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Title: Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in 'Azospirillum brasilense'
Contributor(s): McMillan, Mary  (author)orcid ; Pereg, Lily  (author)
Publication Date: 2014
Open Access: Yes
DOI: 10.1371/journal.pone.0098162Open Access Link
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Abstract: 'Azospirillum brasilense' is a nitrogen fixing bacterium that has been shown to have various beneficial effects on plant growth and yield. Under normal conditions 'A. brasilense' exists in a motile flagellated form, which, under starvation or stress conditions, can undergo differentiation into an encapsulated, cyst-like form. Quantitative RT-PCR can be used to analyse changes in gene expression during this differentiation process. The accuracy of quantification of mRNA levels by qRT-PCR relies on the normalisation of data against stably expressed reference genes. No suitable set of reference genes has yet been described for A. brasilense. Here we evaluated the expression of ten candidate reference genes ('16S' 'rRNA', 'gapB', 'glyA', 'gyrA', 'proC', 'pykA', 'recA', 'recF', 'rpoD', and 'tpiA') in wild-type and mutant 'A. brasilense' strains under different culture conditions, including conditions that induce differentiation. Analysis with the software programs BestKeeper, NormFinder and GeNorm indicated that 'gyrA', 'glyA' and 'recA' are the most stably expressed reference genes in 'A. brasilense'. The results also suggested that the use of two reference genes ('gyrA and glyA') is sufficient for effective normalisation of qRT-PCR data.
Publication Type: Journal Article
Source of Publication: PLoS One, 9(5), p. 1-8
Publisher: Public Library of Science (PLoS)
Place of Publication: United States of America
ISSN: 1932-6203
Field of Research (FOR): 060504 Microbial Ecology
060501 Bacteriology
060503 Microbial Genetics
Socio-Economic Outcome Codes: 970106 Expanding Knowledge in the Biological Sciences
Peer Reviewed: Yes
HERDC Category Description: C1 Refereed Article in a Scholarly Journal
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