Shuttle vectors derived from pN315 for study of essential genes in Staphylococcus aureus

Biological & Pharmaceutical Bulletin
Miki MatsuoKazuhisa Sekimizu

Abstract

Using the par to rep region of the 24653 bp plasmid pN315, which is present in Staphylococcus aureus strain N315, we constructed three vectors that can be shuttled between Escherichia coli and S. aureus and maintained stably in S. aureus. Due to plasmid incompatibility, the resident plasmid in S. aureus cells can be replaced via transformation with an entering plasmid, which carries a different drug resistance gene. To evaluate the applicability of this plasmid-based approach for identifying genes essential for S. aureus cell growth, the chromosomal mraY gene, which is involved in peptidoglycan biosynthesis, was deleted in cells harboring a resident plasmid with an intact mraY gene. The resultant disruptant was then transformed with an empty vector. Cells with a chromosomal mraY deletion but lacking the plasmid supplying mraY could not be recovered, suggesting that mraY is indispensable for staphylococcal cell growth or viability. In contrast, other two genes were shown to be dispensable by this system. Thus, the pN315-based plasmids appear to be useful for studying genes essential for S. aureus cell growth.

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Citations

Jun 5, 2012·Journal of Microbiological Methods·Shinichiro YamachikaHiroyuki Takase
Nov 18, 2016·Antimicrobial Agents and Chemotherapy·Madhuri SinghKeiichi Hiramatsu
Oct 17, 2012·The Journal of Immunology : Official Journal of the American Association of Immunologists·Dong-Jun JungBok Luel Lee
Aug 18, 2012·Molecular BioSystems·Haihong HaoZonghui Yuan

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