Biokemijska karakterizacija proteina RRAS2 iz spužve Eunapius subterraneus ; Istraživačka nastava kemije

master's thesis

• Title (english): Biochemical characterization of the RRAS2 protein from the sponge Eunapius subterraneus ; Inquiry-Based Chemistry Education
• Author: Leonarda Lazarin
• Mentor: Helena Ćetković (mentor)
• Mentor: Draginja Mrvoš-Sermek (mentor)
• Committee member: Draginja Mrvoš-Sermek (predsjednik povjerenstva)
• Committee member: Jasmina Rokov Plavec (član povjerenstva)
• Committee member: Mirela Sertić Perić (član povjerenstva)
• Committee member: Renata Matoničkin Kepčija (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2023-02-23, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

Rak je jedna od najrasprostranjenijih bolesti današnjice i često se javlja uslijed mutacija ili promjena u ekspresiji gena koji kodiraju za proteine uključene u kontrolu rasta i razvoja stanica. RRAS2 je primjer takvog proteina te pripada skupini malih GTPaza. Često je prekomjerno eksprimiran u različitim tipovima raka kod ljudi. Homolog proteina RRAS2 pronađen je i kod evolucijski puno starijih i jednostavnijih životinja – spužvi. Spužve predstavljaju dobar modelni organizam jer imaju jednostavnu morfologiju, a kompleksan genom, te većina gena iz spužvi pokazuje značajnu očuvanost s homolozima kod kralješnjaka. Kako bismo dobili bolji uvid u biokemijske karakteristike homologa RRAS2 iz spužve Eunapius subterraneus, klonirana je spužvina cDNA za RRAS2 te unesena u odgovarajući ekspresijski vektor. Napravljena je prekomjerna ekspresija rekombinantnog proteina u bakteriji Escherichia coli. Analizirana je i potvrđena intrinzična GTPazna aktivnost spužvinog proteina RRAS2, što ukazuje na očuvanost biokemijskih svojstava ovog onkogena. U metodičkom dijelu ovog rada dan je prijedlog nastavnog sata temeljen na istraživačkoj nastavi, za
odabranu temu (Enzimi). Pregledana je stručna i udžbenička literatura, oblikovani su prikladni didaktičko-metodički materijali te su navedeni mogući problemi koji se mogu javiti tijekom izvedbe nastavnog sata istraživačkog učenja.

Abstract (english)

Cancer is one of the most widespread diseases nowadays and often occurs due to mutations or changes in the expression levels of genes that control cell growth and development. RRAS2 is one of such proteins and a small GTPase. The homologue of the human RRAS2 was found in evolutionarily much older and simpler animals – sponges. Despite their simple morphology, sponges have a complex genome with many genes very similar to their vertebrate homologues. To get a better insight into the
biochemical characteristics of the RRAS2 homologue from the sponge Eunapius subterraneus, the cDNA for RRAS2 was cloned and inserted into an appropriate expression vector. E. coli cells were chemically transformed for the expression of the RRAS2 recombinant protein. The GTPase activity of the sponge RRAS2 was analyzed and confirmed, which indicates that the biochemical properties of this oncogene are evolutionary conserved. In the methodological part, a proposal of inquiry-based lesson, for the selected topic (Enzymes) is given. Professional and textbook literature was reviewed, appropriate didactic-methodical materials were designed, and possible problems that may arise during the implementation of the inquiry-based
learning were highlighted.

• Keywords: GTPaze; istraživačka nastava; rak; RRAS2; spužve
• Keywords (english): cancer; GTPases; inquiry-based education; RRAS2; sponges
• Language: croatian
• URN:NBN: urn:nbn:hr:217:021274
• Project: Number: IP-2019-04-5382; Title: Geni spužvi povezani s nastankom raka; Title: Cancer-related genes in sponges; Acronym: CanSponGE; Leader: Helena Ćetković; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Study programme: Title: Integrated undergraduate and graduate university study of Biology and Chemistry Education; specializations in: Integrated undergraduate and graduate university study of Biology and Chemistry Education; Course: Integrated undergraduate and graduate university study of Biology and Chemistry Education; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra edukacije biologije i kemije (magistar/magistra edukacije biologije i kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2025-02-23
• Created on: 2023-06-14 13:21:11