Table 1 Strains and plasmids used in this study
From: Functional genomic analysis of bile salt resistance in Enterococcus faecium
Strain or plasmid | Relevant characteristic(s) | Source or reference | |
|---|---|---|---|
E . faecium | Â | Â | Â |
E1162 | Clinical isolate (bloodstream infection), isolated in France, 1996 | [23] | |
ΔgltK | Markerless deletion mutant of gltK gene of E1162 | This study | |
ΔgspA | Markerless deletion mutant of gspA gene of E1162 | This study | |
ΔgltK+gltK | Complementation strain of ΔgltK; ΔgltK harboring pEF25-gltK | This study | |
ΔgspA+gspA | Complementation strain of ΔgspA; ΔgspA harboring pEF25-gspA | This study | |
E. coli strains | Â | Â | Â |
DH5α | E. coli host strain for routine cloning | Invitrogen | |
EC1000 | MC1000 glgB::repA; host strain for pWS3 derived vectors | [24] | |
Plasmids | Â | Â | Â |
pWS3 | Gram-positive thermosensitive origin of replication; Spcr | [29] | |
pDEL3a | pWS3 carrying the 5′ and 3′ flanking regions of gene gltK for mutant construction | This study | |
pDEL4a | pWS3 carrying the 5′ and 3′ flanking regions of gspA gene cluster for mutant construction | This study | |
pDEL3b | pDEL3a with a Genr cassette which was flanked by lox66- and lox71-sites cloned between the 5′ and 3′ flanking regions | This study | |
pDEL4b | pDEL4a with a Genr cassette which was flanked by lox66- and lox71-sites cloned between the 5′ and 3′ flanking regions | This study | |
pWS3-Cre | pWS3 derivative expressing Cre in E. faecium | [22] | |
pEF25 | Shuttle plasmid pAT18 with spectinomycin resistance cassette cloned in the EcoRI site; Spcr Eryr | [30] | |
pEF25-gltK | Complementation plasmid for ΔgltK; pEF25 carrying gltK | This study | |
pEF25-gspA | Complementation plasmid for ΔgspA; pEF25 carrying gspA | This study | |