Unit:
Department of BiologyLibrary of the School of Science
Author:
TATOULI MARIA-PARASKEVI
Supervisors info:
Διαμάντης Σίδερης, Αναπληρωτής Καθηγητής, Τμήμα Βιολογίας, ΕΚΠΑ
Original Title:
ΠΡΟΣΔΙΟΡΙΣΜΟΣ ΣΤΟΙΧΕΙΩΝ ΤΟΥ ΥΠΟΚΙΝΗΤΉ ΤΗΣ ΑΝΘΡΏΠΙΝΗΣ RNase κ
Translated title:
Study of the promoter of the human RNase κ gene
Summary:
The purpose of this thesis is to identify elements of the gene promoter that encodes human RNASEK and to control their activity both qualitatively and quantitatively. Three regions downstream of the RNASEK gene were selected to be studied for further analysis. There regions were used to construct three expression plasmids to control gene expression. They were cloned upstream of the reporter gene encoding the green fluorescent protein (TurboGFP). To control the experiments, control plasmids were constructed and used. Two plasmids were used as positive controls, carrying upstream of the reporter gene one CMV viral promoter and one endogenous GAPDH gene promoter. The plasmid without promoter upstream of the TurboGFP reporter gene was used as a negative control. The above gene constructs were used to transfect human eukaryotic cells. The analysis of gene expression was followed by the emission fluorescence by green fluorescent protein. Qualitative data analysis was initially performed by observing the fluorescence levels of TurboGFP under a fluorescence microscope. Quantitative control of expression levels was followed by lysis of eukaryotic cells into which the plasmids were inserted, measurement of GFP fluorescence levels by fluorescence and then measurements of total protein by the Bradford method to normalize the result.
Main subject category:
Science
Keywords:
RNase κ, promoter, enhancer, reporter gene GFP, cell lines HEK293
File:
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ΜΕΛΕΤΗ ΤΟΥ ΥΠΟΚΙΝΗΤΗ ΤΗΣ ΑΝΘΡΩΠΙΝΗΣ RNase κ.pdf
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