| Porcine circovirus (PCV), according to serotypes, was divided into porcine circovirus type1(PCV1) and porcine circovirus type2(PCV2). PCV1was first found as a cell contaminants, but the virus was not pathogenic in pigs. Morever, PCV2was widely recognized as an important pathogen, and it was associated with post-weaning multisystemic wasting syndrome (PMWS). PCV2contains two main open reading frames including ORFl and ORF2. And ORF2encodes major cap protein of PCV2. Cap protein could induce immune response and specific antibodies, it became a goal of diagnostic techniques and immune mechanism about PCV2.According to current studies, Cap protein was consisted of amino acids233-235, which included a plurality of linear and conformational epitopes, and some of which were commoned with some epitopes of PCV1. This study is based on current research about Cap protein epitopes to identify and analyze.Firstly, recombinant vector pET-Cap was built and Cap protein was expressed by prokaryotic expression system. According to comparison between Cap protein expressed by prokaryotic and Cap protein expressed by yeast, Cap protein expressed by yeast was better. Immune BALB/c mice with Cap protein expressed by yeast.2strains of hybridoma against Cap protein were selected, and named4E2,1E7. However, another hybridoma against His tag was selected, and named5D1.Secondly, Cap protein was truncated expression into19peptides, which were named JD1-JD19, after removing the nuclear localization signal of Cap protein. There were two peptides reacted with monoclonal antibodies. Further precise position were77NDFLPPG83and138YDPYVNY144. Currently,138YDPYVNY144is a newly discovered epitopes.Last, biological software was used to draw PCV2virions and Cap protein map, in order to identifie and analyze the position and function.In summary, this study prepared monoclonal antibodies for anti-Cap protein. According to peptide scanning method,77NDFLPPG83and138YDPYVNY144were identified. |
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