:: Volume 6, Issue 4 (11-2012) ::
Iran J Virol 2012, 6(4): 13-19 Back to browse issues page
Over Expression of Influenza Virus M2 Protein in Prokaryotic System
MA Alavi-Esfahani , F Fotouhi-Chahooki * , M Saleh , R Tavakoli , B Farahmand , A Ghaemi , M Tavassoti-Kheiri
Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran
Abstract:   (6143 Views)
Background and Aims: Influenza A virus of Orthomyxoviridae family is able to create pandemic influenza. Vaccination is the most effective way to prevent influenza virus infection. Matrix protein 2 (M2) is a homotetramer ion channel with 97 amino acids length and highly conserved among influenza viruses and is considered for development of a universal influenza vaccine. Materials and Methods: We present here cloning and expression of influenza A virus M2 protein as a fusion with 6-His tag in Escherichia coli BL21 strain. The gene was amplified by PCR and ligated into the prokaryotic expression vector pET28a. The expression of M2 protein was induced by IPTG and confirmed by SDS-PAGE and western blotting. The desired protein was purified with affinity chromatography on a Ni-TED resin column and has to be evaluated in animal models for further studies. Results: The results of sequencing showed that M2 gene was cloned in pET28a properly in frame to histidine tag and the product was confirmed by xpreimmune reaction of monoclonal anti-M2 antibody to recombinant M2 in western blotting. Conclusion: This study might provide a basis for production of a universal and broad-spectrum human influenza vaccine.
Keywords: M2 Protein, Influenza Virus, Vaccine, pET28
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Type of Study: Original article | Subject: General
Received: 2014/11/7 | Accepted: 2014/11/7 | Published: 2014/11/7

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Volume 6, Issue 4 (11-2012) Back to browse issues page