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Abstract C3N protein of Rift Valley Fever Virus (ZH-SOl strain) was expressed in Spodoptera frugiperda (Sf-9) insect cell line using recombinant baculovirus carrying C3N gene of RVFY. RVFV was characterized using RT-peR and sequencing of the internal part of the M genomic segment of the virus which revealed that the virus was related genetically to the Egyptian lineage. The full length ~ gene was amplified using RI-peR, cloned into baculovirus transfer vector pBlueBac4.S/VS-His- TOPO® and the recombinant plasmid was checked for correct orientation by peR assay. The cloned C3N gene was introduced into the genome of Autographa californica nuclear polyhydrosis virus (AcMNPV) under control of the polyhedron promoter, through a process of homologous recombination between the recombinant transfer vector identified for correct orientation and a linearized replication-defective baculovirus DNA (Bac-N-Blue™). Recombinant baculovirus was purified using plaque assay and checked for recombination and purity using peR assay. The pure recombinant baculovirus carrying ~ gene of R VFV was propagated in Sf-9 cells for production of high titer (P2) virus stock which was titrated using plaque assay and used in expression of ~ protein. The expressed protein was characterized by immunofluorescence, solid phase ELISA, SDS-PAGE and Western blot assays which revealed that the protein was expressed at a high level especially after 96 hours of infection of the insect cells with the recombinant baculovirus. The expressed protein will be used in future studies in the field of diagnosis in development of new diagnostic kits and in the development of subunit vaccine to RVFV. |