TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant feature or sequence | Sequence | Source or reference |
|---|---|---|---|
| Strains | |||
| M. aeruginosa PCC 7806 | Microcystin-producing cyanobacterium | UNSW culture collection | |
| E. coli DH5α | Cloning host | UNSW culture collection | |
| E. coli BL-21(DE3) | Expression host | UNSW culture collection | |
| Plasmids | |||
| pGEM-T Easy | Cloning vector | Promega | |
| pET-30a | Expression vector | Novagen | |
| Primers | |||
| NF | ntcA degenerate | CAGTGTTTTTGGGGTGYT | Timothy Downing, University of Port Elizabeth |
| NR | ntcA | GTTTCAATCATCATTTCCGT | Timothy Downing, University of Port Elizabeth |
| NF/1 | ntcA pan-handle | CGAGAGTAATTCTACCGGAG | This study |
| NF/2 | ntcA | GATCCCTCCGCAATAATCCC | This study |
| NF/3 | ntcA | TTTCTACCTGCTCGATCGG | This study |
| NF/4 | ntcA | AGTGAACAGATAGCTGTGC | This study |
| NF/5 | ntcA | CTGACCAAACGTGAACCCAT | This study |
| NF/6 | ntcA | CGAGTGTCACCTCTATTAAACAC | This study |
| NTCANDE1F | ntcA cloning | TCCATATGGACTTATCATTAATACAAGATAAAC | This study |
| NTCAXHO1R | ntcA cloning | TCCTCGAGAGTAAATTGTTGACTGAGAGCG | This study |
| T7 term | pET-30a | GCTAGTTATTGCTCAGCGGT | Novagen |
| T7 prom | pET-30a | TAATACGACTCACTATAGGG | Novagen |
| MPF | pGEM-T Easy | CCCAGTCACGACGTTGTAAAACG | Promega |
| MPR | pGEM-T Easy | AGCGGATAACAATTTCACACAGG | Promega |
| T7Pr1 | Adaptor primer | CCCCTATCCACCCTTACACCTATC | 15 |
| M4F | NtcA binding site in mcyA/D promoter | CGAATTCTAATGATTTTTACTAATTTATTGGG | This study |
| M4R | NtcA binding site in mcyA/D promoter | CCGCCGGCGAATTGTTCTGAGCCTCGACAT | This study |
| M5F | NtcA binding site in mcyA/D promoter | CGAATTCCCGTCGGGTTTCCTGT | This study |
| M5R | NtcA site in mcyA/D promoter | CCGCCGGCGCATTGCTGTTCTAACTTTTTCC | This study |
| M6F | ntcA promoter | CGAATTCCAATAGCCGACCCCAGCG | This study |
| M6R | ntcA promoter | CCGCCGGCGTTTTTATTATCCAACGAGTGTCA | This study |
| NtcArealF | ntcA reverse transcription-PCR short product | CATTTCCGTTTGCAGAATCC | This study |
| NtcArealR | ntcA reverse transcription-PCR short product | TGTTTTTGGGGTGCTATCCT | This study |
| RpoC1F | rpoc1 reverse transcription-PCR short product | CCTCAGCGAAGATCAATGGT | This study |
| RpoC1R | rpoc1 reverse transcription-PCR short product | CCGTTTTTGCCCCTTACTTT | This study |
| McyBrealF | mcyB reverse transcription-PCR short product | CTGAGGGGATTACGGATTGA | This study |
| McyBrealR | mcyB reverse transcription-PCR short product | ACCATATAAGCGGGCAGTTG | This study |