KEY RESOURCES TABLE
| Bacillus subtilis strains used in this study | ||
|---|---|---|
|
| ||
| Strain | Genotype or description | Reference |
|
| ||
| PY79 | Prototrophic derivative of B. subtilis 168 | Youngman et al. (1984) |
| JB103 | ΔspoIVA::neo thrC::spoIVAD97A spec | This study |
| JB168 | ΔspoIVA::erm thrC::spoIVAT70A,T71A spec ΔspoVID::neo | This study |
| JB171 | ΔspoIVA::erm DspoVID::neo thrC::spoIVA spec | This study |
| JB174 | ΔspoIVA::erm DspoVID::neo thrC::spoIVA spec amyE::spoVID cat | This study |
| JB175 |
ΔspoIVA::erm thrC::spoIVA spec ΔspoVID::neo amyE::spoVID1−545 cat |
This study |
| JB176 | ΔspoIVA::erm ΔspoVID::neo thrC::spoIVAT70A,T71A spec amyE::spoVID cat | This study |
| JB177 |
ΔspoIVA::erm ΔspoVID::neo thrC::spoIVAT70A,T71A
spec amyE:: spoVID1−545 cat |
This study |
| JB217 | amyE::gfp-spoVID cat | This study |
| JB242 |
amyE::spoVIDT532G
cat ΔspoIVA::erm ΔspoVID::neo thrC:: spoIVAT70A,T71A spec |
This study |
| JB280 |
ΔspoIVA::erm thrC:: spoIVAD97A
spec ΔspoVID::neo |
This study |
| JB281 |
amyE::spoVIDT532G
cat ΔspoIVA::erm ΔspoVID::neo thrC::spoIVA spec |
This study |
| JB293 |
amyE::spoVID cat ΔspoIVA::erm ΔspoVID::neo thrC:: spoIVAD97A spec |
This study |
| JB294 | ΔspoIVA::erm thrC:: spoIVAD97A ΔspoVID::neo spec amyE::spoVIDQ546stop cat | This study |
| JB295 |
amyE::spoVIDT532G
cat ΔspoIVA::erm ΔspoVID::neo thrC::spoIVAD97A spec |
This study |
| JPC156 | ΔspoIVA::neo amyE::spoIVA cat thrC::GFPspoIVA spec | Castaing et al. (2013) |
| JPC221 | ΔspoIVA::neo thrC:: spoIVAT70A,T71A spec | Castaing et al. (2013) |
| JPC243 | ΔspoIVA::neo amyE:: spoIVAT70A,T71A cat thrC::GFP-spoIVAT70A,T71A spec | Castaing et al. (2013) |
| KP73 | ΔspoIVA::kan | Price and Losick (1999) |
| KR394 | ΔspoIVA::neo thrC::spoIVA spec | Ramamurthi and Losick (2008) |
| TD100 | ΔspoVID::neo | This study |
| TD1000 | ΔspoIVA::erm thrC::spoIVA spec ΔspoVID::neo amyE::GFPspoVID514−575 cat | This study |
| TD1001 | ΔspoIVA::erm thrC::spoIVA spec ΔspoVID::neo amyE::GFPspoVID515−575 cat | This study |
| TD1002 | ΔspoIVA::erm thrC::spoIVA spec ΔspoVID::neo amyE::GFPspoVID564−575 cat | This study |
| TD1029 | amyE::GFP-spoVID501−575 cat | This study |
| TD1030 | amyE:: GFP-spoVID501-575(T532G) cat | This study |
| TD1031 | amyE::GFP-spoVID511−575 cat | This study |
| TD1032 | amyE:: GFP-spoVID511−575(T532G) cat | This study |
| TD1056 |
ΔspoIVA::erm ΔspoVID::neo thrC:: spoIVAD97A spec amyE::PspoVID-FLAG-GFP cat |
This study |
| TD1070 | ΔspoVID::neo amyE::GFPspoVID501−575 cat | This study |
| TD1074 | ΔspoIVA::erm thrC::spoIVA spec ΔspoVID::neo amyE::gfp-spoVID cat pyrD::mcherry-spoIVA cat::tet | This study |
| TD1085 | ΔspoVID::neo sacA::GFP- spoVID501−575 cat::tet | This study |
| TD1086 | ΔspoVID::neo amyE:: GFP- spoVID501−575 cat sacA::GFP- spoVID501−575 cat::tet | This study |
| TD1139 | ΔspoIVA::neo thrC::spoIVA spec amyE::spoVIDT532G cat | This study |
| TD1140 | ΔspoIVA::erm thrC::spoIVAT70A,T71A spec amyE::spoVIDT532G cat | This study |
| TD1141 | ΔspoIVA::erm thrC::spoIVAD97A spec amyE::spoVIDT532G cat | This study |
| TD139 |
ΔspoIVA::erm thrC::spoIVA spec
ΔspoVID::neo amyE::gfp-spoVID cat |
This study |
| TD142 |
ΔspoIVA::erm thrC::spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID cat |
This study |
| TD144 |
ΔspoIVA::erm thrC::spoIVAT70A,T71A
spec ΔspoVID::neo amyE::gfp-spoVID cat |
This study |
| TD154 | ΔspoIVA::erm thrC::spoIVA spec ΔspoVID::neo amyE::gfpspoVIDQ546Stop cat | This study |
| TD172 | ΔspoIVA::erm ΔspoVID::neo amyE::gfpspoVID cat | This study |
| TD251 |
ΔspoIVA::erm thrC::spoIVA spec ΔspoVID::neo amyE::gfp-spoVIDT532G cat |
This study |
| TD300 |
ΔspoIVA::neo amyE::spoIVA cat
ΔspoVID::erm thrC::GFP-spoIVA spec |
This study |
| TD302 | ΔspoIVA::neo amyE:: spoIVAT70A,T71A cat ΔspoVID::erm thrC::GFPspoIVAT70A,T71A spec | This study |
| TD931 |
ΔspoIVA::erm thrC::spoIVA spec
ΔspoVID::neo amyE::GFP-spoVID 501−575 |
This study |
| TD933 |
ΔspoIVA::erm thrC:: spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID501−575 |
This study |
| TD953 |
ΔspoIVA::erm DspoVID::neo thrC:: spoIVAD97A spec amyE::gfpspoVIDT532G cat |
This study |
| TD954 |
ΔspoIVA::erm thrC:: spoIVAT70A,T71A
spec ΔspoVID::neo amyE::gfp- spoVIDT532G cat |
This study |
| TD955 | ΔspoIVA::erm DspoVID::neo thrC::spoIVA spec amyE::gfp-spoVID501−575(T532G) cat | This study |
| TD957 | ΔspoIVA::erm ΔspoVID::neo thrC::spoIVAD97A spec amyE::gfpspoVID501–575(T532G) cat | This study |
| TD961 |
ΔspoIVA::erm thrC::spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID511−575 |
This study |
| TD962 |
ΔspoIVA::erm thrC::spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID521−575 |
This study |
| TD979 |
ΔspoIVA::erm thrC:: spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID512−575 |
This study |
| TD980 |
ΔspoIVA::erm thrC:: spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID513−575 |
This study |
| TD981 |
ΔspoIVA::erm thrC:: spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID514−575 |
This study |
| TD982 |
ΔspoIVA::erm thrC:: spoIVAD97A
spec ΔspoVID::neo amyE::gfp-spoVID515−575 |
This study |
| TD983 |
ΔspoIVA::erm thrC:: spoIVAD97A
spec ΔspoVID::neo amyE:: gfp-spoVID516−575 |
This study |
| TD996 |
ΔspoIVA::erm thrC::spoIVA spec
ΔspoVID::neo amyE::gfp-spoVID 511−575 |
This study |
| TD997 |
ΔspoIVA::erm thrC::spoIVA spec
ΔspoVID::neo amyE::gfp-spoVID 521−575 |
This study |
| TD998 |
ΔspoIVA::erm thrC::spoIVA spec
ΔspoVID::neo amyE::gfp-spoVID 512−575 |
This study |
| TD999 |
ΔspoIVA::erm thrC::spoIVA spec
ΔspoVID::neo amyE::gfp-spoVID 513−575 |
This study |
|
| ||
| Escherichia coli strains used in this study | ||
|
| ||
| Strain | Genotype and description | Reference |
|
| ||
| BL21(DE3) | pKR168 (pET28a backbone, PT7-GFP-His6) | Ramamurthi and Losick (2008) |
| BL21(DE3) |
pTD352 (pET28a backbone, P
T7
-GFP-
SpoVID501–575-His6) |
This study |
| BL21(DE3) | pTD354 (pET28a backbone, P T7 -GFPspoVID 501−575 -FLAG) | This study |
| BL21(DE3) | pTD436 (pET28a backbone, P T7 - GFPspoVID501–545-His6) | This study |
|
| ||
| Item | Source | Catalog number |
|
| ||
| Antisera | Purified antigen used | |
|
| ||
| Rabbit anti-GFP | Ramamurthi lab; Covance/LabCorp | Purified GFP-His6 |
| Rabbit anti-SpoIVA | Ramamurthi lab; Covance/LabCorp | Purified His6-SpoIVA |
| Rabbit anti-SpoVID | Ramamurthi lab; Covance/LabCorp | Purified His6-SpoVID |
|
| ||
| Microscopy | ||
|
| ||
| FM4–64 | Thermo Fisher Scientific | T13320 |
| 35 mm Dish, No. 1.5 Coverslip | Mat Tek | P35GC-1.5–14-C |
|
| ||
| Protein purification and detection | ||
|
| ||
| Ni-NTA agarose beads | Qiagen | 30250 |
| Pierce Anti-DYKDDDDK Magnetic Agarose | Thermo Fisher Scientific | A36798 |
| QuikChange Lightning Site-Directed Mutagenesis Kit |
Agilent | 210519 |
| StarBright Blue 700 Goat Anti-Rabbit IgG | BioRad | 12004161 |