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HA Gene of H9N2 sub-type avian influenza virus from three strains in different times was amplified, purified and then sequenced.The resuits of its sequence analysis showed that the whole length of the amplified HA Gene was 1 683 bp, encoding 560 amino acids.The amino acid sequence of three virulent strains at cleavage site was R-S-S-R, which was low-pathogenicity strain.According to the amino acid sequence of the isolated strains, there were 7 potential glycosylation sites, and the receptor-binding site was the specific sequence of the avian-derived influenza virus.Amino acids on the left edge of receptor-binding site were all NGQQG, while amino acids on the right edge of receptor-binding site were GTSKA.From the comparative sequence analysis of HA Gene from some referenced strains, the results indicated that nucleotids and amino acid homology between isolated strains and referenced strains was higher.Evolutionary tree analysis showed that three strains were all Eurasian species, and there was a close relationship with the representative strains of A / duck / Hong Kong/Y280/97.