الفهرس 
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Abstract
The VP6 and VP7 genes of BRV were individually cloned and expressed in Spodoptera frugiperda (Sf9) insect cells using recombinant baculovirus vectorsBRV was propagated in MA - 104 cells and identified by VP6 Mabs based ELISAThe viral RNA was extracted from the tissue culture supernatant of infected cells and the full length of VP6 and VP7 genes was amplified using one step RT - PCRThe PCR products were cloned into the baculovirus shuttle vector ; pBlueBac45 / V5 - His TOPO and the recombinant plasmids carrying VP6 and VP7 genes in correct orientation were identified using PCRThe cloned genes were inserted in the genome of Autographa californica nuclear polyhydrosis virus (AcMNPV) under control of the polyhedrin promoter ; through homologous recombination between the shuttle vector carrying the cloned genes and a linearized baculovirus DNA (Bac - N - Blue\{u2122\}) using liposome mediated transfection techniqueRecombinant baculoviruses carrying VP6 and VP7 genes of BRV were selected by plaque purification ; verified for the presence of target genes using PCR that utilize specific primers for polyhedrin gene and propagated for generation of high - titer viral stockThe recombinant baculoviruses carrying VP6 and VP7 genes of BRV revealed high - level of expression when screened by immunofluoresent test and solid phase ELISA using BRV - specific polyclonal antiserumThe VP6 expressed protein was detected by Coomassie blue staining of SDS - PAGE separated proteins and produced a detectable band in western blot assaywhereas the VP7 expressed protein was not detected by Coomassie blue staining of SDS - PAGE separated proteins but produced a faint band in Western blot assay.