TABLE 1.
Strains, plasmids and oligonucleotides used in this study
| Strain, plasmid, or oligonucleotide | Descriptiona | Derivation or reference |
|---|---|---|
| E. coli DH5α | φ80lacZΔM15 recA1 endA1 gyrA96 thi-1 hsdR17 (rK− mK+) supE44 relA1 deoR Δ(lacZYA-argF)U169 | Laboratory strain |
| B. subtilis | ||
| CU1065 | W168 trpC2 attSPβ | Laboratory strain |
| HB5345 | CU1065 yoeB::MLS | LFH-PCR → CU1065 |
| HB5364 | CU1065 thrC::PxylA-yoeB | pLS20 → CU1065 |
| HB5365 | CU1065 yoeB::MLS thrC::PxylA-yoeB | pLS20 → HB5345 |
| HB5366 | CU1065 amyE::PyoeB-lacZ (+18) | pLS16 → CU1065 |
| HB5370 | CU1065 amyE::PyoeB-lacZ (−22) | pLS15 → CU1065 |
| HB5371 | CU1065 amyE::PyoeB-lacZ (−42) | pLS14 → CU1065 |
| HB5372 | CU1065 amyE::PyoeB-lacZ (−68) | pLS13 → CU1065 |
| HB5377 | CU1065 amyE::PyoeB-lacZ (−514) | pLS08 → CU1065 |
| HB5384 | CU1065 amyE::yoeB-FLAG | pLS22 → CU1065 |
| Plasmids | ||
| pXT | Cloning vector for overexpression | 9 |
| pDG1661 | Vector for integration of lacZ fusions at amyE locus | 12 |
| pLS08 | PyoeB-lacZ (−514) in pDG1661 | This work |
| pLS13 | PyoeB-lacZ (−68) in pDG1661 | This work |
| pLS14 | PyoeB-lacZ (−42) in pDG1661 | This work |
| pLS15 | PyoeB-lacZ (−22) in pDG1661 | This work |
| pLS16 | PyoeB-lacZ (+18) in pDG1661 | This work |
| pLS20 | yoeB cloned into pXT | This work |
| pLS22 | yoeB-FLAG in pDG1730 | This work |
| Oligonucleotides | ||
| 1589 | 5′-GCAGCAGGAGAACCAGCAGTTCC-3′ | |
| 1713 | 5′-CCGTAAGCTTGTCCTATCAG-3′ | |
| 1714 | 5′-GCAGGATCCGCTCATGGTGTTCC-3′ | |
| 1995 | 5′-CCCAAGCTTCCGTAATTATTGTAACATTTGTAAC-3′ | |
| 2002 | 5′-GAGGGTTGCCAGAGTTAAAGGATCTCATTATCGCTCTCTCCTTC-3′ | |
| 2004 | 5′-CCCAAGCTTTTGTAACATAAGAGAAAGAGAT-3′ | |
| 2005 | 5′-CGGAATTCTTCACAAAAAGACAACAGGC-3′ | |
| 2006 | 5′-GTTGGCATTATCCGCTAGAG-3′ | |
| 2007 | 5′-CTTTCCGTTTTGCAGAAGGG-3′ | |
| 2087 | 5′-CGATTATGTCTTTTGCGCAGTCGGCACTGTCCCTACATTAGACGG-3′ | |
| 2088 | 5′-GCATATTGTAGAGCCGATTCGC-3′ | |
| 2132 | 5′-CCCAAGCTTGCAAAAGTATTGTAATCTATCCG-3′ | |
| 2133 | 5′-CCCAAGCTTGAGTAAAAGAATAAAAAATAATTTATGC-3′ | |
| 2404 | 5′-GCGGATCCATCTCTTTCTCTTATGTTACAAATG-3′ | |
| 2430 | 5′-CTGCACGGCCGTATAACTGCGTCAAATTGATTG-3′ | |
| 2640 | 5′-ACAGTACGCGTAGTGCAACC-3′ | |
| 2641 | 5′-CGCCACTTCAACATCAACGG-3′ |
a
Restriction sites are underlined.