TABLE 1.
DNA primers used for construction of plasmids
| Primer | Sequencea | Restriction enzyme |
|---|---|---|
| Primers for gene deletions | ||
| MC044-NqrA-F | AAGAAGGAGATATACCATGGGCAGCAGCCATC | NcoI |
| MC045-NqrA-R | CAGAATTCTTACCCTTCTTTCTCGATCTTG | EcoRI |
| MC046-NqrF-F | TTCCCGGGATGTCTACTATTATTTTTGG | XmaI |
| MC048-NqrEF-F | TTCCCGGGATGGAACATTATATTAGTCT | XmaI |
| MC050-NqrC-R | TTCCCGGGTTAGTTAAGACCTCCGTCAC | XmaI |
| RD-NQRb-Vc | CGATATACATATGGGCCTTAAAAAGTTTCTTG | NdeI |
| Seq-Vc-NQRc-R | CAGACGTGGTTCAATGCTTTTA | |
| TH001-deltaNqrC-F | ATTTGGTACCATGTCTAGCGCAAAAGAGCT | KpnI |
| TH002-deltaNqrC-R | ATTTGGTACCTTATTGCTTGCCGTAGCGCG | KpnI |
| TH003-deltaNqrB-F | GGTACCATGGCAAGCAATAACGATAG | KpnI |
| TH004-deltaNqrB-R | GGTACCTTACCCTTCTTTCTCGATCT | KpnI |
| Flanking primers for site-directed mutagenesis | ||
| NQR-F | GTATGAAGCAGCGGCGACGCTGTTCTATAC | |
| NQR-R | TTGGTATCTGAGCCAATCACGTTAAACAGT | |
| NqrB-Trp227Leu | ||
| NQR3-F | CTCAGGTGACCTAGTActGACgGCcGCAGATGGCTACTCAGGC | EagI |
| NQR3-R | GCCTGAGTAGCCATCTGCgGCcGTCagTACTAGGTCACCTGAG | EagI |
| NqrB-Phe186Ala | ||
| NQR5-1F | GCACTGGGTATTACCgcgGGTGTTGTGGTTGCC | |
| NQR5-1R | GGCAACCACAACACCcgcGGTAATACCCAGTGC | |
| NQR5-2F | TGACCTAGTATGGACgGCcGCAGATGGCTACTCAGGC | EagI |
| NQR5-2R | GCCTGAGTAGCCATCTGCgGCcGTCCATACTAGGTCA | EagI |
a Sequences are given from 5′ to 3′. Restriction sites are underlined, and nucleotides substituted for mutagenesis are designated by lower case letters.