Table 2. Strains and plasmids used in this study.
Unless stated otherwise, all strains were made in the BSB1 wild type background [38]. Genes responsible for resistance to antibiotics are abbreviated as follows: bla (ampicillin), cat (chloramphenicol), erm (erythromycin), kan (kanamycin), phleo (phleomycin), spc (spectinomycin), tet (tetracycline).
| Strain | Relevant features or genotype | Construction, source or reference |
| B. subtilis | ||
| BSB1 | trp+ | [38] |
| AH7 | amyE::Physp-gfp spc | A. Henderson |
| BV2096 | mecA::kan PcomG-comK spc, comG-gfp cat | [7] |
| H121G | trpC2 amyE::Pxyl-gfp-comK spc, comK::kan | H. Strahl |
| PG341 | PcomG-comK spc | BV2096 DNA → BSB1 |
| PG342 | comG-gfp cat | BV2096 DNA → BSB1 |
| PG343 | mecA::kan | BV2096 DNA → BSB1 |
| PG349 | amyE::Pxyl-gfp-comK spc | H121G → BSB1 |
| PG368 | mecA::tet | This study |
| PG389 | amyE::PcomG-lacZ-gfp+ cat | pPG40 → BSB1 |
| PG401 | amyE::PcomG-lacZ-gfp+ cat, PcomG-comK spc, mecA::tet | [PG341+PG368] DNAs → PG389 |
| PG401-Tn4 | amyE::PcomG-lacZ-gfp+ cat, PcomG-comK spc, mecA::tet, kre:TnYLB1#4 | This study |
| PG405 | amyE::PcomG-mcherry spc | pPG38 → BSB1 |
| PG406 | kre:TnYLB1#4 kan | PG401-Tn4 DNA → BSB1 |
| PG425 | comG-gfp cat, kre:TnYLB1#4 kan | PG401-Tn4 DNA → PG342 |
| PG433 | amyE::PcomG-lacZ-gfp+ cat, kre:TnYLB1#4 | PG401-Tn4 DNA → PG389 |
| PG435 | amyE::PcomC-lacZ-gfp+ cat | pPG55 → BSB1 |
| PG436 | amyE::PcomF-lacZ-gfp+ cat | pPG56 → BSB1 |
| PG437 | amyE::PaddAB-lacZ-gfp+ cat | pPG57 → BSB1 |
| PG438 | amyE::PnucA-lacZ-gfp+ cat | pPG58 → BSB1 |
| PG442 | amyE::Pxyl-msfGFP-kre spc | pPG54 → BSB1 |
| PG445 | amyE::Pxyl-mGFPmut1 spc | pHJS103 → BSB1 |
| PG447 | comK::phleo | This study |
| PG448 | amyE::PcomC-lacZ-gfp+ cat, kre:TnYLB1#4 kan | PG401-Tn4 DNA → PG435 |
| PG449 | amyE::PcomF-lacZ-gfp+ cat, kre:TnYLB1#4 kan | PG401-Tn4 DNA → PG436 |
| PG450 | amyE::PaddAB-lacZ-gfp+ cat, kre:TnYLB1 #4 kan | PG401-Tn4 DNA → PG437 |
| PG455 | aprE::PcomG-comK spc | This study |
| PG457 | amyE::PnucA-lacZ-gfp+ cat, kre:TnYLB1#4 kan | PG401-Tn4 DNA → PG438 |
| PG458 | aprE::PcomG-comK spc, amyE::PcomG-lacZ-gfp | PG455 DNA → PG389 |
| PG459 | aprE::PcomG-comK spc, amyE::PcomG-lacZ-gfp, kre:TnYLB1#4 kan | PG455 DNA → PG433 |
| PG461 | aprE::PcomG-comK spc, mecA::tet, comK::phleo, amyE::PcomG-lacZ-gfp cat | PG447 and PG368 DNAs → PG458 |
| PG463 | aprE::PcomG-comK spc, mecA::tet, comK::phleo, amyE::PcomG-lacZ-gfp cat, kre:TnYLB1#4 kan | PG447 and PG368 DNAs → PG459 |
| PG474 | amyE::Physp-kre spc | pPG59 → BSB1 |
| PG475 | amyE::Pxyl-gfp-comK spc, kre:TnYLB1#4 kan | PG406 DNA → PG349 |
| PG477 | amyE::Pxyl-msfGFP-kre spc, kre:TnYLB1#4 kan | PG406 DNA → PG442 |
| PG479 | kre::erm | This study |
| PG482 | amyE::Pxyl-msfGFP-kre spc, comG-gfp cat | PG342 DNA → PG442 |
| PG485 | amyE::Pxyl-msfGFP-kre spc, kre:TnYLB1#4 kan, comG-gfp cat | PG342 → PG477 |
| PG488 | amyE::PcomG-lacZ-gfp+ cat, kre::erm | PG479 DNA → PG389 |
| PG490 | comG:PcomG-gfp cat, amyE::Physp-kre spc | PG474 DNA → PG342 |
| PG491 | comG:PcomG-gfp cat, amyE::Physp-kre spc, kre::erm | [PG474+PG479] DNAs → PG342 |
| PG494 | amyE::Pxyl-kre-mSFgfp | pPG61 → BSB1 |
| PG500 | amyE:: cat Pveg-lacZ-gfp+ | pPG60 → BSB1 |
| PG501 | amyE:: cat Pkre-lacZ-gfp+ | pPG62 → BSB1 |
| PG512 | amyE:: cat Pveg-lacZ-gfp+, kre::erm | PG479 DNA → PG500 |
| PG505 | amyE::Pxyl-gfp-comK spc; kre:TnYLB1#4 kan, comK::phleo, mecA::tet | [PG447+PG368] DNAs → PG475 |
| PG508 | amyE::Pxyl-gfp-comK spc kan, comK::phleo, mecA::tet | [PG447+PG368] DNAs → PG349 |
| PG535 | amyE::Pxyl-mGFPmut1 spc, kre:TnYLB1#4 kan | PG406 DNA → PG445 |
| PG537 | amyE::Pxyl-mGFPmut1 spc, comK::phleo, mecA::tet | [PG447+PG368] DNAs → PG445 |
| PG538 | amyE::Pxyl-mGFPmut1 spc, kre:TnYLB1#4 kan, comK::phleo, mecA::tet | [PG447+PG368] DNAs → PG535 |
| PG548 | amyE::Physp-kre* | pPG59*fs → BSB1 |
| PG604 | kre:Pkre-gfpmut1 cat Pxyl-Pkre-kre | pPG66 → BSB1 (Campbell) |
| PG678 | kre:Pkre-gfpmut1 cat Pxyl-Pkre-kre, comK::phleo | PG447 DNA → PG604 |
| PG688 | kre:Pkre-gfpmut1 cat Pxyl-Pkre-kre, amyE::PcomG-mcherry spc | PG405 DNA → PG604 |
| PG710 | PcomG: cat PcomG-luc+ | pPG118 → BSB1 (Campbell) |
| PG724 | PcomG: cat PcomG-luc+, kre::erm | PG710 DNA → PG479 |
| PG746 | aprE::PcomG-lacZ-gfp+ kan | pPG63 → BSB1 |
| PG747 | amyE::PcomG-kre spc | pPG126 → BSB1 |
| PG753 | aprE::PcomG-lacZ-gfp+ kan, amyE::PcomG-kre spc | PG747 DNA → PG746 |
| PG755 | aprE::PcomG-lacZ-gfp+ kan, amyE::PcomG-kre spc, kre::erm | PG479 DNA → PG746 |
| PG763 | amyE:: cat Pkre-lacZ-gfp+, mecA::kan | PG343 DNA → PG501 |
| PG764 | amyE:: cat Pkre-lacZ-gfp+, comK::phleo | PG447 DNA → PG501 |
| PG765 | amyE:: cat Pkre-lacZ-gfp+, mecA::kan, comK::phleo | [PG343+PG447] DNAs → PG501 |
| PG811 | amyE::PpksA-lacZ-gfp+ cat | pPG136 → BSB1 |
| PG815 | amyE::PpksA-lacZ-gfp+ cat, kre::erm | PG479 DNA → PG811 |
| PG820 | amyE::Physp-gfp spc | AH7 DNA → BSB1 |
| PG821 | amyE::Physp-gfp spc, kre::erm | AH7 DNA → PG479 |
| E. coli | ||
| DH5α | F-, φ80lacZΔM15, Δ(lacZYAargF)U196, recA1, endA1, hsdR17, (rK-, mK+), phoA, supE44, λ-, thi-1, gyrA96, relA1 | Laboratory stock |
| Plasmid | Relevant features or genotype | Construction, source or reference |
| pDR111 | bla, amyE3', spc, lacI, Physp, amyE5' | [37] |
| pMarB | bla erm P ctc Himar1 kan (TnYLB-1) | [35] |
| pSG1164 | bla, cat, Pxyl-gfpmut1 | [61] |
| pMutin-GFP+ | bla erm Pspac-gfp+ lacI | [55] |
| pUC18Cm::luc | bla cat luc+ | [36] |
| pAWC3 | bla, aprE5', kan, lacI, lacZ-gfp+ aprE3' | Gamba et al., in preparation |
| pHJS103 | bla, amyE3', spc, Pxyl-mgfpmut1 | [61] and H. Strahl |
| pHJS105 | bla, amyE3', spc, Pxyl-mSFgfp, amyE5' | [61] and H. Strahl |
| pPG2 | bla, amyE3', spc, gfpmut1, amyE5' | [56] |
| pPG20 | bla, erm, lacI, gfp+ | This study |
| pPG21 | bla, erm, lacI, mcherry | This study |
| pPG22 | bla, amyE3', spc, gfp+, amyE5' | This study |
| pPG23 | bla, amyE3', spc, mcherry, amyE5' | This study |
| pPG34 | bla, amyE3', spc, PcomG-gfp+, amyE5' | This study |
| pPG35 | bla, amyE3', spc, PcomG-lacZ-gfp+, amyE5' | This study |
| pPG38 | bla, amyE3', spc, PcomG-mcherry, amyE5' | This study |
| pPG40 | bla, amyE3', cat, PcomG-lacZ-gfp+, amyE5' | This study |
| pPG49 | bla, amyE3', spc, mSFgfp, amyE5' | This study |
| pPG54 | bla, amyE3', spc, Pxyl-mSFgfp-kre, amyE5' | This study |
| pPG55 | bla, amyE3', cat, PcomC-lacZ-gfp+, amyE5' | This study |
| pPG56 | bla, amyE3', cat, PcomF-lacZ-gfp+, amyE5' | This study |
| pPG57 | bla, amyE3', cat, PaddAB-lacZ-gfp+, amyE5' | This study |
| pPG58 | bla, amyE3', cat, PnucA-lacZ-gfp+, amyE5' | This study |
| pPG59 | bla, amyE3', spc, lacI, Physp-kre amyE5' | This study |
| pPG59*fs | bla, amyE3', spc, lacI, Physp-kre*fs amyE5' | This study |
| pPG60 | bla, amyE3', cat, Pveg-lacZ-gfp+, amyE5' | This study |
| pPG61 | bla, amyE3', spc, Pxyl-kre-msfGFP, amyE5' | This study |
| pPG62 | bla, amyE3', cat, Pkre-lacZ-gfp+, amyE5' | This study |
| pPG63 | bla, aprE5', kan, lacI, PcomG-lacZ- gfp+, aprE3' | This study |
| pPG66 | bla Pxyl-Pkre-gfpmut1 cat | This study |
| pPG118 | bla cat PcomG-luc+ | This study |
| pPG126 | bla, amyE3', spc, PcomG-kre, amyE5' | This study |
| pPG136 | bla, amyE3', cat, PpksA-lacZ-gfp+, amyE5' | This study |