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| Status |
Public on May 31, 2010 |
| Title |
P. aeruginosa PA14 gene expression in artificial sputum medium (ASMDM) |
| Platform organism |
Pseudomonas aeruginosa |
| Sample organism |
Pseudomonas aeruginosa UCBPP-PA14 |
| Experiment type |
Expression profiling by array
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| Summary |
Pseudomonas aeruginosa airway infection is the leading cause of morbidity and mortality in cystic fibrosis (CF) patients. In vitro models that closely mimic CF sputum are needed to improve understanding of the pathobiology of P. aeruginosa in the CF airway. We developed an artificial sputum medium (ASMDM) that more closely resembles the composition of CF sputum than current media. In order to validate the utility of ASMDM, we used GeneChip microarrays to compare expression data of P. aeruginosa UCBPP-PA14 (PA14) in ASMDM with published data for this strain grown under the same conditions in an artificial medium containing 10% (v/v) CF sputum. Thirty-seven of 39 nutrition-related genes were differentially expressed in the same manner in both media. However, 24 quorum-sensing (QS) genes, 23 Type III secretion system and several anaerobic respiration genes were more highly expressed in ASMDM than in sputum-containing medium. When grown to stationary phase in ASMDM, PA14 differentially expressed about 50 biologically significant genes compared to stationary phase growth in Luria Broth; genes involved in iron acquisition (pfeA, fepC) and in assimilatory nitrate reduction (nasC, nirD) were upregulated, while 24 QS genes, including the regulator rhlR, lasA, rsaL, aprADEI and phenazine genes phzC2DD2EG2 were downregulated. Downregulation of QS-regulated virulence genes has been noted in chronic P. aeruginosa infection. ASMDM thus appears highly suitable for studies on gene expression of (i) P. aeruginosa strains from acutely and chronically infected CF patients and (ii) established biofilms that are a hallmark of advanced CF lung disease.
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| Overall design |
PA14 was grown in four different ways:
1. Logarithmic growth for early gene expression
Cells were grown in MOPS-Glucose and separately in ASMDM. The average of two biological duplicates in each case was compared to the other to determine differential gene expression.
2. Stationary phase growth for gene expression
Cells were grown in Luria Broth and separately in ASMDM. The average of two biological duplicates in each case was compared to the other to determine differential gene expression.
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| Contributor(s) |
Manos J, Fung C, Naughton S, Arthur J, Turnbull L, Whitchurch C, Rose B, Hassett DJ, Harbour C |
| Citation(s) |
20522626 |
| Submission date |
Oct 16, 2009 |
| Last update date |
Jul 06, 2016 |
| Contact name |
Jim Manos |
| E-mail(s) |
jim.manos@sydney.edu.au
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| Phone |
+612 9351 8942
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| Organization name |
University of Sydney
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| Street address |
Camperdown
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| City |
Sydney |
| State/province |
NSW |
| ZIP/Postal code |
2006 |
| Country |
Australia |
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| Platforms (1) |
| GPL84 |
[Pae_G1a] Affymetrix Pseudomonas aeruginosa Array |
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| Samples (8)
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| GSM462061 |
Stationary phase gene expression profile of P. aeruginosa PA14 grown in ASMDM (replicateA) |
| GSM462062 |
Stationary phase gene expression profile of P. aeruginosa PA14 grown in ASMDM (replicateB) |
| GSM462063 |
Stationary phase gene expression profile of P. aeruginosa PA14 grown in Luria broth(replicateA) |
| GSM462064 |
Stationary phase gene expression profile of P. aeruginosa PA14 grown in Luria Broth (replicateB) |
| GSM462352 |
Gene expression profile of P. aeruginosa PA14 grown to early log phase in MOPS-Glucose(replicateA) |
| GSM462353 |
Gene expression profile of P. aeruginosa PA14 grown to early log phase in MOPS-Glucose(replicateB) |
| GSM462354 |
Gene expression profile of P. aeruginosa PA14 grown to early log phase in ASMDM (replicateA) |
| GSM462355 |
Gene expression profile of P. aeruginosa PA14 grown to early log phase in ASMDM (replicateB) |
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| Relations |
| BioProject |
PRJNA121469 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE18594_RAW.tar |
6.1 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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