TABLE 5.
Bacterial strains, plasmids, cell line, and plasmids used in this study
| Strain, plasmid, cell line, or primer | Genotype, relevant characteristic(s), or sequence | Reference no. or source |
|---|---|---|
| S. aureus and E. coli strains, plasmids, and cell line | ||
| S. aureus RN6390 | Wild type | 6 |
| S. aureus QT7 | RN6390 (tet38::cat) mutant | 6 |
| S. aureus RN6390 or QT7 (pLI50-tet38) | tet38 overexpressor; Cmr | 6 |
| S. aureus RN6390 or QT7 (pLI50-tet38-ΔL1) | tet38(L1-ΔDDL) overexpressor; Cmr | This study |
| S. aureus RN6390 or QT7 (pLI50-tet38-ΔL7) | tet38(L7-ΔFPG) overexpressor; Cmr | This study |
| S. aureus RN6390 or QT7 (pLI50-tet38-R106C) | tet38(R106C) overexpressor; Cmr | This study |
| S. aureus RN6390 or QT7 (pLI50-tet38-G151C) | tet38(G151C) overexpressor; Cmr | This study |
| E. coli BL21 | F− ompT gal dcm lon hsdSB(rB−mB−) [malB+]K-12(λS) | Novagen |
| pLI50 | Shuttle plasmid E. coli-S. aureus; Cmr | 6 |
| pTrcHisA | Cloning plasmid for protein expression | Invitrogen |
| pTrcHisA-tet38 | Tet38 protein expression | 4 |
| pTrcHisA-tet38-ΔL1 | Tet38-ΔL1 protein expression | This study |
| pTrcHisA-tet38-ΔL7 | Tet38-ΔL7 protein expression | This study |
| A549 | Human lung adenocarcinoma cells | ATCC |
| Primers for site-directed mutagenesis | ||
| Tet38-1F | Loop L1; 5′-AACATATCTGTAACGACAGTAAGT-3′ | |
| Tet38-1R | Loop L1; 3′-AAATTGATTTCTGTAGCCATTGCT-5′ | |
| Tet38-7F | Loop L7; 5′-AATGATGCATTAAGTTCACATTTC-3′ | |
| Tet38-7R | Loop L7; 3′-CTAATCGATTTTAAAGCATTAAAT-5′ | |
| Tet38-R106CF | R106C; 5′-CTTTTAGTTGGACGTATTATTCAAACT-3′ | |
| Tet38-R106CR | R106C; 3′-ATTTTCCAACATCAATTCCCATTA-5′ | |
| Tet38-R151CF | G151C; 5′-TCATTAGTTATCGGTACATTATCAGGT-3′ | |
| Tet38-R151CR | G151C; 3′-TTAAGTACGAGCAGTTATTCCTTG-5′ |