Table 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristicsa | Source or reference |
|---|---|---|
| Strains | ||
| Bacillus subtilis | ||
| JH642 | trpC2 pheA1 | Laboratory stock |
| L43 | JH642 fer::Kmr | This study |
| L43C | JH642 fer::KmramyE::Plac-fer Cmr | This study |
| LSC9 | JH642 ykuNOP::Pspac-ykuNOP PykuNOP-lacZ Ermr Lmr | This study |
| LSC28 | L43 ykuNOP::Pspac-ykuNOP PykuNOP-lacZ Kmr Ermr Lmr | This study |
| LSC29 | LSC28 thrC::Pxyl-ykuN Kmr Ermr Lmr Spr | This study |
| LSC30 | LSC28 thrC::Pxyl-ykuP Kmr Ermr Lmr Spr | This study |
| Escherichia coli DH5α | supE44 thi-1 ΔlacU169(φ80lacZΔM15) endA1 recA1 hsdR17gyrA96relA1 trp6 cysT329::lacinmλpI(209) | Laboratory stock |
| Plasmids | ||
| pCR-Blunt II-TOPO | E. coli cloning vector; Kmr | Invitrogen |
| pGEMT-Easy | E. coli cloning vector; Ampr | Promega |
| pJM114 | Integrative vector used for gene disruption; Kmr | 17 |
| pJM114F1 | pJM114 with fragment F1 cloned into the SacI/BamHI site | This study |
| pJM114F1F2 | pJM114F1 with fragment F2 cloned into the XhoI/KpnI site | This study |
| pLM1 | pGEMT Easy containing a 609-bp fragment corresponding to the 5′ end of the fer gene | This study |
| pLM2 | pGEMT Easy containing a 555-bp fragment corresponding to the 3′ end of the fer gene | This study |
| pLM3 | pGEM-T Easy containing a 308-bp fer gene | This study |
| pDR67 | amyE integration plasmid with the IPTG-inducible Pspac promoter; Cmr | 17 |
| pDR67Cfd | Contains the fer gene cloned into HindIII/BglII sites in pDR67 under the control of the Pspac promoter | This study |
| pJM116 | Integrative vector to construct transcriptional fusions to lacZ; integrates at the amyE locus of B. subtilis; Cmr | 17 |
| pMutin | Integrates at a specific locus of B. subtilis; Ermr Lmr | 28 |
| pLSC1 | pCR-Blunt II-TOPO containing a 549-bp fragment corresponding to the 5′ end of the ykuNOP operon | This study |
| pLSC9 | Pspac::ykuNOP-lacZ cloned into pMutin; Ermr Lmr | This study |
| pDG1731 | Integrates at the thrC locus of B. subtilis; Spr | Laboratory stock |
| pGS247 | Xylose promoter cloned into pDG1731; Spr | Laboratory stock |
| pLSC10 | ykuN cloned under the control of Pxyl in pGS247; Kmr Ermr Lmr Spr | This study |
| pLSC11 | ykuP cloned under the control of Pxyl in pGS247; Kmr Ermr Lmr Spr | This study |
a
Cmr, Tetr, Kmr, Ermr, Lmr, and Ampr denote resistance to chloramphenicol, tetracycline, kanamycin, 0.5 μg erythromycin ml−1, 12.5 μg lincomycin ml−1, and ampicillin, respectively.