Characterization of Monoclonal Antibody Against Replication-Associated Protein of Porcine Circovirus

Abstract

The replication-associated (Rep) protein of porcine circovirus (PCV) was suggested to play an essential role in the replication and translation of viral DNA. In this study, one monoclonal antibody (mAb) specific for Rep protein of porcine circovirus type 1 (PCV1), two mAbs against Rep protein of porcine circovirus type 2 (PCV2), and five mAbs to both Rep protein of PCV1 and PCV2 were generated using, respectively, Rep protein of PCV1 and PCV2 expressed in Escherichia coli as an immunogen. Western blot analysis showed that native Rep protein of PCV2 virions appeared in two forms with different molecular weight in PCV2-infected cells. Laser confocal analyses further exhibited that Rep protein distributed mainly in the cellular nucleoplasm at the early stage of PCV2 infection, and moved to the nuclear periphery and the cytoplasm at the last stage of PCV2 infection. The results from this study confirmed that Rep protein of PCV2 distributed in both nucleus and cytoplasm, and provided an mAb tool to further analyze replications of PCV1 and PCV2 in vitro and in vivo.