Identification And Preliminary Evaluation Of Immunology Of P23-33 Gene Recombinant Adenoviral Vector Of Theileria Sergenti

Theileria sergenti is a haemoparasitic disease which causes hyperpyrexia, hyphemia, athrepsy and lymphadenectasis in cattles.It spreads widely in resent years, causes serious economic losses in the cattle industry,the morbidity and mortality are increasing and becoming a serious disease in cattle.P23 and P33 have good immunogenicity playing an important function in countering the desease of Theileria sergenti.In order to utilize the P23 and P33 very well, in this study,using PMD18-T-P23-P33 as template,design of specific primers, Theileria sergenti P23-33 fusion gene by PCR cloning,enzyme P23-33 fusion gene, subclone into the adenovirus shuttle vector PCR 259.The recombinant shuttle plasmid PCR259-P23-33 convert to the adenovirus backbone plasmid containing Transpose-Ad294 of E.coli HighQ-1 competent cells,By homologous recombination getting the recombinant plasmid pAd-P23-33.by PCR,restriction enzyme analysis correctly transfectes HEK-293 cells,Adenovirus packages,detection of gene expression by RT-PCR and IPMA.The results indicated,successfully constructed recombinant adenovirus vector pAd-P23-33 with 1 135bp,detected in fusion gene and was expressed in BHK-293 cells by RT-PCR and IPMA(Immunoperodase monolayer assay).Determination of adenovirus titer 6.3x107IU/ml,immuned BALB/c mice by recombinant adenovirus pAd-P23-33,serum antibody levels and cytokines IL-4,IFN-y were significantly higher than that of the recombinant plasmid PVAX-P23-P33 immune group and the PBS control group (P<0.01).The test esterblished the basis for the bovine theileriosis sergenti-depth study of recombinant adenovirus vaccine.