Izolacija plazmida pBAD-HheC, pUC-GW-RadH i pUC-GW-NoxV iz velikog volumena kulture bakterije Escherichia coli

undergraduate thesis

• Title (english): Isolation of plasmids pBAD-HheC, pUC-GW-RadH and pUC-GW-NoxV from a large volume of Escherichia coli culture
• Author: Emma Karolina Vasung
• Mentor: Marina Svetec Miklenić (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biology and Microbial Genetics); Zagreb
• Defense date and country: 2022-09-16, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Plazmidi su iznimno važni u genetičkom inženjerstvu, gdje se upotrebljavaju kao vektori, tj. molekule DNA koje omogućuju umnožavanje kloniranog fragmenta DNA u stanici domaćinu. U genetičkom inženjerstvu E. coli se koristi kao univerzalan domaćin za kloniranje. Cilj ovog rada bio je izolirati plazmide pBAD-HheC, pUC-GW-RadH i pUC-GW-NoxV iz velikog volumena bakterije Escherichia coli. Nakon izolacije i pročišćavanja plazmidne DNA, provedena je gel-elektroforeza radi provjere uspješnosti izolacije. Stvarna struktura izoliranih plazmida provjerena je restrikcijskom analizom pomoću dva restrikcijska enzima. Veličina dobivenih fragmenata nakon cijepanja bila je u skladu s restrikcijskim mapama plazmida.

Abstract (english)

Plasmids play an important role in genetic engineering; they are used as vectors, i.e., DNA molecules which allow the cloned DNA fragment to be propagated in the host cell. In genetic engineering, E. coli is used as a universal host for cloning. The aim of this work was to isolate the plasmids pBAD-HheC, pUC-GW-RadH and pUC-GW-NoxV from a large volume of E. coli culture. Following the isolation and purification of the plasmid DNA, gel-electrophoresis was used to determine whether the DNA isolation was successful. The actual structure of the isolated plasmids was confirmed using restriction enzyme analysis. The size of the cleaved fragments matched the restriction maps of the plasmids.

• Keywords: Escherichia coli; plazmid; izolacija; ekspresijski vektor
• Keywords (english): Escherichia coli; plasmid; isolation; expression vector
• Language: croatian
• URN:NBN: urn:nbn:hr:159:045114
• 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: 2024-04-09 12:42:41