TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Anabaena sp. strainsa | ||
| PCC 7120 | Wild type | Pasteur Culture Collection |
| 216 | hetR(S179N) | 4 |
| UHM100 | PpetE-hetN ΔpatS | 3 |
| UHM101 | ΔpatA | 20 |
| UHM103 | ΔhetR | 3 |
| UHM109 | ΔhetR ΔpatA | 20 |
| UHM130 | ΔhetF | This study |
| UHM131 | PpetE-hetN ΔpatS ΔhetF | This study |
| UHM132 | ΔhetR ΔhetF | This study |
| UHM134 | ΔhetR ΔpatA ΔhetF | This study |
| UHM135 | ΔpatA ΔhetF | |
| Plasmids | ||
| pAM504 | Shuttle vector for replication in E. coli and Anabaena; Kmr Nmr | 23 |
| pAM505 | Same as pAM504 with multiple cloning site inverted | 23 |
| pAM1951 | pAM505 with PpatS-gfp | 29 |
| pAM1956 | pAM505 with promoterless gfp | 29 |
| pRL277 | Suicide vector; Smr Spr | 2 |
| pET21a | Expression vector; Apr | Novagen |
| pPROEX-1 | Expression vector for making polyhistidine epitope-tagged proteins; Apr | Life Technologies |
| pSMC127 | pAM504 carrying PhetR-gfp | 6 |
| pSMC138 | pAM505 carrying PpetE-gfp | This study |
| pSMC148 | pPROEX-1 carrying hetR | This study |
| pAM1951 | Shuttle vector carrying PpatS-gfp | 28 |
| pDR204 | pAM505 carrying Pnir-hetRH6 | 21 |
| pDR216 | pAM505 Pnir-hetR | 21 |
| pDR226 | pAM505 Pnir-hetRH6(S179N) | 21 |
| pDR255 | pET21a carrying hetFH6 | This study |
| pDR256 | Suicide plasmid based on pRL277 used to delete hetF | This study |
| pDR266 | pAM1956 with PhetF fused to gfp | This study |
| pDR267 | pAM505 carrying PhetF-hetF | This study |
| pDR281 | pAM505 carrying PhetF-hetFH6 | This study |
| pDR282 | pAM505 carrying PhetF-hetF (H201Y) | This study |
| pDR284 | pAM505 carrying PhetF-hetF (C246A) | This study |
| pDR293 | pAM504 PpetE-hetR-gfp | This study |
| pDR302 | pAM505 carrying PhetF-hetF and Pnir-hetRH6 | This study |
| pDR309 | pAM505 carrying Pnir-hetRH6 and PpetE-gfp | This study |
| pDR310 | pAM505 carrying PhetF-hetF and PpetE-gfp | This study |
| pDR311 | pAM504 carrying Pnir-lacZ | This study |
| pDR312 | pAM504 carrying PhetF-lacZ | This study |
| pDR324 | pAM1956 with Pnir fused to gfp | This study |
| pDR350 | pAM504 carrying PpetE-lacZ | This study |
| pPetHetR | Shuttle vector carrying PpetE-hetR | 5 |
a
For the Anabaena strains constructed in this study, mutations were introduced in the order indicated.