The aim of the present study was to investigate the physical-chemical and biological characteristics of Cyprinid herpesvirus 2 (CyHV-2) and its ultrastructural morphogenesis in vitro. By using the newly established cell line derived from gibel carp brain, designated GiCB, which was highly permissive to the infection of CyHV-2, the physical-chemical and biological characteristics, and growth curve of CyHV-2 were investigated; meanwhile, the susceptibility of available fish cell lines to CyHV-2 infection was compared and the ultrastructural morphogenesis was conducted in this study. The results indicated that the CyHV-2 is sensitive to the treatment of heat, acid (pH 3.0), alkali (pH10.0), chloroform, ether and frozen-thawing. CyHV-2 could not propagate in fish cell lines EPC, RTG-2, Koi-Fin, CIK, CCK and PF-Fin, etc., which were verified by CPE observation and CyHV-2 DNA Nest-PCR assay; According to the growth curve of CyHV-2 in GiCB cells, a log phase increase in virus production was followed by an eclipse period of about 12 h. Virus production peaked around 96 h (10^7.52±0.26 TCID₅₀/mL). The CyHV-2 ultrastructural morphogenesis in GiCB cells presented in three main stages, including virus adsorption and entry, replication and assembly, maturation and budding. This study provided useful information for the further studies on CyHV-2 infection mechanisms, pathogenicity and the control methods of the disease caused by the virus.