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. Author manuscript; available in PMC: 2019 Jun 8.
Published in final edited form as: ACS Infect Dis. 2018 Mar 23;4(6):1007–1018. doi: 10.1021/acsinfecdis.8b00017

Figure 3. The pTarKO system enables various chromosomal modifications for reverse genetic analysis in S. aureus.

Figure 3

(a) Schematic of the method and targeting plasmid used for allelic exchange of a target locus in S. aureus. (b) Table summarizing chromosomal modification of various targets in S. aureus. Days required to obtain some mutants using the pKFC vector is shown for comparison. (c) Schematic of the pathway for biosynthesis of lipoteichoic acid. (d) Genes involved in the lipoteichoic acid pathway are synthetically sick or lethal with the WTA pathway. A ΔltaA strain is partially viable in the presence of 1 μg/mL tunicamycin (tuni), which inhibits TarO in the WTA pathway. (e) The C-terminus of UgtP was fused to GFP protein in the chromosome. (f) The native promoter of SAOUHSC_01050 was swapped with the inducible tetR promoter that is controlled with anhydrotetracycline. In the presence of 1 μg/mL amsacrine (AMSA), which inhibits D-alanylation, the mutant is only viable if gene expression is induced.

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