Original title: Attempts on chromatin immunoprecipitation with \kur{C. elegans} nuclear receptor NHR-25
Translated title: Attempts on chromatin immunoprecipitation with \kur{C. elegans} nuclear receptor NHR-25
Authors: POSPĚCH, Alexandr
Document type: Master’s theses
Year: 2010
Language: cze
Abstract: The aim of the work presented in this thesis was to establish chromatin immunoprecipitation method in our laboratory as a tool to study target genes of the nuclear receptor NHR-25 in C. elegans. Once the method is established, it will be also useful for studies of other DNA binding proteins. ChIP was performed in transiently transfected cells HEK293 and analyzed using PCR and qPCR. Although ChIP is typically used to find authentic target genes in the cell or in organisms, testing protein-DNA interactions by ChIP in transient transfection system (by transfecting both the expression vector of the protein of interest and a vector containing potential binding sequence/promoter of the protein) can be useful as it serves as a relatively quick tool to confirm the direct binding. Since the detection is by PCR, this method is sensitive yet less costly non radioactive method to analyze protein-DNA interaction. For the first step towards ChIP in C. elegans; pulling down tagged protein directly from the worm was also performed as a preparation for in vivo analysis of NHR-25 regulated genes.
Keywords: Caenorhabditis elegans; Chromatin immunoprecipitation; Nuclear receptor
Citation: POSPĚCH, Alexandr. Attempts on chromatin immunoprecipitation with \kur{C. elegans} nuclear receptor NHR-25. České Budějovice, 2010. diplomová práce (Mgr.). JIHOČESKÁ UNIVERZITA V ČESKÝCH BUDĚJOVICÍCH. Přírodovědecká fakulta

Institution: University of South Bohemia in České Budějovice (web)
Document availability information: Fulltext is available in the Digital Repository of University of South Bohemia.
Original record: http://www.jcu.cz/vskp/9750

Permalink: http://www.nusl.cz/ntk/nusl-80493


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Universities and colleges > Public universities > University of South Bohemia in České Budějovice
Academic theses (ETDs) > Master’s theses