|
E. coli
|
|
|
| DH10B |
F– mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 |
Laboratory |
|
recA1 endA1 araD139 Δ(ara leu)7697 galU galK rpsL nupG |
collection |
| S17-1λpir |
TpR SmR, recA, thi, pro, hsdR-M+RP4∶2-Tc:Mu: Km Tn7 λpir |
Laboratory |
|
|
collection |
|
P. aeruginosa
|
|
|
| PAO1 |
PT5, Wild type |
Laboratory |
|
|
collection |
| PA14 |
Wild type |
[38]
|
| PAO1 |
AHQ biosensor strain, CTX pqsA::luxCDABE
|
[36]
|
|
pqsA::lux
|
|
|
| PAO1-W5 |
3.5 kbp deletion in PA2228-pslA region, pTS400 |
This study |
| PT1617 |
PAO-W5 after spontaneous loss of pTS400 |
This study |
| PT466 |
PAO1ΔlasI::Tc |
[22]
|
| PT498 |
PAO1ΔlasR::Tc |
[22]
|
| PT1850 |
PAO1-UW vqsM::Tn5-Tc |
[39]
|
| PT1833 |
PAO1-UW pqsA::Tn5-Tc |
[39]
|
| PT1842 |
PAO1ΔvqsM
|
This study |
|
Plasmids
|
|
|
| pEX18T |
Suicide vector; Tet |
[47]
|
| pRK2013 |
RK2, helper plasmid; Km |
[28]
|
| pSB1075 |
lasR, lasI-luxCDABE, ColE1; Ap |
[33]
|
| pTS400 |
lasB-lacZ translational fusion; Ap |
[35]
|
| pEX1.8 |
expression vector, lacIQ; Ap |
[48]
|
| pJPP8 |
pEX1.8, ptac::rhlR; Ap |
[48]
|
| pIApX2 |
pUCP20 derivative containing promoter pX2 and gfp; Ap |
Ina Attree |
| pLIGF1 |
lasI-gfp promoter fusion; Ap |
This study |
| ppqsA1 |
pqsA-gfp promoter fusion; Ap |
This study |
| pME3872 |
lasR on mini-Tn7 gene delivery vector; Ap Gm |
[25]
|
| pME3078 |
Mobilizable suicide vector, ColE1; Tc |
[27]
|
| pHO1.1 |
1,383 bp PCR fragment harboring PA2228, cloned in BamHI- |
This study |
|
HindIII cleaved pIApX2; Ap |
|
| pHO2.1 |
1,483 bp PCR fragment harboring PA2227 (vqsM) cloned in |
This study |
|
BamHI-HindIII cleaved pIApX2; Ap |
|
| pHO3.1 |
1,698 bp PCR fragment harboring PA2229-PA2230 cloned in |
This study |
|
BamHI-HindIII cleaved pIApX2; Ap |
|
| pHO4.1 |
6,023 bp PCR fragment harboring PA2227-PA2222 cloned in |
This study |
|
BamHI-HindIII cleaved pIApX2, Ap |
|
| pHO6.1 |
4,059 bp DNA fragment harboring PA2227-PA2225, obtained |
This study |
|
by XbaI digest and religation of pHO4.1; Ap |
|
| pHO9.1 |
2,000 bp PCR fragment harboring PA2226-PA2225 cloned in |
This study |
|
BamHI-HindIII cleaved pIApX2; Ap |
|
| pHO12.1 |
646 bp PCR fragment harboring PA2225 cloned in BamHI- |
This study |
|
HindIII cleaved pIApX2; Ap |
|
| pHO13.1 |
765 bp PCR fragment harboring PA2226- cloned in BamHI- |
This study |
|
HindIII cleaved pIApX2; Ap |
|
|
Primers
|
5′- 3′ sequence |
|
| PA2225-Bam |
AGCTGGATCCAGATCAGGGTGGCGTCAGAGTA
|
This study |
| PA2225-Hind |
AGCTAAGCTTCATTGAGCATGAGCTGGAAAA
|
This study |
| PA2226-Bam |
AGCTGGATCCGAGCAATTGCACTGCTACTGAA
|
This study |
| PA2226-Hind |
AGCTAAGCTTCCGATCAGACTCAGCAATACCA
|
This study |
| PA2228-Bam |
CAGTGGATCCTGAAATGCCTTGATCTTCACC
|
This study |
| PA2228-Hind |
CAGTAAGCTTGGCGCCATTCAAAATCAAATA
|
This study |
| PA2228-Bam2 |
CAGTGGATCCAGGGATATGCCTGGCTCATTT
|
This study |
| PA2222-Hind |
CAGTAAGCTTGAACTTGTGCGCATATTTGG
|
This study |
| vqsM-Bam |
CAGTGGATCCTGAGTGGGTCAAGGCATCAAC
|
This study |
| vqsM-Hind |
CAGTAAGCTTCGCTAATTAGTCCAGCAAGCA
|
This study |
| PA2229-Bam |
CAGTGGATCCGTAAGCACCGTAGCGTGCAA
|
This study |
| PA2230-Hind |
CAGTAAGCTTCTCGTCGAAGGCGGAAGAG
|
This study |
| PA2228-1 |
GTATGGGCCCCTCACAGTTTA
|
This study |
| PA2228-2 |
CGACCGCCATGAAGAAATAAT
|
This study |
| PA2222-1 |
ATGCAATCCTGGCTTTGTCTTC
|
This study |
| PA2222-2 |
AACGTTGCAAACACGTTCCTTT
|
This study |
| vqsM-1 |
CGTCAGCGAATCGAGTTATTCC
|
This study |
| vqsM-2 |
TCTTTGCGGGCCTTATCTACAA
|
This study |
| PA0439-Bgl |
CGCAGATCTATCCAGGTCGACGGACAGATG
|
This study |
| PA0439-Hind |
GCGAAGCTTGTAGTCGAAGGCGGGTCGAT
|
This study |
| vqsM-upR-Gm |
TCAGAGCGCTTTTGAAGCTAATTCGGCGTCATTCCACTC
|
This study |
|
TGGCTTA |
|
| vqsM-dnF-Gm |
AGGAACTTCAAGATCCCCAATTCGCGTCAGCGAATCGAG
|
This study |
|
TTATTCC |
|
| ppqsA-Hind |
ACACAAGCTTGATTTCAACAGGGAAGCCTGC
|
This study |
| ppqsA-Xba |
ACACTCTAGAGAAATCGAGGCGGAACAGAAC
|
This study |
