High-Yield Expression of M2e Peptide of Avian Influenza Virus H5N1 in Transgenic Duckweed Plants

Molecular Biotechnology
Aleksey FirsovSergey Dolgov

Abstract

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 F...Continue Reading

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Citations

Jul 22, 2016·Plant Biotechnology Journal·Naila Shahid, Henry Daniell
Oct 19, 2016·International Journal of Molecular Sciences·Young Hee JoungHyun-Soon Kim
Jul 9, 2020·Plants·Balamurugan ShanmugarajWaranyoo Phoolcharoen
Sep 26, 2020·Frontiers in Immunology·P P M HeenatigalaHongwei Hou
Oct 17, 2020·Frontiers in Veterinary Science·Naila ShahidAbdul Qayyum Rao
Jan 17, 2021·Biomolecules·Gui-Li YangAi-Juan Tan
May 1, 2021·3 Biotech·Pavel KhvatkovSergey Dolgov

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