Expression stability of 13 housekeeping genes during carbon starvation of Pseudomonas aeruginosa

Alqarni, B; Colley, B; Klebensberger, J; McDougald, D; Rice, SA

Expression stability of 13 housekeeping genes during carbon starvation of Pseudomonas aeruginosa

Alqarni, B Colley, B Klebensberger, J McDougald, D

Rice, SA

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Publication Type:: Journal Article
Citation:: Journal of Microbiological Methods, 2016, 127 pp. 182 - 187
Issue Date:: 2016-08-01

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Field	Value	Language
dc.contributor.author	Alqarni, B	en_US
dc.contributor.author	Colley, B	en_US
dc.contributor.author	Klebensberger, J	en_US
dc.contributor.author	McDougald, D https://orcid.org/0000-0001-5827-8441	en_US
dc.contributor.author	Rice, SA https://orcid.org/0000-0002-9486-2343	en_US
dc.date.available	2016-06-09	en_US
dc.date.issued	2016-08-01	en_US
dc.identifier.citation	Journal of Microbiological Methods, 2016, 127 pp. 182 - 187	en_US
dc.identifier.issn	0167-7012	en_US
dc.identifier.uri	http://hdl.handle.net/10453/49139
dc.description.abstract	© 2016 Elsevier B.V. Quantitative real-time polymerase chain reaction (qRT-PCR) is a reliable technique for quantifying mRNA levels when normalised by a stable reference gene/s. Many putative reference genes are known to be affected by physiological stresses, such as nutrient limitation and hence may not be suitable for normalisation. In this study of Pseudomonas aeruginosa, the expression of 13 commonly used reference genes, rpoS, proC, recA, rpsL, rho, oprL, anr, tipA, nadB, fabD, ampC, algD and gyrA, were analysed for changes in expression under carbon starvation and nutrient replete conditions. The results showed that rpoS was the only stably expressed housekeeping gene during carbon starvation. In contrast, other commonly used housekeeping genes were shown to vary by as much as 10-100 fold under starvation conditions. This study has identified a suitable reference gene for qRT-PCR in P. aeruginosa during carbon starvation. The results presented here highlight the need to validate housekeeping genes under the chosen experimental conditions.	en_US
dc.relation	http://purl.org/au-research/grants/arc/DP140102192
dc.relation.ispartof	Journal of Microbiological Methods	en_US
dc.relation.isbasedon	10.1016/j.mimet.2016.06.008	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Pseudomonas aeruginosa	en_US
dc.subject.mesh	Carbon	en_US
dc.subject.mesh	Rec A Recombinases	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	RNA, Messenger	en_US
dc.subject.mesh	Reverse Transcriptase Polymerase Chain Reaction	en_US
dc.subject.mesh	Gene Expression Regulation, Bacterial	en_US
dc.subject.mesh	Genes, Essential	en_US
dc.title	Expression stability of 13 housekeeping genes during carbon starvation of Pseudomonas aeruginosa	en_US
dc.type	Journal Article
utslib.citation.volume	127	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	1108 Medical Microbiology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	closed_access
pubs.publication-status	Published	en_US
pubs.volume	127	en_US

Abstract:

© 2016 Elsevier B.V. Quantitative real-time polymerase chain reaction (qRT-PCR) is a reliable technique for quantifying mRNA levels when normalised by a stable reference gene/s. Many putative reference genes are known to be affected by physiological stresses, such as nutrient limitation and hence may not be suitable for normalisation. In this study of Pseudomonas aeruginosa, the expression of 13 commonly used reference genes, rpoS, proC, recA, rpsL, rho, oprL, anr, tipA, nadB, fabD, ampC, algD and gyrA, were analysed for changes in expression under carbon starvation and nutrient replete conditions. The results showed that rpoS was the only stably expressed housekeeping gene during carbon starvation. In contrast, other commonly used housekeeping genes were shown to vary by as much as 10-100 fold under starvation conditions. This study has identified a suitable reference gene for qRT-PCR in P. aeruginosa during carbon starvation. The results presented here highlight the need to validate housekeeping genes under the chosen experimental conditions.

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http://hdl.handle.net/10453/49139