%0 Journal Article %1 ISI:000380624800030 %A Alqarni, Budoor %A Colley, Brendan %A Klebensberger, Janosch %A McDougald, Diane %A Rice, Scott A. %C PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS %D 2016 %I ELSEVIER SCIENCE BV %J JOURNAL OF MICROBIOLOGICAL METHODS %K Carbon Housekeeping genes normalisation} starvation; {qRT-PCR; %P 182-187 %R 10.1016/j.mimet.2016.06.008 %T Expression stability of 13 housekeeping genes during carbon starvation of Pseudomonas aeruginosa %V 127 %X 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. (C) 2016 Elsevier B.V. All rights reserved.

@article{ISI:000380624800030, abstract = {{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. (C) 2016 Elsevier B.V. All rights reserved.}}, added-at = {2017-05-18T11:32:12.000+0200}, address = {{PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}}, affiliation = {{Rice, SA (Reprint Author), 60 Nanyang Dr,SBS 01N-27, Singapore, Singapore. Alqarni, Budoor; Colley, Brendan; Rice, Scott A., Univ New South Wales, Ctr Marine Bioinnovat, Earth \& Environm Sci, Sydney, NSW, Australia. Alqarni, Budoor; Colley, Brendan, Univ New South Wales, Sch Biotechnol \& Biomol Sci, Earth \& Environm Sci, Sydney, NSW, Australia. Rice, Scott A., Univ New South Wales, Sch Biol Earth \& Environm Sci, Sydney, NSW, Australia. Klebensberger, Janosch, Univ Stuttgart, Inst Tech Biochem, Allmandring 31, D-70569 Stuttgart, Germany. McDougald, Diane, Univ Technol Sydney, Inst i3, Sydney, NSW 2007, Australia. McDougald, Diane; Rice, Scott A., Nanyang Technol Univ, Singapore Ctr Environm Life Sci Engn, Singapore, Singapore. McDougald, Diane; Rice, Scott A., Nanyang Technol Univ, Sch Biol Sci, Singapore, Singapore.}}, author = {Alqarni, Budoor and Colley, Brendan and Klebensberger, Janosch and McDougald, Diane and Rice, Scott A.}, author-email = {{RSCOTT@ntu.edu.sg}}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2c0cfcf1a8c0e9a3ae78877b059427ac8/hermann}, doi = {{10.1016/j.mimet.2016.06.008}}, eissn = {{1872-8359}}, funding-acknowledgement = {{Australian Research Council {[}DP140102192]; Singapore Centre for Environmental Life Sciences Engineering; National Research Foundation Singapore; Ministry of Education; Nanyang Technological University; National University of Singapore, under its Research Centre of Excellence Program}}, funding-text = {{Funding for this work was provided by the Australian Research Council (DP140102192). We acknowledge financial support from the Singapore Centre for Environmental Life Sciences Engineering, whose research is supported by the National Research Foundation Singapore, Ministry of Education, Nanyang Technological University and National University of Singapore, under its Research Centre of Excellence Program.}}, interhash = {c07c202f68160ef92571a83f16f6d4ea}, intrahash = {c0cfcf1a8c0e9a3ae78877b059427ac8}, issn = {{0167-7012}}, journal = {{JOURNAL OF MICROBIOLOGICAL METHODS}}, keywords = {Carbon Housekeeping genes normalisation} starvation; {qRT-PCR;}, keywords-plus = {{REAL-TIME-PCR; RT-PCR; CYSTIC-FIBROSIS; OPPORTUNISTIC PATHOGEN; ESCHERICHIA-COLI; QUANTITATIVE PCR; TARGET GENES; RESISTANCE; QUANTIFICATION; RNA}}, language = {{English}}, month = {{AUG}}, number-of-cited-references = {{54}}, pages = {{182-187}}, publisher = {{ELSEVIER SCIENCE BV}}, research-areas = {{Biochemistry \& Molecular Biology; Microbiology}}, times-cited = {{0}}, timestamp = {2017-05-18T09:32:12.000+0200}, title = {{Expression stability of 13 housekeeping genes during carbon starvation of Pseudomonas aeruginosa}}, type = {{Article}}, volume = {{127}}, web-of-science-categories = {{Biochemical Research Methods; Microbiology}}, year = {{2016}} }