The Ebola hemorrhagic fever caused by Ebola virus is an extremely dangerous disease， and effective therapeutic agents are still lacking. Platforms for the efficient production of vaccines are crucial to ensure quick response against an Ebola virus outbreak. Ebola virus glycoprotein (EbolaGP) on the virion surface is responsible for membrane binding and virus entry， thus becoming the key target for vaccine development. However， heterologous expression of this protein still faces engineering challenges such as low production levels and insoluble aggregation. Here， the authors design and compare various fusion strategies， attaching great importance to the solubility-enhancing effect， and tag removal process. It is found that a C-terminal intein-based tag greatly enhances the solubility of EbolaGP and allows one-step chromatographic purification of the untagged EbolaGP through thiol-catalyzed self-cleavage. The purified untagged EbolaGP alone or with Freund's adjuvant are highly immunogenic， as confirmed in a mouse model. Consequently， the present study puts forward a new strategy for the efficient and soluble expression of untagged immunogenic EbolaGP. The intein-based protein fusion approach may be of importance for the large-scale production of Ebola virus subunit vaccine.