TABLE 1.
Strains and plasmids used in this study
| Strain, plasmid, or primer | Relevant characteristic(s) or 5′-to-3′ sequence | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH10B | F− mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80dlacZΔM15 lacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK ΔrpsL endA1 nupG | Gibco/BRL |
| C. perfringens | ||
| NCTC 8798 | Acute food poisoning strain | R. Labbe |
| SM101 | Electroporation-efficient derivative of strain NCTC 8798 | 18 |
| SM102 | Electroporation-efficient derivative of strain NCTC 8798 | 18 |
| SM124 | SM101 with mutation in gene CPR_1831 | This study |
| SM127 | SM102 with mutation in gene CPR_2104 | This study |
| Plasmids | ||
| pGEM-T Easy | Cloning vector | Promega |
| pKRAH1 | Contains bgaR-PbgaL and polylinker; chloramphenicol resistance | 13 |
| pHLL58 | Contains the CPR_1831 coding sequence in pGEM-T Easy | This study |
| pHLL59 | Contains the CPR_1831 coding sequence in pKRAH1 | This study |
| pHLL60 | Contains the CPR_2104 coding sequence in pGEM-T Easy | This study |
| pHLL61 | Contains the CPR_2104 coding sequence in pKRAH1 | This study |
| Primers | ||
| OHL141 | GTCGACGAACTCATTTTATTTAACTTACTACGGAGGG | This study |
| OHL142 | GGATCCCACAAAATACTAGCATGTGAAATTACTC | This study |
| OKM13 | GTCGACTCTATATTAAAGTTGTTTAGAAGAGGG | This study |
| OKM14 | GGATCCTTATCTAGCTTTACCCTTTATATTTTTTATTG | This study |