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undergraduate thesis
Ispitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina u staničnu stijenku kvasca Saccharomyces cerevisiae
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2021. urn:nbn:hr:159:832036
Orešković, Nikolina
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

Cite this document

Orešković, N. (2021). Ispitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina 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:159:832036

Orešković, Nikolina. "Ispitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina u staničnu stijenku kvasca Saccharomyces cerevisiae." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2021. https://urn.nsk.hr/urn:nbn:hr:159:832036

Orešković, Nikolina. "Ispitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina u staničnu stijenku kvasca Saccharomyces cerevisiae." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2021. https://urn.nsk.hr/urn:nbn:hr:159:832036

Orešković, N. (2021). 'Ispitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina u staničnu stijenku kvasca Saccharomyces cerevisiae', Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 05 November 2024, https://urn.nsk.hr/urn:nbn:hr:159:832036

Orešković N. Ispitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina u staničnu stijenku kvasca Saccharomyces cerevisiae [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2021 [cited 2024 November 05] Available at: https://urn.nsk.hr/urn:nbn:hr:159:832036

N. Orešković, "Ispitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina u staničnu stijenku kvasca Saccharomyces cerevisiae", Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:159:832036

Metadata
TitleIspitivanje utjecaja N-glikozilacije i endocitoze na ugradnju heterolognih proteina u staničnu stijenku kvasca Saccharomyces cerevisiae
Title (english)Effect of N-glycosilation and endocytosis on incorporation of heterologus proteins in the Saccharomyces cerevisiae cell wall
AuthorNikolina Orešković
MentorRenata Teparić (mentor)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Chemistry and Biochemistry)
(Laboratory for Biochemistry)
Zagreb
Defense date and country2021-09-16, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Izlaganje proteina na staničnoj površini (˝surface display˝) predstavlja poboljšanu tehniku imobilizacije proteina te omogućuje kontinuiranu sintezu rekombinantnih proteina zahvaljujući fuziji gena koji kodira za ispitivani enzim i gena koji kodiraju za nativne proteine stanične stijenke. U ovom radu rekombinantni protein β-laktamaza (gen bla) fuzioniran je sa proteinom Pir2, odnosno sa dijelom proteina Ccw12. Pir2 veže se za stijenku svojim Nterminalnim dijelom. Dio gena CCW12, korišten u ovom radu, kodira za C-terminalnu signalnu sekvencu na koju se veže GPI-sidro, preko kojeg se protein veže u stijenku. Najveći je nedostatak ovakvog pristupa mali prinos imobiliziranih proteina u stijenku. S pretpostavkom da mutacije u N-glikozilaciji i endocitozi povećavaju učinkovitost imobilizacije ispitivan je utjecaj mutacija u N-glikozilaciji (och1 mutant i mnn mutanti) te mutacija u endocitozi (vam4 mutant i end mutanti) na efikasnost ugradnje rekombinantnog proteina u stijenku mjerenjem aktivnost β-laktamaze, što omogućuje procjenu količine imobiliziranog rekombinantnog proteina u staničnoj stijenci.
Abstract (english)
„Surface display“ system provides enhanced immobilization technique that enables countinuous synthesis of recombinant protein. This is possible because of the genetic fusion between genes encoding for native cell wall proteins and enzyme that is reasearched. In this study the enzyme β-lactamase (gene bla) and Pir2 or a fragment of Ccw12 are fuzed. Pir2 is linked to the cell wall on its N-terminal end, while Ccw12 is linked to the cell wall by GPI anchor on C- terminal region. One of the major disadvantages of this approach is low yield in protein immobilization. With the assumption that mutation in N-glycosylation and endocytosis enhances surface display system efficiency, this system was examined in mutants in Nglycosylation (och1 mutant and mnn mutants) and in mutants in endocytosis (vam4 mutant and end mutants). By measuring β-lactamase activity, it is possible to estimate the amount of recombinant protein immobilized in cell wall.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:832036
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 on2021-09-29 09:15:20