Ispitivanje utjecaja ekspresije humanog proteina Tau na vijabilnost stanica kvasca Saccharomyces cerevisiae s delecijom gena SSD1

undergraduate thesis

• Title (english): Influence of human Tau protein expression on the viability of Saccharomyces cerevisiae yeast cells with deletion of the SSD1 gene
• Author: Petra Kruljac
• 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: 2022-09-19, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Agregacija proteina Tau povezana je s odumiranjem neurona i nastankom bolesti, no uzroci toksičnosti nedovoljno su poznati. U svrhu boljeg razumijevanja uzroka toksičnosti, cilj rada bio je ispitati je li Tau protein čovjeka toksičan kada se eksprimira u kvascu Saccharomyces cerevisiae kojemu nedostaje Ssd1, protein važan u odgovoru na temperaturni stres. Tehnikama rekombinantne DNA konstruiran je plazmid s genom za Tau pod kontrolom Z-promotora koji se inducira progesteronom. Plazmid je transformiran u delecijski mutant kvasca ssd1Δ, u čiji genom je prethodno integriran konstrukt za ekspresiju transkripcijskog faktora potrebnog za aktivaciju Z-promotora, te je ekspresija proteina Tau provjerena western blotom. Toksičnost proteina Tau određena je ispitivanjem utjecaja ekspresije Tau na vijabilnost stanica, metodom testa rasta. Rezultati pokazuju da ekspresija proteina Tau ne utječe na vijabilnost mutanta ssd1Δ uzgojenog na optimalnoj i povišenoj temperaturi, što ukazuje na to da navedeni uvjeti ne dovode do toksičnosti proteina Tau.

Abstract (english)

Tau protein aggregation has been associated with neuronal death and disease, but the underlying mechanisms of toxicity are unclear. To elucidate the causes of Tau toxicity, the aim of this study was to test whether human Tau protein is toxic when expressed in yeast Saccharomyces cerevisiae lacking Ssd1, a protein involved in heat-stress response. Using recombinant DNA techniques, Tau-gene was placed on a plasmid under the control of the progesterone-inducible Z-promoter and transformed into ssd1Δ deletion mutant yeast, which had been modified by a construct expressing a progesteroneinducible transcription factor. Tau protein expression was verified by Western blot. Tau-mediated toxicity was investigated by examining the effect of Tau expression on cell viability using a growth test. The results show that Tau does not affect viability of ssd1Δ mutant grown at a normal and elevated temperature, indicating that heat-shock in the absence of functional Ssd1 does not lead to Tau toxicity.

• Keywords: protein Tau; SSD1; kvasac Saccharomyces cerevisiae; temperaturni stres
• Keywords (english): Tau protein; SSD1; yeast Saccharomyces cerevisiae; temperature stress
• Language: croatian
• URN:NBN: urn:nbn:hr:159:310872
• Study programme: Title: 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 resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-04-15 12:24:46