. "DNA undergoes conformational transitions which substantially change its biophysical and molecular biological properties. The mostly used method of research into DNA conformational transitions is CD spectroscopy, Which nevertheless is not suitable for mapping the transitions along segments of genomic DNA, as it provides a picture of the whole molecule behaviour. In this project we intend to develop two complementary methods based on the fact that UV-induced DNA damage is DNA conformation dependent. We will detect photoproducts via primer extension method, which gives information at single nucleotide resolution, but in segments of hundreds nucleotides only. To detect the transitions along kilobase DNA molecules, the fact that damaged DNA inhibits restrictases cleavage will be used. These methods will be applied to the research into the conformational in plasmid pUC19-preferentially for B-A transition and for negative and positive superhelicity induced transitions."@en . . . . "DNA vykazuje konforma\u010Dn\u00ED p\u0159echody, kter\u00E9 v\u00FDrazn\u011B m\u011Bn\u00ED jej\u00ED biofyzik\u00E1ln\u00ED a molekul\u00E1rn\u011B biologick\u00E9 vlastnosti. Nej\u010Dast\u011Bji pou\u017E\u00EDvanou metodou ke studiu konforma\u010Dn\u00EDch p\u0159echod\u016F v DNA je CD spektroskopie. Ta ale nen\u00ED vhodn\u00E1 pro mapov\u00E1n\u00ED konforma\u010Dn\u00EDch p\u0159echod\u016F pod\u00E9l dlouh\u00FDch \u00FAsek\u016F genomov\u00E9 DNA, proto\u017Ee poskytuje integr\u00E1ln\u00ED obraz o chov\u00E1n\u00ED cel\u00E9 molekuly. V tomto projektu chceme vyvinout dv\u011B komplement\u00E1rn\u00ED metody zalo\u017Een\u00E9 na skute\u010Dnosti, \u017Ee po\u0161kozen\u00ED vyvolan\u00E9 na DNA UV sv\u011Btlem z\u00E1vis\u00ED na konformaci DNA. K detekci fotoprodukt\u016F vyu\u017Eijeme metodu primer extension, kter\u00E1 d\u00E1v\u00E1 informace s nukleotidov\u00FDm rozli\u0161en\u00EDm, ale jen v \u00FAsec\u00EDch o d\u00E9lce stovek nukleotid\u016F. K detekci konforma\u010Dn\u00EDch p\u0159echod\u016F pod\u00E9l kilob\u00E1zov\u00FDch molekul DNA vyu\u017Eijeme skute\u010Dnost, \u017Ee po\u0161kozen\u00ED DNA blokuje jej\u00ED \u0161t\u011Bpen\u00ED restrikt\u00E1zami. Tyto metody vyu\u017Eijeme ke studiu konforma\u010Dn\u00EDch p\u0159echod\u016F v molekule plasmidu pUC19. P\u0159ednostn\u011B se zam\u011B\u0159\u00EDme na p\u0159echor B-A a na p\u0159echody indukovan\u00E9 negativn\u00EDm a pozitivn\u00EDm nad\u0161roubovicov\u00FDm vinut\u00EDm."@cs . "751"^^ . "0"^^ . "1"^^ . "3"^^ . "Conformational Transitions in Plasmid DNA"@en . . . "3"^^ . "conformational transitions; UV light; plasmid DNA; supercoiling; photoproducts; primer extension; restrictases; CD spectroscopy"@en . "751"^^ . . . . . "Konforma\u010Dn\u00ED p\u0159echody v plasmidov\u00E9 DNA"@cs . "0"^^ . .