TABLE 2.
Primers for generating fragments to be cloned into recombinant plasmids in efflux pump inactivation and gyrA mutation studies
| Primer name | Sequence (5′-3′) | Product size (bp) | Target gene/cluster |
|---|---|---|---|
| Efflux pump inactivation studya | |||
| 0035cF | CTGCACTTTCTCATGTAGAA | 809 | Cj0035c |
| 0035cR | GGCATTTACAAGTAAAATCA | ||
| 0309cF | GCGTGGTGATCATGAAAGAA | 228 | Cj0309c |
| 0309cR | CAGCGATTAAAAAAAGTTTC | ||
| 0309c-10cF | CGCTGTACAAGTAATAATCA | 591 | Cj0309c-Cj0310c |
| 0309c-10cR | TTAGGTGGAATTATAGAGTG | ||
| 0366cF | GCAATGATGCAGTTCCAATT | 1,166 | Cj0366c (cmeB) |
| 0366cR | GCCTGTGATACTTAATCCAG | ||
| 0560F | CTATAGGATCAGCAGTGCAA | 944 | Cj0560 |
| 0560R | TGCCTTATTAAAAGCTGTCA | ||
| 0619F | GGGATTTGCTTTCTAAGTAT | 1,210 | Cj0619 |
| 0619R | CCACGCAAAAACTCATCTGC | ||
| 1033F | CGACCTATTACCGTGCTAAT | 1,023 | Cj1033 |
| 1033R | GCAAGCAATGAGAGTTGTAC | ||
| 1173F | TTGCTGCTATTGTATTTGAA | 259 | Cj1173 |
| 1173R | CCAACGATAGAAAGAACAAT | ||
| 1173-4F | TTGCTGCTATTGTATTTGAA | 584 | Cj1173-Cj1174 |
| 1173-4R | CTGCTATTATCACGATAATG | ||
| 1241F | TGGCAGGAGTTTTGGTTGAT | 1,050 | Cj1241 |
| 1241R | AAGCAGGGCTTACAAAAAGT | ||
| 1257cF | TGGCTTATGCTTTAGGAACT | 545 | Cj1257c |
| 1257cR | CCCTCCAAGTCCTACACCTA | ||
| 1687F | CGGATTTAAAGCAGCTGTGT | 1,110 | Cj1687 |
| 1687R | ACAAGACCCATTTGTCTAAC | ||
| 16S 1a | AATACATGCAAGTCGAACGA | 1,004 | 16s rRNA |
| 16S 1b | TTAACCCAACATCTCACGAC | ||
| gyrA mutation studyb | |||
| upF | TGCTCTGCTTTTGTGAATTA | 850 | Cj1028c and gyrA |
| 86R | AAACTGCTGTATCTCCATGT | (ACA, Thr-86) | |
| upF | TGCTCTGCTTTTGTGAATTA | 850 | Cj1028c and gyrA |
| 86mR | AAACTGCTATATCTCCATGT | (ATA, Ile-86) |
a
Boldface primers were used for efflux gene RT-PCR only.
b
Point mutation sites of interest are underlined.