TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype and genetic markersa | Comment(s) | Reference or source |
|---|---|---|---|
| Strains | |||
| B. subtilis 168 | trpC2 | Wild type | 36 |
| B. subtilis PB100 | bpr | bpr-deficient B. subtilis 168 | This study |
| B. subtilis DB104 | nprE aprE | nprE- and aprE-deficient B. subtilis 168 | 4 |
| B. subtilis SB300 | nprE aprE epr | epr-deficient DB104 | This study |
| B. subtilis PB300 | nprE aprE bpr | bpr-deficient DB104 | This study |
| B. subtilis DB428 | nprE aprE epr bpr | Four-exoprotease gene-deficient B. subtilis 168 | 4 |
| B. subtilis LB700 | nprE aprE epr bpr mpr nprB wprA F−araD139 Δ(ara-leu)7696 galE15 | Seven-exoprotease gene-deficient B. subtilis 168 | 15 |
| E. coli MC1061 | galK16 ΔlacX74 rpsL (Strr) hsdR2 (rK− mK+) mcrA mcrB1 | Host for DNA manipulation | 25 |
| Plasmids | |||
| pSM704 | Kanr; Bacillus ori | Shuttle expression vector with subtilisin promoter and signal sequence | 15 |
| pSP704 | Kanr Ampr; Bacillus ori, E. coli ori | Shuttle expression vector with subtilisin promoter | This study |
| pMpr | Kanr Ampr; Bacillus ori, E. coli ori | Mpr secretion plasmid | This study |
| pMprH6 | Kanr Ampr; Bacillus ori, E. coli ori | C-terminally His6-tagged Mpr secretion plasmid | This study |
| pMprH6(S93E) | Kanr Ampr; Bacillus ori, E. coli ori | pMprH6 plasmid harboring S93E mutation | This study |
| pBpr | Kanr Ampr; Bacillus ori, E. coli ori | Bpr secretion plasmid | This study |
| pUC19 | Ampr; E. coli ori | Cloning vector | 41 |
| pUCTV2 | Kanr Ampr; Bacillus ts-ori, E. coli ori | Temperature sensitive suicide vector | 37 |
| pDEPR | Tcr Ampr; Bacillus ts-ori, E. coli ori | Gene replacement vector with epr::tet | This study |
| pDBPR | Tcr Ampr; Bacillus ts-ori, E. coli ori | Gene replacement vector with bpr::tet | This study |
a
ori, origin of replication; ts-ori, temperature-sensitive ori.