Prokaryotic Expression Of The BovIFN-γ Gene And The Region D Of Fibronectin-binding Protein A Gene Of Staphylococcus Aureus

Staphylococcus aureus (S. aureus) is the most common etiologic agent of mastitis in bovines and a major cause of economic losses in the dairy industry. S. aureus mastitis has been particularly difficult to treat, and antibiotic-resistant strains have become common. A vaccine to protect against infection would therefore be beneficial. Interferon-gamma (IFN-γ) is a pleiotropic cytokine characteristically produced bynatural killer cells and T helper 1 type cells in response to antigenic or mitogenic stimulus. Apart from its capacity to directly exert anti-viral activity and anti-tumor activity, it plays many critical roles in the modulation of immune responses. Due to its important biological activity, IFN-γis a focus among the medical and biological fields. In recent years, animal studies have pointed out that IFN-γin the direct preparations for the treatment or as a vaccine adjuvant for use of certain animal diseases show results. The present report describes a newly fusion protein, named IFN-γ-FnBPA that was constructed by fusion of the IFN-γgenes from bovine and FnBPA of S. aureus and expressed in E. coli. This study provides the basis for the vaccine development of cow mastitis caused by drug fast S. aureus.The result of the present study as following:(i) IFN-γgenes from bovine was successfully cloned. Amino acids sequence analysis revealed that RhIFN-gamma shares a homology of 100% with IFN-γgene (GenBank database accession no. M29867)(ii) DNA recombinant technology is used to construct recombinant plasmid containing IFN-γ-FnBPA fusion gene, named pET32a-IFN-γ-FnBPA.(iii) The recombinant plasmid was highly expressed in E.coli. BL21(DE3) after induction by IPTG and the fusion protein was purified further. Fusion protein activity was confirmed through a series of immunological detection.