TABLE 1.
Bacterial strains and plasmids used in this study
| Strain, plasmid, or oligonucleotide | Relevant markers and characteristics | Reference or origin |
|---|---|---|
| Strains | ||
| E. coli TG1 | supE Δthi(lac-proAB) hsdD5 (F′+ traD36 proAB lacIqZDM15) | 14 |
| L. lactis | ||
| MG1363 | Plasmid-free derivative of L. lactis subsp. cremoris NCDO712, Lac− Prt− | 13 |
| Wg2 | Wild-type L. lactis subsp. cremoris strain harboring pWV05, prtP1 | 27 |
| SK11 | Wild-type L. lactis subsp. cremoris strain harboring pSK111, prtP3 | 9 |
| Plasmids | ||
| pBSSK+ | Ampr, M13ori pBR322ori | Stratagene |
| pGEM-T Easy | Ampr, M13ori pBR322ori, linear T-overhang vector | Promega |
| pGhost 8 | Tetr, thermosensitive replicative plasmid in L. lactis | 34 |
| pJIM2366 | Emr, promoter probe vector containing luxAB genes | 47 |
| pJIM2374 | Emr, integrative promoter probe vector containing luxAB genes | 7 |
| Integrative plasmids carrying lux fusion | ||
| pJIM3029 | PpepC-lux, XbaI fusion of pJIM2374 and pBSSK+ containing PpepC on a 654-bp fragment | This work |
| pJIM3030 | PpepN-lux, XbaI fusion of pJIM2374 and pBSSK+ containing PpepN on a 767-bp fragment | This work |
| pJIM3031 | PpepX-lux, XbaI fusion of pJIM2374 and pBSSK+ containing PpepX on a 687-bp fragment | This work |
| pJIM3032 | PpepF22-lux, XbaI fusion of pJIM2374 and pBSSK+ containing PpepF22 on a 654-bp fragment | This work |
| pJIM3063 | PpepF21-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PpepF21 on a 772-bp fragment | This work |
| pJIM3066 | PpepA-lux, BamHI fusion of pJIM2374 and pGEM-T Easy containing PpepA on a 575-bp fragment | This work |
| pJIM3064 | PoppD-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PoppD on a 587-bp fragment | This work |
| pJIM3065 | PoppA-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PoppA on a 688-bp fragment | This work |
| pJIM3080 | PpepDA1-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PpepDA1 on a 763-bp fragment | This work |
| pJIM3081 | PpepM-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PpepM on a 913-bp fragment | This work |
| pJIM3082 | PpepP-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PpepP on a 778-bp fragment | This work |
| pJIM3106 | PpepQ-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PpepQ on a 760-bp fragment | This work |
| pJIM3118 | PpepT-lux, SpeI fusion of pJIM2374 and pGEM-T Easy containing PpepT on a 866-bp fragment | This work |
| Replicative plasmids carrying lux fusion | ||
| PJIM3120 | PprtP1-lux, pJIM2366 carrying PprtP1 on a 1,051-bp SpeI-NotI fragment from strain Wg2 | This work |
| pJIM3119 | PprtP3-lux, pJIM2366 carrying PprtP3 on a 1,056-bp SpeI-NotI fragment from strain SK11 | This work |
| Oligonucleotide pairs used for amplification or promoter and probe fragments | ||
| PpepA | GGGAATTCTAAAGTTTTGGCATT/CGGGATCCATAAATCCGACCTCAT | X81089 |
| PpepC | GGGAATTCCTTTGATTATTCATC/CGGGATCCAGATTGTTTTTGATTC | M86245 |
| PpepDA1 | TGTAGCAGACCTTGTATCAGC/GACATCGCAACATTAGCACTA | AE005176 |
| PpepF21 | GGGAATTCCAATTGAGCGAACTA/CGGGATCCTTGAGACGCTTAATA | X99710 |
| PpepF22 | GGGAATTCTAGTTTTATTCGTCA/CGGGATCCTTGATACTTTGCTTCTC | X99710 |
| PpepM | ACCTTTCCAGATGACCCGTTA/GTGCCTACTTTGGCTTGTTCA | AE005176 |
| PpepN | GGGAATTCTTGTTTTAAATGATGG/CGGGATCCTTCTTCTTCATCAAAT | M87840 |
| PpepP | TGGTTCGTCGTGAAGCACCAT/TCATCAGCAATTGCACAGGCT | Y08842 |
| PpepT | GCTATTGAGCCTTTTACT/GTTGTTCCATCAGTATGA | L27596 |
| PpepQ | TCTCCATCGATTGTTCCGACT/TCATCTGCTGACTTAATCAAA | AE005176 |
| PpepX | GGGAATTCCAAACATCGCACCTA/CGGGATCCCAATCTAATTCTATA | M35865 |
| PoppD | GGGAATTCTGCTTTTATTATTTCCT/CGGGATCCTACTTGTTCTAAAA | L18760 |
| PoppA | GGGAATTCTTTGGGAACAATGATAA/CGGGATCCGTTACTTCTGAACCA | L18760 |
| PprtP | CGGAATTCCTTGTACAAGCTACT/CGGAATTCAGCTTATTGTAATCA | M26310 |
| pepQ | ATCCGACAACTCTAAATTATCT/TAGCAGCGCGAGCTCCTGATA | AE005176 |
| oppA | AGCAGCTACACTCCTAAGTGCT/CGGGATCCGTTACTTCTGAACCA | L18760 |
| oppD | AGTCTTAAACGTGGTGAAGT/CGGGATCCTACTTGTTCTAAAA | L18760 |
| PpepDA2 | TGAGGTTTCTGTCGCAGTCAT/CGGTCAGCAATGGTTACCTTA | AE005176 |
| dtpT | TGACTCACGTCGTGACACTGGAT/TCCGTTCAAGAGACCTGGAAGT | U05215 |
| dtpP | ACAGCACTCTATGCAAGGGTCATA/AGTCGCTGGATCAACCGCACGTT | AE005176 |