الفهرس 
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Abstract
In the present study, The VP3 gene of Chicken Anemia Virus (CAV) was cloned and expressed in Spodopetra Forugiperda (Sf9) insect cells using recombinant baculovirus vectors. Chicken anemia virus H91/2000 local strain was inoculated in one-day-old SPF chicks. The histopathological effect of virus was observed in lymphoid heamopiotic organs (liver, spleen, thymus, bursa, proventriculus, spleen, pancrease) along 7 days post inoculation. CAV antibodies in sera of the inoculated SPF chicks were detected by ELISA. Also, CAV-DNA genome was detected in thymus and liver by PCR. Full length of VP3 gene was amplified by PCR and was cloned into the baculovirus shuttle vector, pBlue Bac 4.5/V5-His TOPO. Recombinant plasmids carrying the VP3 gene in correct orientation was identified using colony PCR assay. The cloned gene was inserted in the genome of Autographa California nuclear polyhydrosis virus (AcMNPV) under control of the polyhedron promoter, through homologous recombination between the shuttle vector carrying the cloned gene and a linearized baculovirus DNA (Bac-N-BlueTM ) using liposome mediated transfection technique. Recombinant baculoviruses were selected by plaque purification, verified the presence of VP3 gene of CAV using PCR and propagated for generation of high-titer viral stock. The recombinant baculoviruses carrying VP3 gene of CAV revealed high-level of expression when screened by immunofluorescent test, solid phase ELISA and Western blot assay. In conclusion, this study is the first detailed report on cloning and expression of VP3 gene of CAV in Egypt. The recombinant expressed VP3 protein (apoptin) is represents a potential anti-tumor agent in transformed and malignant cells. Also, the expressed apoptin protein can be used as a diagnostic tool for detection of individuals with an increased risk for hereditary cancer and premalignant lesions.