Table 1.
Bacterial strains or plasmids used in this study.
| Strain or Plasmid | Relevant genotype or description | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| EC101 | E. coli JM101(supEthi (lacproAB) (F’ traD36 proABlacIq ZΔM15) with repA from pWV01 integrated in chromosome | Laboratory stock (Law et al., 1995) |
| C2992I | E. coli DH5α F̀ proA+B+ lacIq Δ(lacZ)M15 zzf::Tn10/ fhuA2Δ(argF-lacZ)U169 phoA glnV44 Φ80Δ(lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17, tetr | New England Biolabs Inc., |
| EcASCtsR | E. coli EC101 carrying pSIPSYNCtsR | This study |
| Ecsyn | E. coli EC101 carrying pSIPSYN | (Darsonval et al., 2016) |
| BL21 | E. coli F− dcm ompT ΔhsdS (rB- mB-) gal [malB+]K-12(λS) | Thermo Fisher Scientific |
| BL21-CtsR | BL21 carrying pETctsR25 | This study |
| BL21-Lo18 | BL21 carrying pEThsp18 | This study |
| DHT1 | F−glnV44(AS) recA1 endA1 gyrA96 (nalr) thi-1 hsdR17 spoT1 rfbD1 cya−854ilv-691::Tn10 | (Dautin et al., 2000) |
| DHT1 zip-zip | E. coli DHT1 harboring pT25-zip and pT18-zip encoding the GCN4 leucine zipper | (Karimova et al., 2000) |
| Oenococcus oeni | ||
| O. oeni ATCC BAA-1163 | wild type strain, vanr | Laboratory stock |
| OoASctsR | O. oeni ATCC BAA-1163, pSIPSYNASCtsR | This study |
| Oosyn | O. oeni ATCC BAA-1163, pSIPSYN | (Darsonval et al., 2016) |
| Bacillus subtilis | ||
| 168 | Wild type, trpC2 | Laboratory stock |
| QB4991 | trpC2 amyE::(‘lacZ aphA3) ΔctsR | (Derré et al., 1999) |
| WT | trpC2 amyE ::(hps18’-bgaB cat) | pDLhsp18 →168 (Grandvalet et al., 2005) |
| ΔctsR | trpC2 ΔctsR amyE ::(hps18’-bgaB cat) | pDLhsp18 → QB4991 (Derré et al., 2000; Grandvalet et al., 2005) |
| ΔctsR-XTOoctsR pXTOoCtsR | trpC2, ΔctsR amyE::(hps18’-bgaB cat), thrC::(pxylA-ctsR-O. oeni-spc) | pXTctsR25 → ΔctsR-hsp18’ |
| Plasmids | ||
| pSIPSYN | Replicative and low-copy number plasmid allowing gene expression in O. oeni under the control of PSY N promoter, eryR | (Darsonval et al., 2016) |
| pSIPSYNASCtsR | pSIPSYN derivative encoding the ctsR ORF in antisense orientation under the control of PSY N promoter | This study |
| pET28a(+) | Vector for expression of N-terminal His-tagged proteins in BL21 under the control of pT7 promoter, kanR | Novagen |
| pETctsR | pET28a(+) derivative carrying the O. oeni ctsR coding sequence, kanR | This study |
| pEThsp18 | pET28a(+) derivative carrying the O. oeni hsp18 coding sequence. | This sudy |
| pXT | pDG1728 derivative allowing transcriptional fusion with PxylA xylose inducible promoter and integration into the B. subtilis thrC locus, specR, ampR | (Derré et al., 2000) |
| pXTctsR25 | pXT derivative carrying the O. oeni ctsR coding sequence under the control of PxylA xylose inducible promoter | This study |
| pDL | Integrative plasmid for constructing transcriptional fusions with the G. stearothermophilus bgaB gene and integration into the B. subtilis amyE locus, cmR | (Yuan and Wong, 1995) |
| pDLhsp18 | pDL derivative with transcriptional fusion hsp18’-bgaB | (Grandvalet et al., 2005) |
| pKT25 | BACTH vector designed to express in DHT1 a given polypeptide fused in frame at its N-terminal end with T25 fragment, kanR | (Karimova et al., 2000) |
| pKNT25 | BACTH vector designed to express in DHT1 a given polypeptide fused in frame at its C-terminal end with T25 fragment, kanR | (Karimova et al., 2000, 2005) |
| pUT18C | BACTH vector designed to express in DHT1 a given polypeptide fused in frame at its N-terminal end with T18 fragment, ampR | (Karimova et al., 2000) |
| pUT18 | BACTH vector designed to express in DHT1 a given polypeptide fused in frame at its C-terminal end with T18 fragment, ampR | (Karimova et al., 2000) |
| pKT25-ctsR | Full-length ctsRORF cloned into pKT25 | This study |
| pKT25-clpC | Full-length clpCORF cloned into pKT25 | This study |
| pKT25-clpL1 | Full-length clpL1ORF cloned into pKT25 | This study |
| pKT25-clpL2 | Full-length clpL2 ORF cloned into pKT25 | This study |
| pKNT25-ctsR | Full-length ctsR ORF without stop codon cloned into pKNT25 | This study |
| pKNT25-clpC | Full-length clpC ORF without stop codon cloned into pKNT25 | This study |
| pKNT25-clpL1 | Full-length clpL1 ORF without stop codon cloned into pKNT25 | This study |
| pKNT25-clpL2 | Full-length clpL2 ORF without stop codon cloned into pKNT25 | This study |
| pUT18C-ctsR | Full-length ctsR ORF cloned into pUT18C | This study |
| pUT18C-clpC | Full-length clpC ORF cloned into pUT18C | This study |
| pUT18C-clpL1 | Full-length clpL1 ORF cloned into pUT18C | This study |
| pUT18C-clpL2 | Full-length clpL2 ORF cloned into pUT18C | This study |
| pUT18-ctsR | Full-length ctsR ORF without stop codon cloned into pUT18 | This study |
| pUT18-clpC | Full-length clpC ORF without stop codon cloned into pUT18 | This study |
| pUT18-clpL1 | Full-length clpL1 ORF without stop codon cloned into pUT18 | This study |
| pUT18-clpL2 | Full-length clpL2 ORF without stop codon cloned into pUT18 | This study |
vanr, vancomycin resistance; eryr, erythromycin resistance; cmr, chloramphenicol resistance; specr, spectinomycin resistance; ampr, ampicillin resistance; kanr, kanamycin resistance; tetR, tetracyclin resistance.