Labelling of nucleosides and oligonucleotides by solvatochromic 4-aminophthalimide fluorophore for studying DNA-protein interactions
2012
Abstrakt
Solvatochromic fluorescent 4-aminophthalimide and 4-(N,N-dimethylamino)phthalimide were attached covalently to 2'-deoxycytidine or -adenosine via non-conjugated propargyl linker by Sonogashira cross-coupling reactions. The nucleosides were phosphorylated to triphosphates and enzymatically incorporated to oligonucleotides by DNA polymerases.
API-labelled DNA was used for detection of DNA protein interactions with p53 protein or single strand binding protein. Both proteins changed the polarity around the fluorophore and increased the intensity of API fluorescence.