Usporedba razine ekspresije gena koji kodiraju za nekovalentno vezane proteine stanične stijenke kvasca Saccharomyces cerevisiae u logaritamskoj i stacionarnoj fazi rasta

master's thesis

• Title (english): Comparison of expression levels of genes encoding yeast Saccharomyces cerevisiae noncovalently bound cell wall proteins in logarithmic and stationary growth phase
• Author: Ida Kovačević
• Mentor: Igor Stuparević (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2018-10-16, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Stanična stijenka kvasca Saccharomyces cerevisiae izvanstanična je organela koja stanici osigurava mehaničku i osmotsku stabilnost. GraĎena je od polisaharidne mreţe sastavljene od β-1,3-glukana, β-1,6-glukana i hitina na koju su kovalentno ili nekovalentno vezani proteini. Molekularni sastav i struktura stanične stijenke variraju tijekom različitih faza staničnog ciklusa i rasta, zbog čega se očekuje da dolazi i do promjena u razini ekspresije gena koji kodiraju za proteine stanične stijenke. U ovom radu praćena je razina ekspresije gena koji kodiraju za nekovalentno vezane proteine stanične stijenke tijekom logaritamske i stacionarne faze rasta. Konstruirani su sojevi kvasca kojima su geni BGL2, SCW10 i SCW11 obiljeţeni hemaglutininskom sekvencom kako bi bilo omogućeno detektiranje obiljeţenih proteina specifičnim antitijelima. Razina ekspresije ovih gena praćena je na razini transkripta, semi-kvantitativnom PCR metodom, i na razini proteina, western blot metodom. Pokazana je smanjena razina ekspresije ispitivanih gena u stacionarnoj fazi u odnosu na logaritamsku fazu rasta.

Abstract (english)

Yeast Saccharomyces cerevisiae cell wall is an extracellular organelle that provides mechanical and osmotic stability. The yeast cell wall is constructed of a polysaccharide network composed of β-1,3-glucan, β-1,6-glucan and chitin to which proteins are covalently or non-covalently attached. Molecular composition and structure of the cell wall vary throughout the cell cycle and growth phases. Therefore, changes in expression level of genes encoding cell wall proteins are also expected. In this work, differences in the expression level of genes encoding non-covalently bound cell wall proteins was monitored during the logarithmic and stationary growth phase. Yeast strains in which BGL2, SCW10 and SCW11 genes were tagged with a haemagglutinin sequence were constructed to enable detection of tagged proteins with specific antibodies. The expression level of these genes was monitored at the transcript level by semi-quantitative PCR method and at the protein level by western blot method. Performed analysis revealed a decrease in expression level of examined genes in the stationary phase compared to the logarithmic growth phase.

• Keywords: kvasac Saccharomyces cerevisiae; faze rasta; stanična stijenka; proteini Scw; protein Bgl2
• Keywords (english): yeast Saccharomyces cerevisiae; growth phases; cell wall; Scw proteins; Bgl2 protein
• Language: croatian
• URN:NBN: urn:nbn:hr:159:913429
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2019-11-27 13:52:19