TABLE 1.
Strain and plasmid descriptions
| Strain or plasmid | Lab strain designation | Descriptiona | Gene (norA, qacC, and/or qacA) present or other descriptionb | Reference |
|---|---|---|---|---|
| S. aureus strains | ||||
| RN4220 | DSM1935 | Wild type | norA | 62 |
| RN4220 ΔnorA | DSM1936 | S. aureus RN4220 with a clean deletion of norA | None | This study |
| RN4220 N1 | DSM1937 | RN4220 with a norA promoter mutation in the −10 element (TACAAT to TATAAT) | norA+ | This study |
| RN4220 N2 | DSM1938 | RN4220 with a norA promoter mutation in the 5′ untranslated region (TTTTT to TTCTT) | norA+ | This study |
| RN4220 (pCN38) | DSM1715 | RN4220 carrying cloning vector pCN38 | norA | 47 |
| RN4220 (pCN38-qacC [pSK41]) | DSM1939 | RN4220 carrying pCN38 containing qacC from pSK41 (accession number AF051917.1) | norA, qacC+ | This study |
| RN4220 (pCN38-qacC [pSK89]) | DSM1940 | RN4220 carrying pCN38 containing qacC from pSK89 (accession number M37889.1) | norA, qacC | This study |
| RN4220 (pCN38-qacA qacR) | DSM1941 | RN4220 carrying pCN38 containing qacA and qacR from pC02 (accession number CP012121.2) | norA, qacA | This study |
| RN4220 (pCN38-qacA qacR qacC [pSK41]) | DSM1942 | RN4220 carrying pCN38 containing qacA, qacR, and qacC from pSK41 | norA, qacA, qacC+ | This study |
| RN4220 (pCN38-qacA qacR qacC [pSK89]) | DSM1943 | RN4220 carrying pCN38 containing qacA, qacR, and qacC from pSK89 | norA, qacA, qacC | This study |
| RN4220 ΔnorA (pCN38-qacA qacR) | DSM1944 | RN4220 ΔnorA carrying pCN38 containing qacA and qacR | qacA | This study |
| RN4220 N1 (pCN38-qacA qacR) | DSM1945 | RN4220 N1 carrying pCN38 containing qacA and qacR | norA+, qacA | This study |
| RN4220 (pCN51) | DSM1946 | RN4220 carrying pCN51 | norA | This study |
| RN4220 ΔnorA (pCN51) | DSM1947 | RN4220 ΔnorA carrying pCN51 | None | This study |
| RN4220 ΔnorA (pCN51-norA) | DSM1948 | RN4220 ΔnorA carrying pCN51 containing norA from S. aureus RN4220 | norA | This study |
| RN4220 (pCN51-qacC) | DSM1950 | RN4220 carrying pCN51 containing qacC from pSK41 (accession number AF051917.1) | norA, qacC | This study |
| RN4220 (pCN51-qacA) | DSM1949 | RN4220 carrying pCN51 containing qacA from pC02 (accession number CP012121.2) | norA, qacA | This study |
| 2014.C02 (also known as C02) | DSM1418 | Clinical isolate carrying qacA-positive pC02 (accession number CP012121.2) | norA, qacA | 18 |
| 1969.N | DSM1504 | Clinical isolate carrying qacA-positive p1969.Nb (accession number CP016860.2) | norA, qacA | 18 |
| 2148.C01 | DSM1499 | Clinical isolate carrying qacA-positive p2148.C01b (accession number CP017095.2) | norA, qacA | 18 |
| 5118.N | DSM1491 | Clinical isolate carrying qacA-positive p5118.Nb (accession number CP016854.2) | norA, qacA | 18 |
| E. coli strains | ||||
| DH5 α (pCN38) | DSM1595 | DH5 α carrying pCN38 | 47 | |
| TOP10 (pCN38-qacC [pSK41]) | DSM1972 | TOP10 carrying pCN38 containing qacC from pSK41 (accession number AF051917.1) | This study | |
| TOP10 (pCN38-qacC [pSK89]) | DSM1973 | TOP10 carrying pCN38 containing qacC from pSK89 (accession number M37889.1) | This study | |
| TOP10 (pCN38-qacA qacR) | DSM1974 | TOP10 carrying pCN38 containing qacA and qacR from pC02 (accession number CP012121.2) | This study | |
| TOP10 (pCN38-qacA qacR qacC [pSK41]) | DSM1975 | TOP10 carrying pCN38 containing qacA, qacR, and qacC from pSK41 | This study | |
| TOP10 (pCN38-qacA qacR qacC [pSK89]) | DSM1976 | TOP10 carrying pCN38 containing qacA, qacR, and qacC from pSK89 | This study | |
| DH5 α (pCN51) | DSM1933 | DH5 α carrying pCN51 | This study | |
| TOP10 (pCN51-norA) | DSM1977 | TOP10 carrying pCN51 containing norA from S. aureus RN4220 | This study | |
| TOP10 (pCN51-qacC) | DSM1978 | TOP10 carrying pCN51 containing qacC from pSK41 (accession number AF051917.1) | This study | |
| TOP10 (pCN51-qacA) | DSM1979 | TOP10 carrying pCN51 containing qacA from pC02 (accession number CP012121.2) | This study | |
| TOP10 (pKOR1-ΔnorA) | DSM1930 | TOP10 carrying pKOR1 containing a fused upstream and downstream region of norA | This study | |
| TOP10 (pKOR1-N1) | DSM1931 | TOP10 carrying pKOR1 containing norA promoter with the N1 mutation | This study | |
| TOP10 (pKOR1-N2) | DSM1932 | TOP10 carrying pKOR1 containing norA promoter with the N2 mutation | This Study | |
| Plasmids | ||||
| pCN38 | Shuttle vector for E. coli and S. aureus | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus | 67 | |
| pCN38-qacC [pSK41] | pCN38 containing qacC from pSK41 | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus | This study | |
| pCN38-qacC [pSK89] | pCN38 containing qacC from pSK89 | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus | This study | |
| pCN38-qacA qacR | pCN38 containing qacA and qacR from S. aureus 2014.C02 | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus | This study | |
| pCN38-qacA qacR qacC [pSK41] | pCN38 containing qacA and qacR from S. aureus 2014.C02 and qacC from pSK41 | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus | This study | |
| pCN38-qacA qacR qacC [pSK89] | pCN38 containing qacA and qacR from S. aureus 2014.C02 and qacC from pSK89 | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus | This study | |
| pCN51 | Shuttle vector for E. coli and S. aureus. Plasmid multicloning site is located downstream of a cadmium-inducible promoter. | Encodes ampicillin resistance in E. coli and erythromycin resistance in S. aureus | 67 | |
| pCN51-norA | pCN51 containing norA from S. aureus RN4220 | Encodes ampicillin resistance in E. coli and erythromycin resistance in S. aureus | This study | |
| pCN51-qacA | pCN51 containing qacA from S. aureus 2014.C02 | Encodes ampicillin resistance in E. coli and erythromycin resistance in S. aureus | This study | |
| pCN51-qacC | pCN51 containing qacC from pSK41 | Encodes ampicillin resistance in E. coli and erythromycin resistance in S. aureus | This study | |
| pKOR1-ΔnorA | pKOR1 containing the ΔnorA construct | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus. Plasmid used to create S. aureus RN4220 ΔnorA. | This study | |
| pKOR1-N1 | pKOR1 containing the N1 norA promoter construct | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus. Plasmid used to create S. aureus RN4220 N1. | This study | |
| pKOR1-N2 | pKOR1 containing the N2 norA promoter construct | Encodes ampicillin resistance in E. coli and chloramphenicol resistance in S. aureus. Plasmid used to create S. aureus RN4220 N2. | This study |
a
Accession numbers are from GenBank.
b
“+” indicates increased expression.