Abstract: Objecuiwe To consuruct a non-replicating vaccinia virus expressing HPV16 L1,L2E7 proteins as a candidate vaccine for cervical cancer. Methods Using vaccinia virus vector,we generated a strain of non-replicating recombinant vaccinia virus vaccine expressing HPV16 L1,L2E7 proteins by homologous recombination and identified by PCR and Western blot. Results We demonstrated that the L1,L2E7 gene of HPV16 were integrated into vaccinia genosome and could express L1,L2E7 protein stably when infected the CEF using PCR and Western blot assay. Conclusion 　NTVJL1/ L2 E7 can express L1, L2 E7 protein of HPV16 and can be taken as a candidate vaccine for HPV16 associated diseases.