Selection of suitable reference genes for RT-qPCR analyses in cyanobacteria

Cyanobacteria are a group of photosynthetic prokaryotes that have a diverse morphology, minimal nutritional requirements and metabolic plasticity that has made them attractive organisms to use in biotechnological applications. The use of these organisms as cell factories requires the knowledge of th...

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Bibliographic Details
Published in:PloS one 2012-04, Vol.7 (4), p.e34983-e34983
Main Authors: Pinto, Filipe, Pacheco, Catarina C, Ferreira, Daniela, Moradas-Ferreira, Pedro, Tamagnini, Paula
Format: Article
Language:English
Subjects:
Algorithms
Biology
Biosynthesis
Carotenoids
Cyanobacteria
Cyanobacteria - genetics
Cyanobacteria - isolation & purification
Data processing
Gene expression
Gene Expression Profiling
Genes
Light
Lyngbya
Metabolism
Morphology
Nitrogen
Nostoc
Nutritional requirements
Organisms
Photosynthesis
Plasticity
Polymerase chain reaction
Prochlorococcus
Prokaryotes
Real-Time Polymerase Chain Reaction - methods
Reference Standards
Stability analysis
Studies
Synechococcus
Synechocystis
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Summary:Cyanobacteria are a group of photosynthetic prokaryotes that have a diverse morphology, minimal nutritional requirements and metabolic plasticity that has made them attractive organisms to use in biotechnological applications. The use of these organisms as cell factories requires the knowledge of their physiology and metabolism at a systems level. For the quantification of gene transcripts real-time quantitative polymerase chain reaction (RT-qPCR) is the standard technique. However, to obtain reliable RT-qPCR results the use and validation of reference genes is mandatory. Towards this goal we have selected and analyzed twelve candidate reference genes from three morphologically distinct cyanobacteria grown under routinely used laboratory conditions. The six genes exhibiting less variation in each organism were evaluated in terms of their expression stability using geNorm, NormFinder and BestKeeper. In addition, the minimum number of reference genes required for normalization was determined. Based on the three algorithms, we provide a list of genes for cyanobacterial RT-qPCR data normalization. To our knowledge, this is the first work on the validation of reference genes for cyanobacteria constituting a valuable starting point for future works.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0034983