Expression and Immunogenicity of M2e Peptide of Avian Influenza Virus H5N1 Fused to Ricin Toxin B Chain Produced in Duckweed Plants

Frontiers in Chemistry
Aleksey FirsovSergey Dolgov

Abstract

The amino acid sequence of the extracellular domain of the virus-encoded M2 matrix protein (peptide M2e) is conserved among all subtypes of influenza A strains, enabling the development of a broad-range vaccine against them. We expressed M2e from avian influenza virus A/chicken/Kurgan/5/2005 (H5N1) in nuclear-transformed duckweed plants for further development of an avian influenza vaccine. The 30-amino acid N-terminal fragment of M2, including M2e (denoted M130), was selected for expression. The M2e DNA sequence fused in-frame to the 3' end of ricin toxin B chain (RTB) was cloned under control of the CaMV 35S promoter into pBI121. The resulting plasmid was used for duckweed transformation, and 23 independent transgenic duckweed lines were obtained. Asialofetuin-binding ELISA of protein samples from the transgenic plants using polyclonal anti-RTB antibodies confirmed the expression of the RTB-M130 fusion protein in 20 lines. Quantitative ELISA of crude protein extracts from these lines showed RTB-M130 accumulation ranging from 0.25-2.5 μg/g fresh weight (0.0006-0.01% of total soluble protein). Affinity chromatography with immobilized asialofetuin and western blot analysis of protein samples from the transgenic plants showed exp...Continue Reading

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Citations

Sep 26, 2020·Frontiers in Immunology·P P M HeenatigalaHongwei Hou
Jan 17, 2021·Biomolecules·Gui-Li YangAi-Juan Tan
May 1, 2021·3 Biotech·Pavel KhvatkovSergey Dolgov

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Methods Mentioned

BETA
transgenic
PCR
ELISA
protein assay
electrophoresis
glycosylation

Software Mentioned

Statistica
StatSoft

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