TABLE 1.
List of strains and plasmids used in this work
| Strain or plasmid | Relevant characteristicsa | Source or referenceb |
|---|---|---|
| Strainsc | ||
| Escherichia coli XL1-Blue | endA1 gyrA96 thi hsdR17(rK− mK+) supE44 relA1 lac/F′ proAB+ lacIqlacZΔM15 Tn10 | Stratagene, Inc. |
| B. subtilis JH642 | pheA1 trpC2 | BGSCd |
| Δqox | trpC2 ΔqoxABCD::kan | 32 |
| Δspo0A | pheA1 trpC2 Δspo0A::erm | 13 |
| 168A | trpC2 | Laboratory stock |
| Strains derived from 168A | ||
| LUH14 | ΔqoxABCD::kan | Δqox→168A |
| LUH15 | ΔctaCD::ble | 33 |
| LUH17 | ΔqoxABCD::kan ΔctaCD::ble | 33 |
| LUW10 | ΔcydABCD::cat | 33 |
| LUW20 | ΔcydABCD::tet | 33 |
| LUW22 | ΔcydABCD::tet qox::pSPOX | pSPOX→LUW20 |
| LUW23 | ΔctaCD::ble ΔcydABCD::tet | LUW20→LUH15 |
| LUW24 | ΔctaCD::ble ΔcydABCD::tet, qox::pSPOX | pSPOX→LUW23 |
| LUW29 | ΔcydABCD::cat ΔqoxABCD::kan | LUW10→LUH14 |
| LUW32 | qox::pSPOX | pSPOX→168A |
| LUW33 | ΔctaCD::ble ΔcydABCD::tet ΔqoxABCD::kan | LUH14→LUW23 |
| LUW42 | ΔctaCD::ble qox::pSPOX | pSPOX→LUH15 |
| LUW122 | ΔythAB::tet | 2 |
| LUW128 | ΔcydCD::cat | pCYD24→168A |
| 1A1 | trpC2 | BGSCd |
| Strains derived from 1A1 | ||
| LUW34 | ΔcydABCD::cat | LUW10→1A1 |
| LUW46 | ΔqoxABCD::kan | LUH14→1A1 |
| LUW112 | Δspo0A::erm | Δspo0A→1A1 |
| LUW137 | ΔythAB::tet | LUW122→1A1 |
| LUW138 | ΔqoxABCD::kan ΔythAB::tet | LUW122→LUW46 |
| LUW142 | ΔctaCD::ble | LUH15→1A1 |
| LUW143 | ΔctaCD::ble ΔqoxABCD::kan | LUW46→LUW142 |
| LUW145 | ΔctaCD::ble ΔcydABCD::cat | LUW34→LUW142 |
| LUW147 | ΔctaCD::ble ΔythAB::tet | LUW122→LUW142 |
| LUW148 | ΔctaCD::ble ΔqoxABCD::kan ΔythAB::tet | LUW46→LUW147 |
| LUW196 | ΔctaCD::ble ΔcydABCD::cat ΔythAB::tet | LUW10→LUW147 |
| LUW198 | ΔcydABCD::cat ΔythAB::tet | LUW10→LUW137 |
| Plasmids: | ||
| pDH88 | Cmr Amr | 11 |
| pSPOX | qoxA′ in pDH88 | This work |
| pHP13 | Cmr Emr | 8 |
| pCYD13 | cydA′ and cydD′ in pHV32 | 33 |
| pCYD22 | cydCD in pHP13 | 33 |
| pCYD23 | cydABCD and the cyd promoter in pHP13 | 33 |
| pCYD24 | cydC′ and cydD′ in pHV32 | This work |
| pCYD25 | cydCD and the cyd promoter in pHP13 | This work |
a
Amr, Cmr, and Emr indicates resistance to ampicillin, chloramphenicol, and erythromycin, respectively.
b
Arrows indicate transformation and point from donor to recipient.
c
All B. subtilis strains derived from the parental strains 168A and 1A1 contain the trpC2 mutation. During this work, strain 168A was found to be oligosporogenic, which affects growth in liquid media. Therefore, doubling times of oxidase mutants were calculated for strains derived from the parental strain 1A1.
d
Bacillus Genetic Stock Center. Department of Biochemistry, Ohio State University, Columbus.