Table 1 Strains and plasmids used in this study

S. aureus

   Newman

Clinical isolate (ATCC 25904), rsbU⁺

[64]

   RN4220

NCTC8325-4 r^- m⁺

[65]

   CQ33

NewmanΔsa2056

This study

   CQ39

Newman pME2, Tc^r, Mc^r

This study

   CQ65

NewmanΔsa2339

This study

   CQ66

NewmanΔsecDF

This study

   CQ69

NewmanΔsecDF pME2, Tc^r, Mc^r

This study

   CQ85

Newman pCN34, Km^r

This study

   CQ86

Newman pCN34 pME2, Km^r, Tc^r, Mc^r

This study

   CQ87

NewmanΔsecDF pCN34, Km^r

This study

   CQ88

NewmanΔsecDF pCN34 pME2, Km^r, Tc^r, Mc^r

This study

   CQ89

NewmanΔsecDF pCQ27, Km^r

This study

   CQ90

NewmanΔsecDF pCQ27 pME2, Km^r, Tc^r, Mc^r

This study

E. coli

   DH5α

Cloning strain, [F-Φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rk-, mk+) phoA supE44 thi-1 gyrA96 relA1 λ-]

Invitrogen

Plasmid

Relevant genotype or phenotype

Reference or source

   pCN34

S. aureus-E. coli shuttle vector, pT181-cop-wt repC aphA-3 ColE1 Km^r

[56]

   pCQ27

pCN34 derivative carrying secDF and its promoter (Newman), Km^r

This study

   pCQ30

pKOR1 derivative carrying 1 kb fragments of the region up- and downstream of sa2056 amplified from Newman, ligated together with EcoRI and recombined at the attP sites, Ap^r, Cm^r

This study

   pCQ31

pKOR1 derivative carrying 1 kb fragments of the region up- and downstream of sa2339 amplified from Newman, ligated together with HindIII and recombined at the attP sites, Ap^r, Cm^r

This study

   pCQ32

pKOR1 derivative carrying 1 kb fragments of the region up- and downstream of secDF amplified from Newman, ligated together with HindIII and recombined at the attP sites, Ap^r, Cm^r

This study

   pKOR1

E. coli-S. aureus shuttle vector used to create markerless deletions; repF(Ts) cat attP ccdB ori ColE1 bla P _xyl /tetO secY570, Ap^r, Cm^r

[23]

   pME2

pBUS1 derivative carrying mecA and its promoter (COLn), Tc^r, Mc^r

[28]