Synthesis of 2'-O-propargyl nucleoside triphosphates for enzymatic oligonucleotide preparation and "click" modification of DNA with Nile red as fluorescent probe.

摘要: Uridine, adenosine, guanosine, and cytidine that carry a propargyl group attached to the 2′-oxygen were converted efficiently corresponding nucleoside triphosphates (pNTPs). Primer extension experiments revealed pUTP, pATP, pGTP can be successfully incorporated in oligonucleotides so-called 9°N Therminator DNA polymerases. Most importantly, ethynyl as single 2′-modification of enzymatically prepared applied for postsynthetic labeling. This was representatively shown by PAGE analysis after “click”-type cycloaddition with fluorescent nile red azide. These results show 2′-position one most important modification sites is now accessible not only synthetic, but also enzymatic oligonucleotide preparation.