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Ocena vpliva G-kvadrupleksa v promotorju gena TNFSF11 na njegovo aktivnost = The assessment of the G-quadruplex effect within the TNFSF11 promoter region on its activity : magistrska naloga ; enoviti magistrski študij farmacijeGošnjak, TejaOsteoporoza je najpogostejša presnovna bolezen kostnine, za katero je značilno izguba kostne mase in propad kostnega tkiva. Posledično so kosti šibkejše in bolj nagnjene k zlomom. Vzrok za nastanek ... osteoporoze je neravnovesje med izgradnjo in razgradnjo kostnine. Proces kostne prenove je med drugimi uravnan preko sistema RANKL/RANK/OPG, ki nadzoruje proces osteoklastogeneze. V promotorski regiji gena TNFSF11 najdemo tudi strukturo imenovano G-kvadrupleks. G-kvadrupleksi so sekundarne prostorske strukture nukleinskih kislin, ki jih najdemo v zaporedjih, bogatih z gvanini. Narejenih je bilo že veliko študij, ki so pokazale, da ima struktura G-kvadrupleksa pomemben vpliv na izražanje številnih genov. Namen magistrske naloge je bil preučiti, ali spremembe v zaporedju G-kvadrupleksa promotorske regije gena TNFSF11 vplivajo na izražanje proteina med procesom prepisovanja ali med procesom prevajanja. To smo preverili tako, da smo v človeško celično linijo HeLa (celice raka materničnega vratu) vnesli reporterske plazmide s promotorsko regijo z G-kvadrupleksom. Pri delu smo uporabili reporterski plazmidni vektor pGL3 z delom zapisa promotorske regije gena TNFSF11, pri katerem smo v G-kvadrupleks vnesli zamenjavo baze gvanin v bazo timin na različnih mestih. Plazmide smo s transfekcijo vstavili v celice HeLa. Vpliv spremembe zaporedja G-kvadrupleksa na proces prepisovanja smo določili z analizo mRNA prepisane regije z metodo kvantitativne verižne reakcije s polimerazo (qPCR), vpliv na proces prevajanja pa smo določili z merjenjem aktivnosti promotorja gena TNFSF11 s testi luciferazne aktivnosti. Rezultate plazmidov s spremenjenim zaporedjem G-kvadrupleksa smo primerjali z rezultati izvornega plazmida. Na podlagi rezultatov smo ugotovili, da imajo vse testirane spremembe v G-kvadrupleksu promotorja gena TNFSF11 vpliv na izražanje proteina RANKL. Ugotovili smo tudi, da imajo spremembe večji vpliv na proces prevajanja kot na proces prepisovanja. Ne moremo pa zagotovo trditi, da imajo spremembe vpliv samo na enega izmed teh procesov, za to bi bile potrebne nadaljnje študije, s katerimi bi lahko bolj podrobno opazovali vpliv sprememb v G-kvadrupleksu na proces prepisovanja.Type of material - master's thesis ; adult, seriousPublication and manufacture - Ljubljana : [T. Gošnjak], 2023Language - slovenianCOBISS.SI-ID - 169776131
Author
Gošnjak, Teja
Other authors
Lovšin, Nika
Topics
Osteoporoza |
RANKL |
TNFSF11 |
G-kvadrupleks |
ligand receptorja za aktivacijo jedrnega dejavnika κB |
naddružina ligandov tumorje nekrotizirajočih dejavnikov
Library/institution |
City | Acronym | For loan | Other holdings |
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Faculty of Pharmacy, Ljubljana | Ljubljana | FFALJ |
reading room 1 cop.
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Gošnjak, Teja | |
Lovšin, Nika | 19649 |
Source: Personal bibliographies
and: SICRIS
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