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undergraduate thesis
Ispitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2020. urn:nbn:hr:159:050974
Matičević, Ana
University of Zagreb
Faculty of Food Technology and Biotechnology
Department of Chemistry and Biochemistry
Laboratory for Biochemistry

Institutional repository: Repository of the Faculty of Food Technology and Biotechnology

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Matičević, A. (2020). Ispitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae (Undergraduate thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:050974

Matičević, Ana. "Ispitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2020. https://urn.nsk.hr/urn:nbn:hr:159:050974

Matičević, Ana. "Ispitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2020. https://urn.nsk.hr/urn:nbn:hr:159:050974

Matičević, A. (2020). 'Ispitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae', Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 21 December 2024, https://urn.nsk.hr/urn:nbn:hr:159:050974

Matičević A. Ispitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2020 [cited 2024 December 21] Available at: https://urn.nsk.hr/urn:nbn:hr:159:050974

A. Matičević, "Ispitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae", Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2020. Available at: https://urn.nsk.hr/urn:nbn:hr:159:050974

Metadata
TitleIspitivanje načina vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae
Title (english)Examination of the way of binding of the mutated Scw4 protein into the cell wall of yeast Saccharomyces cerevisiae
AuthorAna Matičević
MentorRenata Teparić (mentor)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Chemistry and Biochemistry)
(Laboratory for Biochemistry)
Zagreb
Defense date and country2020-07-10, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Protein Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae može se vezati nekovalentno i kovalentno, no i dalje nije poznato koji dio proteina je odgovoran za kovalentno povezivanje. U prethodnom istraživanju uvedena je mutacija u jednu od regija koja je slična Pir repetitivnim regijama za koje se pretpostavlja da bi mogle biti odgovorne za kovalentno povezivanje. Također je utvrđeno kako se nativni Scw4 u staničnoj stijenci pojavljuje u tri forme različite veličine – neprocesiranoj, procesiranoj sa Kex2 proteazom te procesiranoj sa japsinskim proteazama. U ovom radu ispitivan je način vezanja mutiranog proteina Scw4 u staničnu stijenku kvasca Saccharomyces cerevisiae pri uzgoju na pH 4 i pH 7. Prethodno izraĎen konstrukt sa mutacijom umnožen je u bakteriji Escherichia coli, izoliran te su njime transformirane kvaščeve stanice. Kvasci su uzgajani u selektivnoj podlozi za uzgoj u uvjetima indukcije GAL promotora. Nakon uzgoja provedena je izolacija i analiza načina vezanja proteina stanične stijenke metodama SDS elektroforeze te metodom western blota. Na osnovu dobivenih rezultata može se zaključiti da je ispitivana regija u sekvenci Scw4 proteina bitna za njegovo kovalentno vezanje u staničnu stijenku.
Abstract (english)
Protein Scw4 can be bound into the cell wall of yeast Saccharomyces cerevisiae non-covalently and covalently. However, the part of the protein responsible for the covalent bond remains unknown. According to previous research, a mutation has been introduced into one of the regions similar to Pir repetitive regions which are supposed to be responsible for the covalent bonding. It has also been established that the Scw4 protein can appear in the cell wall in three different sized forms – unprocessed, processed with Kex2 protease, and processed with yapsin proteases. In this particular research, the way of binding mutated Scw4 protein grown on pH 4 and pH 7 has been examined. A previously made mutated construct has been replicated in Escherichia coli, isolated, and used to transform yeast cells which have been grown on selective media under the induction of the promotor. Cell wall proteins have been isolated and analysed by SDS PAGE and by western blotting. According to the results obtained, it can be concluded that the examined region in the Scw4 protein sequence is important for its covalent binding to the cell wall.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:050974
Study programmeTitle: Biotechnology
Study programme type: university
Study level: undergraduate
Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) icon
Created on2020-07-22 09:02:20