TABLE 1.
B. subtilis strains and plasmids
| Strain or plasmid | Relevant genotype or features of plasmid | Construction or referenced |
|---|---|---|
| Strains | ||
| 168 | trpC2 | 48 |
| BSA46 | PY22 SPβ ctc::lacZ cat86 erma | 11 |
| BSA70 | PY22 rsbU::kan SPβ ctc::lacZa | 11 |
| BSA73 | PY22 rsbU::kan rsbV312 SPβ ctc::lacZ | 11 |
| BSA159 | PY22 rsbV312 rsbX::pWH25-spcb | 65 |
| BSM21 | trpC2 rsbU::kan | BSA70⇒168 |
| BSM22 | trpC2 rsbU::kan rsbV312 | BSA73⇒168 |
| BSM24 | trpC2 rsbV312 rsbX::pWH25-spc | BSA159⇒168 |
| BSM29 | trpC2 sigBΔ2::spc | FSB5⇒168 |
| BSM30 | trpC2 rsbP::spc | BSM201⇒168 |
| BSM149 | trpC2 rsbP::spc rsbU::kan | BSM30⇒BSM21 |
| BSM151 | trpC2 SPβ ctc::lacZ | BSA46⇒168 |
| BSM152 | trpC2 rsbP::spc SPβ ctc::lacZ | BSA46⇒BSM30 |
| BSM153 | trpC2 rsbP::spc rsbU::kan SPβ ctc::lacZ | BSA46⇒BSM149 |
| BSM154 | trpC2 rsbU::kan SPβ ctc::lacZ | BSA46⇒BSM21 |
| BSM155 | trpC2 rsbV312 rsbX::pWH25-spc SPβ ctc::lacZ | BSA46⇒BSM24 |
| BSM156 | trpC2 sigBΔ2::spc SPβ ctc::lacZ | BSA46⇒BSM29 |
| BSM201 | PY22 rsbP::spc | pAV01⇒PY22 |
| BSM202 | PY22 rsbP::spc rsbU::kan | BSA70⇒BSM201 |
| BSM227 | PY22 amyE::pGK30-gsiB::gfp cat86c | pGK30⇒PY22 |
| BSM267 | trpC2 rsbP::spc rsbU::kan rsbV312SPβ ctc::lacZ | BSA73⇒BSM152 |
| BSM269 | trpC2 amyE::pGK30-gsiB::gfp cat86c | BSM227⇒168 |
| BSM275 | trpC2 rsbV312 rsbX::pWH25-spc amyE::pGK30-gsiB::gfp | BSM227⇒BSM24 |
| cat86c | ||
| BSM276 | trpC2 sigBΔ2::spc amyE::pGK30-gsiB::gfp cat86c | BSM227⇒BSM29 |
| BSM277 | trpC2 rsbU::kan amyE::pGK30-gsiB::gfp cat86c | BSM227⇒BSM21 |
| BSM278 | trpC2 rsbP::spc amyE::pGK30-gsiB::gfp cat86c | BSM227⇒BSM30 |
| BSM279 | trpC2 rsbP::spc rsbU::kan rsbV312 | BSM202⇒BSM22 |
| BSM280 | trpC2 rsbP::spc rsbU::kan rsbV312 amyE::pGK30-gsiB::gfp cat86c | BSM227⇒BSM279 |
| FSB5 | trpC2 pheA1 sigBΔ2::spc | 59 |
| PY22 | trpC2 | P. Youngman, University of Georgia |
| Plasmids | ||
| pAV01 | AprrsbP::spc | This work |
| pDG1726 | Apr Spcr | 29 |
| pGK30 | AprgsiB::gfp cat86c | G. Kuhnke and U. Völker, unpublished data |
| pFSB79 | Low-copy-number gfp transcriptional fusion vector for integration into the amyE locus | F. Spiegelhalter and E. Bremer, unpublished data |
a
The ctc-lacZ fusion in the SPβ prophage is linked to the chloramphenicol (cat86) and erythromycin (erm) resistance genes.
b
The integrative plasmid pWH25 contains a 2-kb EcoRI-SphI fragment, including the 3′ end of rsbX and 1.9 kb downstream of rsbX.
c
The strain contains part of plasmid pGK30 inserted into the amyE locus of the chromosome via a double-crossover event, thus linking the transcriptional gsiB-gfp fusion to the chloramphenicol resistance gene (cat86).
d
The arrow indicates the construction of the strain by transformation.