Table 1.
Plasmids and strains used in this study.
| Plasmid or straina | Relevant genotype |
|---|---|
| Sporulation, competence, and colony architecture | |
| KB19 | amyE::Pspank-(empty vector) spc |
| KB26 | amyE::Pspank-rap60 spc |
| KB27 | amyE::Pspank-rap60-phr60 spc |
| KB28 | amyE::Pspank-phr60 spc |
| KG1780 | pTA1060 |
| KG1782 | pTA1060-Δphr60 |
| KG1781 | pTA1060-Δrap60Δphr60 |
| KG1897 | DS2569 pTA1060 |
| KG1899 | DS2569 pTA1060-Δphr60 |
| KG1898 | DS2569 pTA1060-Δrap60Δphr60 |
| KG37 | 3610 Δspo0A::cm |
| Sporulation and cannabilism gene promoter-lacZ fusions | |
| KB45 | lacA:PspoIIA-lacZ tet amyE:: Pspank-(empty vector) spc |
| KB46 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60 spc |
| KB47 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60-phr60 spc |
| KB48 | lacA::PspoIIA-lacZ tet amyE:: Pspank-phr60 spc |
| KB63 | lacA:PspoIIA-lacZ tet amyE:: Pspank-rapC spc |
| KB138 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rapB spc |
| KG1766 | amyE::PspoIIA-lacZ neo pTA1060 |
| KG1771 | amyE::PspoIIA-lacZ neo pTA1060-Δrap60Δphr60 |
| KG1772 | amyE::PspoIIA-lacZ neo pTA1060-Δphr60 |
| KG1911 | amyE::Pskf-lacZ spc pTA1060 |
| KG1912 | amyE::Pskf-lacZ spc pTA1060-Δphr60 |
| PH62 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60(F23A) spc |
| PH28 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60(R26A) spc |
| PH63 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60(R26P) spc |
| PH64 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60(N27A) spc |
| PH65 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60(Q46A) spc |
| PH66 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60(Q46N) spc |
| PH67 | lacA::PspoIIA-lacZ tet amyE:: Pspank-rap60(L66A) spc |
| Matrix gene promoter-lacZ fusions | DS2569 amyE::PepsA-lacZ cat pTA1060 |
| KG1928 | DS2569 amyE::PepsA-lacZ cat pTA1060-Δphr60 |
| KG1929 | DS2569 amyE::PtapA-lacZ cat pTA1060 |
| KG1930 | DS2569 amyE::PtapA-lacZ cat pTA1060-Δphr60 |
| KG1931 | DS2569 amyE::PabrB-lacZ spc pTA1060 |
| KG1903 | DS2569 amyE::PabrB-lacZ spc pTA1060-Δphr60 |
| KG1905 | |
| Competence gene promoter-lacZ fusions | |
| KB22 | thrC::PsrfA-lacZ erm amyE:: Pspank-(empty vector) spc |
| KB23 | thrC::PsrfA-lacZ erm amyE:: Pspank-rap60 spc |
| KB24 | thrC::PsrfA-lacZ erm amyE:: Pspank-rap60-phr60 spc |
| KB25 | thrC::PsrfA-lacZ erm amyE:: Pspank-phr60 spc |
| KB137 | thrC::PsrfA-lacZ erm amyE:: Pspank-rapC spc |
| KB53 | lacA::PsrfA-lacZ tet amyE::Pspank-(empty vector) spc |
| KB54 | lacA::PsrfA-lacZ tet amyE::Pspank-rap60 spc |
| KB58 | lacA::PsrfA-lacZ tet amyE::Pspank-rap60 spcΔspoOK::erm |
| KG1759 | amyE::PsrfA-lacZ neo pTA1060 |
| KG1764 | amyE::PsrfA-lacZ neo pTA1060-Δrap60Δphr60 |
| KG1765 | amyE::PsrfA-lacZ neo pTA1060-Δphr60 |
| KG1793 | amyE::PcomK-lacZ neo pTA1060 |
| KG1796 | amyE::PcomK-lacZ neo pTA1060 Δspo0A::cat |
| KG1795 | amyE::PcomK-lacZ neo pTA1060-Δphr60 |
| PH55 | lacA::PsrfA-lacZ tet amyE:: Pspank-rap60(F23A) spc |
| PH9 | lacA::PsrfA-lacZ tet amyE:: Pspank-rap60(R26A) spc |
| PH56 | lacA::PsrfA-lacZ tet amyE:: Pspank-rap60(R26P) spc |
| PH57 | lacA::PsrfA-lacZ tet amyE:: Pspank-rap60(N27A) spc |
| PH58 | lacA::PsrfA-lacZ tet amyE:: Pspank-rap60(Q46A) spc |
| PH59 | lacA::PsrfA-lacZ tet amyE:: Pspank-rap60(Q46N) spc |
| PH60 | lacA::PsrfA-lacZ tet amyE:: Pspank-rap60(L66A) spc |
| Protein purificationb | |
| KB98 | pBADAp18-Nhis6-KinA |
| KG1878 | pBADAp18-Nhis6-Spo0F |
| KG1879 | pBADCm33-Nhis6-Spo0B |
| KG1880 | pBADCm33-Nhis6-Spo0A |
| KB95 | pBADCm33-Nhis6-Rap60 |
| PH537 | pBADCm33-Nhis6-Rap60(F23A) |
| PH538 | pBADCm33-Nhis6-Rap60(R26A) |
| PH539 | pBADCm33-Nhis6-Rap60(R26P) |
| PH540 | pBADCm33-Nhis6-Rap60(N27A) |
| PH541 | pBADCm33-Nhis6-Rap60(Q46A) |
| PH542 | pBADCm33-Nhis6-Rap60(Q46N) |
| PH543 | pBADCm33-Nhis6-Rap60(L66A) |
| KG1881 | pBADCm33-Nhis6-RapA |
| KG1882 | pBADCm33-Nhis6-RapB |
| KG1883 | pBADAp18-Nhis6-RapC |
| KG1958 | pBADAp18-Nhis6-ComP(CD) |
| KG1959 | pBADAp18-Nhis6-ComA |
| KG1884 | pET21a–Nhis6-SUMO-Spo0F |
| KG1885 | pET21a–Nhis6-SUMO-Spo0B |
| KG1960 | pET21a–Nhis6-SUMO-ComA |
a
All B. subtilis strains are derivatives of JH642 and contain mutations in trpC and pheA (not shown) unless otherwise noted. Strain DS2569 is a pBS32 plasmid-cured derivative of the prototrophic strain NCIB3610.
b
Plasmids used for protein purification are in strain DH5α.