Synthesis and recognition of novel isonucleoside triphosphates by DNA polymerases

Isonucleosides have been attracting a lot of attention in recent years due to the chemical and enzymatic stability and potential anticancer and antiviral activities. We have reported some of the isonucleosides which exhibited significant anticancer activity and found that the oligonucleotide incorporated with isonucleoside could increase the enzymatic stability against the degradation by phosphodiesterase. In this paper, we investigated the recognition of the isonucleoside triphosphates 1–6 by Taq, Vent(exo−), DeepVent(exo−), 9° Nm, and Therminator DNA polymerases by a non-radioactivity method. We found that most of the isonucleoside triphosphates can be recognized by various DNA polymerase and act as terminators. Isonucleoside triphosphates 2 and 6 can be incorporated as substrates into the primer at 3′ terminus to lengthen the chain dependent on a DNA template by Vent(exo−) and DeepVent(exo−) DNA polymerases.

Isonucleoside triphosphates 2 and 6 can be incorporated as substrates into the primer at 3′ terminus to lengthen the chain on a DNA template by Vent(exo−) and DeepVent(exo−) DNA polymerases.Figure optionsDownload as PowerPoint slide