Table 1.
Bacterial strains and plasmids
| Strain | Reference or source | Description |
|---|---|---|
| Strains | ||
| SB1003 | 54 | R. capsulatus WT, Rifr |
| B10 | 24 | R. capsulatus WT, Rifs |
| DE442 | ?a | R. capsulatus RcGTA overproducer, Rifr |
| Y262 | 13 | R. capsulatus RcGTA overproducer, Rifr |
| DH5α | 27 | E. coli, general cloning strain |
| S17-1 λ pir | 28 | E. coli pir, plasmid conjugation |
| TEC5 | 29 | E. coli, plasmid conjugation |
| BL21(DE3) | Invitrogen | E. coli, protein overexpression |
| SB555 | 14 | R. capsulatus SB1003 Δrcc00555 Rifr Kanr |
| DE555 | This work | R. capsulatus DE442 Δrcc00555 Rifr Kanr |
| SB1003 cckA | 16 | R. capsulatus SB1003 ΔcckA Rifr Kanr |
| DEKOcckA | This work | R. capsulatus DE442 ΔcckA Rifr Kanr |
| Plasmids | ||
| pUC19 | Invitrogen | Cloning vector, Ampr |
| TOPO TA | Invitrogen | Cloning vector, Ampr |
| pZJD29A | J. Jiang and C. E. Bauer, unpublished data | Suicide plasmid, sacB Gmr |
| pRK415 | 31 | Broad-host-range vector, Tetr |
| pRCckA | This work | CckA complementation plasmid, Tetr |
| pR555 | 14 | rcc00555 complementation plasmid, Tetr |
| pIND4 | 55 | Inducible expression plasmid, Kanr |
| pIND4Gm | This work | Inducible expression plasmid, Gmr |
| pI556 | This work | rcc00556 complementation plasmid, inducible, Kanr |
| pI556Gm | This work | rcc00556 complementation plasmid, inducible, Gmr |
| pXCA601 | 32 | Reporter plasmid, lacZ Tetr |
| pXCA-555 | This work | rcc00555 promoter::lacZ fusion plasmid, Tetr |
| pET28a(+) | Novagen | E. coli T7 expression vector, inducible, Kanr |
| pET-555C | This work | rcc00555 expression plasmid, C-terminal 6-His tag, inducible, Kanr |
| p601-g65 | 15 | ORF g1 promoter::lacZ fusion plasmid, Tetr |
| p601-g15 | This work | Truncated ORF g1 promoter::lacZ fusion plasmid, no start codon, Tetr |
| p601-g64 | This work | Truncated ORF g1 promoter::lacZ fusion plasmid lacking the 5′-most 5′ end and the −10/−35 sequence, Tetr |
| p601-g61 | This work | Truncated ORF g1 promoter::lacZ fusion plasmid lacking both 5′ ends and the −10/−35 sequence, Tetr |
a
The strain is of uncertain provenance but is a crtD mutant probably derived from Y262 (B. Marrs, personal communication).