Konstrukcija i mjerenje aktivnosti rekombinantnog proteina Pir2bla_O izloženog na površini stanične stijenke

undergraduate thesis

• Title (english): Construction and activity determination of recombinant protein Pir2bla_O displayed on the yeast cell surface
• Author: Stefani Prekpalaj
• Mentor: Bojan Žunar (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2023-09-14, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Stanice kvasca Saccharomyces cerevisiae mogu proteine koje izlučuju vezati na svoju staničnu stijenku. Pritom nativni proteini kvasca S. cerevisiae često sadrže nestrukturiranu spojnicu, tj. dio polipeptidnog lanca koji se ne smata u unaprijed predodređenu konformaciju. Cilj ovog rada bio je ispitati kako na efikasnost izlaganja na staničnu stijenku proteina izvjestitelja Pir2bla_L utječe njegova nestrukturirana spojnica. U tu svrhu je iz plazmida pRSⅠⅠ423-pir2bla_L uklonjen dio koji kodira za nestrukturiranu spojnicu proteina izvjestitelja, čime je konstruiran plazmid pRSⅠⅠ423-pir2bla_O, koji kodira za protein izvjestitelj Pir2bla_O. Novokonstruirani plazmid zatim je transformiran u stanice kvasca S. cerevisiae te je efikasnost izlaganja proteina Pir2bla_O na površini stanične stijenke pomoću nitrocefinskog testa uspoređena s efikasnošću izlaganja izvornog proteina Pir2bla_L. Dobiveni rezultati pokazali su da je uklanjanje spojnice smanjilo efikasnost izlaganja proteina izvjestitelja na staničnoj stijenci (102 % naspram 152 %), ali da navedena razlika nije statistički značajna (p = 0,06).

Abstract (english)

Yeast Saccharomyces cerevisiae can bind the proteins it secretes to its cell wall. The native proteins of the yeast S. cerevisiae often contain an unstructured linker, a part of the polypeptide chain that does not fold into a predetermined conformation. The aim of this work was to examine how the unstructured linker affected the cell wall display efficiency of the reporter protein Pir2bla_L. For this purpose, the sequence encoding unstructured linker was removed from the plasmid pRSII423-pir2bla_L, thus constructing the plasmid pRSII423-pir2bla_O, which was then transformed into yeast S. cerevisiae. Using the nitrocefin test, the cell wall display efficiency of the newly constructed protein Pir2bla_O and original protein Pir2bla_L was compared. The results showed that the removal of the linker reduced the efficiency of the display of the reporter protein on the cell wall (102% vs. 152%), but the difference was not statistically significant (p = 0.06).

• Keywords: Pir2 protein; β-laktamaza; rekombinantni proteini; proteini stanične stijenke; kvasac Saccharomyces cerevisiae
• Keywords (english): Pir2 protein; β-lactamase; recombinant proteins; cell wall proteins; yeast Saccharomyces cerevisiae
• Language: croatian
• URN:NBN: urn:nbn:hr:159:768089
• 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: 2024-04-29 13:23:34