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. 2007 Aug 1;81(20):11170–11178. doi: 10.1128/JVI.01217-07

FIG. 5.

FIG. 5.

Manipulation of potential glycosylation site motifs in the HA of a recent virus. (A) Sequence analysis of HA from A/England/26/99 determined that it contained two new potential glycosylation site motifs at positions 122 to 124 and 133 to 135 that were not found in strains from the previous influenza season or from older viruses such as A/Victoria/3/75. Using site-directed mutagenesis, the new potential glycosylation site motif at 122 to 124 was destroyed by mutating the motif from NES to TEG to create G1Δ. Similarly, the new potential glycosylation site motif at 133 to 135 was destroyed by mutating the motif from NGT to NGG to create G2Δ. (B) Western blot analysis of HA proteins from purified virions obtained following infection of MDCK cells with recombinant influenza viruses bearing HA and NA genes of A/England/26/99 or HA modified as illustrated in panel A to alter the glycosylation motifs at amino acids 122 to 124 (G1Δ) or 133 to 135 (G2Δ).