TABLE 1.
Strains, plasmids, and primers
| Strain, plasmid, or primer | Relevant genotype, phenotype, and/or sequence | Reference or source |
|---|---|---|
| M. xanthus strains | ||
| DK1622 | Wild type | 8 |
| DK10410 | ΔpilA | 24 |
| DK10404 | DK 1622::pilR-Ω3163 | 24 |
| DK10415 | ΔpilS | 23 |
| DK12600 | DK10410::pLOJ7 | This study |
| DK12601 | DK10410::pLOJ7/pLOJ6 | This study |
| DK12602 | DK1622::pLOJ5 | This study |
| DK12603 | DK1622::pLOJ5/pLOJ6 | This study |
| DK12604 | DK1622::pLOJ5/pLOJ28 | This study |
| DK12612 | DK10404::pLOJ7 | This study |
| E. coli strains | ||
| TOP10 | Invitrogen | |
| C600 | Wild-type lacZ gene | |
| Plasmids | ||
| pBluescriptII KS(+) | Cloning vector, Ampr | Stratagene |
| pBGS18 | Cloning vector, Kanr | 17 |
| pSWU30 | Cloning vector, Mx8 attP Tcr | 22 |
| pET28 | Cloning vector, lacIq gene, Kanr | Novagen |
| pLOJ1 | pilA promoter, Ampr | This study |
| pLOJ2 | Leader with one lac operator, Kanr | This study |
| pLOJ4 | Inducible pilA promoter, Kanr | This study |
| pLOJ5 | Inducible lacZ gene, one operator, Kanr | This study |
| pLOJ6 | lacIq, Mx8 attP Tcr | This study |
| pLOJ7 | Inducible pilA gene, Kanr | This study |
| pLOJ15 | pilA promoter, Ampr | This study |
| pLOJ16 | pilA promoter-lacI, Ampr | This study |
| pLOJ28 | pilA promoter-lacI, Tcr | This study |
| Primersa | ||
| 1 | 5′-GGCCCAAGCTTGATGGCACCGTCATGTTGGACG-3′ | |
| 2 | 5′-CGGGATCCCCCGCGGATGGGATTAGCACTA-3′ | |
| 3 | 5′-CGGGATCCGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGAGGTACCCC-3′ | |
| 4 | 5′-GGGGTACCTCTCCTTCTTAAAGTTAAACAAAATTATTTCTAGAGGGGAATTGTTATCCGCTCACAATTCCCGGATCCCG-3′ | |
| 5 | 5′-GTACCATGCGCGTCTCGCGATTCAAC-3′ | |
| 6 | 5′-GGAATTCGGCCCGAAGAACAGTGGACC-3′ | |
| 7 | 5′-CGGGGTACCATGACCATGATTACGGATTCACTGGC-3′ | |
| 8 | 5′-CCTGCCCGGTTATTATTATTTTTGACACC-3′ | |
| 10 | 5′-GGGATATCCTCAGAGAAGGTTGCAACGGGG-3′ | |
| 11 | 5′-TCCCCCGGGCATGTGAAACCAGTAACGTTATACGATGTCGC-3′ | |
| 12 | 5′-CGGAATTCCGCACCTGTCCTACGAGTTGC-3′ | |
| 13 | 5′-CAGTTGGTTAGAACGCCGGC-3′ | |
| 14 | 5′-CTGATTCTCCCGAGCCCCAC-3′ | |
| 15 | 5′-CAGCACCTTGCGCTCACGTC-3′ | |
| 16 | 5′-GGGCAAAACCCGAGCCTCTC-3′ |
a
Use of the primers is described in Materials and Methods.