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undergraduate thesis
Uvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode
Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology, 2016. urn:nbn:hr:159:596211
Vučenović, Ivan
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

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Vučenović, I. (2016). Uvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode (Undergraduate thesis). Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology. Retrieved from https://urn.nsk.hr/urn:nbn:hr:159:596211

Vučenović, Ivan. "Uvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2016. https://urn.nsk.hr/urn:nbn:hr:159:596211

Vučenović, Ivan. "Uvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode." Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, 2016. https://urn.nsk.hr/urn:nbn:hr:159:596211

Vučenović, I. (2016). 'Uvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode', Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, accessed 05 July 2024, https://urn.nsk.hr/urn:nbn:hr:159:596211

Vučenović I. Uvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode [Undergraduate thesis]. Zagreb: University of Zagreb, Faculty of Food Technology and Biotechnology; 2016 [cited 2024 July 05] Available at: https://urn.nsk.hr/urn:nbn:hr:159:596211

I. Vučenović, "Uvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode", Undergraduate thesis, University of Zagreb, Faculty of Food Technology and Biotechnology, Zagreb, 2016. Available at: https://urn.nsk.hr/urn:nbn:hr:159:596211

Metadata
TitleUvođenje mutacije u gen SCW4 iz kvasca Saccharomyces cerevisiae pomoću PCR metode
Title (english)Introducing mutation in Saccharomyces cerevisiae gene SCW4 using PCR method
AuthorIvan Vučenović
MentorRenata Teparić (mentor)
GranterUniversity of Zagreb
Faculty of Food Technology and Biotechnology
(Department of Chemistry and Biochemistry)
(Laboratory for Biochemistry)
Zagreb
Defense date and country2016-09-20, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Food Technology
Abstract
PCR (lančana reakcija polimeraze) je metoda koja omogućava selektivno, brzo i osjetljivo umnažanje željenog slijeda nukleotida DNA. Kroz tri stupnja reakcije dobije se kao produkt željena sekvenca DNA u velikom broju kopija. U ovom radu je PCR korišten za uvođenje mutacija u gen SCW4 kvasca Saccharomyces cerevisiae na način da su konstruirana dva fragmenta od kojih je jedan sadržavao sekvence uzvodno, a drugi nizvodno od mjesta mutacije. Mutacija se odnosila na deleciju dijela gena koji kodira za 126 aminokiseline na N-terminalnom kraju proteina Scw4. Dobiveni fragmenti su zatim drugim krugom PCR-a spojeni u jedan konstrukt budući da su međusobno komplementarni u dijelu gena neposredno uzvodno i nizvodno od deletiranog dijela u nativnom genu. Dobiveni konstrukt je zatim TA-ligacijom unesen u plazmid kojim je transformirana bakterija E. coli u kojoj je umnožen i iz koje je uspješno izoliran.
Abstract (english)
PCR (polymerase chain reaction) is a method enabling rapid, selective and sensitive amplification of the DNA sequence. As a result, after three reaction steps, PCR product contains large number of copies of the target DNA sequence. In this experiment, PCR was used to introduce mutation in the SCW4 gene of the yeast Saccharomyces cerevisiae. First PCR was used for creating two fragments, one fragment contained sequence upstream of mutation position in gene sequence, and the second fragment contained sequence downstream of mutation position. Mutation consisted of deletion of the part of the gene encoding for 126 aminoacids in N-terminal part of Scw4 protein. Two products of first PCR were combined into one final construct, since they are complementary in sequence upstream and downstream of the deleted region. This fragment is then introduced into the plasmid using TA-ligation method and plasmid obtained was used for transformation of E. coli. In the end, the plasmid containing mutated form of SCW4 gene was successfully multiplied and isolated from transformed bacteria.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:159:596211
Study programmeTitle: Food Technology
Study programme type: university
Study level: undergraduate
Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka prehrambene tehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka prehrambene tehnologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
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Created on2017-04-24 11:08:00