TABLE 1.
Bacterial strains and plasmids
| Bacterial strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| Strain | ||
| E. coli ER2275 | NEBb | |
| E. coli OneShot chemically competent TOP10 | Invitrogen | |
| C. acetobutylicum ATCC 824 | ATCCc | |
| M5 | AADC− CoAT− | 4 |
| Plasmid | ||
| pAN1 | Cmr; carries the φ3T I gene | 17 |
| pFNK6 | Ampr MLSr; ace operon | 18 |
| pFNK7 | Ampr MLSrctfA ctfB | 18 |
| pSOS95 | ace2 operon | 30 |
| pSOS95del | Ampr MLSr; thl promoter | This study |
| pADC38AS | Ampr MLSr; thl promoter; antisense adc38 | This study |
| pADC68AS | Ampr MLSr; thl promoter; antisense adc68 | This study |
| pADC100AS | Ampr MLSr; thl promoter; antisense adc100 | This study |
| pCTFA2AS | Ampr MLSr; thl promoter; antisense ctfa2 | This study |
| pCTFB1AS | Ampr MLSr; thl promoter, antisense ctfb1 | This study |
| pCOAT11AS | Ampr MLSr; thl promoter; combined antisense ctfb1 and antisense ctfa1 | This study |
adc, AADC gene; ctfA, CoAT subunit A gene; ctfB, CoAT subunit B gene; Cmr, chloramphenicol resistant; φ3T I, Bacillus subtilis phage φ3T I methyltransferase gene; Ampr, ampicillin resistant; MLSr, macrolide-lincosamide-streptogramin B resistant; ace operon, synthetic operon which contains the three homologous acetone formation genes (adc, ctfA, and ctfB) transcribed from the adc promoter; ctfA, gene encoding for the A subunit of CoAT; ctfB, gene encoding for the B subunit of CoAT; ace2 operon, synthetic operon which contains the three homologous acetone formation genes (adc, ctfA, and ctfB) transcribed from the thl promoter; thl promoter, promoter region for the thiolase gene (thl) of C. acetobutylicum strain ATCC 824; adc38, DNA fragment containing approximately 38% of the sequence of the adc transcript; adc68, DNA fragment containing approximately 68% of the sequence of the adc transcript; adc100, DNA fragment containing approximately 100% of the sequence of the adc transcript; ctfa2, DNA fragment containing approximately 79% of the ctfA structural gene; ctfb1, DNA fragment containing approximately 39% of the ctfB structural gene; ctfa1, DNA fragment containing approximately 53% of the ctfA structural gene.
NEB, New England Biolabs (Beverly, Mass.).
ATCC, American Type Culture Collection (Manassas, Va.).