Rock bream iridovirus (RBIV) is a causative agent of epizootics among cultured rock bream (0plegnathus fasciatus) in Korea. Despite the importance of virus infection in Korean aquacultural industry, researches on vaccines for RBIV have been limited and there are only few studies about antigen analysis of fish iridoviruses. In this study, transmembrane protein genes of the ORF 017L, ORF 049L and ORF 107L and ankyrin repeats gene of ORF 112L from RBIV were tested to develop protein vaccines against RBIV. Genes of RBIV were cloned into expression vector of pGEX-4T-1. The recombinant proteins were successfully expressed using E. coli BL21(DE3). The soluble recombinant RBIV proteins were applied to affinity column for the purification of the proteins. BALB/c mice were immunized by the injection with purified recombinant proteins emulsified with an equal volume of Freund's adjuvant to produce polyclonal antibodies. Western blot was carried out to identify the immune reaction abilities of polyclonal antibodies to recombinant proteins. The effects of vaccines and antigenic properties of these proteins were evaluated by using neutralization test with BF-2 cells. In neutralization test, BF-2 cells infected with the mixture of RBIV and antisera obtained by anti-049L and anti-112L polyclonal antibody were healthy showing few CPE similar with the negative control (without RBIV). These studies suggest that it may be possible to develop an expressed genes-based vaccine and diagnostic kit against RBIV.