TABLE 1.
Primers used in this study
| Primer | Sequence (including restriction enzyme sequence) | Position | Primer combination | Template DNAa | PCR product size (bp) |
|---|---|---|---|---|---|
| P1L | 5′-GGGGGTACCGCTACTACTAAAAAGAAACCTA-3′ (after bp 136 including EcoRI site in fbe) | 59-80 in fbe | P1L-P1R | HB | 654 (fbe1; 527 after digestion) |
| P1R | 5′-CGCGGATCCGGCGGCTGACTTGTTGTT-3′ (BamHI site underlined) | 697-712 in fbe | |||
| P2L | 5′-GAGCTGCAGCGCCGACGCTTAAACATTCA-3′ (PstI site underlined) | 2059-2077 in fbe | P2L-P2R | HB | 1,179 (fbe2) |
| P2R | 5′-CCCAAGCTTGCGCGTGCCTTTAGAGCCATAAT-3′ (HindIII site underlined) | 3218-3238 in fbe | |||
| P3L | 5′-AGGTCATAATGAGGCCAAAGC-3′ | 169-189 in fbe::ermB | P3L-P3R | HBmut | 1,672 |
| P3R | 5′-CTGTGGTATGGCGGGTAAGA-3′ | 1822-1841 in fbe::ermB | HB | 0 (none) | |
| P4L | 5′-ATTGTCCGAGAGTGATTGGTC-3′ | 922-942 in fbe::ermB | P4L-P4R | HBmut | 919 |
| P4R | 5′-CTGTGGTATGGCGGGTAAGA-3′ | 1822-1841 in fbe::ermB | HB | 0 (none) | |
| P5L | 5′-ACTTACCCGCCATACCACAG-3′ | 1822-1841 in fbe::ermB | P5L-P5R | HBmut | 1,574 |
| P5R | 5′-CGTTTCCCTAATAATAACGCTCC-3′ | 3374-3396 in fbe::ermB | P3L-P5R | HB | 0 (none) |
| HBmut | 2,227 (for fbe::ermB sequencing) | ||||
| P6L | 5′-TAAACACCGACGATAATAACCAAA-3′ | 306-329 in fbe | P6L-P6R | HBmut | 0 (none) |
| P6R | 5′-GGTCTAGCCTTATTTTCATATTCA-3′ | 776-800 in fbe | HB | 495 |
a
HB, wild type; HBmut, mutant.