Abstract
Staphylococcus aureus (S. aureus) is a versatile bacterium which exhibits multiple antibiotic resistances. To ameliorate the undesirable diseases causing potential, there is a need to design a protective vaccine capable of stimulating immune response against this pathogen. In a similar study in our laboratory, reverse vaccinology approach was used to nominate potential vaccine candidate genes against S. aureus. Zinc Metalloproteinase Aureolysin (aur) gene was one of the nominated genes based on that previously published in-silico study. The objective of this study is the cloning, expression, purification of aur gene and testing the aur protein reactivity with serum antibodies collected from groups of human patients with confirmed Staphylococcal disease. Cloning was done in pH6HTN His6HaloTag® vector and it was expressed in E. coli BL21 (DE3) using these conditions; 0.5 mM Isopropyl β-D-1-thiogalactopyranoside (IPTG) for 4 h at 37°C. Purification was carried out by Immobilized Metal Affinity Chromatography (IMAC). The his-tag aur protein was detected at ~86 KDa as a single band after western blot assay and was successfully reacted with antibodies obtained from humans infected with S. aureus. The results encourage further testing of aur protein as a potential vaccine candidate for S. aureus. Key words: Staphylococcus aureus, Zinc Metalloproteinase Aureolysin (aur), cloning, expression.
Highlights
Staphylococcus aureus has been indicated as a causative microorganism a lot of diseases, including osteomyelitis, septic arthritis and Necrotizing pneumonia
The aims of the current study are cloning, expression, purification of aur and demonstration of the reactivity of the purified aur against antibodies obtained from human infected with S. aureus
The increasing rate of prevalence of antibiotic resistance in S. aureus necessitated the research into the development of a protective vaccine against the organism, but all the clinical trials that were carried out to date failed as reported by Proctor (2015)
Summary
Staphylococcus aureus has been indicated as a causative microorganism a lot of diseases, including osteomyelitis, septic arthritis and Necrotizing pneumonia. Olaniyi et al (2017) reported that S. aureus is responsible for most of the skin and soft tissue infections. MethicillinResistant S. aureus (MRSA) has been implicated as the cause of most nosocomial infections and is reported with high prevalence especially in the hospitals with significant mortality and morbidity as reported by Toleman et al, (2019), Marlieke et al,(2011). It was reported that multidrug resistant S. aureus was increasingly detected globally with fewer antibiotics remaining for effective treatment as reported by Harik et al (2016), Chong et al (2015) and Bendary et al (2016).
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