Expression of the nucleotide sequence for the M2e peptide of avian influenza virus in transgenic tobacco plants

Abstract

The M2e peptide of the H5N1 A/Chicken/Kurgan/05/2005 avian influenza virus was successfully synthesized in transgenic tobacco plants. The amino-terminal segment of the M2 protein, comprised of 22, 30, or 43 amino acids, including the M2e peptide (M122, M130, and M143 variants, respectively), was translationally fused with the N-terminus of β-glucuronidase in the pBI121 plant expression vector. The nucleotide sequence of the target fragment was synthesized by ligation from synthetic oligonucleotides; its codon composition was adapted for expression in plants. Tobacco plants were successfully transformed with the obtained vectors (pBIM122, pBIM130, and pBIM143, respectively). In the plants transformed with pBIM143, the Ml43-β-glucuronidase fusion protein was not produced, probably due to the presence of the M2 protein transmembrane domain (25–43 aa of M2) in this construct. In the pBIM122- and pBIMl30-transformed plants, the target M2e peptide was expressed as a component of the Ml22-β-glucuronidase and M130-β-glucuronidase fusion proteins, respectively, as was detected by Western blot analysis. These proteins were detected as bands of the expected size without apparent degradation. As a result, the M2e peptide of the H5N1 avian influenza virus was successfully synthesized for the first time in nuclear-transformed transgenic plants. The results obtained in this study will be used for developing a transgenic plant-based edible antiinfluenza vaccine.