TABLE 1.
Plasmids and bacterial strains used in this study
| Plasmid or strain | Description | Source or reference |
|---|---|---|
| Plasmids | ||
| pGEM-T Easy | PCR product TA cloning vector; Ampr | Promega |
| pGBKB | pGEM-T Easy carrying the 1,973-bp PCR product BmoUP-Kan-BmoDN (bmoX partially deleted and Kanr inserted into the deleted region) | This study |
| pBluescript II SK | Cloning vector, 3.0 kb; AprlacZ′ | Stratagene |
| pBSbmoxyb | pBluescript carrying a 3.68-kb bmoXYZ PCR product cloned with KpnI | This study |
| Strains | ||
| P. butanovora | ATCC 43655; n-butane-assimilating bacteria | 30 |
| PBKB | Mutant strain with bmoX partially deleted and Kanr inserted into the deleted region; created by homologous recombination of BmoUP-Kan-BmoDN from pGBKB with P. butanovora | This study |
| Rev WT | Butane-oxidizing control strain used for comparison of P. butanovora mutants containing single amino-acid substitutions in BMOH-α; created by homologous recombination of bmoXYB from pBSbmoxyb with P. butanovora PBKB replacing Kanr with wild-type bmoX sequence | This study |
| E. coli | ||
| JM109 | Cloning host strain; Ampr | 41 |
| DH5α | Cloning host strain; Ampr | Invitrogen |