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Vpliv elektroporacije na vnos antibiotikov in iztekanje DNA pri bakterijah
ID Lovšin, Žana (Author), ID Kotnik, Tadej (Mentor) More about this mentor... This link opens in a new window, ID Klančnik, Anja (Comentor)

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Abstract
Kombiniranje elektroporacije s protimikrobnimi snovmi omogoči, da dosežemo višjo stopnjo inaktivacije bakterij, kot če bi uporabili le eno metodo. Naš namen je bil preveriti, kako na potenciacijo inaktivacije bakterij s takšnim kombiniranim pristopom vpliva način delovanja antibiotika, ter kakšen vpliv ima sestava celične stene grampozitivnih in gramnegativnih bakterij. Za boljše temeljno razumevanje te potenciacije smo vzpostavili bolj sistematičen pristop, ki nam je omogočil ločitev vplivov posameznih parametrov. Elektroporacija je najbolj povečala učinkovitost antibiotikov, ki potrebujejo transport preko celičnih ovojnic, da dosežejo svojo tarčo. Pri bakterijah Escherichia coli je bil to ampicilin, ki mora preiti le preko zunanje membrane, pri bakterijah Lactiplantibacillus plantarum pa tetraciklin, ki mora preiti celotno celično steno. V manjši meri potenciacijo z elektroporacijo dosežemo tudi za antibiotike, ki ne potrebujejo transporta preko celičnih ovojnic, torej so poleg povečanega transporta antibiotika pomembne tudi poškodbe membrane in iztekanje celične vsebine. Preverili smo tudi vpliv električnih pulzov na strukturo plazmidne in genomske DNA. Izpostavitev plazmidne DNA električnim pulzom je povzročila predvsem odvitje iz dodatno zvitega kovalentno zaprtega kroga v obliko odprtega kroga, delež teh sprememb je bil odvisen od dovedene energije. Izpostavitev genomske DNA električnim pulzom pa je vodila v njeno fragmentacijo, vendar delež teh sprememb ni bil odvisen od parametrov elektroporacije ali dovedene energije.

Language:Slovenian
Keywords:elektroporacija, antibiotiki, sinergija, Escherichia coli, Lactiplantibacillus plantarum, inaktivacija bakterij, dinamika rasti, iztekanje DNA, fragmentacija DNA
Work type:Doctoral dissertation
Typology:2.08 - Doctoral Dissertation
Organization:BF - Biotechnical Faculty
Publisher:[Ž. Lovšin]
Year:2024
PID:20.500.12556/RUL-159314 This link opens in a new window
UDC:602.621:577.18(043.3)
COBISS.SI-ID:200955907 This link opens in a new window
Publication date in RUL:06.07.2024
Views:237
Downloads:75
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Secondary language

Language:English
Title:The effect of bacterial electroporation on antibiotic uptake and DNA release
Abstract:
Combining electroporation with antimicrobials allows to achieve higher bacterial inactivation than if only one method is used. Our purpose was to determine the effect of antibiotic’s mode of action on its potentiation by electroporation and the effect of gram-positive and gram-negative bacteria cell wall structure. To gain an improved understanding of the mechanisms underlying this potentiation, we established a more systematic approach, which enabled us to separate the influence of individual parameters. Highest potentiation by electroporation was achieved for antibiotics that require transport across the cell wall envelopes to reach their target. For Escherichia coli bacteria, this was ampicillin, which only has to pass through the outer membrane, and for Lactiplantibacillus plantarum bacteria, this was tetracycline, which has to pass across the entire cell wall. To a lesser extent, potentiation by electroporation was also achieved for antibiotics that do not require transport across the cell envelopes, which means that in addition to increased antibiotic transport, membrane damage and leakage of cell contents are also important. We also determined the influence of electric pulses on the structure of plasmid and genomic DNA. Exposing the plasmid DNA to electric pulses mainly caused unwinding from an additionally coiled covalently closed circle to an open circle shape, the proportion of these changes depended on the delivered energy. Exposing the genomic DNA to electric pulses led to its fragmentation, however, the proportion of these changes did not depend on the electroporation parameters or the delivered energy.

Keywords:electroporation, antibiotics, synergy, Escherichia coli, Lactiplantibacillus plantarum, bacterial inactivation, growth dynamic, DNA leakage, DNA fragmentation

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