TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| Strains | ||
| B. subtilis | ||
| 168 | trpC2 | C. Anagnostopoulosa |
| HØR18 | trpC2 pucR::pBOER | 15 |
| HØR26 | trpC2 pucR::Pspac-pucR | Transformation of 168 by pIMut4 selecting for Err |
| LCC26 | trpC2 tnrA62::Tn917 | Transformation of 168 with DNA from SF416R selecting for Err (2) |
| E. coli | ||
| MC1061 | F−araD139 Δ(ara-leu)7697 galE15 galK16 Δ(lac)74 rpsL (Strr) hsdR2 (r− m+) mcrA mcrB1 | Laboratory stock |
| Plasmids | ||
| pDG268neo | Apr (E. coli) Neor (B. subtilis); vector used for construction of transcriptional lacZ fusion designed to integrate into the amyE gene of B. subtilis | 11 |
| pIMut4 | Lnr, Err, Pspac-pucR, LacZ−; pMUTIN4 digested with EcoRI-SacI, ligated with PCR-generated fragment containing RBS and 211 bp of the 5′-end of pucR; integration into the pucR locus results in a 3′ truncated PucR copy expressed from the pucR promoter and an intact PucR copy expressed from Pspac | This work |
| pMUTIN4 | Integrational plasmid used for gene inactivation by recombination and for generation of transcriptional lacZ fusions | 17 |
| pBOE335 | Apr (E. coli) Cmr (B. subtilis); integration vector, pUC19 containing cat gene cloned into KasI site | 11 |
| pBOER | Cmr; pBOE335 containing an internal part of the pucR transcriptional unit; integration of pBOER results in pucR inactivation | 15 |
a
Centre National de la Recherche Scientifique, Jouy-en-Josas, France.