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Název:
Utilisation of enzymatic labelling with 4-aminophtalimide and 4-hydroxybenzylideneimidazolinone fluorescent derivates for monitoring of DNA-protein interaction
Autoři: Orság, Petr ; Pivoňková, Hana ; Riedl, Jan ; Hocek, Michal ; Fojta, Miroslav
Typ dokumentu: Příspěvky z konference
Konference/Akce: Symposium on Chemistry of Nucleic Acid Components /16./, Český Krumlov (CZ), 2014-06-08 / 2014-06-13
Rok: 2014
Jazyk: eng
Abstrakt: The 5’-substituted deoxycytosine triphosphates with conjugated solvatochromic derivates of 4-aminophtalimide (API) and derivates of the green fluorescent protein, 4-hydroxybenzylideneimidazolinone (HBI) were synthetized and successfully tested for enzymatic incorporation using primer extension assay. Site specifically labelled oligonucleotide probes were prepared and tested for interaction with p53 and SSB proteins, displaying distinct DNA-binding properties. The incorporation of multiple fluorescent labels did not interfere with natural protein binding and protein interaction leaded in both cases the to the gradual ratiometric increase of the fluorescence intensity moreover accompanied with the changes of the fluorescence emission spectra profile. Neither effect was observed after incubation with BSA, non-DNA binding protein, confirming the specificity of the interaction. Modified nucleoside triphosphates with conjugated fluorescence labels derivates of API and HBI can be used as substrates for preparation of the specific oligonucleotide labelled probes and provide the novel tool for studying and monitoring the DNA-protein interaction.
Klíčová slova: API; DNA-protein interaction; HBI
Číslo projektu: GBP206/12/G151 (CEP), GAP301/11/2076 (CEP), CZ.1.07/2.3.00/30.0019
Poskytovatel projektu: GA ČR, GA ČR, MŠMT
Zdrojový dokument: Chemistry of Nucleic Acid Components. 16th Symposium, ISBN 978-80-86241-50-0
Instituce: Ústav organické chemie a biochemie AV ČR (web)
Informace o dostupnosti dokumentu: Dokument je dostupný v příslušném ústavu Akademie věd ČR.
Původní záznam: http://hdl.handle.net/11104/0234541
Trvalý odkaz NUŠL: http://www.nusl.cz/ntk/nusl-174783
Záznam je zařazen do těchto sbírek:
Věda a výzkum > AV ČR > Ústav organické chemie a biochemie
Konferenční materiály > Příspěvky z konference
Autoři: Orság, Petr ; Pivoňková, Hana ; Riedl, Jan ; Hocek, Michal ; Fojta, Miroslav
Typ dokumentu: Příspěvky z konference
Konference/Akce: Symposium on Chemistry of Nucleic Acid Components /16./, Český Krumlov (CZ), 2014-06-08 / 2014-06-13
Rok: 2014
Jazyk: eng
Abstrakt: The 5’-substituted deoxycytosine triphosphates with conjugated solvatochromic derivates of 4-aminophtalimide (API) and derivates of the green fluorescent protein, 4-hydroxybenzylideneimidazolinone (HBI) were synthetized and successfully tested for enzymatic incorporation using primer extension assay. Site specifically labelled oligonucleotide probes were prepared and tested for interaction with p53 and SSB proteins, displaying distinct DNA-binding properties. The incorporation of multiple fluorescent labels did not interfere with natural protein binding and protein interaction leaded in both cases the to the gradual ratiometric increase of the fluorescence intensity moreover accompanied with the changes of the fluorescence emission spectra profile. Neither effect was observed after incubation with BSA, non-DNA binding protein, confirming the specificity of the interaction. Modified nucleoside triphosphates with conjugated fluorescence labels derivates of API and HBI can be used as substrates for preparation of the specific oligonucleotide labelled probes and provide the novel tool for studying and monitoring the DNA-protein interaction.
Klíčová slova: API; DNA-protein interaction; HBI
Číslo projektu: GBP206/12/G151 (CEP), GAP301/11/2076 (CEP), CZ.1.07/2.3.00/30.0019
Poskytovatel projektu: GA ČR, GA ČR, MŠMT
Zdrojový dokument: Chemistry of Nucleic Acid Components. 16th Symposium, ISBN 978-80-86241-50-0
Instituce: Ústav organické chemie a biochemie AV ČR (web)
Informace o dostupnosti dokumentu: Dokument je dostupný v příslušném ústavu Akademie věd ČR.
Původní záznam: http://hdl.handle.net/11104/0234541
Trvalý odkaz NUŠL: http://www.nusl.cz/ntk/nusl-174783
Záznam je zařazen do těchto sbírek:
Věda a výzkum > AV ČR > Ústav organické chemie a biochemie
Konferenční materiály > Příspěvky z konference
Záznam vytvořen dne 2014-07-10, naposledy upraven 2021-11-24.