ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2009, Vol. 40 ›› Issue (9): 1363-1369.

• 预防兽医 • Previous Articles     Next Articles

Construction and Immunogenicity of a Recombinant Adenovirus Expressing HA Gene of H5N1 Subtype Swine Influenza Virus

WU Yunpu 1,2, QIAO Chuanling 1*, YANG Huanliang 1, CHEN Yan 1, ZHAN Xiaoguo 1，XIN Xiaoguang 1, CHEN Hualan 1

  

  1. 1.Animal Influenza Laboratory of the Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China; 2. Liaoning Center for Animal Disease Control and Prevention, Shenyang 110164, China
  • Received:2009-03-12 Revised:1900-01-01 Online:2009-09-24 Published:2009-09-24
  • Contact: QIAO Chuanling

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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.

Key words: swine influenza virus, H5N1 subtype, HA gene, recombinant adenovirus