Table 1.
Bacterial strains, plasmids and primers
| Description | Reference | |
|---|---|---|
| Strains | ||
| CF10230 | nic+ derivative of DY329 (20) | Michael Cashel, unpublished data |
| UM285 | As CF10230 but ΔcysQ, kmR | (15) |
| UM341 | As CF10230 but orn under control of P LtetO-1, TetR, kmR | This study |
| DH5αZ1 | tetR under control of PN25, lacI under control of Placiq, SpR | (19) |
| BSF66 | B. subtilis 168, ytqI::pMUTIN2MCS, EmR | (21) |
| DB430 | B. subtilis 168, deficient in multiple proteases | (22) |
| Plasmids | ||
| pBAD18 | Vector, Para, pBR replicon, ApR | (45) |
| pGEM-T Easy | Vector for TA cloning | Promega |
| pZE21-MCS1 | Vector, P LtetO-1, ColE1 replicon, KmR | (19) |
| pUM404 | As pBAD18, CysQ with C-terminal his-tag | (15) |
| pUM408 | As pBAD18, Orn with C-terminal his-tag | (15) |
| pUM412 | As pBAD18, YtqI with C-terminal his-tag | This study |
| Primers | ||
| UM153 | 5′GATTGCGGCCGCGCTTGGATTCTCACCAATAAA3′ | |
| UM154 | 5′GCGTTTTTTATTGGTGAGAATCCAAGCGCGGCCGCAATCATAAAAAA TTTATTTGCTTTCAGGAAAATTT3′ | |
| UM155 | 5′GATACACGCAATTGTGGATCAATATAGTCACTGTGAATGGGTGGAAA ATAGCGTCGACGGCGGATTTGTCCTACTCAGG3′ | |
| UM156 | 5′CCGGTCATCTCAAGATCGATCCAAATCAGGTTGTTTTCATTGGCACTCA TGGTACCTTTCTCCTCTTTAATGAATTCGG3′ | |
| UM158 | 5′TTCCACATCAGCGTGCATAC3′ | |
| UM159 | 5′GCAACGAACAGGTCACTATC3′ | |
| UM160 | 5′CCTGCGCCATCAGATCCTTG3′ | |
| UM161 | 5′TCAGCGCCAGCTGTTCATCA3′ | |
| UM172 | 5′GTGCTGCAAGGCGATTAAGT3′ | |
| UM173 | 5′CGCCAAGCTATTTAGGTGAC3′ | |
| UM175 | 5′GGGGAATTCACCATGAAAACAGAATTGATCAGAACCATATC3′ | |
| UM176 | 5′GGGGCTCGAGCTCGTGTTCTTTACATAATGTTTCCAGATC3′ |