diplomsko delo
Petra Luzar (Author), Janez Plavec (Mentor)

Abstract

Večje število gvaninskih ponovitev v promotorju onkogena mdm2, ki je vključen v razvoj tumorskega fenotipa pri liposarkomih, nakazuje, da je možen nastanek različnih štirivijačnih struktur DNA ali G-kvadrupleksov. V okviru diplomskega dela smo določili zmožnost tvorbe štirivijačnih struktur DNA v inducibilnem promotorju mdm2 in določiti pogoje, kjer bo prisotna ena sama struktura, kar bi omogočilo njeno podrobno strukturno karakterizacijo. Poleg celotnega nukleotidnega zaporedja mdm2-15, ki vsebuje pet gvaninskih ponovitev, smo opazovali tudi dve skrajšani zaporedji mdm2-14 in mdm2-24, kjer pri mdm2-14 opazujemo prve štiri gvaninske ponovitve, pri mdm2-25 pa zadnje štiri gvaninske ponovitve. S pomočjo 1D 1H NMR eksperimentov smo določili število G-kvartetov v nastalih G-kvadrupleksih in s CD spektroskopijo opisali topološke značilnosti (prisotnost paralelnih, antiparalelnih in (3+1) zvrsti). Preverili smo, kako prisotnost K+ in Na+ kationov vpliva na nastanek G-kvadrupleksov in ugotovili, da so kalijevi ioni uspešnejši pri stabilizaciji G-kvadrupleksov, ki jih tvorijo z gvanini bogata zaporedja iz onkogena mdm2. Pokazali smo, da je najugodnejši pH za zvijanje oligonukleotidov nevtralen.

Keywords

gvanin;G-kvadrupleksi;oligonukleotidi;onkogen mdm2;jedrska magnetna resonanca;NMR;CD spektroskopija;diplomska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.11 - Undergraduate Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [P. Luzar]
UDC: 577.113(043.2)
COBISS: 31516163 Link will open in a new window
Views: 479
Downloads: 143
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Other data

Secondary language: English
Secondary title: Structural properties of guanine rich sequence in inducibile promoter mdm2
Secondary abstract: Large number of guanine tracts in oncogene mdm2, which are responsible for development of tumor fenotype in liposarcomas, suggest formation of different G-quadruplexes is possible. The major aim of this diploma thesis was to determine the possibility of a G-quadruplex formation within inducibile promoter mdm2 and to determine the conditions where single structure would be formed. In addition to nucleotide sequence mdm2-15 (contains five guanin repetitions), we also studied two shorter sequences mdm2-14 and mdm2-25, which are shortend for the first guanine repetition from 3'- or 5'-end. With 1D 1H NMR experiments we defined the number of different G-quartets in G-quadruplexes and in combination with CD spectroscopy aimed at description of topological determinants (parallel, antiparallel and (3+1) topology). Two different cations, K+ in Na+, were tested for their ability to stabilize G-quadruplex structures and the former was recognized as more successful at stabilization of G-quadruplexes of mdm2. Among acidic, basic and neutral pH, the latter was found to be most suited for promoting formation of predominant species.
Secondary keywords: nuclear magnetic resonance;G-quadruplex;CD spectroscopy;
Type (COBISS): Bachelor thesis/paper
Study programme: 1000373
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, UNI Kemija
Pages: 32 str.
ID: 12033030