Double-Stranded DNA-Templated Copper Nanoclusters for Detection of DNA Polymerase Activity

Abstract

DNA polymerase plays an important role in the proliferation and survival of all forms of life and the variety of biotechnological applications. In this study, DNA-templated copper nanoclusters were utilized to detect Klenow fragment exo- (KF-) activity and monitor its inhibition in the presence of 5-fluorouracil by measurement of fluorescence intensities. This study showed that the fluorescence intensities increased continuously for KF- concentrations ranging from 0 to 5 U/mL. In addition, inhibition of KF- activity was measured at 5-fluorouracil concentrations ranging from 0 to 150 mu M. Using nonlinear regression, the corresponding IC50 was determined to be 6.23 mu M in the presence of 5.0 U/mL KF-.