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. 2021 Feb 5;7:571999. doi: 10.3389/fvets.2020.571999

Figure 1.

Figure 1

Gene construction and expression of M2e–HA2 recombinant protein in Escherichia coli Rosetta Blue (DE3)pLysS. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis and evaluation of recombinant M2e–HA2 protein by indirect ELISA with positive control sera. (A) Synthetic gene constructs coding for M2e (2–24 AA) and HA2 conserved regions (N-terminal 1–38 AA and Long α-helix 76–130 AA) with chicken dendritic cell binding peptides were designed by placing glycine linkers (GGG) in between them. (B) SDS-PAGE analysis of recombinant M2e–HA2 protein by Coomassie blue stain and (C) Western blot analysis of recombinant M2e–HA2 protein (38 kDa) showed high reactivity with polyclonal avian influenza virus (AIV) serum. (D) Indirect ELISA with recombinant M2e–HA2 fusion protein showed high reactivity with different clades (2.2 and 2.3.2.1) of H5N1 AIV, H9N2 AIV but failed to react with NDV and control chicken serum.