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Original title:
Utilisation of enzymatic labelling with 4-aminophtalimide and 4-hydroxybenzylideneimidazolinone fluorescent derivates for monitoring of DNA-protein interaction
Authors: Orság, Petr ; Pivoňková, Hana ; Riedl, Jan ; Hocek, Michal ; Fojta, Miroslav
Document type: Papers
Conference/Event: Symposium on Chemistry of Nucleic Acid Components /16./, Český Krumlov (CZ), 2014-06-08 / 2014-06-13
Year: 2014
Language: eng
Abstract: 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.
Keywords: API; DNA-protein interaction; HBI
Project no.: GBP206/12/G151 (CEP), GAP301/11/2076 (CEP), CZ.1.07/2.3.00/30.0019
Funding provider: GA ČR, GA ČR, MŠMT
Host item entry: Chemistry of Nucleic Acid Components. 16th Symposium, ISBN 978-80-86241-50-0
Institution: Institute of Organic Chemistry and Biochemistry AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0234541
Permalink: http://www.nusl.cz/ntk/nusl-174783
The record appears in these collections:
Research > Institutes ASCR > Institute of Organic Chemistry and Biochemistry
Conference materials > Papers
Authors: Orság, Petr ; Pivoňková, Hana ; Riedl, Jan ; Hocek, Michal ; Fojta, Miroslav
Document type: Papers
Conference/Event: Symposium on Chemistry of Nucleic Acid Components /16./, Český Krumlov (CZ), 2014-06-08 / 2014-06-13
Year: 2014
Language: eng
Abstract: 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.
Keywords: API; DNA-protein interaction; HBI
Project no.: GBP206/12/G151 (CEP), GAP301/11/2076 (CEP), CZ.1.07/2.3.00/30.0019
Funding provider: GA ČR, GA ČR, MŠMT
Host item entry: Chemistry of Nucleic Acid Components. 16th Symposium, ISBN 978-80-86241-50-0
Institution: Institute of Organic Chemistry and Biochemistry AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0234541
Permalink: http://www.nusl.cz/ntk/nusl-174783
The record appears in these collections:
Research > Institutes ASCR > Institute of Organic Chemistry and Biochemistry
Conference materials > Papers
Record created 2014-07-10, last modified 2021-11-24