From: Fnr (EtrA) acts as a fine-tuning regulator of anaerobic metabolism in Shewanella oneidensisMR-1
Strain/plasmid/primer/Probe | Relevant genotype or sequencea,b | Source |
---|---|---|
Escherichia coli K-12 | ||
β2155 | DAP auxotroph | [61] |
Shewanella oneidensis | ||
MR-1 (ATCC 700550T) | wild type | [62] |
EtrA7-1 | As MR-1 but ΔetrA::loxP | This study |
EtrA7-1 complement | EtrA7-1 complemented with the etrA gene (SO2356) cloned into | This study |
pCM62 | ||
EtrA7-1 with pCM62 | EtrA7-1 harboring the pCM62 as a negative control for complementation | This study |
Plasmids | ||
pCM62 | Tcr; traJ' trfA oriT oriV | [63] |
pCM157 | Tcr; trfA oriT oriV ColE1 ori; lacZp-cre | [46] |
pCM184 | Apr Tcr; loxP-kan-loxP | [46] |
pKNOCK-Gm | Gmr; oriR6K oriTRP4 | [64] |
pCCG02 | As pKNOCK-Gm but etrA'-loxP-kan-loxP-'etrA | This study |
pCCG03 | As pCM62 but lacZp-etrA | This study |
Primersc | ||
etrAN Fwd | GCCGCGGTCATGTCGGTTCTCAAGT | |
etrAN Rev | CGAGCTCCGACAGCTATCTGTTAGTCT | |
etrAC Fwd | CGAATTCAAATCACCGCTTTTAACTTG | |
etrAC Rev | GCATATGCCAGATAAATCACACCTTTT | |
etrAScreenout | AATTCTTCAGGCATTTGACTCG | |
Fwd | ||
etrAScreenout | GGCCGTATCTTGAGTTATACCC | |
Rev | ||
etrAcomp Fwd | GGATCCAGGTGTGATTTATCTGGCG | |
etrAcomp Rev | GAATTCCCGACATGACAATAGAGCAGA | |
23SRT Fwd | TAGCGAAATTCCTTGTCGGG | |
23SRT Rev | GAGACAGCGTGGCCATCATT | |
23Stemp Rev | GTATCAGTTAGCTCAACGCCTC | |
napART Fwd | AGAAAGCCCTGTTAACCGTGG | |
napART Rev | TCATCCGCAGCAATGGTGT | |
napAtemp Rev | GATCGAAGCTACGGTTCTCG | |
nrfART Fwd | GCCACATGTATGCCGTGACT | |
nrfART Rev | TTTACAGCTCCAGCAAGCCA | |
nrfAtemp Rev | ACGTTTCATACTCGGGATGC | |
Probes | ||
23SRTProbe | AGTTCCGACCTGCACGAATGGCG | |
napARTProbe | CTGTATTAAAGGTTACTTCCTGTCGAAAATCATGTACGG | |
nrfARTProbe | CGTAATACCTTGCGTACTGGCGCGC |