TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant markers, phenotypes, and characteristics | Reference or source |
|---|---|---|
| E. coli strains | ||
| LLB1 | supE Δthi (lac-proAB) hsdD5 (F′+traD36 proAB lacIqZΔM15) pcnB | 47 |
| ER2566 | F− λ−fhuA2 [lon] ompT lacZ::T7 gene1 gal sulA11 Δ(mcrC-mrr) 114::IS10 R(mcr-73::miniTn10-TetS)2 R(zgb-210::Tn10)(TetS) endA1 [dcm] | NEB |
| JIM8551 | Apr, ER2566 containing pJIM5740 for FhuR protein expression | This work |
| L. lactis strains | ||
| IL1403 | L. lactis subsp. lactis, His− Iso− Leu− Val− | 14 |
| JIM8475 | Emr, IL1403 containing pJIM1062 integrated at the PpurH locus, His− Iso− Leu− Val− | This work |
| JIM8494 | IL1403 ΔfhuR after homologous recombination of pJIM5702, His− Iso− Leu− Val− | This work |
| JIM8499 | Emr, IL1403 containing pJIM5706 integrated at the PmetB2 locus, His− Iso− Leu− Val− | This work |
| JIM8500 | Emr, IL1403 containing pJIM5707 integrated at the PfhuR locus, His− Iso− Leu− Val− | This work |
| JIM8539 | Emr, IL1403 containing pJIM5724 integrated at the PyriD locus, His− Iso− Leu− Val− | This work |
| JIM8543 | Emr, IL1403 containing pJIM5728 integrated at the PyjgC locus, His− Iso− Leu− Val− | This work |
| JIM8545 | Emr, IL1403 containing pJIM5737 integrated at the PyhcE locus, His− Iso− Leu− Val− | This work |
| JIM8546 | Emr, IL1403 metB2 after homologous recombination of pJIM5732, His− Iso− Leu− Val− | This work |
| JIM8547 | Emr, IL1403 cysK after homologous recombination of pJIM5733, His− Iso− Leu− Val− | This work |
| JIM8657 | Emr, IL1403 containing pJIM5751 integrated at the PcysM locus, His− Iso− Leu− Val− | This work |
| JIM8658 | Emr, IL1403 containing pJIM5752 integrated at the PplpA locus, His− Iso− Leu− Val− | This work |
| JIM8659 | Emr, IL1403 containing pJIM5754 integrated at the PcysD locus, His− Iso− Leu− Val− | This work |
| JIM8662 | Emr, IL1403 yhcE after homologous recombination of pJIM5750, His− Iso− Leu− Val− | This work |
| JIM8681 | Emr, IL1403 containing pJIM5760 integrated at the PcysE locus, His− Iso− Leu− Val− | This work |
| Plasmids | ||
| pGEM-T easy | Apr, M13ori pBR322ori, linear T-overhangs vector | Promega |
| pTYB1 | Apr, expression vector used in the IMPACT-CN protein purification system | NEB |
| pGhost 8 | Tetr, thermosensitive replicative plasmid in L. lactis | 51 |
| pGhost 9 | Emr, thermosensitive replicative plasmid in L. lactis | 51 |
| pJIM2242 | Emr, ori+ ΔrepA, integrative vector in L. lactis | 32 |
| pJIM2374 | Emr, ori+ ΔrepA, integrative promoter probe vector containing luxAB genes | 15 |
| Plasmid for protein expression | ||
| pJIM5740 | 942-bp fragment carrying the fhuR gene cloned into NdeI-SapI sites of pTYB1 | This work |
| Plasmid for gene deletion | ||
| pJIM5702 | SalI fusion of pGhost 9 and pGEM-T easy containing 713-bp fragment upstream and 746-bp fragment downstream fhuR gene ligated in BamHI | This work |
| Plasmids for promoter-lux fusion | ||
| pJIM1062 | PpurH-lux, SalI fusion of pJIM2374 and pGEM-T easy containing PpurH on a 442-bp fragment | This work |
| pJIM5706 | PmetB2-lux, SalI fusion of pJIM2374 and pGEM-T easy containing PmetB2 on a 680-bp fragment | This work |
| pJIM5707 | PfhuR-lux, SalI fusion of pJIM2374 and pGEM-T easy containing PfhuR on a 471-bp fragment | This work |
| pJIM5724 | PyriD-lux, SalI fusion of pJIM2374 and pGEM-T easy containing PyriD on a 715-bp fragment | This work |
| pJIM5728 | PyjgC-lux, NcoI fusion of pJIM2374 and pGEM-T easy containing PyjgC on a 692-bp fragment | This work |
| pJIM5737 | PyhcE-lux, SalI fusion of pJIM2374 and pGEM-T easy containing PyhcE on a 675-bp fragment | This work |
| pJIM5751 | PcysM-lux, SalI fusion of pJIM2374 and pGEM-T easy containing PcysM on a 691-bp fragment | This work |
| pJIM5752 | PplpA-lux, NcoI fusion of pJIM2374 and pGEM-T easy containing PplpA on a 829-bp fragment | This work |
| pJIM5754 | PcysD-lux, SalI fusion of pJIM2374 and pGEM-T easy containing PcysD on a 723-bp fragment | This work |
| pJIM5760 | PcysE-lux, SphI fusion of pJIM2374 and pGEM-T easy containing PcysE on a 697-bp fragment | This work |
| Plasmids for gene inactivation | ||
| pJIM5732 | SalI fusion of pJIM2242 and pGEM-T easy containing 668-bp metB2 internal fragment | This work |
| pJIM5733 | SalI fusion of pJIM2242 and pGEM-T easy containing 602-bp cysK internal fragment | This work |
| pJIM5750 | SalI fusion of pJIM2242 and pGEM-T easy containing 694-bp yhcE internal fragment | This work |