TABLE 1.
Strains and plasmids used in the study
| Strain or plasmid | Genotype | Reference or source |
|---|---|---|
| E. coli K-12 strains | ||
| JM109 | recA1 supE44 endA1 hsdR17 gyrA96 relA1 thi Δ68(lac-proAB) F′ [traD36 proAB+ lacIq lacZΔM15] | 49 |
| BTH101 | F− cya-99 araD139 galE15 galK16 rpsL1 (Strr) hsdR2 mcrA1 mcrB1 | 20 |
| C43(DE3) | F′ hsdS gal1 DE3; spontaneous mutation of BL21(DE3) | 50 |
| B. subtilis 168 strains and plasmids | ||
| IMW610 | dctB::Specr | This study |
| pBQ200 | B. subtilis/E. coli shuttle vector, expression of genes via the degQh promoter, Ampr in E. coli, Eryr in B. subtilis | 51 |
| pDG1726 | Plasmid with Specr for long flanking homology PCR | 52 |
| pMW1927 | dctA-His6 expression plasmid, pBQ200 derivative, Ampr in E. coli, Eryr in B. subtilis | This study |
| Plasmids for mSPINE and mHPINE | ||
| pASK-IBA3+ | Expression plasmid for proteins with a C-terminal Strep tag, AHT-inducible promoter, Ampr | IBA, Göttingen, Germany |
| pET28a | Expression plasmid for proteins with an N- or C-terminal His6 tag, T7 promoter, Kanr | Novagen |
| pBAD33 | Expression plasmid for proteins under the control of the arabinose-inducible pBAD promoter, Camr | 53 |
| pMW1788 | dctS-Strep expression plasmid, pASK-IBA3+ derivative (Ampr) | This study |
| pMW1791 | dctB-His expression plasmid, pET28a derivative (Kanr) | This study |
| pMW1793 | dctA-His expression plasmid, pET28a derivative (Kanr) | This study |
| pMW1795 | dctB-His expression plasmid, pBAD33 derivative (Camr) | This study |
| pMW1797 | dctA-His expression plasmid, pBAD33 derivative (Camr) | This study |
| pMW1800 | His6-dctP expression plasmid, pET28a derivative (Kanr) | This study |
| pMW1974 | dctA-Strep expression plasmid, pASK-IBA3+ derivative (Ampr) | This study |
| pMW1975 | dctA-His and dctB-His expression plasmid, pBAD33 derivative (Camr) | This study |
| Plasmids for BACTH measurements | ||
| pKNT25 | Plasmid for the expression of C-terminal T25-fusion proteins (Kanr) | 20 |
| pKT25 | Plasmid for the expression of N-terminal T25-fusion proteins (Kanr) | 20 |
| pUT18 | Plasmid for the expression of C-terminal T18-fusion proteins (Ampr) | 20 |
| pUT18C | Plasmid for the expression of N-terminal T18-fusion proteins (Ampr) | 20 |
| pKT25-Zip | T25-Zip expression plasmid, pKT25 derivative (Kanr) | 19 |
| pUT18C-Zip | T18-Zip expression plasmid, pUT18C derivative (Ampr) | 19 |
| pMW1695 | DctS-T18 expression plasmid, pUT18 derivative (Ampr) | This study |
| pMW1696 | T18-DctS expression plasmid, pUT18C derivative (Ampr) | This study |
| pMW1697 | T25-DctS expression plasmid, pKT25 derivative (Kanr) | This study |
| pMW1699 | DctR-T18 expression plasmid, pUT18 derivative (Ampr) | This study |
| pMW1704 | T18-DctA expression plasmid, pUT18C derivative (Ampr) | This study |
| pMW1705 | T25-DctB expression plasmid, pKT25 derivative (Kanr) | This study |
| pMW1708 | T18-DctB expression plasmid, pUT18C derivative (Ampr) | This study |