Table 4.
Strains and plasmids used in this study
| Strain/plasmid | Description | Source or reference |
|---|---|---|
| Strain | ||
|
E. coli DH5α |
supE44 Δ(lacZYA-argF) U169 (Φ80lacZ ΔM15) hsdR17 recA endA1 gyrA96 thi-1 relA1 |
Invitrogen |
|
E. coli BW25113 |
lacIqrrnBT14 ΔlacZWJ16hsdR514 ΔaraBADAH33 ΔrhaBADLD78 |
28 |
|
E. coli BW25113 (ΔaroKΔaroL) |
E. coli BW25113, ΔaroKΔaroL |
This study |
|
E. coli BW25113 (ΔaroKΔaroL, P
pps
::P
lacQ1
, P
csrB
::P
lacQ1
) |
E. coli BW25113, ΔaroKΔaroL, replacement of the native promoter of the pps and csrB gene with the P
lacQ1
promoter |
This study |
|
E. coli BW25113 (ΔaroKΔaroL, P
pps
::P
lacQ1
, P
csrB
::P
lacQ1
, P
T5
-pps, P
T5
-csrB) |
E. coli BW25113(ΔaroKΔaroL, P
pps
::P
lacQ1
, P
csrB
::P
lacQ1
) with an additional chromosomal copy of the pps and csrB genes under the control of the T5 promoter |
This study |
| SA110 |
CIChE strain of the aroG
fbr
, tktA, aroB and aroE gene cluster resistance to 2 μM triclosan from E. coli BW25113 (ΔaroKΔaroL, P
pps
::P
lacQ1
, P
csrB
::P
lacQ1
), ΔrecA |
This study |
| SA112 |
SA110 with an additional chromosomal copy of the tktA gene under the control of the 5Ptac promoter (5 tandem repeats of the core-tac-promoter) |
This study |
| SA114 |
SA112 with an additional chromosomal copy of the pntAB genes under the control of the 5Ptac promoter |
This study |
| SA116 |
SA112 with an additional chromosomal copy of the nadK gene under the control of the 5Ptac promoter |
This study |
| Plasmid | ||
| pHKKF3T5b |
CIChE integration expression vector, attPHK site, Kanr |
16 |
| pHKKT5b |
Integration expression plasmid, attPHK site, PT5 promoter, Kanr |
33,34 |
| pP21KT5b |
Integration expression plasmid, attPP21 site, PT5 promoter, Kanr |
33,34 |
| pAH69 |
Helper plasmid expressing phage HK022 Int, Ampr |
31 |
| pAH121 |
Helper plasmid expressing phage P21 Int, Ampr |
31 |
| pCP20 |
pSC101 repliconts Flp(λRp) cI857, Cmr, Ampr |
28 |
| pKD3 |
oriRγ, FRT::cat::FRT template plasmid, Cmr, Ampr |
28 |
| pSIM6 |
pSC101 repliconts PL-gam-bet-exo cI857, Ampr |
29 |
| p5TG |
pSC101 repliconts, 5Ptac promoter, Spcr |
32 |
| pMP5 |
P5TG derivative, pSC101 ori, Constitutive expression, Spcr |
This study |
| pBAD24 |
pMB1 ori, PBAD L-arabinose inducible, Ampr |
30 |
| pBAD-csrB-pps |
pBAD24 derivative containing the csrB and pps genes |
Lab storage |
| pBEB |
pBAD24 derivative containing the aroB and aroE genes |
This study |
| pBEBG |
pBAD24 derivative containing the aroG
fbr
, aroB and aroE genes |
This study |
| pHKEBG |
pHKKF3T5b derivative containing aroG
fbr
, aroB and aroE genes |
This study |
| pHKEBGT |
pHKKF3T5b derivative containing aroG
fbr
, tktA, aroB and aroE genes |
This study |
| pMPsthA |
pMP5 derivative containing sthA gene |
This study |
| pMPpntAB |
pMP5 derivative containing pntAB genes |
This study |
| pMPnadK |
pMP5 derivative containing nadK gene |
This study |
| pP21KT5b-csrB-pps |
pP21KT5b derivative containing csrB and pps genes |
This study |
| pHKKT5b-tktA |
pHKKT5b derivative containing tktA gene |
This study |
| pHKK5Tacb |
Integration expression plasmid, attPHK site, 5tac promoter, Kanr |
This study |
| pP21K5Tacb |
Integration expression plasmid, attPP21 site, 5tac promoter, Kanr |
This study |
| pP21K5Tacb-pntAB |
pHK5Tacb derivative containing pntAB genes |
This study |
| pP21K5Tacb-nadk | pHK5Tacb derivative containing nadK gene | This study |