Cloning of the Gene Encoding M2e of Influenza Virus in B. subtilis

Background and Aims: The ectodomain of matrix protein of influenza virus is a weak immunogen that is highly conserved among all subtypes of influenza A virus. Tandem repeats of these genes along with linker were used to enhance immunogenicity of M2e protein and so it can be served as a universal vaccine in both humans and livestock. Materials and Methods: In this study, the sequences of extra-domain of matrix protein of influenza A registered in NCBI was converted into codons compatible for Bacillus subtilis using JAVA codon adaptation tool software. Results: A cassette consist of four repeats of this codon optimized sequence, spaced by appropriate linkers and flanked by BamHI and HindIII restriction sites was designed and thoroughly used for the synthesis. The cassette then was cloned into pMR12 shuttle expression vector. Conclusion: Two kinds of prokaryotic host, E. coli BL21and Bacillus subtilis WB600 were transformed by pMR12+4M2e. The fidelity of the construct in both transformants was confirmed by enzymatic analysis and PCR.