Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| E. coli BL21(DE3) | F− ompT hsdSB(rB− mB−) gal(λcI857 ind1, Sam7 nin5 lacUV5-T7 gene1) dcm(DE3) | Novagen |
| B. subtilis 168 | trpC2 | Laboratory stock |
| S. aureus | ||
| FDA209P | Wild type | MCRFa |
| N315 | Methicillin sensitive | 13 |
| MS16526 | Methicillin resistant | MCRF |
| Streptomyces sp. strain MK851-mF8 | Signermycin B producing | This study |
| E. faecalis | ||
| JCM5803 | Wild type | MCRF |
| NCTC12201 | Vancomycin resistant | MCRF |
| S. mutans UA159 | Wild type | ATCC |
| Plasmids | ||
| pHT10 | Expression vector for E. coli and B. subtilis; Apr Cmr | MoBi Tec |
| pET-21a(+) | Expression vector, Apr, C-terminal His tag | Novagen |
| pET-22b(+) | Expression vector, Apr, C-terminal His tag | Novagen |
| pBsG-full | WalK(Bs) (B. subtilis 168 aa 1 to 611) cloned into pHT10 | This study |
| pETYycG-tru | WalK(Bs) (B. subtilis 168 aa 207 to 611) cloned into pET-21a(+) | 23 |
| pBsGsubA | WalK(Bs) (B. subtilis 168 aa 366 to 611) cloned into pET-21a(+) | This study |
| pBsGsubB | WalK(Bs) (B. subtilis 168 aa 437 to 611) cloned into pET-21a(+) | This study |
| pABF | B. subtilis 168 WalR under control of Pspac | 13 |
| pYycG Sa | WalK(Sa) (S. aureus N315 aa 204 to 608) cloned into pET-21a (+) | 31 |
| pETEfWalK | WalK(Efa) (E. faecalis JCM5803 aa 203 to 611) cloned into pET-21a(+) | This study |
| pETSMVicK31-450 | WalK(Sm) (VicK of S. mutans UA159, aa 31 to 450) cloned into pET22b(+) | 8 |
a
MCRF, Microbial Chemistry Research Foundation, Tokyo, Japan.