Erns is a highly glycosylated envelope protein of classical swine fever virus (CSFV) with RNase activity. Erns can induce neutralizing antibodies and provide immune protection against CSFV infection. In this study, the RNase domain of the Erns was produced in Escherichia coli. Its reactivity with CSFV-positive sera and its ability to induce antibodies and to provide protective immunity were then investigated. The serological tests showed that the prokaryotically expressed RNase domain of the Erns retained its antigenicity and induced high titers of humoral responses. However, only partial protection and a limited amount of neutralizing antibodies were demonstrated by an in vitro neutralization test and an immunization/challenge test. The results suggest that other essential factors rather than simply enhancing the immunogenicity of Erns should be taken into consideration when Erns is enrolled as one of the components of a candidate vaccine.