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Study of nucleic-acid structure and interactions by resonance Raman scattering
Klener, Jakub ; Štěpánek, Josef (advisor)
Despite the decades of intensive research, nucleic acids represent still a permanent object of structural studies. Within the framework of the doctoral work, the apparatus for measurement of UV excited resonance Raman spectra (UV RRS) was built up and optimized. A realistic and complex interpretation table was prepared based on analysis of published data and extensive series of UV RRS measurements on NA model structures, mononucleotides, and polynucleotides. The established methodology was verified when applied in several structural studies of nucleic acids, mainly the study of the influence of magnesium ions on the equilibrium between duplexes and triplexes formed by PolyA and PolyU homopolynucleotides, a study of temperature-induced structural changes in DNA double helix and DNA hairpin, and investigation of slow structural transitions of guanine quadruplexes induced by the presence of potassium ions. The results of the test measurements and the above-mentioned studies have shown that the created methodology for studying UV RRS of nucleic acids brings most of the expected benefits of the resonance excitation: the possibility of Raman scattering measurements at the same concentrations as in the case of UV absorption, high sensitivity to fine temperature-induced structural changes and good interpretability...
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Study of nucleic-acid structure and interactions by resonance Raman scattering
Klener, Jakub ; Štěpánek, Josef (advisor) ; Michl, Martin (referee) ; Vlčková, Blanka (referee)
Despite the decades of intensive research, nucleic acids represent still a permanent object of structural studies. Within the framework of the doctoral work, the apparatus for measurement of UV excited resonance Raman spectra (UV RRS) was built up and optimized. A realistic and complex interpretation table was prepared based on analysis of published data and extensive series of UV RRS measurements on NA model structures, mononucleotides, and polynucleotides. The established methodology was verified when applied in several structural studies of nucleic acids, mainly the study of the influence of magnesium ions on the equilibrium between duplexes and triplexes formed by PolyA and PolyU homopolynucleotides, a study of temperature-induced structural changes in DNA double helix and DNA hairpin, and investigation of slow structural transitions of guanine quadruplexes induced by the presence of potassium ions. The results of the test measurements and the above-mentioned studies have shown that the created methodology for studying UV RRS of nucleic acids brings most of the expected benefits of the resonance excitation: the possibility of Raman scattering measurements at the same concentrations as in the case of UV absorption, high sensitivity to fine temperature-induced structural changes and good interpretability...
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