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master's thesis
Ciljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9
2019. urn:nbn:hr:166:822193
Vuković, Mila
University of Split
Faculty of Science
Department of Physics

Cite this document

Vuković, M. (2019). Ciljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9 (Master's thesis). Split: University of Split, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:166:822193

Vuković, Mila. "Ciljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9." Master's thesis, University of Split, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:166:822193

Vuković, Mila. "Ciljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9." Master's thesis, University of Split, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:166:822193

Vuković, M. (2019). 'Ciljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9', Master's thesis, University of Split, Faculty of Science, accessed 14 November 2024, https://urn.nsk.hr/urn:nbn:hr:166:822193

Vuković M. Ciljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9 [Master's thesis]. Split: University of Split, Faculty of Science; 2019 [cited 2024 November 14] Available at: https://urn.nsk.hr/urn:nbn:hr:166:822193

M. Vuković, "Ciljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9", Master's thesis, University of Split, Faculty of Science, Split, 2019. Available at: https://urn.nsk.hr/urn:nbn:hr:166:822193

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Metadata
TitleCiljana inaktivacija gena GFP u E. coli metodom CRISPR/Cas9
Title (english)Targeted inactivation of the GFP gene in E. coli by the CRISPR/Cas9 method
AuthorMila Vuković
MentorJasna Puizina (mentor)
MentorIvica Aviani (komentor)
Committee memberJasna Puizina (predsjednik povjerenstva)
Committee memberIvica Aviani (član povjerenstva)
Committee memberIvica Šamanić (član povjerenstva)
GranterUniversity of Split
Faculty of Science
(Department of Physics)
Split
Defense date and country2019-10-22, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Physics
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
U radu je prikazana primjena metode CRISPR/Cas9 za ciljanu inaktivaciju gena koji kodira za zeleni fluorescentni protein (GFP) u bakteriji E. coli. Eksperiment je proveden tako da su se primjenom bioinformatičkih alata dizajnirale vodeće RNA molekule čiji su odgovarajući DNA oligonukleotidi uneseni u plazmid BPK764. Ovaj plazmid je specifičan po tome što sadrži gen za kaspazu Cas9 i insercijsko mjesto za unos molekule sgRNA. Drugi ključni plazmid je pGLO jer on sadrži gen GFP čija je inaktivacija cilj ovog rada. Ova inaktivacije gena uspješno je provedena primjenom sustava CRISPR/Cas9 što je pokazano vizualizacijom kontrolnih i eksperimentalnih skupina bakterija na UV transiluminatoru. Bakterije obiju skupina snimljene su mikroskopom atomskih sila (AFM) primjenom kvantitativnog načina rada (QI) i invertnim optičkim mikroskopom s fluorescentnim filterom na kojem je također potvrđena uspješnost provedenog eksperimenta.
Abstract (english)
This thesis presents the application of the CRISPR/Cas9 method for targeted inactivation of the gene encoding green fluorescent protein (GFP) in E. coli. The experiment was performed by designing single guide RNA molecules (sgRNA) using bioinformatics tools, whose corresponding DNA oligonucleotides were inserted into the BPK764 plasmid. This plasmid is specific because it contains the caspase 9 gene (cas9) and an insertion site for DNA oligonucleotide that correspond to sgRNA molecule. Another important plasmid is pGLO because it contains GFP gene whose inactivation is the goal of this thesis. This gene inactivation was successfully performed using the CRISPR/Cas9 system as evidenced by the visualization of control and experimental groups of bacteria on the UV transilluminator. The bacteria of both groups were imaged by an atomic force microscope (AFM) using a quantitative imaging mode (QI) and observed by an inverted optical microscope with the fluorescence option, which also showed the success of the experiment.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:166:822193
Study programmeTitle: Physics; specializations in: major in teaching, major in Astrophysics, major in Biophysics, major in Enviromental physics, major in Computational physics
Course: major in Biophysics
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra fizike (magistar/magistra fizike)
Type of resourceText
File originBorn digital
Access conditionsAccess restricted to authenticated users
Terms of useLicence In copyright icon
Created on2019-11-06 11:26:12