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Strukturne lastnosti z gvanini bogatega zaporedja v inducibilnem promotorju mdm2
Luzar, Petra (Author), Plavec, Janez (Mentor) More about this mentor... This link opens in a new window

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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.

Language:Slovenian
Keywords:NMR, G-kvadrupleks, CD spektroskopija
Work type:Bachelor thesis/paper (mb11)
Tipology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2020
COBISS.SI-ID:31516163 This link opens in a new window
Views:270
Downloads:128
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Secondary language

Language:English
Title:Structural properties of guanine rich sequence in inducibile promoter mdm2
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.

Keywords:NMR, G-quadruplex, CD spectroscopy

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