TABLE 1.
Strain/Plasmid/Primer | Description | Source/Purpose |
P. aeruginosa | ||
PAK | Wild type strain of Pseudomonas aeruginosa | David Bradley |
ΔKH-S1 | PAK with pnp (KH and S1) deletion | Chen et al., 2016 |
ΔKH-S1 /Tn7T-pnp | PAKΔKH-S1 with pnp inserted on chromosome with mini-Tn7T insertion; | Chen et al., 2016 |
PAKΔPA0614 | PAK deleted of PA0614 | This study |
PAKΔPA0629 | PAK deleted of PA0629 | This study |
PAKΔprtN | PAK deleted of prtN | This study |
ΔKH-S1ΔPA0614 | PAKΔKH-S1 deleted of PA0614 | This study |
ΔKH-S1ΔPA0629 | PAKΔKH-S1 deleted of PA0629 | This study |
ΔKH-S1ΔprtN | PAKΔKH-S1 deleted of prtN | This study |
PAK/pMMB67EH | PAK containing plasmid pMMB67EH | This study |
ΔKH-S1/pMMB67EH | PAKΔKH-S1 containing plasmid pMMB67EH | This study |
PAK/pMMB67EH-prtR | PAK containing plasmid pMMB67EH-prtR | Chen et al., 2017 |
Plasmid | ||
pEX18Tc | Gene replacement vector; Tcr, oriT+, sacB+ | Hoang et al., 1998 |
pUC18T-mini-Tn7T-Tc | mini-Tn7 base vector from insertion into chromosome attTn7 site; Tcr | Choi and Schweizer, 2006 |
pUC18T-mini-Tn7T-PprtR-lacZ | prtR promoter of PAK fused to promoterless lacZ on pUC18T-mini-Tn7T | Sun et al., 2014 |
pMMB67EH | Expression vector with tac promoter;Apr | Furste et al., 1986 |
pUCP20 (no promoter) | Escherichia-Pseudomonas shuttle vector; no promoter; Ampr | This study |
pUCP20(no promoter) -pRkaraRed(43)-PrtR-His | 6 × His-tagged PrtR driven by the PBAD promoter with 43 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
pUCP20(no promoter) -pRkaraRed(15)-PrtR-His | 6 × His-tagged PrtR driven by the PBAD promoter with 15 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
pUCP20(no promoter) -pRkaraRed(43)-GFP | GFP driven by the PBAD promoter with 43 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
pUCP20(no promoter) -pRkaraRed(15)-GFP | GFP driven by the PBAD promoter with 15 bp of the 5′-UTR sequence on pUCP20(no promoter) | This study |
Primer | Sequence (5′→3′) | Function |
PA0636-RT-S | TGGAAGACCCGGCAGAAG | RT-PCR |
PA0636-RT-AS | CGTTGAGCTTGGACAGATCCT | RT-PCR |
PA0614-RT-S | CGCTGCCTGCCAAGGA | RT-PCR |
PA0614-RT-AS | ATCAGTACCCAGAGCGGCATT | RT-PCR |
PA0629-RT-S | GTGGAGAACCTCAATTACAG | RT-PCR |
PA0629-RT-AS | TAGGTGTTGTCGGCAATC | RT-PCR |
prtR-RT-S | GATGCGCAACCTGAAGCA | RT-PCR |
prtR-RT-AS | TGAATGGTGTTCTGCGAAACC | RT-PCR |
prtN-RT-S | CGACGATAGCCACAAG | RT-PCR |
prtN-RT-AS | GGATGCGATGCTGTC | RT-PCR |
lexA-RT-S | AATCCCGCCTTCTTCAAT | RT-PCR |
lexA-RT-AS | AATGCCGATGTCCTTCAT | RT-PCR |
recA-RT-S | ATATCAAGAACGCCAACT | RT-PCR |
recA-RT-AS | TAGAACTTCAGTGCGTTA | RT-PCR |
BamHI-PprtR-lacZ-S# | CGCGGATCC GAGCCAGGACCAGTTCGTTGGC | Transcriptional fusion |
HindIII-lacZ-AS | ATTATAAAGCTT TTATTTTTGACACCAGACCAACTGG | Transcriptional fusion |
SacI-PBAD-S | CCAAGAGCTC TTATGACAACTTGACGGC | Translational fusion |
HindIII-prtR-AS | ATTATAAAGCTT TCAGTGGTGGTGGTGGTGGTGACCTCCCC GCACCAGGGACGGGCCGC | Translational fusion |
XhoI- prtR(43)-GFP S | CCGCTCGAG TAGGCTCTTTACAGAAAATCCATCGGTCTGTAGA TTGCCGAGCATGAGTAAAGGAGAAGAACTTTTCACTG | Translational fusion |
XhoI- prtR(15)-GFP S | CCGCTCGAG TGTAGATTGCCGAGCATGAGTAAAGGAGAAGAA CTTTTCACTG | Translational fusion |
HindIII –GFP-AS | CCCAAGCTT TTATTTGTATAGTTCATCCATGCCATG | Translational fusion |
#The endonuclease cutting sites are underlined.