coding for VP6 protein] of NCDV were inserted into a baculovirus expression vector. A recombinant baculovirus carrying the VP6 gene was constructed through homologous recombination between the baculovirus recombinant plasmid carrying the VP6 gene and Autographa californica nuclear polyhedrosis virus [AcNPV] under the control of the polyhedrin promotor. Infection of Spodoptera frugiperda [Sf9] cells with the recombinant baculovirus expressing VP6 protein revealed a high-level of expression when tested by immunoflurescence and solid phase ELISA tests using BRV-specific polyclonal antibodies. The VP6 expressed protein was detected in Coomassie blue stained SDS-PAGE and produced a detectable band in Western blot assay. The high degree of reactivity with BRV-specific polyclonal antibodies confirmed that the antigenic determinants of the expressed protein were unaltered. The use of the in vitro expressed VP6 protein in the field diagnosis and vaccine development to control rotavirus infection is of considerable intere