Abstract:

To construct recombinant adenovirus shuttle plasmid pDC315H5HAEGFP, HA gene of A/Swine/Fujian/1/2001(H5N1) was amplified by RTPCR and then inserted into adenovirus shuttle plasmid pDC315. A replicationdefective recombinant adenovirus expressing HA gene (rAdH5HAEGFP) was generated by cotransfecting HEK293 cells with the recombinant shuttle plasmid pDC315H5HAEGFP and the genomic plasmid pBHGlox△E1, E3Cre. The recombinant adenovirus was confirmed by PCR, RTPCR and Western blot assay. These results demonstrated that HA protein was properly expressed by the rAdH5HAEGFP in HEK293 cell with natural biological activities. The TCID50 of the rAdH5HAEGFP was evaluated as 2.26×1010 mL1 after propagation and purification. And the results of immunization in BALB/c mice indicated rAdH5HAEGFP could induce HI antibodies and inhibit virus replication in mice lungs.