TABLE 1.
Bacteria and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| Strains | ||
| C. sakazakii | ||
| BAA-894 | Isolated from powdered infant formula associated with a NICU outbreak, obtained from the ATCC (Manassas, VA) | 16 |
| BAA894NA | BAA-894 nalidixic acid-resistant spontaneous mutant | This study |
| BAA894Δcpa | BAA-894 cpa isogenic mutant | This study |
| BAA894Δcpa/pMMB66EH::cpa | BAA894Δcpa complemented with pMMB66EH::cpA | This study |
| E. coli | ||
| BL21 | F−dcm ompT hsdS(rB− mB−) gal [malB+]K-12(λS) | Novagen |
| DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 (rK− mK+) phoA supE44 λ−thi-1 gyrA96 relA1 | Invitrogen |
| DH5α/pMMB66EH | DH5α containing plasmid pMMB66EH | This study |
| DH5α/pMMB66EH::cpa | DH5α containing pMMB66EH::cpa | This study |
| DH5α λpir | DH5α containing λpir | 52 |
| SM10 λpir | thi thr leu tonA lacY supE recA::RP4-2-Tc::Mu Km λpir | 4 |
| Plasmids | ||
| pET30(+) | Expression vector, T7 promoter-driven system | Novagen |
| pET30::cpa | Plasmid expressing His6-CpA fusion protein | This study |
| pCVD442 | Suicide vector, R6K ori, mobRP4, bla, sacB | 4 |
| pMMB66EH | Broad-host-range Ptac expression vector | 10 |
| pMMB66EH::cpa | cpa cloned into pMMB66EH | This study |
| pCVD442::Δcpa | Δcpa and flanking region cloned into pCVD442 | This study |
a
NICU, neonatal intensive care units; ATCC, American Type Culture Collection.