Table 2 Strains and oligonucleotides used in this study
From: Phosphate uptake by the phosphonate transport system PhnCDE
Strain | Relevant phenotype or sequence | Reference |
|---|---|---|
MG1655 derivatives | ||
MG1655 | E. coli K-12 phnE | Lab collection |
RI57 | MG1655 ΔpitA ΔugpBAEC ΔpstSCAB-phoUphnE+ (spontaneous phnE+ revertant) | This Study |
RS02 | MG1655 ΔphnCDE::Km | This Study |
RS03 | MG1655 phnE+ (P1 transduction from BL21 to MG1655) | This study |
RS04 (pitA+3 Δ) | MG1655 ΔphnCDE ΔugpBAEC ΔpstSCAB-phoU (pitA+) | This Study |
RS05 (ugp+3 Δ) | MG1655 ΔphnCDE ΔpitA ΔpstSCAB-phoU (ugpBAEC+) | This Study |
RS06 (pst+3 Δ) | MG1655 ΔpitA ΔphnCDE ΔugpBAEC (pstSCAB-phoU+) | This Study |
RS07 (phn+3 Δ) | MG1655 ΔpitA ΔugpBAEC ΔpstSCAB-phoU (phnE+) | This Study |
RS08 (phn+4 Δ) | MG1655 ΔpitA ΔpitB ΔugpBAEC ΔpstSCAB-phoU (phnE+) | This Study |
Other Strains | ||
BL21 | E. coli B phnE+ | |
KM32 | ΔrecBCD::Ptac-gam-bet-exo cat | [28] |
KM44 | ΔrecBCD::Ptac-gam-bet-exo kan | [28] |
Oligonucleotides | ||
pitA_mut_Fow | TCAAAATGGCGTAACGTCCTATGCTACATTTGTTTGCTGGGTGTAGGCTGGAGCTGCTTC | |
pitA_mut_Rev | GCTCGTTTTGGTGCGTACGATTACAGGAACTGCAAGGAGACATATGAATATCCTCCTTAG | |
pitA_ver_Fow | TTGTAAAGATTCCTCAGTGGTC | |
pitA_ver_Rev | TGATGCGCTACGCTTATCAG | |
pitB_mut_Fow | GAAATGCCCGATCGCCAGGACCGGGCATTTTCAGGAAGGGTGTAGGCTGGAGCTGCTTC | |
pitB_mut_Rev | TGCGTCCGTTCGTAAATTCAAAATGGCGTAATCTAATATCCATATGAATATCCTCCTTA | |
pitB_ver_Fow | TTAACCAGTGGAATACCTGTG | |
pitB_ver_Rev | CGAGGAAGATTGCCATAACG | |
phn_mut_Fow | GGATGTGTAGACAAGTGCATAGATATCAATGCCTCGCTTAGTGTAGGCTGGAGCTGCTTC | |
phn_mut_Rev | ATCCGCCACGATGGAGCCACTTTTTTAGGGAGGCTGCATCCATATGAATATCCTCCTTAG | |
phn_ver_Fow | TTTGCGGCTATCTCTTGATAGC | |
phn_ver_Rev | GTCACAATAATCCGCCACGA | |
phn_seq_Fow | GTCACAATAATCCGCCACGA | |
pst_mut_Fow | GTCTGGTGAATTATTTGTCGCTATCTTTCCCCGCCAGCAGTGTGTAGGCTGGAGCTGCTTC | |
pst_mut_Rev | AGGAGACATTATGAAAGTTATGCGTACCACCGTCGCAACTCATATGAATATCCTCCTTAG | |
pstS_S_Fow: | TGTAATTGACTGAATATCAACGCT | |
phoU_S_Rev: | GCTCGCAGTTATTAACTTTGTG | |
ugp350 | AACGATGAAACCGTTACATTATACAGCTTCAGCACTGGCGGTGTAGGCTGGAGCTGCTTC | |
ugp4480 | CCAGATGCAGCCACAGCGTGCTGCCTGCCGTCGGGCGCTCCATATGAATATCCTCCTTAG | |
ugp858 | ACCGCCTTGTCATCTTTCTG | |
ugp5210 | CTCGTTGTCCTGTTTCACC |