20.500.12556/RUL-183563 Kloniranje in izražanje podenote 8B rastlinskih RNA polimeraz II, IV in V Cloning and expression of subunit 8B plant RNA polymerases II, IV and V Jod predstavlja enega od esencialnih elementov za vse vretenčarje in je v zmernih količinah koristen tudi za rastline, čeprav njegova vloga pri rastlinah še ni dobro raziskana. Ključno vlogo pri vretenčarjih ima kot sestavni del ščitničnih hormonov T3 in T4. Čeprav so v rastlinah odkrili analoge ščitničnih hormonov, mehanizem njihovega nastanka in pomen za rastlino še nista razjasnjena. Na podlagi značilnosti parov jodiranih tirozinskih aminokislinskih ostankov v tiroglobulinu, ki tvorijo ščitnične hormone, smo izbrali proteine navadnega repnjakovca (Arabidopsis thaliana), ki vsebujejo pare tirozinskih aminokislinskih ostankov z ustreznimi lastnostmi ter zapisa za dva od izbranih proteinov poskusili vstaviti v ekspresijski vektor za izražanje v E. coli. Zapis za podenoto 8B od DNA odvisnih RNA polimeraz II, IV in V (Rpb8) smo uspešno klonirali in rekombinantni protein izrazili ter očistili z Ni$^{2+}$ afinitetno kromatografijo. Očiščeni protein smo kemijsko jodirali, pri čemer je prišlo do agregacije proteina. Iodine is considered an essential element for all vertebrates and is also beneficial in moderate concentrations for plants, although its role in plants has not yet been sufficiently studied. For vertebrates, it is a crucial part of the thyroid hormones T3 and T4. Even though the presence of thyroid hormone analogues in plants has been confirmed, the mechanism of their synthesis and their role in plants are not yet fully understood. Considering the characteristics of hormonogenic pairs of iodinated tyrosine amino acid residues in thyroglobulin, we chose plant proteins from the mouse-ear cress (Arabidopsis thaliana) that contain pairs of tyrosine amino acid residues with similar characteristics. We tried to insert sequences encoding two of these plant proteins into an expression vector for expression in E. coli. Sequence encoding DNA-directed RNA polymerases II, IV and V subunit 8B (Rpb8) was successfully cloned, and the recombinant protein was expressed and purified via Ni$^{2+}$ affinity chromatography. The purified protein was chemically iodinated, which caused it to aggregate. jod ščitnični hormoni tirozin Arabidopsis thaliana iodine thyroid hormones tyrosine Arabidopsis thaliana true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2026-06-15 10:45:02 2026-06-15 10:45:12 2026-07-08 10:53:31 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 25639 284028419 Cankar_Nina_-_Kloniranje_in_izrazanje_podenote_8B_rastlinskih_RNA_polimeraz_II,_IV_in_V.pdf Cankar_Nina_-_Kloniranje_in_izrazanje_podenote_8B_rastlinskih_RNA_polimeraz_II,_IV_in_V.pdf 1 E5B67C31FEE8E2AD37FDF80600708B85 0aa0c59c03c5f3f795762b9032034a4d933270e0c782ee159df81a51e8a9869c 7a3f2306-6896-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=235631 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0