الفهرس | Only 14 pages are availabe for public view |
Abstract In the present study, recombinant baculovirus-expressing BoHV-l glycoprotein gE was antigenically and molecularly characterized to be used in the development of diagnostic tools. The P- 2 stock of recombinant baculoviruses was prepared and titrated in Sf 9 by plaque assay using X gal for preparation and characterization of gE expressed protein. PCR analysis was carried out and demonstrated the size of recombinant gE protein at 2182 bp (1866+316) and 1866 bp by using PH primers and specific gE gene primers, respectively. The IF studies demonstrated the intracellular distribution of the recombinant glycoprotein in infected Sf9 with rBaclBoHV-lgE virus. The characterization of gE recombinant protein by SDS-PAGE and western blot assays revealed that gE recombinant protein expressed at approximately 80 KDa that had the same mobility in polyacrylamide gel stained with Coomassie blue stain. gE recombinant protein was used as a coating antigen in an indirect ELISA to detect antibody against BoHV -1 in serum samples to distinguish the vaccinated cattle with marker vaccine from infected animals. The crude lysate from SF9 cells infected with rBaclBoHV-lgE virus proved to be sensitive and efficient in development of ELISA kit. |