Naslov
Istraživanje interakcija proteina BPM1, DMS3 i RDM1 bimolekulskom fluorescencijskom komplementacijom u stanicama duhana
(Protein interaction analysis of BPM1, DMS3 and RDM1 in tobacco cells using bimolecular fluorescent complementation)

Autori
Rendulić, Toni

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, diplomski rad, diplomski

Fakultet
Prirodoslovno-matematički fakultet

Mjesto
Zagreb

Datum
17.04

Godina
2018

Stranica
39

Mentor
Bauer, Nataša

Ključne riječi
BTB ; MATH ; RdDM ; BiFC ; transformation

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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