TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| B. subtilis 168 strains | ||
| KSS1001 | trpC2 | K. Bott |
| KSS1570 | amyE::pdivIVB mreD-minCD | This study |
| KSS1571 | Ω pLEE100, amyE::pdivIVB mreD-minCD | This study |
| KSS1572 | trpC2, Ω pLEE100 | This study |
| KSS1573 | KSS1571 (pLEE504) | This study |
| MO1099 | amyE::erm | 9 |
| E. coli strains | ||
| DH5α | endA1 gyrA96 hsdR17 ΔlacU169 (φ80lacZΔM15) recA1 supE44 thi-1 | BRLc |
| M15 | StrrlacZΔM15 (pRep4 Kmr) | Qiagen |
| Plasmids | ||
| pCHA2489 | mreC ORFa PCR product cloned into pT7Blue | This study |
| pCHA2506 | NotI (in mreC, blunted)-SalI (vector site) mreC DNA fragment from pCHA2489 cloned into EcoICRI-SalI site of pQE30 | This study |
| pCR2.1 | PCR fragment cloning vector, Apr Kmr | Invitrogen |
| pDG364 | amyE locus integration vector | BGSCd (9) |
| pDG3053 | Shuttle expression plasmid utilizing the S. aureus lactose operon promoter; Apr Kanr | This study |
| pLEE50 | pUS19 with the lacI spac EcoRI-BamHI insert from pSI-1 | This study |
| pLEE100 | MreC-inducible expression integration vector | This study |
| pLEE200 | pDG364 containing pdivIVB mreC-minCD | This study |
| pLEE504 | pDG3053 containing truncated mreCb | This study |
| pQE30 | Six-histidine tag vector, Apr | Qiagen |
| pSI-1 | IPTG-inducible expression shuttle plasmid | D. Henner (2, 33) |
| pT7Blue | PCR product cloning vector, Apr | Invitrogen |
| pUS19 | pUC19 with Spr expressed in B. subtilis | 3 |
a
ORF, open reading frame.
b
Truncated mreC (contains bp 1 to 540).
c
BRL, Bethesda Research Laboratories.
d
BGSC, Bacillus Genetic Stock Center, Ohio State University.