Incorporation of an inducible nucleotide analog into DNA by DNA polymerases

Non-natural nucleotides with diverse functionalities are highly useful in many areas of basic research and practical applications. We have previously developed an efficient method for post-synthetic modifications of 2-amino-6-vinylpurine (AVP)-containing oligonucleotides, which permits conjugations of a variety of useful functional appendages to the AVP moiety in DNA. Here we report an investigation on the ability of various DNA polymerases to use 5′-triphosphate of 2′-deoxyribosyl-2-amino-6-(2-methylthioethyl)purine (a stable precursor of AVP) as the substrate for templated DNA synthesis.

2′-Deoxyribosyl-2-amino-6-(2-methylthioethyl)purine nucleoside 5′-triphosphate (dAVP (SMe)-TP) was examined as a substrate of various DNA polymerases in DNA polymerization reactions to synthesize DNA containing this non-natural functional nucleotide.Figure optionsDownload as PowerPoint slide