Measuring Symbiodinium sp. gene expression patterns with quantitative real-time PCR
Boldt, L., Yellowlees, D., and Leggat, W. (2010) Measuring Symbiodinium sp. gene expression patterns with quantitative real-time PCR. In: Proceedings of the 11th International Coral Reef Symposium, pp. 118-122. From: 11th International Coral Reef Symposium, 7 - 11 July 2008, Fort Lauderdale, FL, USA.
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Quantitative real-time PCR is a popular method for measuring gene expression in many biological organisms. The dinoflagellate Symbiodinium spp. lives in eukaryotic symbioses with reef-building corals and other marine invertebrates and protists. The use of quantitative real-time PCR to measure gene expression levels of the dinoflagellate symbiont has been limited by the lack of validated normalisation genes and the inability to purify mRNA from the alga without significant host contamination. Normalisation genes to correct for errors inherent to this technique are essential and as yet no validated normalisation genes have been identified for use in Symbiodinium gene expression studies. Three commonly used normalisation genes, β-actin, proliferating cell nuclear antigen (PCNA) and 18S rRNA, are tested for use as internal standards in light manipulation experiments of the major light harvesting protein complexes (chlorophyll a chlorophyll c2 peridinin protein complexes - acpPC). Using the algorithm software program geNorm, β-actin and PCNA were found to be more stably expressed than the more highly expressed 18S rRNA genes, and result in similar gene expression profiles when used to normalise gene expression data for three different acpPC genes.
|Item Type:||Conference Item (Refereed Research Paper - E1)|
|Keywords:||qRT-PCR; normalisation genes; Symbiodinium sp; light harvesting protein complexes|
|Date Deposited:||09 May 2011 00:26|
|FoR Codes:||06 BIOLOGICAL SCIENCES > 0601 Biochemistry and Cell Biology > 060102 Bioinformatics @ 100%|
|SEO Codes:||96 ENVIRONMENT > 9603 Climate and Climate Change > 960399 Climate and Climate Change not elsewhere classified @ 100%|
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