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master's thesis
Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana
Zagreb: University of Zagreb, Faculty of Science, 2018. urn:nbn:hr:217:006801
Rendulić, Toni
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

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Rendulić, T. (2018). Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:006801

Rendulić, Toni. "Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana." Master's thesis, University of Zagreb, Faculty of Science, 2018. https://urn.nsk.hr/urn:nbn:hr:217:006801

Rendulić, Toni. "Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana." Master's thesis, University of Zagreb, Faculty of Science, 2018. https://urn.nsk.hr/urn:nbn:hr:217:006801

Rendulić, T. (2018). 'Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana', Master's thesis, University of Zagreb, Faculty of Science, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:006801

Rendulić T. Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2018 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:217:006801

T. Rendulić, "Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2018. Available at: https://urn.nsk.hr/urn:nbn:hr:217:006801

Metadata
TitleIstraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana
Title (english)Protein interaction analysis of BPM1, DMS3 and RDM1 in tobacco cells using bimolecular fluorescent complementation
AuthorToni Rendulić
MentorNataša Bauer (mentor)
Committee memberNataša Bauer (član povjerenstva)
Committee memberŽeljka Vidaković-Cifrek (član povjerenstva)
Committee memberMartina Šeruga Musić (član povjerenstva)
Committee memberDunja Leljak-Levanić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2018-04-17, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Proteini BPM sudjeluju u regulaciji nekoliko transkripcijskih faktora kroz ubikvitinsko-proteasomski put. Proteini DMS3 i RDM1 su važne komponente sustava za RNA-posredovanu metilaciju DNA (RdDM). U preliminarnom istraživanju pokazalo se da su DMS3 i RDM1 potencijalni interakcijski partneri proteina BPM1. U ovom radu su istražene interakcije proteina BPM1, DMS3 i RDM1 pomoću bimolekulske fluorescencijske komplementacije (BiFC). Radi provedbe analize BiFC, generirani su prikladni plazmidni konstrukti tehnikom kloniranja In-Fusion. Stanice epiderme duhana Nicotiana benthamiana, epiderme luka Allium cepa i stanice duhana BY-2 transformirane su generiranim plazmidnim konstruktima pomoću agroinfiltracije ili bombardiranjem mikročesticama zlata. Detekcija proteinskih interakcija provedena je fluorescencijskom mikroskopijom. DMS3 i RDM1 su imali interakciju u svim istraživanim stanicama, dok je formacija dimera DMS3 detektirana u epidermalnim stanicama duhana N. benthamiana i luka A. cepa. Interakcija proteina BPM1 i DMS3 je potvrđena u duhanu N. benthamiana i u stanicama BY-2. Interakcija između BPM1 i RDM1 nije detektirana u ovom radu. Potrebna su dodatna istraživanja kako bi se bolje razumjele pojedinosti interakcije između BPM1 i DMS3 te potencijalni utjecaj BPM1 na RdDM.
Abstract (english)
BPM proteins are known to participate in the regulation of several transcription factors via the ubiquitin proteasome pathway. DMS3 and RDM1 proteins are important components of the RNA-directed DNA methylation (RdDM) machinery. In preliminary research, DMS3 and RDM1 were shown to be potential interactors of BPM1. Here, interactions of BPM1, DMS3 and RDM1 proteins were analyzed via bimolecular fluorescence complementation (BiFC). To perform BiFC analysis, suitable plasmid constructs were generated using In-Fusion cloning technique. Nicotiana benthamiana, Allium cepa and BY-2 cells were transiently transformed with the generated constructs via agroinfiltration or microparticle bombardment. Finally, fluorescence microscopy was used to detect and analyze protein interactions. DMS3 and RDM1 interacted in all examined cells, while DMS3 dimer formation was detected in N. benthamiana and A. cepa epidermal cells. Interaction between BPM1 and DMS3 was proved in N. benthamiana leaf epiderm and in tobacco BY-2 cells. By using BiFC, no interaction was detected between BPM1 and RDM1 in this study. Further research is required to better understand the details of interaction between BPM1 and DMS3 proteins and the potential effect of BPM1 on RdDM.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:006801
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2018-05-03 08:34:57