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Funkce Zinc-finger proteinu 644 (Zfp644) v myším organismu. / Function of Zinc finger protein 644 (Zfp644) in mouse organism.Szczerkowska, Katarzyna Izabela January 2022 (has links)
ZNF644 (Zinc Finger Protein 644) is a C2H2 zinc finger gene encoding a putative transcription regulator, of which a point mutation (S672G) is associated with inherited high myopia in humans. It is also described to be a partner of the G9a/GLP (G9a- euchromatic histone- lysine N-methyltransferase 2, EHMT2; GLP - euchromatic histone-lysine N-methyltransferase 1, EHMT1) complex, known for its essential role in histone methylation, specifically H3K9me1and H3K9me2. It was reported that another transcription factor, WIZ (Widely-Interspaced Zinc Finger-Containing Protein), can bind to this complex and cooperate in gene silencing simultaneously. In order to study Zfp644 impact on myopia, we generated a mouse model, Zfp644S673G that mimics human mutation. In addition, a mouse with a persuasive truncated form of the protein, Zfp644Δ8 was created. Both mouse models went through an examination of retinal function and morphology. Moreover, with use of ultrasonography, different ocular parameters were examined. We conclude, that Zfp644 gene is causative for myopia in mice. Further examinations of Zfp644Δ8 animals show severe symptoms in metabolism and female fertility. To describe the impact of Zfp644 in mouse fertility we performed various experiments including analysis of expression of Zfp644 in reproductive...
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An analysis of the effect of transformation on global– and gene–specific DNA methylation in four cultured cell lines / Jean du ToitDu Toit, Jean January 2010 (has links)
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.
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An analysis of the effect of transformation on global– and gene–specific DNA methylation in four cultured cell lines / Jean du ToitDu Toit, Jean January 2010 (has links)
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.
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