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 |