Abstract: Universal vaccine via intranasal immunization was designed based on antigenicity of the antigen epitope and the mucosal adjuvant of E. coli heatlabile toxin B subunit (LTb). Several epitopes of swine influenza virus (SIV) were designed and linked with LTb gene in C terminal. The recombinant pET30(a)-ep-LTb was constructed and the fusion protein was expressed in E. coli BL21(DE3). SDS-PAGE electrophoresis and Western blot were used for recombinant protein expression and bioactirity analysis. Humoral and mucosal antibody of the mice immunized with the recombinant protein was detected using ELISA and HI test. The recombinant protein was about 38 kDa in SDSPAGE, which the molecular weight was the same with expect. EP-LTB fusion protein can reacted with rabbit anti-CTB antibody and anti His-tag monoclonal antibody. Serum IgG and mucosal IgA in immunized mice were higher than that of the control. Especially the mucosal IgA antibody can be a good protection against the pathogen infection through the mucosa. Serum HI antibody of the mice immunized with the EP-LTB was higher than that of the control. EP-LTB fusion protein has the immunogenicty and reactogenicity of SIV, which may be a good candidate for developing novel universal vaccine．