Regulacija heterologne ekspresije proteina ZmMAB1 u transgeničnim linijama uročnjaka (Arabidopsis thaliana) (L.) Heynh.

master's thesis

• Title (english): The regulation of heterologous ZmMAB1 protein expression in the transgenic (Arabidopsis thaliana(L.) Heynh. ) lines
• Author: Matija Šimac
• Mentor: Dunja Leljak-Levanić (mentor)
• Committee member: Dunja Leljak-Levanić (predsjednik povjerenstva)
• Committee member: Željka Vidaković-Cifrek (član povjerenstva)
• Committee member: Mirjana Kalafatić (član povjerenstva)
• Committee member: Inga Marijanović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2013-10-25, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Za protein MATH-BTB iz kukuruza (ZmMAB1) pokazano je prethodno da ima ulogu u regulaciji i pravilnom odvijanju asimetričnih dioba tijekom razvoja muškog i ženskog gametofita. Dokazana je i njegova uloga u proteasomalnoj razgradnji gdje sudjeluje kao dio kompleksa ubikvitin ligaze E3. U kukuruzu je ekspresija gena ZmMAB1 precizno kontrolirana, a njegov proteinski produkt je kratkoživući protein što otežava istraživanja izražaja ovog gena u homolognom sustavu. Strukturna komponenta mnogih ligaza E3 je kulin. Kako je dokazano da protein CUL3a iz vrste Arabidopsis thaliana (L.) Heynh. stvara komplekse s proteinom MAB1 iz kukuruza, vrsta Arabidopsis thaliana je odabrana za istraživanje regulacije ekspresije proteina MAB1 u heterolognom sustavu. U svrhu daljnjeg razumijevanja uloge i regulacije proteina ZmMAB1, protein ZmMAB1 obilježen fluorescentnim proteinom GFP ugrađen je u genom uročnjaka posredstvom bakterije Agrobacterium tumefaciens. Cilj ovog diplomskog rada bio je odrediti na kojoj se razini nastanka proteina ZmMAB1 (posttranskipcijski ili posttranslacijski) događa regulacija njegova izražaja. Rezultati su pokazali kako se protein ZmMAB1 ubikvitinira te se na taj način regulira njegova razina u stanici.

Abstract (english)

Zea mays MATH-BTB protein (ZmMAB1) has been shown to have a role in regulation and proper asimetric cell divisions during the male and female gametophyte development. Its role has been demonstrated in proteasomal degradation as part of a ubiquitin E3 ligase complex. ZmMAB1 gene expression in Zea mays is precisely regulated, and the protein product is a short-lived protein. Therefore ZmMAB1 gene expression research in a homolougus system is difficult. Cullin is a structural component of many E3 ligases. It has been shown that Cul3a protein from Arabidopsis thaliana (L.) Heynh. forms complexes with MAB1 from Zea mays, so Arabidopsis thaliana has been chosen for expression regulation research of MAB1 in a heterologous system. For the cause of better understanding the role and regulation of ZmMAB1 protein, GFP florescent protein labeled ZmMAB1 protein has been inserted in the genome of A. thaliana with bacterium Agrobacterium tumefaciens. Goal of this thesis was to determine on which level of formation (posttranscriptionaly or posttranslationaly) ZmMAB1 protein is regulated. Results have shown that ZmMAB1 protein is undergoing ubiqutination and that way his levels in the cell are regulated.

• Keywords: MATH-BTB; ubikvitin; kulin; regulacija; proteasom; heterologna ekspresija; asimetrična dioba
• Keywords (english): MATH-BTB; ubiqutin; cullin; regulation of expression; proteasome; heterologous expression; asimetric cell division
• Language: croatian
• URN:NBN: urn:nbn:hr:217:736441
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-28 08:33:44