Opposing roles of σ B and σ B -controlled SpoVG in the global regulation of esxA in Staphylococcus aureus

Bettina Schulthess; Dominik A Bloes; Brigitte Berger-Bächi

doi:10.1186/1471-2180-12-17

BMC Microbiology

Table 1 Strains and plasmids used in this study

From: Opposing roles of σ^B and σ^B-controlled SpoVG in the global regulation of esxA in Staphylococcus aureus

Strain or plasmid	Relevant genotype; phenotype	Reference or source
S. aureus
Newman	Clinical isolate, ATCC 25904, natural saeS constitutive mutant	[21, 22]
BS304	Newman ΔesxA, markerless deletion	This study
SM148	Newman Δ(yabJ-spoVG)::erm(B); Em^r	[8]
IK184	Newman Δ(rsbUVW-sigB)::erm(B); Em^r	[23]
MS64	Newman sigB1(Am); Tc^r	[24]
SM99	Newman ΔarlR::cat; Cm^r	[9]
BS310	Newman Δagr::ermB; Em^r	This study
KS186	Newman Δagr::tet(M); Tc^r	[25]
BS309	Newman ΔsarA::ermB; Em^r	This study
LR15	Newman ΔsarA::tet(L); Tc^r	L. Reutimann
BB1002	Newman mec, MRSA derivative, Mc^r	[26]
BS307	BB1002 ΔesxA, markerless deletion	This study
NM143	Newman GISA derivative, in vitro selected mutant; Te^r	[27]
BS308	NM143 ΔesxA, markerless deletion	This study
*E. coli*
DH5α	F^-Φ80d/acZΔM15 recA1	Invitrogen
Plasmids
pKOR1	E. coli-S. aureus shuttle vector for markerless deletions using the counter selection system	[28]
pAC7	Expression plasmid containing the P_BAD promoter and the araC gene; Cm^r	[29]
pAC7-sigB	pAC7 with a 0.75 kb fragment containing the gene sigB from S. aureus Col; Cm^r	[30]
pBus1	E. coli-S. aureus shuttle plasmid with multicloning site from pBluescript II SK (Stratagene) and the rrnT14 terminator sequence from pLL2443; Tc^r	[31]
pyabJ	pBus1 containing a bacA promoter-yabJ ORF fusion construct; Tc^r	[10]
pspoVG	pBus1 containing a bacA promoter-spoVG ORF fusion construct; Tc^r	[10]
pyabJspoVG	pBus1 containing a bacA promoter-yabJ-spoVG operon fusion construct; Tc^r	[10]
pSP-luc⁺	Firefly luciferase casette vector; Ap^r	Promega
pesxAp-luc⁺	pBus1 containing an esxAp-luc⁺ fusion fragment; Tc^r	This study
pesxApΔσ^A-luc⁺	pesxAp-luc⁺ with deletion of the σ^A promoter	This study
pesxApΔσ^B-luc⁺	pesxAp-luc⁺ with deletion of the σ^B promoter	This study
pSB40N	Promoter probe plasmid; Ap^r	[29]
pasp23p	pSB40N with a 0.6 kb fragment covering the asp23 promoter region fused to the reporter gene lacZα; Ap^r	[30]
pesxAp	pSB40N with a 0.5 kb fragment covering the esxA promoter region fused to the reporter gene lacZα; Ap^r	This study
pyabJp	pSB40N with a 0.4 kb fragment covering the yabJ promoter region fused to the reporter gene lacZα; Ap^r	This study
pSTM07	pSB40N with a 0.37 kb fragment covering the capA promoter region fused to the reporter gene lacZα; Ap^r	[9]

^aAbbreviations are as follows: Ap^r, ampicillin resistant; Cm^r, chloramphenicol resistant; Em^r, erythromycin resistant; Mc^r, methicillin resistant; Tc^r, tetracycline resistant; Te^r, teicoplanin resistant.

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