Molekularna karakterizacija neočekivanih genetičkih događaja nastalih tijekom zamjene gena u kvascu

undergraduate thesis

• Title (english): The molecular characterization of unexpected genetic events arising during gene replacement in yeast
• Author: Valentino Blasina
• Mentor: Ivan-Krešimir Svetec (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: 2012-07-25, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Prilikom transformacije kvasca Saccharomyces cerevisiae linearnom molekulom DNA moguće je usmjeriti integraciju transformirajuće DNA u željeni ciljni lokus ukoliko su krajevi transformirajuće DNA homologni ciljnoj regiji u genomu (ciljanje gena). Ukoliko su krajevi transformirajuće DNA nakon sparivanja s homolognom ciljnom regijom, orijentirani jedan od drugoga, očekivani ishod transformacije je zamjena ciljne regije transformirajućom DNA (rekombinacija krajevi-van). Cilj ovog rada bio je odrediti vrstu i učestalost pojedinih aberantnih genetičkih događaja u transformantima kvasca u određenom eksperimentalnom sustavu u kojem je očekivani ishod zamjena gena HIS3 genom LEU2. Aberantni transformanti prethodno su odabrani na temelju auksonografske analize, a u ovom radu podvrgnuti su molekularnoj karakterizaciji metodom hibridizacije DNA po Southernu. Utvrđeno je da je najčešći aberantni događaj duplikacija ciljnog kromosoma (44.44%).

Abstract (english)

During the transformation of yeast Saccharomyces cerevisiae with a linear DNA molecule it is possible to target the integration of the transforming DNA in the desired target locus if the ends of the transforming DNA are homologous to the target region in the genome (gene targeting). If the ends of the transforming DNA are oriented from one another, then the transformation will result with a gene replacement (ends-out recombination). The aim of this study was to determine the type and frequency of each aberrant genetic event in yeast transformants in a specific experimental system in which the expected genetic event was the replacement of HIS3 gene with LEU2 gene. Based on a previously conducted auxanographic analysis, aberrant yeast transformants were selected and subjected to a molecular characterization by DNA hybridization (Southern blot). The results showed that the most common aberrant genetic event was the duplication of the targeted chromosome (44.44%).

• Keywords: Saccharomyces cerevisiae; homologna rekombinacija; zamjena gena; rekombinacija krajevi-van; genska terapija
• Keywords (english): Saccharomyces cerevisiae; homologous recombination; gene replacement; ends–out recombination; gene therapy
• Language: croatian
• URN:NBN: urn:nbn:hr:159:504341
• 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: Access restricted to students and staff of home institution
• Created on: 2017-05-16 09:26:58