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Background: The PB1-F2 is the 11th protein of influenza A virus, encoded by the +1 alternate open reading frame (ORF) in the PB1 gene and previously reported to be a virulence factor that could contributed to the viral pathogenicity.To understand how the PB1-F2 protein evolved in different suptypes of influenza A viruses in different hosts, molecular characteristics of this protein were analyzed.