Helicobacter pylori is one of the most common infectious pathogenic agents in human being which causes gastritis, peptic ulcer disease, gastric adenocarcinoma and malignant tumour of the stomach. ure B and hsp A gene were cloned and fused into a shuttle vector pADTrack-CMV. The resultant plasmid was linarized by digesting with restriction endonuclease PmeⅠ, and subsequently were contransformed into E.coli. BJ5183 with an adenoviral backbone plasmid PAdEasy-1. The linearized recombinant plasmid was transfected into adenovirus packaging cell lines HEK-293. The recombinant adenoviruses were typically generated within 7 to 12 days. The recombinant adenoviruses titer was monitored by GFP expression. ELISA was used to test whether adenoviruses efficiently express the Ure B-Hsp A antigen protein in HEK-293. To determine the immunogenicity of the recombinant adenovirus, the delayed type serum IgG, secreted IgA, in the immunized mice were tested. The results showed that both humoral and mucusal immune response in mice were induced.