TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or characteristic | Source or reference |
|---|---|---|
| Strains | ||
| BL21(DE3) | F−ompT hsdSB gal dcm (DE3) | Novagen |
| Novablue (DE3) | endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 Δlac [F′ proAB+ lacIqZΔM15::Tn10] (DE3) | Novagen |
| TOP10F′ | F′ (lacIq Tn10 [Tetr]) mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 deoR araD139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| INV F′ | F′ endA1 recA1 hsdR17 supE44 thi-1 gyrA96 relA1 φ80lacZΔM15 (lacZYA-argF)U169 | Invitrogen |
| Verdun | L. interrogans serovar icterohaemorrhagiae strain Verdun (National Reference Center for Leptospira, Institut Pasteur, Paris, France) | |
| RZ11 | L. interrogans serovar pomona strain RZ11 | 24 |
| Plasmids | ||
| pGEM7Zf(+) | Confers resistance to AMP | Promega |
| pET26b(+) | Confers resistance to kanamycin | Novagen |
| pCRII-TOPO | TA vector; confers resistance to AMP and kanamycin | Invitrogen |
| pCR2.1 | TA vector; confers resistance to AMP and kanamycin | Invitrogen |
| pG-PBP1 | ponA gene (XbaI restriction fragment from L. interrogans strain Verdun cosmid library) cloned in XbaI-digested pGEM7Zf(+) | This paper |
| pG-PBP3 | pbpB gene (ClaI restriction fragment from L. interrogans strain Verdun cosmid library) cloned in ClaI-digested pGEM7Zf(+) | This paper |
| pET-PBP3 | Translational fusion of the pelB leader sequence with the predicted periplasmic part of PBP 3 from strain Verdun [PCR product obtained with primers PBP3M and PBP3L cloned in vector pET26b(+)] | This paper |
| p921-1 | Serovar pomona pbpB gene (PCR product using primers 921 and 922) cloned in pCR2.1 vector | This paper |
| p513-3 | Serovar pomona ponA gene (3.1-kb PCR product using primers 155 and 513) cloned in pCR2.1 vector | This paper |
| pKB1 | 5.5-kb BamHI restriction fragment (containing two-thirds of the strain RZ11 ponA gene including the 5′ end) cloned in pBluescript II KS(+) vector | This paper |
| pK127 | 1.3-kb genome walking clone (amplified from RZ11 DNA using the gene-specific primer 173 and linker-specific primer AP1) cloned in pCR2.1 vector | This paper |