Table 1.
Bacterial strains, plasmids and primers used in this study
| Genotype, characteristic(s) or sequences | Reference or source | |
|---|---|---|
| S. aureus strains | ||
| RN4220 | restriction-deficient mutagenized RN450 | 7 |
| MW2 | CA-MRSA (USA 400 lineage) | 7 |
| MW2 tet38 | tet38 partial deletion mutant with cat gene insertion | 7 |
| MW2 (pMSP) | MW2 carrying empty plasmid vector pMSP3535 | this study |
| MW2 tet38 (pMSP) | MW2 tet38 carrying empty plasmid vector pMSP3535 | this study |
| MW2 tet38 (pTet38) | tet38-complemented strain | this study |
| E. coli strains | ||
| E. coli DH5a | general host for pMSP3535 vector | Invitrogen |
| Plasmids | ||
| pMSP3535 | Gram-positive bacterial shuttle vector, ERYRa | 20 |
| pMSP::tet38 (pTet38) | pMSP3535 with tet38 gene, ERYR | this study |
| PCR primers (5′–3′) | ||
| tet38 XhoI-Forb | GCTACTCGAGTGGATGCGTATGGGTATTTTAG | this study |
| tet38 BamHI-Rev | GCTAGGATCCTTATTTTTCAGATTGTGTCCAACG | this study |
| pMSP-For | AATGCAGGTTAACCTGGCTTATC | this study |
| pMSP-Rev | TGCATCACCACGCATTACAA | this study |
| qRT–PCR primers (5′–3′) | ||
| gmk-RT For | ACTAGGGATGCGTTTGAAGC | 7 |
| gmk-RT Rev | TCATGACCTTCGTCCATTGT | 7 |
| tet38-RT For | TGACAGGTGTGGCTATTGGT | this study |
| tet38-RT Rev | TTGCCTGGGAAATTTAATGC | this study |
a
ERY, erythromycin. Plasmid selection: 300 mg/L erythromycin for E. coli and 10 mg/L erythromycin for S. aureus.
b
Restriction sites are indicated by underlining.