Konstrukcija mutanta SLX4 kvasca Saccharomyces cerevisiae

undergraduate thesis

• Title (english): Construction of mutant SLX4 yeast Saccharomyces cerevisiae
• Author: Renata Buđa
• 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

Palindromske sekvencije prisutne su u genetičkom materijalu svih živih organizama te su vrlo često uključene u regulaciju fundamentalnih staničnih procesa. Međutim, dulji palindromi mogu u DNA formirati rekombinogene sekundarne strukture koje mogu uzrokovati pucanje kromosoma, genetičke prerasporede i genetičke bolesti kod ljudi. Dosadašnji rezultati sugeriraju da bi za nastanak loma u palindromu u uvjetima in vivo mogao biti odgovoran gen SLX4. Upravo zbog toga je cilj ovoga rada bio provesti inaktivaciju navedenog gena u kvaščevom soju BYUHA-8 koji se otprije koristi za istraživanje rekombinogenosti palindroma. U tu je svrhu najprije lančanom reakcijom polimeraze sintetiziran linearni fragment DNA za inaktivaciju gena SLX4 i tim je fragmentom zatim transformiran kvaščev soj BYUHA-8. Molekularna analiza dobivenih transformanata pokazala je da je u svim transformantima došlo do ilegitimne integracije transformirajuće DNA, a uzrok tome bi mogla biti nedovoljna duljina homologija na krajevima korištenog linearnog fragmenta koja je u ovom radu iznosila 45 parova baza.

Abstract (english)

Palindromic sequences are present in the genetic material of all living organisms and are often involved in the regulation of fundamental cellular processes. However, longer palindromes can form recombinogenic secondary structures that can induce chromosomal breaks, genome rearrangements and cause genetic diseases in humans. Previous results suggest that the SLX4 gene product could be responsible for the introduction of breaks in palindromic sequences in vivo. Therefore, the goal of this paper was to inactivate the SLX4 gene in the yeast strain BYUHA-8, which is used to study palindrome recombinogenicity. To achieve this goal, we synthesized the SLX4 disruption cassette by PCR, and used it to transform yeast strain BYUHA-8. Molecular analysis of the transformants demonstrated that in all cases the disruption cassette integrated at random sites in the genome, which could be due to insufficent length of homology on the ends of the disruption cassette used for transformation.

• Keywords: palindromi; „ends-out“; ilegitimna inegracija; gen SLX4; homologne regije
• Keywords (english): palindromes; „ends-out“; illegitimate integration; SLX4 gene; homologous regions
• Language: croatian
• URN:NBN: urn:nbn:hr:159:025295
• 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 10:27:28