Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in Azospirillum brasilense
Identifieur interne : 000A93 ( Main/Exploration ); précédent : 000A92; suivant : 000A94Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in Azospirillum brasilense
Auteurs : Mary Mcmillan ; Lily PeregSource :
- PLoS ONE [ 1932-6203 ] ; 2014.
Abstract
Url:
DOI: 10.1371/journal.pone.0098162
PubMed: 24841066
PubMed Central: 4026395
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in <italic>Azospirillum brasilense</italic>
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<author><name sortKey="Mcmillan, Mary" sort="Mcmillan, Mary" uniqKey="Mcmillan M" first="Mary" last="Mcmillan">Mary Mcmillan</name>
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<author><name sortKey="Pereg, Lily" sort="Pereg, Lily" uniqKey="Pereg L" first="Lily" last="Pereg">Lily Pereg</name>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Evaluation of Reference Genes for Gene Expression Analysis Using Quantitative RT-PCR in <italic>Azospirillum brasilense</italic>
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<author><name sortKey="Mcmillan, Mary" sort="Mcmillan, Mary" uniqKey="Mcmillan M" first="Mary" last="Mcmillan">Mary Mcmillan</name>
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<author><name sortKey="Pereg, Lily" sort="Pereg, Lily" uniqKey="Pereg L" first="Lily" last="Pereg">Lily Pereg</name>
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<series><title level="j">PLoS ONE</title>
<idno type="eISSN">1932-6203</idno>
<imprint><date when="2014">2014</date>
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<front><div type="abstract" xml:lang="en"><p><italic>Azospirillum brasilense</italic>
is a nitrogen fixing bacterium that has been shown to have various beneficial effects on plant growth and yield. Under normal conditions <italic>A. brasilense</italic>
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 <italic>A. brasilense</italic>
. Here we evaluated the expression of ten candidate reference genes (<italic>16S rRNA, gapB, glyA, gyrA, proC, pykA, recA, recF, rpoD, and tpiA</italic>
) in wild-type and mutant <italic>A. brasilense</italic>
strains under different culture conditions, including conditions that induce differentiation. Analysis with the software programs BestKeeper, NormFinder and GeNorm indicated that <italic>gyrA</italic>
, <italic>glyA</italic>
and <italic>recA</italic>
are the most stably expressed reference genes in <italic>A. brasilense</italic>
. The results also suggested that the use of two reference genes (<italic>gyrA</italic>
and <italic>glyA</italic>
) is sufficient for effective normalisation of qRT-PCR data.</p>
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<tree><noCountry><name sortKey="Mcmillan, Mary" sort="Mcmillan, Mary" uniqKey="Mcmillan M" first="Mary" last="Mcmillan">Mary Mcmillan</name>
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