TABLE 1.
Bacterial strains and plasmids
| Strain, plasmid, or phage | Relevant characteristics | Construction | Source or reference |
|---|---|---|---|
| Strains | |||
| RM4606 | sup0 F+ | R. Maurer | |
| WU620 | As RM4606, except nlpI::cm | MO102(f1R189) × RM4606 | This study |
| WU62 | As RM4606, except nlpI::cm | WU620(P1) × RM4606 | This study |
| WU63 | As RM4606, except nlpI::cm | WU620(P1) × RM4606 | This study |
| MO101 | DH5α carrying plasmid pBU142 | This study | |
| MO102 | JM109 carrying pBIP3-nlpI::cm | This study | |
| DH5α | supE44 ΔlacU169(φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA | Bethesda Research Laboratories | |
| JM109 | F′ traD36 proA+B+ lacIq Δ(lacZ)M15/recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 Δ(lac-proAB) e14− (Mcr−) | New England Biolabs | |
| INVαF′ | F′ endA1 recA1 hsdR17 supE44 thi-1 gyrA96 relA1 φ80lacZΔM15 Δ(lacZYA-argF)U169 | Invitrogen | |
| Plasmids | |||
| pCR2.1 | TA cloning vector; Ampr Kmr | Invitrogen | |
| pBAD24 | Arabinose regulation; Ampr | 10 | |
| pBIP3 | Suicide vector; KmrsacB (sensitivity to sucrose) | R. Maurer | |
| pCU11 | nlpI cloned into pCR2.1 | PCR product (primers U1F1 and U1R) | This study |
| pCU21 | nlpI plus 195 bp immediately upstream of the translational start codon cloned into pCR2.1 | PCR product (primers U1F2 and U1R) | This study |
| pBU142 | nlpI subcloned into pBAD24 | EcoRI-HindIII fragment of pCU11 subcloned into pBAD24 | This study |
| Phages | |||
| f1R189 | Helper phage for transduction (gene II amber mutant that will not replicate in sup+ target strains) | R. Maurer | |
| P1 | Generalized transduction | Laboratory stock |