Preparation and identification of polyclonal antibody to serotype I Marek＇s disease virus sorf2 protein

Abstract:

Abstract:［Objective］To obtain mice and rabbit polyclonal antibody of serotype I Marek＇s disease virus (MDV) sorf2 protein with higher titer and to identify the specificity．［Methods］ Using serotype I MDV GX0101 as template，we amplified sorf2 gene and then cloned it into pET-28a (+) and pET-32a (+) respectively．The recombinant plasmid pET-28a-sorf2 and pET-32a-sorf2 was separately transformed into Escherichia coli BL21 (Rosetta) competent cell which was induced with isopropylthio-β-D-galactoside (IPTG)．After purification，immuned 6－8 Balb/c mice and adult New Zealand white rabbit with the purified fusion protein and the anti-sorf2 polyclonal antibody were prepared．The specificity of the serum was detected by Western blot and the indirect immunofluorescence assay (IFA) method．［Results］Serotype I MDV sorf2 protein was expressed successfully in the recombinant E coli． Mice and rabbit anti-sorf2 polyclonal antibody with higher titer could react with sorf2 protein specifically．［Conclusion］The prepared anti-sorf2 polyclonal antibody could identify MDV sorf2 gene deletion strain specifically．In addition，it could be used for differential of MDV vaccine poison HVT and serotype I MDV，which was useful for the separation and identification of clinical MDV.