Abstract
Staphylococcus aureus is a major problem in both the clinical setting and within the community. S. aureus can quickly develop resistance to a wide range of antibiotics through a number of different mechanisms, of which, using transporters located in the cell membrane to pump antibiotics out of the cell is the most serious concern. In staphylococcal species, QacA, one such important transporter, is encoded by qacA. QacA is 55kD in size and has 14 transmembrane segments (TMS) (TMS 1-TMS 14). This research describes the mutation process of the amino acid residues in TMS 11 of QacA using site-direct PCR. In this research, 15 primers were successfully designed for site-directed mutagenesis PCR. The site-mutagenesis PCR was successfully conducted to create 15 qacA mutants. These mutants will be used in further functional research of QacA.
Highlights
Staphylococcus aureus is a major problem in both the clinical setting and within the community (Kumar et al, 2020)
S. aureus has a wide range of efflux transporters including TetA(K), which is the efflux pump that confers resistance to tetracycline, as well as the permeases NorA, NorB, and QacA/B that are all involved in mediating resistance to a diverse spectrum of functional and structural substrates (Nikaido, 2009)
The aim of this research was to apply sitedirected mutagenesis PCR to mutate 15 codons in qacA encoding for 15 amino acids that are involved in transmembrane segments (TMS) 11 of QacA
Summary
Staphylococcus aureus is a major problem in both the clinical setting and within the community (Kumar et al, 2020). S. aureus has the ability to quickly acquire and spread antibiotic resistance genes through horizontal gene transfer on mobile genetic elements such as plasmids. This has resulted in many strains having high levels of resistance, limiting the Nguyen Thi Hang et al (2019). S. aureus can develop resistance to a wide range of antibiotics through a number of different mechanisms. These mechanisms include enzyme inactivation of the antibiotic, alteration of the antibiotic target, alteration of metabolic pathways, inhibition of drug uptake via altered permeability of the cell wall, and increased efflux of the antibiotics out of the cell (Radestock & Forrest, 2011). S. aureus has a wide range of efflux transporters including TetA(K), which is the efflux pump that confers resistance to tetracycline, as well as the permeases NorA, NorB, and QacA/B that are all involved in mediating resistance to a diverse spectrum of functional and structural substrates (Nikaido, 2009)
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