tailieunhanh - báo cáo khoa học: " Selection of reference genes for quantitative gene expression normalization in flax (Linum usitatissimum L.)"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Selection of reference genes for quantitative gene expression normalization in flax (Linum usitatissimum L.) | Huis et al. BMC Plant Biology 2010 10 71 http 1471-2229 10 71 BMC Plant Biology RESEARCH ARTICLE _ Open Access Selection of reference genes for quantitative gene expression normalization in flax Linum usitatissimum L. Rudy Huis Simon Hawkins and Godfrey Neutelings Abstract Background Quantitative real-time PCR qRT-PCR is currently the most accurate method for detecting differential gene expression. Such an approach depends on the identification of uniformly expressed housekeeping genes HKGs . Extensive transcriptomic data mining and experimental validation in different model plants have shown that the reliability of these endogenous controls can be influenced by the plant species growth conditions and organs tissues examined. It is therefore important to identify the best reference genes to use in each biological system before using qRT-PCR to investigate differential gene expression. In this paper we evaluate different candidate HKGs for developmental transcriptomic studies in the economically-important flax fiber- and oil-crop Linum usitatissimum L . Results Specific primers were designed in order to quantify the expression levels of 20 different potential housekeeping genes in flax roots internal- and external-stem tissues leaves and flowers at different developmental stages. After calculations of PCR efficiencies 13 HKGs were retained and their expression stabilities evaluated by the computer algorithms geNorm and NormFinder. According to geNorm 2 Transcriptional Elongation Factors TEFs and 1 Ubiquitin gene are necessary for normalizing gene expression when all studied samples are considered. However only 2 TEFs are required for normalizing expression in stem tissues. In contrast NormFinder identified glyceraldehyde-3-phosphate dehydrogenase GADPH as the most stably expressed gene when all samples were grouped together as well as when samples were classed into different sub-groups. qRT-PCR was then used to investigate the relative .