The classical dengue fever (DF) and dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) caused by dengue virus (DV) are the most important infection diseases mediated by arthropod-borne viruses. The pathogenesis of DV infection is not completely clear and there is no licensed vaccine or specific antiviral drug available currently. Envelope (E) protein of DV plays an important role in pathogenesis and immunity response. In this study, the recombinant E/pGEX-6P-1 plasmid expressing E protein of dengue virus serotype 2 (DV2) was constructed. After optimizing the expression conditions, including the optimum temperature and incubation time, high purity of E protein could be obtained and it could induce high titre of specific antibodies against DV2 in BALB/c mice. This study will provide useful information to further understand the function and application of DV E protein.