TABLE 2.
Strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Bacterial strains | ||
| C. acetobutylicum | ||
| 824 | ATCC 824, wild-type strain | ATCC |
| ΔccpA | ccpA::intron | 14 |
| 824-P | ATCC 824 carrying the pIMP1-Pthl plasmid | This study |
| 824-ccpA | ATCC 824 carrying the pIMP1-Pthl-ccpA plasmid | This study |
| 824-ccpA-m | ATCC 824 carrying the pIMP1-Pthl-ccpAmut plasmid | This study |
| 824 m-P-ccpA | ΔccpA carrying the pIMP1-PccpA-ccpA plasmid | This study |
| 824 m-Pm-ccpA | ΔccpA carrying the pIMP1-PccpAmut-ccpA plasmid | This study |
| creP/824 | ATCC 824 carrying the pIMP1-PccpA-lacZ plasmid | This study |
| creP-m/824 | ATCC 824 carrying the pIMP1-PccpAmut-lacZ plasmid | This study |
| creORF/824 | ATCC 824 carrying the pIMP1-PccpA-ccpA-lacZ plasmid | This study |
| creORF-m/824 | ATCC 824 carrying the pIMP1-PccpA-ccpAmut-lacZ plasmid | This study |
| creORF/ΔccpA | ΔccpA carrying the pIMP1-PccpA-ccpA-lacZ plasmid | This study |
| creORF-m/ΔccpA | ΔccpA carrying the pIMP1-PccpA-ccpAmut-lacZ plasmid | This study |
| E. coli | ||
| Top10 | General cloning host strain | Invitrogen |
| ER2275 | Strain used to methylate the expression vector | New England Biolabs |
| Rosetta(DE3) | Strain used for protein overexpression | Novagen |
| Plasmids | ||
| pIMP1-Pthl | Vector with thl promoter | This study |
| pIMP1-Pthl-ccpA | ccpA overexpression vector derived from pIMP1-Pthl | This study |
| pIMP1-Pthl-ccpAmut | ccpAmut overexpression vector derived from pIMP1-Pthl | This study |
| pIMP1-PccpA-ccpA | ccpA overexpression vector derived from pIMP1-PccpA | This study |
| pIMP1-PccpAmut-ccpA | ccpA overexpression vector derived from pIMP1-PccpAmut | This study |
| pIMP1-PccpA-lacZ | lacZ reporter gene driven by ccpA promoter | This study |
| pIMP1-PccpAmut-lacZ | lacZ reporter gene driven by mutant ccpA promoter | This study |
| pIMP1-PccpA-ccpA-lacZ | Fusion of a segment of the ccpA gene and lacZ reporter driven by the ccpA promoter | This study |
| pIMP1-PccpA-ccpAmut-lacZ | Fusion of a segment of the ccpAmut gene and lacZ reporter driven by the ccpA promoter | This study |
| pIMP1-PccpAmut-ccpAmut-lacZ | Fusion of a segment of the ccpAmut gene and lacZ reporter driven by the ccpAmut promoter | This study |