title: Synthesis of 2'-O-propargyl nucleoside triphosphates for enzymatic oligonucleotide preparation and "click" modification of DNA with Nile red as fluorescent probe.
abstract: Uridine, adenosine, guanosine, and cytidine that carry a propargyl group attached to the 2'-oxygen were converted efficiently to the corresponding nucleoside triphosphates (pNTPs). Primer extension experiments revealed that pUTP, pATP, and pGTP can be successfully incorporated in oligonucleotides in the so-called 9°N and Therminator DNA polymerases. Most importantly, the ethynyl group as single 2'-modification of the enzymatically prepared oligonucleotides can be applied for postsynthetic labeling. This was representatively shown by PAGE analysis after the "click"-type cycloaddition with the fluorescent nile red azide. These results show that the 2'-position as one of the most important modification sites in oligonucleotides is now accessible not only for synthetic, but also for enzymatic oligonucleotide preparation.
authors: Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), D-76131, Karlsruhe, Germany.
authors: Ulrike
authors: U
authors: Wenge
authors: Thomas
authors: T
authors: Ehrenschwender
authors: Hans-Achim
authors: HA
authors: Wagenknecht
mesh_terms: D000255:Adenosine Triphosphate / Q000138:chemical synthesis* / Q000235:genetics; D057930:Click Chemistry / Q000379:methods*; D004247:DNA / Q000138:chemical synthesis* / Q000235:genetics; D005456:Fluorescent Dyes / Q000138:chemical synthesis; D006160:Guanosine Triphosphate / Q000138:chemical synthesis* / Q000235:genetics; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000235:genetics; D010078:Oxazines / Q000138:chemical synthesis*; D014544:Uridine Triphosphate / Q000138:chemical synthesis* / Q000235:genetics
chemical_list: D005456:Fluorescent Dyes; D009841:Oligonucleotides; D010078:Oxazines; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D004247:DNA; C044808:nile red; D014544:Uridine Triphosphate