20.500.12556/RUL-171445 Priprava rekombinantnih proteinov navadnega repnjakovca, ki predstavljajo potencialni vir rastlinskih analogov hormonov T3 in T4 Production of recombinant proteins from Arabidopsis thaliana representing potential sources of plant analogs of hormones T3 and T4. Vloga joda v rastlinah je še vedno slabo raziskana, vendar novejše raziskave kažejo, da joda rastline ne le kopičijo, temveč ga vključujejo v organske spojine, tudi proteine. Identifikacija rastlinskih analogov ščitničnih hormonov (T3 in T4) postavlja vprašanja, kako ti hormoni v rastlini nastanejo in kakšna je njihova fiziološka vloga. V tem delu smo želeli identificirati proteine iz proteoma rastline Arabidopsis thaliana, ki bi lahko, na podlagi svojih strukturnih značilnosti, predstavljali ogrodje za nastanek molekul T3 in T4. Nabor proteinov smo z uporabo bioinformatskih orodij zožili na pet kandidatov, primernih za izražanje v bakterijskem ekspresijskem sistemu: Q9M8Z7, Q9C6Z3, Q501D5, F4J9A0 in O81888. Zapise za te proteine smo poskušali prenesti v izbrani ekspresijski vektor, jih izraziti v bakterijskih celicah Escherichia coli in očistiti z nikljevo afinitetno kromatografijo. Dva proteina smo izrazili v bakterijah, vendar se je izkazalo, da se pri izbranih pogojih proteina nahajata v netopni frakciji. Da bi proteine pridobili v zadostni količini za nadaljnje raziskave, bi bila potrebna optimizacija pogojev izražanja. The role of iodine in plants remains poorly understood. Recent studies suggest that plants not only accumulate iodine, but also incorporate it into organic compounds, including proteins. The identification of plant-derived thyroid hormone analogs (T3 and T4) raises fundamental questions about the biosynthesis of these molecules and their potential physiological roles in plants. In this study, we aimed to identify proteins from the Arabidopsis thaliana proteome, that could, based on their structural features, serve as potential scaffolds for the synthesis of T3 and T4 analogs. Using bioinformatic tools, we narrowed the candidate list to the following five proteins, suitable for recombinant expression in a bacterial system: Q9M8Z7, Q9C6Z3, Q501D5, F4J9A0 and O81888. The corresponding coding sequences were cloned into a selected expression vector, expressed in Escherichia coli and purified via nickel affinity chromatography. Two proteins were expressed in bacteria, however, under the tested conditions, both localized to the insoluble fraction. To obtain sufficient quantities of soluble protein for further analysis, optimization of expression conditions would be required. jod rastlinski analogi hormonov T3 in T4 molekulsko kloniranje izražanje in čiščenje proteinov iodine plant-derived thyroid hormone analogs (T3 and T4) molecular cloning protein expression protein purification true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2025-08-26 13:55:06 2025-08-26 13:55:15 2025-09-02 03:39:16 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 25558 247096323 Hvalec_Jan_-_Priprava_rekombinantnih_proteinov_navadnega_repnjakovca,_ki_predstavljajo_potencial.pdf Hvalec_Jan_-_Priprava_rekombinantnih_proteinov_navadnega_repnjakovca,_ki_predstavljajo_potencial.pdf 1 D9590E2FDC658D0931324DB2FFC03629 7917f80b475824fbb188e7e6b0a1ea67e820edf6ac92e0b0d37cce3bee51ba9e 6b1b2d2e-8273-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=214769 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0