TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Strains | ||
| B. subtilis | ||
| 168 | trpC2 | BGSCa |
| 3G18Δ8 | ade met trpC2 ΔhemH::ble | 16 |
| 168Δ8 | trpC2 ΔhemH::ble | This work |
| 168S54A | trpC2 hemHS54A | This work |
| 168Q63A | trpC2 hemHQ63A | This work |
| E. coli | ||
| JM109 | recA1 supE44 endA1 hsdR17 gyrA96 relA1 thi Δ(lac-proAB) (F′ traD36 proAB+lacIqlacZΔM15) | 43 |
| BL21(DE3) | hsdS gal (λcIts857 ind1 Sam7 nin5 lacUV5-T7 gene 1) | 37 |
| Plasmids | ||
| pLUG185 | bla; 2.8-kb SphI fragment of pLUG1301 in pUC18; contains hemH flanked by 50% of hemE and 90% of hemY | 14 |
| pLUGS54Aa1 | pLUG185 derivative, contains a T-to-G point mutation in bp 3197b of hemH | This work |
| pLUGQ63Aa1 | pLUG185 derivative, contains a CA-to-GC point mutation in bp 3224 and 3225b of hemH | This work |
| pLUGT7-H | bla; B. subtilis hemH cloned downstream of the T7 promoter of pBluescript II KS(−) | 17 |
| pT7S54Ac1 | pLUGT7-H derivative, contains a T-to-G point mutation in bp 3197b of hemH | This work |
| pT7Q63Ab2 | pLUGT7-H derivative, contains a CA-to-GC point mutation in bp 3224 and 3225b of hemH | This work |
a
BGSC, Bacillus Genetic Stock Center, Department of Biochemistry, Ohio State University, Columbus.
b
Base pair numbering is according to that of a previous study (14).