Fotodinamička inaktivacija u bakterije Escherichia coli

master's thesis

• Title (english): Photodynamic inactivation in Escherichia coli
• Author: Nataša Gašpar
• Mentor: Antonio Starčević (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Section for Bioinformatics); Zagreb
• Defense date and country: 2014-12-16, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Cilj ovoga rada je bio analizirati E.coli K-12 mutante u fotodinamičkoj inaktivaciji. Korišteni sojevi potječu iz Keio kolekcije i imaju inaktiviran jedan gen za sintezu dijela lipopolisaharidnog omotača (rfaC, rfaE, lpcA). Glavni cilj rada je bila konstrukcija duplih mutanata koji imaju u isto vrijeme inaktiviran za jedan od spomenutih gena i inaktivaciju za transportni protein TolC. Dotični geni su se pokazali posebno osetljivima u fotodinamičkoj inaktivaciji s klorinom E6. Budući da spomenuti mutanti imaju fenotip ˝deep rough˝ i ne mogu biti inficirani s P1 bakteriofagom, standardna genetička transdukcija posredovana P1 bakteriofagom se ne može koristiti za konstrukciju mutanata, već alternativna metoda bazirana na sustavu λ-crvene rekombinaze. Koristeći dobivene rezultate, odnos između bakterijske stanične površine (lipopolisaharidni omotač) i njegov utjecaj na preživljavanje u fotodinamičkoj inaktivaciji, trebao bi biti razjašnjen. Rezultati ovoga rada trebali bi unaprijediti metodu fotodinamičke inaktivacije u borbi protiv gram-negativnih bakterija.

Abstract (english)

The aim of this thesis is to analyze lipopolysaccharide (LPS) E. coli K-12 mutants in photodynamic inactivation (PDI). The used strains are from the Keio collection and have single knockout genes for the lipopolysaccharide bilayer (rfaC, rfaE, lpcA). Goal of this thesis was to create double knockout mutants which have a knockout for one of these mentioned LPS genes and at the same time a tolC deletion. Those three genes are particularly sensitive in PDI with chlorine E6 (CE6). Since the LPS mutants have a ˝deep rugh˝ phenotype and can`t be infected with P1 bacteriophage, the standard genetic transduction couldn`t be used and different methods had to be performed, the λ-red recombinase system. Using the obtained results, the relationship between the composition of the bacterial cell surface and the survival in the PDI should be clarified. The results of this work should improve the PDI method against the Gram-negative bacteria.

• Keywords: lipopolisaharidni omotač; fotodinamička inaktivacija; klorin E6; Keio kolekcija; sustav λ- crvene rekombinaze
• Keywords (english): Lipopolysaccharide bilayer; Photodynamic inactivation; Chlorine E6; Keio collection; λ-red recombinase system
• Language: english
• URN:NBN: urn:nbn:hr:159:941055
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-26 12:15:57