Abstract: The main purpose of this research is to construct a model cell which can form tumor in experimental animals, and the model cell and model animal can be used to study the vaccine of HPV16L1 as the main target in vitro and in vivo. First of all, HPV16L1 gene sequences were obtained by polymerase chain reaction (PCR) amplification and full-length open reading frame (ORF) was constructed in-frame into the pcDNA 3.1/blasticidin expression plasmid. The cell line of stable expression of HPV16L1 was obtained by transfection of recombinant plasmid into TC-1 cell line under the resistant screening of blasticidin. The addition of exogenous gene HPV16L1 did not cause significant changes in the characteristics of cells through the comparison of proliferation and tumorigenesis between TC-1 and TC-1-HPV16L1. TC-1-HPV16L1 can be killed by the spleen lymphocytes of mice immunized by HPV16L1 target vaccine in vitro test. Experiment of tumor challenge proved that the vaccine had protective effect on TC-1-HPV16L1 in animals.