Table 1.
Bacterial strains and plasmids used during this study.
| Name | Description | Source |
|---|---|---|
| Strains | ||
| UAS391 | Prolific biofilm forming MRSA USA300 strain isolated from a patient with an abscess in a Belgian hospital. | [18,19] |
| UAS391-EryS | Erythromycin-sensitive variant (loss of ermC gene) of S. aureus UAS391 obtained by plasmid curing through growth at 44 °C. | [17] |
| JE2 | Plasmid-cured derivative of MRSA USA300 LAC, isolated from a skin and soft tissue infection in a detainee from the Los Angeles County jail. | [16] |
| RN0450 (NRS135) | MSSA strain derived by successive cycles of UV treatment of S. aureus strain NCTC8325, curing it of phages Φ11, Φ12 and Φ13. | NARSA repository |
| RN0451 (NRS136) | MSSA strain derived from S. aureus strain RN0450, lysogenic for phage Φ11. | NARSA repository |
| RN4220 | Generated through UV and chemical mutagenesis of S. aureus strain RN0450 and selected for transformability with DNA from E. coli (restriction deficient through mutation in sau1 hsdR). | NARSA repository |
| DH5α | Escherichia coli cloning strain with multiple mutations (fhuA2 lacΔU169 phoA glnV44 Φ80’ lacZΔM15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17) that enable efficient transformation. | Thermo Fisher Scientific Inc., Waltham, MA, USA |
| ATCC® 6538™ | Positive quality control for biofilm formation of S. aureus under no flow conditions. | [20] |
| 5374 | Negative quality control for biofilm formation of S. aureus under no flow conditions. | [18] |
| ATCC® 25923™ | PIA/PNAG-dependent biofilm producing strain used as positive quality control during fluorescent staining and enzymatic treatment of biofilm matrix. | [21] |
| NE106 (NR-46649) | JE2 Tn mutant (insertion position: 943092) in argininosuccinate lyase (argH, SAUSA300_0863; 942072-943451). | NARSA repository |
| NE134 (NR-46677) | JE2 Tn mutant (insertion position: 2290057) in arginase (rocF, SAUSA300_2114; 2289284-2290192). | NARSA repository |
| NE427 (NR-46970) | JE2 Tn mutant (insertion position: 1985575) in fumarate hydratase, class II (fumC, SAUSA300_1801; 1984212-1985597). | NARSA repository |
| NE491 (NR-47034) | JE2 Tn mutant (insertion position: 1799416) in isocitrate dehydrogenase, NADP-dependent (icd, SAUSA300_1640; 1798291-1799559). | NARSA repository |
| NE569 | JE2 Tn mutant (insertion position: 1247122) in succinyl-CoA synthetase, beta subunit (sucC, SAUSA300_1138; 1246832-1247998). | NARSA repository |
| NE594 (NR-47137) | JE2 Tn mutant (insertion position: 1800430) in citrate synthase II (gltA, SAUSA300_1641; 1799608-1800729). | NARSA repository |
| NE626 (NR-47169) | JE2 Tn mutant (insertion position: 1145819) in succinate dehydrogenase, flavoprotein subunit (sdhA, SAUSA300_1047; 1145459-1147225). | NARSA repository |
| NE808 (NR-47351) | JE2 Tn mutant (insertion position: 1147490) in succinate dehydrogenase iron-sulfur subunit (sdhB, SAUSA300_1048; 1147225-1148040). | NARSA repository |
| NE861 (NR-47404) | JE2 Tn mutant (insertion position: 1367722) in aconitate hydratase (acnA, SAUSA300_1246; 1367131-1369836). | NARSA repository |
| Plasmids | ||
| pALC2073 | Contains the pSK236 vector, with the tetR-gene and the xyl/tetO promotor, originating from pWH35. | [22] |
| pGV5990 | argH gene amplified with primers ArgH-1 and ArgH-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | This study |
| pGV5992 | gltA gene amplified with primers GltA-1 and GltA-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV5994 | icd gene amplified with primers Icd-1 and Icd-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV5996 | sdhB gene amplified with primers SdhB-1 and SdhB-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV5998 | sucC gene amplified with primers SucC-1 and SucC-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV5999 | gltA & icd genes (operon of two genes; 1st gene gltA & 2nd gene icd) amplified with primers GltA-1 and Icd-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV6000 | sucC genes (operon of two genes) amplified with primers SucC-1 and SucC-3 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV6001 | sdhA gene amplified with primer SdhA-1 and SdhA-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV6002 | sdhA (2nd gene in operon of 3 genes) & sdhB (3rd gene in operon of 3 genes) genes amplified with primers SdhA-1 and SdhB-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV6003 | rocF gene amplified with primers RocF-1 and RocF-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV6005 | fumC gene amplified with primers FumC-1 and FumC-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |
| pGV6007 | acnA gene amplified with primers AcnA-1 and AcnA-2 and cloned in the EcoRI site of pALC2073 via Gibson cloning. | |