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Construction of DNA Vaccine Expressing H1 Subtype Swine Flu Virus HA Gene and Its Immunogenicity in Balb/c Mice |
LIU Chun-guo1 ,LIU Ming1,LI Hong-tao1,2,DU Jin-ling1,ZHANG Xin-tao1,SHI Wei-lin2 |
1.Animal Influenza Center of the Ministry of Agriculture and the National Avian Influenza Reference Laboratory, State Key Laboratory of Veterinary Biotechnology , Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China
2.Northeast Agricultural University Animal Medical School, Harbin 150003, China |
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Abstract The HA gene of subtype H1 swine influenza virus (SIV) A/Swine/Guangdong/LM/2004(H1N1) was amplified by RT-PCR , then it was cloned into eukaryotic expression vector pCI-neo and successfully constructed a recombinant expression plasmid pCI-HA. The pCI-HA and pCI-neo were transfected into vero E6 cells and 293T cells respectively to detect the expression of HA protein by Immunoperoxidase monolayer assay (IPMA),indirect immunofluorescence assay(iIFA) and Western blot(WB).The results showed that the HA protein expressed efficiently in vero E6 cells and 293T cells and with good biological activity. ELISA results showed that anti-H1 specific antibody response was elicited after immunizing Balb/c mice of 8 weeks old with DNA vaccine pCI-HA. This laid a foundation for H1 subtype swine influenza DNA vaccine research.
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Received: 16 February 2009
Published: 29 October 2009
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