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undergraduate thesis
Konstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2018. urn:nbn:hr:159:999781
Rešetar, Josip
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

Rešetar, J. (2018). Konstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris (Undergraduate thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:999781

Rešetar, Josip. "Konstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2018. https://urn.nsk.hr/urn:nbn:hr:159:999781

Rešetar, Josip. "Konstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2018. https://urn.nsk.hr/urn:nbn:hr:159:999781

Rešetar, J. (2018). 'Konstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris', Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 23 July 2024, https://urn.nsk.hr/urn:nbn:hr:159:999781

Rešetar J. Konstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2018 [cited 2024 July 23] Available at: https://urn.nsk.hr/urn:nbn:hr:159:999781

J. Rešetar, "Konstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris", Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2018. Available at: https://urn.nsk.hr/urn:nbn:hr:159:999781

Metadata
TitleKonstrukcija plazmidnog vektora za transformaciju kvasca Pichia pastoris
Title (english)Construction of shuttle vector for the transformation of yeast Pichia pastoris
AuthorJosip Rešetar
MentorRenata Teparić (mentor)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Chemistry and Biochemistry)
(Laboratory for Biochemistry)
Zagreb
Defense date and country2018-07-16, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Pichia pastoris često je korišteni mikroorganizam u biotehnološkoj proizvodnji heterolognih proteina. Zbog mogućnosti rasta do visokih staničnih gustoća važna je mogućnost njegove primjene u dobivanju kemikalija na ekološki prihvatljiv i ekonomski isplativ način. U ovom radu konstruiran je plazmidni vektor pogodan za transformaciju kvasca Pichia pastoris koji nosi gen za rekombinantni protein Ccw12XR iz kvasca Saccharomyces cerevisiae. Rekombinantni gen se sastoji od dijela koji kodira za enzim ksiloza reduktazu (XR) i dijela koji kodira za protein Ccw12 stanične stijenke kvasca S. cerevisiae. Rekombinantni gen umnožen je PCR metodom i ugrađen TA ligacijom u plazmidni vektor pGEM®-T Easy. Plazmid je umnožen u kompetentnim stanicama E. coli, nakon čega je iz plazmida izrezan rekombinantni gen CCW12XR korištenjem SapI restrikcijskog enzima. Naposljetku je CCW12XR gen ligiran s lineariziranim plazmidom pBSY3Z. Uspješnost ligacije je, nakon umnažanja plazmidnog vektora u kompetentnim stanicama E. coli, provjerena restrikcijom pomoću odabranih restrikcijskih enzima i elektroforezom na agaroznom gelu.
Abstract (english)
Pichia pastoris is a commonly used host in the biotechnological production of heterologous proteins. Since it can grow to very high cell densities, Pichia pastoris is applicable in the eco-friendly and economical production of chemicals. This thesis describes the construction of the shuttle vector, suitable for transformation of Pichia pastoris carrying the gene for the Ccw12XR recombinant protein from Saccharomyces cerevisiae. The recombinant gene encodes both the xylose reductase (XR) and the Ccw12 cell wall protein of S. cerevisiae. The recombinant gene is amplified using PCR and ligated into pGEM®-T Easy. The plasmid is replicated in competent Escherichia coli cells, following which the recombinant CCW12XR gene is restricted from the constructed plasmid using SapI. The isolated CCW12XR gene is then ligated with the linearized pBSY3Z plasmid. Finally, following the replication of the shuttle vector in component E. coli cells, the ligation efficiency is verified via restriction analysis and DNA electrophoresis.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:999781
Study programmeTitle: Nutrition
Study programme type: university
Study level: undergraduate
Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) nutricionizma (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) nutricionizma)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2018-07-19 09:20:28