Table 1

Strains and plasmids used in this study

Strain or plasmid

Description

Reference or source


E. coli strains


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


King et al. BMC Microbiology 2012 12:8   doi:10.1186/1471-2180-12-8

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