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| Status |
Public on Apr 06, 2016 |
| Title |
Next generation sequencing of Pseudomonas aruginosa PAO1 in the presence of Free Nitrous Acid stress |
| Organism |
Pseudomonas aeruginosa |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Purpose: Free nitrous acid (FNA) has recently been demonstrated as an antimicrobial agent to a range of microorganisms. However, its antimicrobial mechanism is largely unknown. For this study Pseudomonas aeruginosa PAO1 was selected to gain a systematic understanding of the antimicrobial mechanism of FNA.
Methods: PAO1 was grown under anaerobic denitrifying conditions and when FNA was added (0.1 mg-N/L) growth temporarily stopped. From cultures with and without added FNA, RNA was extracted and the abundance of gene transcripts was detected by sequencing. Transcripts exhibiting ≥2.5 or ≤-2.5 fold abundance change in the presence of FNA were designated as differentially expressed.
Results:In response to FNA, 167 genes showed increased transcription while 424 exhibited deceased transcript levels. Respiration was likely inhibited as denitrification activity was severely decreased. Genes coding for enzymes of the tricarboxylic acid cycle were down regulated, while highly increased transcripts were detected for pyruvate fermentation, indicating a possible survival response to FNA. Moreover, genes coding ribosomal proteins had significantly decreased transcript levels, implicating that protein biosynthesis was severely inhibited by FNA. Whereas, genes encoding proteins functioning to preserve ribosome units under stressful conditions, were up-regulated during FNA exposure.
Conclusion: In this study, we performed genome-wide high throughput RNA sequencing (RNA-Seq) analysis on PAO1 in the absence and presence of a bacteriostatic-level of FNA (0.1 mg N/L). By combining the global activated/suppressed gene profiles and detected physiological responses, we detected the microbial response to FNA exposure and revealed potential antimicrobial mechanisms of FNA on this prevalent opportunistic pathogen and environmentally relevant denitrifier.
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| Overall design |
mRNA profiles of P. aeruginosa PAO1 towards FNA stress were generated by next generation sequencing in triplicate via Illumina 2000.
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| Contributor(s) |
Gao S, Bond PL |
| Citation(s) |
27116299 |
| |
| Submission date |
Sep 22, 2015 |
| Last update date |
May 15, 2019 |
| Contact name |
Shuhong Gao |
| E-mail(s) |
gaoshuhong1127@163.com
|
| Phone |
+61 (0) 7 334 67217
|
| Organization name |
The University of Queensland
|
| Department |
Advanced Water Management Centre
|
| Lab |
Gehrmann Laboratories (60#)
|
| Street address |
Research Road, St Lucia
|
| City |
Brisbane |
| State/province |
QLD |
| ZIP/Postal code |
4072 |
| Country |
Australia |
| |
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| Platforms (1) |
| GPL18644 |
Illumina HiSeq 2000 (Pseudomonas aeruginosa) |
|
| Samples (5)
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| Relations |
| BioProject |
PRJNA296610 |
| SRA |
SRP063996 |
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