Karakterizacija proteina DRG1 iz spužve Eunapius subterraneus

master's thesis

• Title (english): Characterization of sponge Eunapius subterraneus DRG1 protein
• Author: Matea Modrić
• Mentor: Helena Ćetković (mentor)
• Committee member: Morana Dulić (predsjednik povjerenstva)
• Committee member: Nives Galić (član povjerenstva)
• Committee member: Željka Soldin (član povjerenstva)
• Committee member: Aleksandra Maršavelski (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2020-10-28, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

Spužve, morfološki jednostavne životinje bez tkiva i organa koje se nalaze na samoj bazi
filogenetskog stabla Metazoa, predstavljaju dobar model za objašnjenje evolucijskih obrazaca
razvoja životinja. Važne su za proučavanje karakteristika homologa kod pretka svih Metazoa
prije odvajanja i specijalizacije ovih gena u razvijenijim životinjama i za razumijevanje
osnovne fiziološke funkcije gena povezanih s nastankom raka kod jednostavnijih životinja. U
okviru diplomskog rada okarakteriziran je protein iz slatkovodne špiljske spužve Eunapius
subterraneus čiji je homolog kod čovjeka DRG1 (engl. Developmentally Regulated GTP
binding protein 1) povezan s nastankom raka. Cilj je dobiti što bolji uvid u evoluciju ovog
supresora metastaziranja. cDNA koja kodira za protein DRG1 uspješno je uklonirana u
ekspresijski vektor pET28b te je napravljena prekomjerna ekspresija koja je potvrđena
western blotting tehnikom. Protein DRG1 ima GTPaznu aktivnost koja je testirana mjerenjem
količine neiskorištenog GTP-a nakon reakcije hidrolize. Osim toga, protein je podvrgnut
metodi gel-filtracije kako bi se procijenila njegova molekularna masa.

Abstract (english)

Sponges, morphologically simple animals without tissues and organs and possibly the earliest
branching animal phylum, represent a good model for explaining the evolutionary patterns of
animal development. They are important for studying homologous characteristics in the
ancestor of all Metazoa prior to separation and specialization of these genes in more complex
animals and for understanding the basic physiological function of cancer-related genes in
simpler animals. The thesis characterizes the protein from the freshwater cave sponge
Eunapius subterraneus, whose homologue in humans DRG1 (Developmentally Regulated
GTP binding protein 1) is associated with cancer. The goal is to gain a better insight into the
evolution of this metastatic suppressor. The cDNA encoding the DRG1 protein was
successfully cloned into the pET28b expression vector and overexpression was performed.
The DRG1 protein has GTPase activity that was tested by measuring the amount of unused
GTP after the hydrolysis reaction. In addition, the protein was subjected to a gel filtration
method to estimate its molecular weight.

• Keywords: DRG1; gel-filtracija; GTPazna aktivnost; spužve
• Keywords (english): DRG1; gel filtration; GTPase activity; sponges
• Language: croatian
• URN:NBN: urn:nbn:hr:217:552357
• Study programme: Title: Graduate university studies in chemistry; specializations in: Research programme; Course: Research programme; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2022-10-28
• Created on: 2020-10-30 11:07:19