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Identifier 000326856
Title Ένα miRNA με πλουραλιστική δράση στα φυτά: miR414
Author Νταντάμη, Ελένη
Thesis advisor Καλαντίδης, Κρίτων
Δελιδάκης, Χρήστος
Abstract microRNAs are a conserved class of small RNAs (~21 nt) found in both plants and animals. They are negative regulators of gene expression preventing the protein production from their target mRNAs. microRNAs have been implicated in various important processes like development, differentiation and stress responses. The initial query of this work was to identify plant miRNAs with multiple targets. This investigation led to the characterization of miR414 which has an astonishingly wide array of potential targets. Our approach consisted of in silico analysis using online databases and miRNA - target prediction programs and molecular biology techniques leading to the production of transgenic plants potentially overexpressing miR414. We searched for miR414 targets in Arabidopsis thaliana and Oryza sativa using a plant specific miRNA target finder program. Then we tested if the potential targets comply with the rules of miRNA-target pairing. The number of the predicted targets was high and although the interactions with miR414 were close to perfect, there was no obvious relation between them. We further investigated the targets in the protein level and we found that the region that is targeted by the miRNA at the transcriptional level, when translated, consists of an Asp- rich motif. More bioinformatic analysis in the upstream sequences of the miR414 precursor resulted in prediction of regulatory elements overrepresented in miRNA promoters and potential transcription start site. In order to investigate the role of miR414 in vivo and verify the in silico predictions, we produced transgenic Arabidopsis thaliana and Nicotiana benthamiana plants. These plants overexpress the miR414 precursor driven by the CaMV 35S constitutive promoter. Recently a work in Physcomitrella patens was published supporting our bioinformatic results predicting many potential miR414 targets.
Language Greek
Issue date 2007-11-28
Collection   Faculty/Department--Faculty of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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