TABLE 5.
Bacterial strains, plasmids, cell line, and primers used in this study
| Strain, plasmid, or primer | Genotype or DNA sequence | Reference or source |
|---|---|---|
| S. aureus strains | ||
| RN6390 | Wild type | 30 |
| QT7 | RN6390 tet38 mutant | 21 |
| QT5 | RN6390 norB::cat | 21 |
| RN6390(pLI50) | Cmr | 20 |
| RN6390(pLI50-tet38) | tet38 overexpressor; Cmr | 20 |
| E. coli strains | ||
| DH10B | F− mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 | 31 |
| ΔlacX74 endA1 recA1 deoR Δ(ara leu)7697 araD139 | ||
| galU galK nupG rpsL λ− | ||
| BL21 | B F– ompT gal dcm lon hsdSB(rB– mB–) [malB+]K-12(λS) | Invitrogen |
| BL21(pTrcHis 2A-tet38) | Protein purification (Tet38) | 23 |
| BL21(pTrcHis 2A-norA) | Protein purification (NorA) | 19 |
| Plasmids | ||
| pTrcHis2A | Expression vector | Invitrogen |
| pLI50 | Shuttle plasmid E. coli-S. aureus; Cmr | 30 |
| Cell line A549 | Human lung adenocarcinoma (ATCC) | 20 |
| Primers | ||
| Synthesis of tet38 overexpressors | ||
| Forward | TCATTGGTGTAGAAGCTTATGATTATGAATa | |
| Reverse | CGCGAATTTATATCTATGGATCCTTATTTTb | |
| Protein expression, tet38 cloning into pTrcHis2A | ||
| Forward, pTrctet38-BamHI | ATCGGGATCCATGAATGTTGAATATTCTAAAATAAb | |
| Reverse, pTrctet38-HindIII | ATCGAAGCTTTTTTTCAGATTGTGTCCAACGATTTAa |
a
The HindIII site is underlined.
b
The BamHI site is underlined.