Cysteine proteinases of C. sinensis are important virulence factors that induce pathological changes associated with larval migration and localized biliary epithelial destruction. This study investigated the immunogenicity and protective efficacy of a DNA vaccine encoding Clonorchis sinensis cysteine proteinase (CsCP). The CsCP cDNA sequence displays significant homology to the mammalian or trematode cathepsin L. Plasmid DNA carrying the CsCP gene (pcDNA3.1-CsCP) was injected into Sprague-Dawley (SD) rats intradermally. Animals injected with pcDNA3.1-CsCP developed CsCP-specific antibodies, which exhibited an IgG2a dominance in sera. In addition, the DNA vaccine elicited the production of IFN-gamma, but not IL-4 in splenocytes, suggesting the induction of a typical Th-1 dominated immune response in rats. The pcDNA3.1-CsCP induced a significant level of protection (31.5%, p<0.05) in SD rats challenged with C. sinensis metacercariae. These results indicate that pcDNA3.1-CsCP induces both humoral and cellular immune responses. The CsCP gene may be a good candidate for use in future studies of vaccination against clonorchiasis.