Successful Application of Site-directed Mutagenesis Polymerase Chain Reaction to Mutate TMS 11 of the Staphylococcal Multidrug Efflux Protein QacA

Date Received: Mar 14, 2019

Date Published: Aug 12, 2020

DOI: https://doi.org/10.31817/vjas.2020.3.1.04

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Issue: Vol. 3 No. 1 (2020): Vietnam Journal of Agricultural Sciences

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ANIMAL SCIENCE – VETERINARY MEDICINE – AQUACULTURE

How to Cite:

Hang, N., Hanh, V., Phai, D., Huong, L., Phuong, N., Truong, L., … Brown, M. (2020). Successful Application of Site-directed Mutagenesis Polymerase Chain Reaction to Mutate TMS 11 of the Staphylococcal Multidrug Efflux Protein QacA. Vietnam Journal of Agricultural Sciences, 3(1), 512–519. https://doi.org/10.31817/vjas.2020.3.1.04

Successful Application of Site-directed Mutagenesis Polymerase Chain Reaction to Mutate TMS 11 of the Staphylococcal Multidrug Efflux Protein QacA

Nguyen Thi Hang (*) 1, 2 , Vu Duc Hanh 3 , Dam Van Phai 1 , Lai Thi Lan Huong 1 , Nguyen Thi Phuong 1 , Le Van Truong 1 , Mohsen Chitsaz 2   , Melissa H Brown 2

  • Corresponding author: hang.phar2012@gmail.com
  • 1 Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi 131000, Vietnam
  • 2 School of Biology, Flinders University, Sturt Road, Bedford Park, South Australia 5042, Australia
  • 3 1Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi 131000, Vietnam

    • Keywords

    S.aureus, gene qacA, protein QacA, site-directed PCR

    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.

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