In this study, a recombinant Pichia pastoris expression system was developed to express HPV16 L1 protein that was driven by a strong AOX1 promoter. HPV16L1 gene was cloned into vector pPICZ%α%B. HPV16 L1 protein expression induced by methanol was screened by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting. The results indicate that the HPV16 L1 protein is secreted by the recombinant P. pastoris, and the purified HPV16 L1 protein can self-assemble into virus-like particles(VLPs), which show a good immunogenicity and induces high-titer antibody in mice. HPV16 L1 protein was induced into methanol was screened by using sodium The results indicate that the HPV16 L1 protein is secreted by the recombinant P. pastoris, and the purified HPV16 L1 protein can self-assemble into virus-like particles (VLPs) , which show a good immunogenicity and induces high-titer antibody in mice.