TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli DH5α | F−endA1 hsdR17(rK− mK+) supE44 thi-1 λ−gyrA96 relA1 Δ(lacZYA argF)U169 (Φ80 lacZΔM15) recA1 | Roche |
| Clostridium cellulolyticum ATCC 35319 | Wild type | 30 |
| Plasmids | ||
| pPSV | pJIR418 derivative carrying the promoterless CAT gene (catP); Emr | 24 |
| pGEM-T Easy | Cloning vector; Apr | Promega |
| pSOS952 | E. coli-Clostridium shuttle vector (ColE1 and pIM13 replicons); Apr Emr; carries the adc-ctfA-ctfB operon transcribed from a modified thl promoter (two lac operators) | 29 |
| pPSVthl | pPSVΩ (SmaI Sos-dir/SosE 259-bp PCR fragment carrying thiolase promoter) | This study |
| pPSV1 | pPSVΩ (SphI/SacI 1,030-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV1ΔCRE | pPSVΩ (SphI/SacI 1,016-bp fragment of cipC ΔCRE promoter cloned in pGEM-T Easy) | This study |
| pPSV2 | pPSVΩ (SphI/SacI 155-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV3 | pPSVΩ (SphI/SacI 701-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV4 | pPSVΩ (SphI/SacI 574-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV5 | pPSVΩ (SphI/SacI 138-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV6 | pPSVΩ (SphI/SacI 250-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV7 | pPSVΩ (SphI/SacI 428-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV8 | pPSVΩ (SphI/SacI 452-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |
| pPSV9 | pPSVΩ (SphI/SacI 540-bp fragment of cipC promoter cloned in pGEM-T Easy) | This study |