Return to search

An analysis of the effect of transformation on global– and gene–specific DNA methylation in four cultured cell lines / Jean du Toit

DNA methylation plays a role in several biological functions, such as gene expression regulation, and several endogenous and exogenous factors affect these DNA methylation patterns in the cell. One such alteration of a cell line's DNA methylation pattern is caused by the insertion of a vector into the cell line. Using the cytosine–extension assay and realtime methylation–specific PCR, alterations of DNA methylation levels on both global and gene–specific levels were investigated. In some cell lines the cellular transformation led to an increase in DNA methylation levels, and in others a decrease in DNA methylation amounts was observed. The same phenomenon was seen in the promoter regions of specific genes, showing that vector–insertion into a cell line caused DNA methylation alterations in many regions of the genome. These alterations in DNA methylation are investigated in this reduced representation study using enrichment of the methylated fraction of fragmented DNA and subsequent GS FLX Titanium sequencing of these methylated fragments. The results of sequence data analysis showed that methylated fragments are distributed over the whole genome, but could be related to only a few specific genes. These results have implications for cell culture work, biotechnological applications and uses in gene therapy. / Thesis (M.Sc. (Biochemistry))--North-West University, Potchefstroom Campus, 2011.

Identiferoai:union.ndltd.org:NWUBOLOKA1/oai:dspace.nwu.ac.za:10394/4560
Date January 2010
CreatorsDu Toit, Jean
PublisherNorth-West University
Source SetsNorth-West University
Detected LanguageEnglish
TypeThesis

Page generated in 0.0022 seconds