Synthesis of the Bovine Rotavirus Major Neutralization Antigen (VP7) Using A Baculovirus Expression System
Synthesis of the Bovine Rotavirus Major Neutralization Antigen (VP7) Using A Baculovirus Expression System
El-Sabagh, I.M.; Hussein, H.A.; Amer, H.M.; El-Sanousi, A.A.; Reda,I.M. and M.A. Shalaby
ABSTRACT
The present study reports the successful production of the major neutralization antigen (VP7) of bovine romvirus Nebraska calf diarrhea virus (NCDV) strain in insect cells. The full-length DNA copies of RNA segment 9 (coding for VP7 protein) of NCDV was inserted into a baculovirus transfer vector under the control of the polyhedrin promotor. A recombinant baculovirus carrying the VP7 gene was constucted through homologous recombination between the baculovirus transfer vector carrying the VP7 gene and Autographa californica Nuclear Polyhedrosis Virus (AcNPV). Infection of Spodoptera frugiperda (SD) cells with Baculovirus recombinants expressing VP7 protein revealed high reactivity with hyperimmune antiserum to BRV when tested by immunoflurescence assay. Using solid phase ELISA, the VP7 expressed protein was detected intracellulary and extracellulary at 48 and 96 hours post-inoculation using polyclonal antibodies against BRV, respectively. The VP7 expressed protein was not detected in Coomassie blue stained SDSPAGE but produced a detectable band in Western blot assay. The reactivity of the VP7 expressed protein with BRV-specific hyperimmune antiserum confirmed that the antigenic deterrninants of the expressed protein were unaltered. The recombinant VP7 expressed protein can provide an effective tool for development of new diagnostic measures and novel vaccine candidate for improved diagnosis and control rotavirus infection in young calves.
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July 2008
Vol. 5, Iss. 1
