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Biochemical analyses of the DNA interaction partners
Valchová, Michaela ; Coufal,, Jan (referee) ; Brázda, Václav (advisor)
This thesis was focused on DNA analysis. The fluorescently labelled oligonucleotides were at first hybridised and subsequently analysed by HRM analysis to determine the melting temperatures of the oligonucleotides depending on the environment. This thesis describes the change of melting temperature of oligonucleotides in environments containing mono and bivalent ions and the influence of protein binding on the stability of these DNA structures. From determined melting points, it was specified whether the ion/protein stabilised or destabilised the oligonucleotide. Furthermore, plasmids were isolated and analyzed by atomic force microscopy.
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Significance, occurrence and determinants of horizontally transmissible colistin resistance in Gram negative bacteria
Kislíková, Karolína ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
Colistin, also known as polymyxin E, is antibiotics active against most of Gram-negative bacteria. In the pas decade, emergency of multidrug-resistant bacteria led to increase of colistin administration as a last resort antibiotic for human infections. The first plasmid-mediated colistin resistance gene mcr-1 was identified in 2015 in animals in China and after first detection, additional mcr genes: mcr-2, mcr-3, mcr-4, mcr-5, mcr-6, mcr-7 a mcr-8 were described throughout the world. The aim of this thesis was to clarify whether there is horizontal transmission colistin resistance encoded by the mcr genes in gram-negative bacteria isolated from the environment, animals and their breeding and food. The mcr-1 gene was detected in 2 strains Escherichia coli isolated from waste water. The mcr-4 gene was detected in 1 strain Shewanella putrefaciens isolate obtained from the lake. The environment is the most important source and way of spreading this type of resistance in the Czech Republic.
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Significance, occurrence and determinants of horizontally transmissible colistin resistance in Gram negative bacteria
Kislíková, Karolína ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
Colistin, also known as polymyxin E, is antibiotics active against most of Gram-negative bacteria. In the pas decade, emergency of multidrug-resistant bacteria led to increase of colistin administration as a last resort antibiotic for human infections. The first plasmid-mediated colistin resistance gene mcr-1 was identified in 2015 in animals in China and after first detection, additional mcr genes: mcr-2, mcr-3, mcr-4, mcr-5, mcr-6, mcr-7 a mcr-8 were described throughout the world. The aim of this thesis was to clarify whether there is horizontal transmission colistin resistance encoded by the mcr genes in gram-negative bacteria isolated from the environment, animals and their breeding and food. The mcr-1 gene was detected in 2 strains Escherichia coli isolated from waste water. The mcr-4 gene was detected in 1 strain Shewanella putrefaciens isolate obtained from the lake. The environment is the most important source and way of spreading this type of resistance in the Czech Republic.
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Biochemical analyses of the DNA interaction partners
Valchová, Michaela ; Coufal,, Jan (referee) ; Brázda, Václav (advisor)
This thesis was focused on DNA analysis. The fluorescently labelled oligonucleotides were at first hybridised and subsequently analysed by HRM analysis to determine the melting temperatures of the oligonucleotides depending on the environment. This thesis describes the change of melting temperature of oligonucleotides in environments containing mono and bivalent ions and the influence of protein binding on the stability of these DNA structures. From determined melting points, it was specified whether the ion/protein stabilised or destabilised the oligonucleotide. Furthermore, plasmids were isolated and analyzed by atomic force microscopy.
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