Objective:To prepare the polyclonal antibody against shortened Fiber-1 (sFiber-1) protein of fowl adenovirus serotype 4 (FAdV-4) ,and to provide a tool for the detection and diagnosis of FAdV-4.Methods:The signal peptide ,hydrophobicity and antigen determination of the Fibre-1 gene sequence of CH/AHMC/2015 isolate of FAdV-4 (MG148335.1:30459-31754) were analyzed ,and the fragments with high immunogenicity were intercepted and the specific primers were designed and synthesized.The DNA of CH/AHMC/2015 isolate was amplified to obtain the sFiber-1 fragment ,which was cloned into pET-32a ,and transformed into the COMPETENT cell BL21 to express sFiber-1 protein.The purified sFiber-1 protein emulsified with adjuvant was used to immunize rats.Polyclond antibody was prepared , the titer of polyclonal antibody was analyzed by ELISA , and the immunogenicity of sFiber-1 was analyzed by Western blot.Results:The sFiber-1 prokaryotic expression vector was constructed to obtain recombinant sFiber-1 protein of FAdV-4.SDS-PAGE showed that sFiber-1 protein was mainly expressed in inclusion bodies with a molecular weight of 52 kDa.The titer of sFiber-1 antibody was 1:213.Western-blot results displayed that recombinant sFiber-1 protein can be recognized by the obtained antibody.Conclusion:In this study ,recombinant sFiber-1 protein of FAdV-4 was successfully expressed and sFiber-1 polyclonal antibody with high immune activity was prepared ,which could lay a foundation for the diagnosis and detection of FAdV-4.
维普
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