Objective To construct adenovirus vector vaccine against H5N1 influenza virus and study on the immunogeuicity. Methods In this study, we amplified hemagglutinin (HA) gene sequence of H5N1 influenza virus (A/Anhui/1/2005), then constructed an adenovirus vector vaccine (Adv-HA), followed by tests in BALB/c mice for the immunogenicity with the vaccine and immunization strategies. Results The recombinate Adv-HA vaccine could effectively induce both humoral and cellular immunity against human H5N1 influenza virus. Conclusion The Adv-HA vaccination against H5N1 influenza is a potential strategy and worthy of further investigation.