Prokaryotic expression and characterization of avian influenza A virus M2 gene as a candidate for universal recombinant vaccine against influenza A subtypes; specially H5N1 and H9N2.
The conserved M2 protein of influenza A virus is considered as a promising candidate target for a broad-spectrum, recombinant influenza A vaccine. In the present study, the open reading frame (ORF) of avian influenza A/chicken/Iran/101/1998 (H9N2) M2 gene was amplified then cloned in pAED4, prokaryotic expression vector. M2 protein was produced through the expression of this recombinant expression vector (pAED4-M2) in E. coli BL21 (DE3) strain. The expressed M2 protein was analyzed on SDS-PAGE. Western blot assay was used to examine the immunoreaction of the expressed protein using commercial polyclonal anti-M2 antibody. The antigenicity and biological activity of the recombinant protein was also qualitatively detected on infected MDCK cells surface by immunofluorescence assay using rabbit's immunized antiserum. So, according to the sequence alignment based on the mentioned isolate and the result of immunoassay reaction, it seems recombinant vaccine based on A/chicken/Iran/101/1998(H9N2) M2 protein isolate might cover majority of influenza A virus strains specially H5 and H9 circulating in Iran and neighbor regions significantly.
Ebrahimi
Seyyed Mahmoud
SM
Department of Biotechnology, Razi Vaccine and Serum Research Institute, Karaj, Tehran, Iran, smebrahimi@shirazu.ac.ir
Tebianian
Majid
M
Aghaiypour
Khosrow
K
Nili
Hassan
H
Mirjalili
Ali
A
D000818:Animals; D001717:Birds / Q000276:immunology / Q000821:virology; D002460:Cell Line; D004285:Dogs; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D015151:Immunoblotting; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000276:immunology*; D053127:Influenza A Virus, H9N2 Subtype / Q000235:genetics / Q000276:immunology*; D007252:Influenza Vaccines / Q000276:immunology*; D005585:Influenza in Birds / Q000276:immunology* / Q000517:prevention & control; D011387:Prokaryotic Cells / Q000378:metabolism*; D014614:Vaccines, Synthetic / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000235:genetics*; D000086082:Viroporin Proteins
D007252:Influenza Vaccines; D014614:Vaccines, Synthetic; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins