Cloning and sequencig of the Influenza A (H9N2) M2e - HSP70 fusion protein in prokaryotic expression vector in order to recombinant vaccine production

*Rigi cheshmehali Garshasb - Mashayekhpoor Saied-Aghaiepour Khosrow

            

Iran-Razi Vaccine and Serum Research Institute

*Corresponding author,s E. mail: garshasbbiotech@gmail.com

Abstract

      One of the concerns about influenza A vaccine based on M2e protein is their limited potency; hence, optimal approaches to enhance immunogenicity of M2e protein immunization remain to be established. It seems by linking this M2e-peptide to an appropriate carrier such as mycobacterium tuberculosis C-terminal 28-kDa domain of HSP70 (HSP70 359-610), we can render it very immunogenic.This study was designed to produce a novel influenza A virus recombinant fusion protein consisted of M2e , a potent immunogenic protein from influenza A virus, fused to C-terminal domain of mycobacterium tuberculosis HSP70, HSP70359-610 , as a carrier and adjuvant. We fused the genes of M2e and HSP70359-610 then inserted in pQE-60, prokaryotic expression vector PQE-60. Identification of the cloned gene was confirmed by PCR, restriction analysis and gene sequencing.Then cloned products were sent for sequencing to check the correct ORF and direct origin of the cloned gene. Comparison of the nocleotid sequence of this gene and sequence of amino acid product of that with registered sequences in genbank shows that this gene with total isolated types has much resemblance and after gene expression in prokaryotic system , it’s protein product can be a good candidate for recombinant vaccine against the Influenza.

 

Keywords: cloning-sequencing-Influenza A-M2e gene-HSP70fusion protein-PQE-60 vector-PCR.recombinant vaccine