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Table 1 Strains and plasmids used in this study

From: Characterisation of a cell wall-anchored protein of Staphylococcus saprophyticus associated with linoleic acid resistance

Strain or plasmid Description Reference or source
E. colistrains   
DH5α F- φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rk- mk+) phoA supE44 λ- thi-1 gyrA96 relA1 Grant et al. [50]
BL21 F- ompT hsdSB(rB- mB-) gal dcm Stratagene
MS2066 DH5α containing pSssFHis This study
MS2067 BL21 containing pSssFHis This study
S. saprophyticus strains   
ATCC 15305 Type strain (genome sequenced) Kuroda et al. [8]
MS1146 Clinical isolate AstraZeneca
MS1146sssF MS1146 isogenic sssF mutant This study
MS1146sssF(pSssF) Complemented MS1146 sssF mutant This study
S. aureus strains   
SH1000 Functional rsbU-repaired derivative of S. aureus 8325-4 Horsburgh et al. [51]
SH1000sasF SH1000 isogenic sasF mutant This study
SH1000sasF(pSKSasF) SH1000 sasF mutant complemented with sasF This study
SH1000sasF(pSKSssF) SH1000 sasF mutant complemented with sssF This study
SH1000sasF(pSK5632) SH1000 sasF mutant with empty pSK5632 vector This study
S. carnosus strains   
TM300 Wild-type SK311 Schleifer & Fischer [52]
TM300(pSssF) TM300 containing pSssF This study
Plasmids   
pBAD/HisB Cloning and protein expression vector, containing N-terminal 6 × His tag; Apr Invitrogen
pNL9164 E. coli/S. aureus TargeTron shuttle vector (temperature sensitive); Apr Emr Sigma
pSK5632 Cloning and expression E. coli/S. aureus shuttle vector; Apr Cmr Grkovic et al. [53]
pPS44 Staphylococcal vector, contains replicon and cat gene of pC194; Cmr Wieland [54]
pSssFHis 1330 bp MS1146 sssF fragment, amplified with primers 873 and 874, digested with EcoRI/XhoI and cloned into EcoRI/XhoI-digested pBAD/HisB, with in-frame N-terminal 6 × His tag; Apr This study
pNK24 pNL9164 shuttle vector retargeted with primers 1001-1003, EBSU to knock out MS1146 sssF (TargeTron system); Apr Emr This study
pNK41 pNL9164 shuttle vector retargeted with primers 2065-2067, EBSU to knock out SH1000 sasF (TargeTron system); Apr Emr This study
pSKSssF 2394 bp fragment, including entire sssF gene from MS1146, amplified with primers 839 and 840 and cloned into the BamHI site of pSK5632; Apr Cmr This study
pSssF 2400 bp BamHI/XbaI fragment, containing sssF gene, subcloned from pSKSssF into BamHI/XbaI-digested pPS44; Cmr This study
pSKSasF 2175 bp fragment, including sasF gene from S. aureus NCTC 8325, amplified with primers 2084 and 2085 and cloned into the HindIII site of pSK5632; Apr Cmr This study