Amino acid sequences of the seven antigenic sites in template and chimaeric HAs constructed in this study. (a) HAs of five vaccine strains, Aichi/68 (Aic68), Yamanashi/77 (Yam77), Fukuoka/85 (Fuk85), Sydney/97 (Syd97), Wyoming/2003 (Wyo03), were used as starting materials. The deduced amino acid sequences showed discrepancies with the reported data (Okada et al., 2010) at six residues, which are highlighted in grey. Since we used these plasmids for the expression of HA the actual sequence determined is indicated in this figure. The name of the chimaeric HA and the amino acids that replaced the original ones are indicated under the template HA. Bars indicate the same amino acids as those of Aichi/68. (b) Location of the amino acids replaced in the chimaeric HA is indicated in the 3D structure of HA by using the examples of the Sydney/97 strain. The amino acids that formed a receptor-binding site were marked in grey. The amino acids replaced in the chimaera were marked as follows: site A, purple (121A), orange (131A) or red (142A); B1, blue; B2, light blue; C1, green; C2, light green; D, pink; E, yellow.