Ispitivanje utjecaja mutacija u ciljenoj regiji gena SCW4 na sposobnost kovalentnog vezanja proteina Scw4 u staničnu stijenku kvasca

undergraduate thesis

• Title (english): Investigation of the effect of a mutation in the target region of the SCW4 gene on the ability to covalently bind the Scw4 protein to the yeast cell wall
• Author: Tea Knežević
• Mentor: Renata Teparić (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2020-09-15, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Food Technology

Abstract

Scw4 protein jedan je od najzastupljenijih proteina u staničnoj stijenci Saccharomyces cerevisiae. U staničnu stijenku veže se kovalentnim i nekovalentnim vezama, no sama priroda te kovalentne veze još uvijek nije poznata. Kako bi se otkrilo koja je regija odgovorna za njegovo kovalentno vezanje, provedeno je istraživanje u kojemu je mutacija uvedena u regiju koja je po svom aminokiselinskom slijedu slična onoj koja se nalazi u Pir proteinima, tzv. V regija. Kvasci su uzgojeni na selektivnim podlogama pri različitim pH vrijednostima, pH 4 i 7. Proteini stanične stijenke su zatim izolirani tretmanom stijenki vrućim SDS-om, odnosno 30 mM NaOH, čime su izolirani nekovalentno vezani odnosno kovalentno vezani proteini. Analiza proteinskih ekstrakata provedena je SDS gel elektroforezom i Western blotom. Iz dobivenih rezultata vidljivo je kako je V regija odgovorna za stvaranje kovalentne veze proteina Scw4 sa glukanom stanične stijenke. U daljnjim istraživanjima potrebno je utvrditi preko koje se aminokiseline unutar te regije ostvaruje kovalentna veza.

Abstract (english)

Scw4 protein is one of the most abundant proteins detected in the cell wall of yeast Saccharomyces cerevisiae. Within the cell wall proteins can be linked by covalent or noncovalent bound, but the nature of this covalent bond is still unknown. In this research project, in order to discover the region responsible for covalent attachment of Scw4, a mutation was introduced in the so-called V region, that has an amino acid sequence similar to that found in the Pir proteins. As part of the project, yeast was cultivated on a selective medium under different pH conditions, namely pH 4 and 7. During isolation cell walls were treated with hot SDS and 30 mM NaOH. The analysis of isolated cell wall proteins was performed by SDS gel electrophoresis and Western blot analysis. The obtained results showed that the ˝V region˝ is responsible for the covalent bonding of Scw4. In the further research exact aminoacid responsible for Scw4 covalent bonding into the cell wall should be detected.

• Keywords: japsinske proteaze; Kex2 proteaza; Saccharomyces cerevisiae; Scw4
• Keywords (english): Kex2 protease; Saccharomyces cerevisiae; Scw4; Yapsins
• Language: croatian
• URN:NBN: urn:nbn:hr:159:405492
• Study programme: Title: Food Technology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka prehrambene tehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka prehrambene tehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-10-26 10:50:37