selection of suitable reference genes for rt-qpcr analyses in cyanobacteria选择合适的参考基因在蓝藻rt-qpcr分析.pdfVIP

Selection of Suitable Reference Genes for RT-qPCR Analyses in Cyanobacteria 1,2 1 1,2¤ 1,3 1,2 Filipe Pinto , Catarina C. Pacheco , Daniela Ferreira , Pedro Moradas-Ferreira , Paula Tamagnini * ˆ 1 IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal, 2 Departamento de Biologia, Faculdade de Ciencias, Universidade do Porto, ˆ ´ Porto, Portugal, 3 ICBAS - Instituto de Ciencias Biomedicas Abel Salazar, Universidade do Porto, Porto, Portugal Abstract 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 n