Table 1.
Bacterial strains and plasmids used in this study.
| Strain/plasmid | Description | Source or Reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| XL10-Gold | Ultra-competent cell for site-directed mutagenesis | Stratagene |
| Genehogs | Electrocompetent cells | Invitrogen |
| S. aureus | ||
| RN4220 | Restriction-deficient derivative of 8325-4 | Novick, 1991 |
| RN6911 | RN6390B agr::tetM (agr-null) | Novick et al., 1993 |
| Z172 | Clinical VISA isolate with spc gene | Chen et al., 2013 |
| CGK5 | Daptomycin-susceptible MRSA | Lee et al., 2010 |
| CGK5mut | MprF L431F derivative of CGK5 | This study |
| CGK5mutR | Reversed derivative of CGK5mut with MprF containing F431L and a new EcoRV site | This study |
| CGK6 | Daptomycin-non-susceptible MRSA | Lee et al., 2010 |
| E. faecium | ||
| 2V076 | Clinical isolate with aadE-sat4-aphA-3 gene cluster | This study |
| Plasmids | ||
| pMAD | pE194ts derivative for gene replacement in Gram-positive bacteria | Arnaud et al., 2004 |
| pMAD-SAT4-tetM | Modified pMAD with sat4 and tetM markers | This study |
| pMprFmut | The mprF fragment amplified from CGK6 and cloned into pMAD-SAT4-tetM for allelic exchange in CGK5 | This study |
| pMprF5 | The mprF fragment amplified from CGK5 and cloned into pMAD-SAT4-tetM for site-directed mutagenesis | This study |
| pMprFmutR | A silent EcoRV site was introduced into the middle of the mprF fragment amplified from pMprF5 and cloned into pMAD-SAT4-tetM for allelic exchange in CGK5mut | This study |
| pluxT2 | pSK5630 derivative containing luxABCDE with T2 terminator | Chen et al., 2014 |
| pluxT2-SPC | pluxT2 with spc marker | This study |
tsStands for thermosensitive.