This work was conducted to provide a useful basis for development of a new subunit vaccine against Chinese giant salamander iridovirus (CGSIV). In order to express major capsid protein (MCP) of CGSIV in baculovirus expression system, the gene of MCP was subcloned into baculovirus transfer vector (pFastBac1). The recombinant plasmid pFastBac-MCP was identified by restriction enzyme digestion and gene sequencing, and then transformed into DH10Bac competent cells containing baculovirus shuttle vectors. After identification by blue/white selection, PCR analysis, and gene sequencing, a recombinant bacmid (rBacmid-MCP) was obtained. Thereafter, the recombinant bacmid was transfected into Sf9 cells with Insect GeneJuice^{?/SUP> Transfection Reagent and recombinant baculoviruses were obtained in Sf9 cells, which could be observed by eletron microscopy in the ultra-thin sections of infected Sf9 cells and were named AcNPV-MCP. The recombinant baculovirus infected Sf9 cells with different multiplicities of infection (MOI=2, 5, or 10) showed the highest production of recombinant protein of MCP. The result of SDS-PAGE analysis showed that the production of expressed MCP could be the highest with MOI=10. By immunofluorescence assay, the presence of MCP was observed on the surface of infected Sf9 cells. The recombinant protein of MCP was purified with the prepared immunomagnetic bead with monoclonal antibodies against CGSIV MCP and its bioactivity was verified by western blot assay with the rabbit antisera against CGSIV MCP. The results of SDS-PAGE and western-blotting assays indicated that the recombinant protein was highly purified and had good immunogenicity, which could specifically react with the rabbit antisera. In conclusion, the recombinant protein of CGSIV MCP was successfully expressed in baculovirus expression system and purified with the immunomagnetic bead assay, which laid a solid foundation for the development of a new subunit CGSIV vaccine in the future.}