Table 2 Strains and plasmids used in this study
Strains | Description | Reference |
|---|---|---|
RN4220 | S. aureus sau1 hsdR laboratory strain used for passaging plasmid DNA | |
USA300 | Community-acquired methicillin resistant S. aureus | NARSA repository [36] |
essB | USA300 carrying an internal deletion of essB | This study |
DH5α | E. coli K12 fhuA2 Δ (argF-lacZ)U169 phoA glnV44 Φ80 Δ (lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17 for cloning | Our collection |
BL21(DE3) | E. coli B F- dcm ompT hsdS (rB-mB-) gal for recombinant protein production | Our collection |
Plasmids | Description | Reference |
pWWW412 | pOS1 derivative carrying the constitutive hprK promoter, CmR | [33] |
pKOR1 | temperature sensitive plasmid for allelic replacement, CmR | [32] |
pET15b | Vector carrying an N-terminal hexa-histidine repeat followed by a thrombin cleavage site and T7 transcription start; AmpR | Novagen |
pGEX-2TK | Vector carrying GST followed by thrombin cleavage and phosphorylation sites, and a taq promoter; AmpR | GE Healthcare |
p essB | pWWW412 expressing wild-type essB codons 1-444 | This study |
p essB N | pWWW412 expressing codons 1–223 of essB | This study |
p essB NM | pWWW412 expressing codons 1–252 of essB | This study |
p essB C | pWWW412 expressing codons 253–444 of essB | This study |
p essB CM | pWWW412 expressing codons 220–444 of essB | This study |
p essB ΔM | pWWW412 expressing essB lacking codons 224-252 | This study |
p his - essB | pET15b expressing histidine tagged essB codons 1-444 | This study |
p his - essB NM | pET15b expressing histidine tagged essB codons 1- 252 | This study |
p his - essB CM | pET15b expressing histidine tagged essB codons 220- 444 | This study |
p his - essB ΔM | pET15b expressing histidine tagged essB lacking codons 224-252 | This study |
p gst - essB N | pGEX-2TK expressing GST fused to codons 1–223 of essB | This study |
p gst - essB C | pGEX-2TK expressing GST fused to codons 253–444 of essB | This study |