Attempts on chromatin immunoprecipitation with \kur{C. elegans} nuclear receptor NHR-25

Bc. Alexandr POSPĚCH

Diplomová práce

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.

Jazyk práce: čeština

Datum vytvoření / odevzdání či podání práce: 5. 1. 2010

Zveřejnit od: 5. 1. 2010

Identifikátor: 9750

Obhajoba závěrečné práce

• Obhajoba proběhla 27. 1. 2010
• Vedoucí: Dr. Masako Asahina-Jindrová, Ph.D.

JIHOČESKÁ UNIVERZITA V ČESKÝCH BUDĚJOVICÍCH

Přírodovědecká fakulta

Magisterský studijní program / obor:
Biologie / Experimentální biologie