Background and purpose: Staphylococcus aureus is a gram-positive cocci that causes many diseases, such as skin infections, food poisoning, risky shocks, and autoimmune disorders. Entrotoxins are the most important toxins of the bacteria. Among them, enterotoxin type B is the most common ones which is a super antigen. The aim of this study was the cloning and recombinant expression of SEB in order to achieve a stable construct for the protein production and use it as a vaccine candidate in future investigations.
Materials and methods: Enzymatic digestion was performed to confirm the presence of the seb gene into pET28a(+) expression vector. Recombinant expression of SEB was induced by IPTG following cloning confirmation. Then, protein purification was done under non-denaturing conditions using a nickel chromatography column. Protein confirmation was performed by Western blotting analysis.
Results: The results showed that the cloning of SEB in pET28a(+) vector was performed in an appropriate position between expected restriction sites. Also, SEB had a good and remarkable expression after induction by IPTG. Expressin confirmation was performed by western blotting.
Conclusion: Stable recombinant construct containing SEB gene was fabricated and transferred to BL21(DE3) host cells and appropriate recombinant expression of the protein was observed. So, this protein could be used in future studies as a vaccine candidate against SEB toxin of Staphylococcus aureus.