GFP-like Fluorophores as DNA Labels for Studying DNA-Protein Interactions

RIEDL, J, P MENOVA, R POHL, Petr ORSÁG, Miroslav FOJTA and Michal HOCEK. GFP-like Fluorophores as DNA Labels for Studying DNA-Protein Interactions. Journal of Organic Chemistry. WASHINGTON: AMER CHEMICAL SOC, 2012, vol. 77, No 18, p. 8287-8293. ISSN 0022-3263. Available from: https://dx.doi.org/10.1021/jo301684b.

Basic information

• Original name: GFP-like Fluorophores as DNA Labels for Studying DNA-Protein Interactions
• Authors: RIEDL, J, P MENOVA, R POHL, Petr ORSÁG, Miroslav FOJTA and Michal HOCEK.
• Edition: Journal of Organic Chemistry, WASHINGTON, AMER CHEMICAL SOC, 2012, 0022-3263.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 4.564
• Doi: http://dx.doi.org/10.1021/jo301684b
• UT WoS: 000308957700049

Abstract

GFP-like 3,5-difluoro-4-hydroxybenzylideneimidazolinone (FBI) and 3,5-bis(methoxy)-4-hydroxy-benzylideneimidazolinone (MBI) labels were attached to dCTP through a propargyl linker, and the resulting labeled nucleotides (dC(MBI)TP and dC(FBI)TP) were used for a facile enzymatic synthesis of oligonucleotide or DNA probes by polymerase-catalyzed primer extension. The MBI/FBI-labeled DNA probes exerted low fluorescence that was increased 2-3.2 times upon binding of a protein. The concept was demonstrated on sequence-specific binding of p53 to dsDNA and on nonspecific binding of single strand binding protein to an oligonucleotide. The FBI label was also used for a time-resolved experiment monitoring a single-nucleotide incorporation followed by primer extension by Vent(exo-) polymerase.