Cloning And Expression Of Pp24 Gene Of CVI988 Strain Of Marek's Disease Virus

pp24 protein, which is encoded bypp24 gene, is a kind of specific protein ofMarek' s Disease Virus (MDV). pp24 protein is suspected to bind with pp38 protein, another kind of specific protein and both of these two protems play an important part in ocongenesis.A pair of primers was designed containing two restriction sites for EcoR I and Sal I. pp24 gene was amplified from CVI988 strain by Polymerase Chain Reaction (PCR). PCR product was cloned to the downstream of GST gene according to the right Open Reading Frame (ORF) in pGEX-6P-l vector, and E.coli BL21 was transformed by the recombinant plasmid for expression. The protein expression was induced by the addition of 1.0mM IPTG at 37℃. The expressed product was identified and purified through SDS-PAGE, and a 43KD fusion protein with GST was found. The specific band was excised from the gel and injected into mice. The antiserum was collected from the immunized mice and used for Indirect Fluorescent Assay (IFA) with Chicken Embryonic Firoblast (CEF) infected by MDV. The positive staining were found in the MDV plaques and localized on the cytoplasme of the infected cell. The result showed that the in vitro expressed protein of pp24 genes has some effective epitopes of MDV.