TABLE 2.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype and/or relevant characteristic(s) | Source |
|---|---|---|
| Strains | ||
| P. alvei CCM 2051T | Wild-type isolate; Kmr | CCM |
| P. alvei CCM 2051 wsfP::Ll.LtrB | P. alvei CCM 2051T carrying a targetron insertion at the wsfP locus; Kmr | This study |
| Escherichia coli DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 (rK− mK−) phoA supE44 thi-1 gyrA96 relA1 λ− | Invitrogen |
| Plasmids | ||
| pNW33N | Geobacillus-Bacillus-E. coli shuttle vector; Cmr | Bacillus Genetic Stock Center |
| pEXALV | pNW33N carrying the sgsE S-layer gene promoter of G. stearothermophilus NRS 2004/3a; Cmr | This study |
| pNW33NΔHindIII | pNW33N without its unique HindIII restriction site; Cmr | This study |
| pJIR750ai | Clostridium perfringens-E. coli shuttle vector carrying alpha-toxin gene (plc) targetron; Cmr | Sigma-Aldrich |
| pJIR750ai_P(SgsE) | pJIR750ai carrying the sgsE S-layer gene promoter of G. stearothermophilus NRS 2004/3a; Cmr | This study |
| pTT_plc | pNW33N without HindIII carrying the sgsE S-layer gene promoter of G. stearothermophilus NRS 2004/3a in front of the plc intron cassette of pJIR750ai; Cmr | This study |
| pTT_wsfP555 | pTT_plc targeted for insertion between positions 555 and 556 from the initial ATG codon of wsfP; Cmr | This study |
| pTT_wsfP654 | pTT_plc targeted for insertion between positions 654 and 655 from the initial ATG codon of wsfP; Cmr | This study |
| pTT_wsfP1176 | pTT_plc targeted for insertion between positions 1176 and 1177 from the initial ATG codon of wsfP; Cmr | This study |
| pEXALV_wsfP | pEXALV carrying the wsfP gene of P. alvei CCM 2051T; Cmr | This study |
| pEXALV_wsaP | pEXALV carrying the wsaP gene of G. stearothermophilus NRS 2004/3a; Cmr | This study |