High-Yield Expression of M2e Peptide of Avian Influenza Virus H5N1 in Transgenic Duckweed Plants.
Avian influenza is a major viral disease in poultry. Antigenic variation of this virus hinders vaccine development. However, the extracellular domain of the virus-encoded M2 protein (peptide M2e) is nearly invariant in all influenza A strains, enabling the development of a broad-range vaccine against them. Antigen expression in transgenic plants is becoming a popular alternative to classical expression methods. Here we expressed M2e from avian influenza virus A/chicken/Kurgan/5/2005(H5N1) in nuclear-transformed duckweed plants for further development of avian influenza vaccine. The N-terminal fragment of M2, including M2e, was selected for expression. The M2e DNA sequence fused in-frame to the 5' end of β-glucuronidase was cloned into pBI121 under the control of CaMV 35S promoter. The resulting plasmid was successfully used for duckweed transformation, and western analysis with anti-β-glucuronidase and anti-M2e antibodies confirmed accumulation of the target protein (M130) in 17 independent transgenic lines. Quantitative ELISA of crude protein extracts from these lines showed M130-β-glucuronidase accumulation ranging from 0.09-0.97 mg/g FW (0.12-1.96 % of total soluble protein), equivalent to yields of up to 40 μg M2e/g plant FW. This relatively high yield holds promise for the development of a duckweed-based expression system to produce an edible vaccine against avian influenza.
Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Prospekt Nauki, 6, Pushchino, Moscow region, Russian Federation, 142290, aleksey_firsov@mail.ru.
Aleksey
A
Firsov
Irina
I
Tarasenko
Tatiana
T
Mitiouchkina
Natalya
N
Ismailova
Lyubov
L
Shaloiko
Alexander
A
Vainstein
Sergey
S
Dolgov
D000818:Animals; D001717:Birds; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology / Q000472:pathogenicity; D007252:Influenza Vaccines / Q000235:genetics* / Q000276:immunology; D005585:Influenza in Birds / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D007251:Influenza, Human / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D059828:Plant Cells; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000086082:Viroporin Proteins
D005966:Glucuronidase; D007252:Influenza Vaccines; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins