Identification of C4-Dicarboxylate Transport Systems in Pseudomonas aeruginosaPAO1
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Bacterial strains and culture conditions.
Strain, bacteriophage, or plasmid | Genotype or relevant characteristicsa | Source or reference |
---|---|---|
Strains | ||
E. coli | ||
DH5α | recA1 endA1 hsdR17 supE44 thi-1 gyrA96 relA1Δ(lacZYA-argF)U169[φ80dlacZΔM15]F−NaIr | 38 |
HB101 | proA2 hsdS20(rB−mB−) recA13 ara-14 lacYI galK2 rpsL20 supE44 xyl-5 mtl-1F− | 38 |
S17-1/λpir | pro thi hsdR recAchromosome::RP4-2 Tc::Mu Km::Tn7/λpir; TprSmr | 27, 41 |
XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR176 supE44 relA1 lac[F′ proAB lacIqZΔM15::Tn10(53)] | Stratagene |
P. aeruginosa | ||
PAO1 | Wild type | 17 |
PAO6358 | PAO1 containing a 900-bp deletion in the rpoNlocus | 16 |
PAO6592 | PAO1 containing a 1,296-bp deletion in the dctAlocus | This study |
PAO6705 | PAO1 containing a 1,798-bp deletion in the PA5165 (dctB) locus | This study |
PAO6706 | PAO1 containing a 1,377-bp deletion in the PA5166 (dctD) locus | This study |
PAO6707 | PAO1 containing a double deletion in the PA5165 locus (1,798 bp) and in the PA5166 locus (1,377 bp) | This study |
PAO6708 | PAO1 containing a 2,899-bp deletion in the PA5167-PA5169 (dctPQM) operon | This study |
PAO6709 | PAO1 containing a double deletion in the dctAlocus (1,296 bp) and in the dctPQM(PA5167-PA5169) operon (2,899 bp) | This study |
PA6710 | PAO6592 with PA5168::Tn5Gm; Gmr | This study |
PA6815 | PAO1 with PA5168::Tn5Gm; Gmr | This study |
Bacteriophage | ||
E79tv-2 | Temperate, transducing variant of E79 | 29 |
Plasmids | ||
pLM1 | Tn5Gm delivery vector; GmrApr | 9 |
pMMB67HE | IncQ expression vector carrying an inducible tacpromoter; Ap/Cbr | 11 |
pME3087 | Suicide vector for allelic replacement; Tcr; ColE1 replicon | 51 |
pME6015 | Cloning vector for translational lacZfusions; Tcr | 39 |
pME6016 | Cloning vector for transcriptional lacZfusions; Tcr | 39 |
pME9506 | Plasmid carrying a transcriptional dctA-lacZfusion | This study |
pME9507 | Plasmid carrying a translational dctA′-′lacZfusion | This study |
pME9508 | Plasmid carrying a translational dctA′-′lacZfusion in which the rpoNputative box of the dctApromoter TGGCAC-N5-CTGCA was replaced with TACCAC-N5-CTTTA | This study |
pME10031 | Suicide construct used for deletion of the PA5165 gene; Tcr | This study |
pME10032 | Suicide construct used for deletion of the PA5166 gene; Tcr | This study |
pME10033 | Suicide construct used for deletion of the PA5167-PA5169 operon; Tcr | This study |
pME10034 | Plasmid carrying a transcriptional dctP-lacZfusion | This study |
pME10035 | Plasmid carrying a translational dctP′-′lacZfusion | This study |
pME10036 | Plasmid carrying a translational dctP′-′lacZfusion in which the rpoNputative box of the dctPQMpromoter TGGCAC-N5-TTGCT was replaced with TACCAC-N5-TTTTT | This study |
pME10037 | pMMB67HE with dctA | This study |
pME10038 | pMMB67HE with PA5165 | This study |
pME10039 | pMMB67HE with PA5166 | This study |
pME10040 | pMMB67HE with PA5167-PA5169 | This study |
pME10041 | Suicide construct used for deletion of the dctAgene; Tcr | This study |
Constructions of plasmids and gene replacement mutants.
β-Galactosidase assays.
Transposon mutagenesis.
Competition experiments.
RESULTS
Identification of the C4-dicarboxylate transporter DctA in P. aeruginosaPAO1.
Search for a second C4-dicarboxylate transporter in a PAO1 ΔdctAbackground.
dctAand dctPQMexpression depends on the growth phase and carbon sources.
The ΔdctAmutant outcompetes the dctQ::Tn5mutant at a low succinate concentration.
The expression of dctAand dctPQMis negatively regulated by DctA and appears not to be regulated by DctPQM.
The expression of dctAand dctPQMis positively regulated by RpoN.
Identification of a two-component system regulating C4-dicarboxylate utilization in P. aeruginosaPAO1.
The expression of dctAand dctPQMis activated by the two-component system PA5165-PA5166.
DISCUSSION
ACKNOWLEDGMENTS
Supplemental Material
REFERENCES
Information & Contributors
Information
Published In
Copyright
History
Contributors
Metrics & Citations
Metrics
Note:
- For recently published articles, the TOTAL download count will appear as zero until a new month starts.
- There is a 3- to 4-day delay in article usage, so article usage will not appear immediately after publication.
- Citation counts come from the Crossref Cited by service.
Citations
If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. For an editable text file, please select Medlars format which will download as a .txt file. Simply select your manager software from the list below and click Download.