A fluorescence-based assay for T4 polynucleotide kinase/phosphatase activity based on a terminal transferase-aided photoinduced electron transfer strategy

Abstract

We have developed a new assay for the detection of T4 polynucleotide kinase/phosphatase activity (T4 PNKP) based on a terminal transferase-aided photoinduced electron transfer (PIET) strategy. The method is highly sensitive and the T4 PNKP detection limit was estimated to be 0.01 U mL⁻¹, which is superior or comparable to previously reported assays. Furthermore, the proposed method was also applied to assay the inhibition of T4 PNKP activity. This approach may offer potential applications in drug screening, clinical diagnostics and some other related biomedical research.