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Identifier 000369984
Title Αλληλεπίδραση του ιοειδούς Potato spindle tuber viroid με πρωτείνες της ντομάτας (Solanum lycopersicum)
Alternative Title Interactions of the viroid Potato spindle tuber viroid with tomato (Solanum lycopersicum) proteins
Author Φραγκούλης, Πέτρος
Thesis advisor Τσαγρή, Ευθυμία
Reviewer Κοκκινίδης, Μιχαήλ
Καραγωγέως, Δόμνα
Abstract Viroids are small, single-stranded circular RNA molecules not known to code for any proteins.They are pathogens for a series of plant hosts. Potato spindle tuber viroid was the first viroid to be discovered and remains a model for the study of viroids. In previous work, a plant protein was isolated during a cDNA screen, using PSTVd RNA as a ligand. This protein, provisionally called Virp2, is further investigated in this thesis. We used electrophoretic mobility shift assays (EMSA), Northwestern blotting assays and dot blots in order to investigate the binding of the tomato protein Virp2 with PSTVd RNA transcripts in vitro. In the in vitro assays that were performed, affinity chromatography purified Virp2 was shown to bind weakly and unspecifically to PSTVd of both polarities, as well as to other DNA and RNA nucleic acids in a dot blot assay test. No interaction could be observed in EMSA or denaturing Northwestern assays. In the second part of this thesis a method was developed that is essentially a combination of native agarose electrophoresis and denaturing SDS-PAGE electrophoresis of crude plant nucleoprotein extracts. Comparison of non infected and infected nucleoprotein extracts by this method showed some differences in the viroid containing fractions, which can be further investigated. This novel, techincally simple method allows the resolution of the nucleic acids of the extract, as well as the separation of proteins both according to their charge and their molecular weight and may therefore be used for the identification of RNA binding proteins in such extracts
Language Greek
Subject Dot blot
Protein-nucleic acid interacion
Αλληλεπίδραση πρωτεΐνης-νουκλεϊκού οξέος
Issue date 2011-11-18
Collection   Faculty/Department--Faculty of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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