MOLECULAR CLONING AND IMMUNOGENICITY EVALUATION OF spa PROTEIN OF METHICILLIN RESISTANT Staphylococcus aureus VACCINE CANDIDATES FOR MASTITIC COWS

Document Type : Research article

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Department of Microbiology, College of Veterinary Medicine, University of Baghdad

Abstract

One of the most prevalent and significant diseases worldwide affecting dairy cows is mastitis. The most prevalent pathogen associated with mastitis is thought to be Staphylococcus aureus. There is a global need for the development of innovative vaccines because current therapeutic techniques to treating S.aureus are not always effective. The objective of this study was to prepare recombinant SpA protein for immunization against mastitis caused by Staphylococcus aureus. and showing Its effect on expression of IL12 and IFN-γ in the spleen of lactating BALB/c mice Methicillin-resistant Staphylococcus aureus (MRSA) they were grown on Luria–Bertani broth (LB) at 37◦C until they were prepared for PCR amplification. The expression of the recombinant proteins was achieved through cloning and using Escherichia coli Rosetta (DE3) and an expression vector( plasmid pET-24a).in addition to expression of IL12 and IFN-γ gene in the spleen of mastitic BALB/c mice model. The SDS-PAGE bands indicate that Protein A (35.8 kDa) was soluble in the lysis buffer for all tested IPTG concentrations. This was observed at an OD600 of 0.6 following an 8-hour induction at 30°C with 0.1 mM IPTG. 100 mg of SpA recombinant protein was obtained. The results indicated that immunization with SpA protein showed significant differences compared to the control group (p < 0.0001). Furthermore, the mice immunized with the SpA protein and adjuvant exhibited a significant increase in IL-12 and IFN-γ levels compared to the control group (p = 0.0001). The immunization with SpA recombinant protein, SpA recombinant protein with incomplete

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