Publication details
Labelling of nucleosides and oligonucleotides by solvatochromic 4-aminophthalimide fluorophore for studying DNA-protein interactions
| Authors | |
|---|---|
| Year of publication | 2012 |
| Type | Article in Periodical |
| Magazine / Source | CHEMICAL SCIENCE |
| Citation | |
| Doi | http://dx.doi.org/10.1039/c2sc20404e |
| Description | Solvatochromic fluorescent 4-aminophthalimide (API) and 4-(N, N-dimethylamino) phthalimide (DAPI) were attached covalently to 2'-deoxycytidine or -adenosine via a non-conjugated propargyl linker by Sonogashira cross-coupling reactions of N-propargylphthalimides with halogenated nucleosides. The nucleosides were phosphorylated to triphosphates and enzymatically incorporated into oligonucleotides by DNA polymerases. API-labelled DNA was used for the detection of DNA protein interactions with either the sequence-specific p53 protein or a non-specific single strand binding (SSB) protein. Both proteins changed the polarity around the fluorophore and increased (2-3 fold) the intensity of API fluorescence. |