F1-V recombinant fusion protein was purified to immunize mice and its immunogenicity and immuno-protection are detected. Different purification strategies were used to purify F1-V recombinant fusion protein. First hydrophobic chromatography, then anion exchange chromatography, and finally size exclusion chromatography. Aluminium hydroxide gel adsorbed F1-V fusion protein was intramuscularly injected into BALB/c mice. Anti-serum titer and protection ratio were detected. The purity of F1-V recombinant fusion protein was more than 90%. The anti-serum titer was high up to 1∶(51 200 ± 800). The protein antigen was capable of inducing an effective immuno-protection against subcutaneous challenge of 400 LD50 virulent Yersinia pestis with the survival rate of 90%. The F1-V recombinant fusion protein was able to protect against subcutaneous challenge of virulent Yersinia pestis, suggesting that the F1-V substance is well suited for developmentas the active ingredient of the next plague vaccine.