A rapid and sensitive method for kinetic study and activity assay of DNase I in vitro based on a GO-quenched hairpin probe

作者：Xu, Wei; Xie, Zhenhua; Tong, Chunyi^*; Peng, Lan; Xiao, Changhui; Liu, Xuanming; Zhu, Yonghua; Liu, Bin^*

来源：Analytical and Bioanalytical Chemistry, 2016, 408(14): 3801-3809.

DOI：10.1007/s00216-016-9474-3

• DNase I
• Graphene oxide
• Hairpin probe
• Real time
• Tumor

As a waste-management endonuclease, DNase I has been suggested to be one of the deoxyribonucleases responsible for DNA fragmentation during apoptosis. We report here an alternative fluorescence method for DNase I assay with high accuracy and sensitivity by applying a DNA/GO (graphene oxide) probe. The method with a detection limit of 1 U mL(-1) was then applied to investigate the effects of external factors including antibiotics and heavy metal ions on DNase I. The results demonstrated that gentamicin sulfate was a strong inhibitor with an IC50 value of 0.57 +/- 0.12 mM. The investigated heavy metal ions showed an inhibitory effect on DNase I activity in a concentration dependent manner with IC50 values of 0.04 mu g/mL (Hg2+), 0.10 mu g/mL (Pb2+), 1.35 mu g/mL (Cd2+), 1.20 mu g/mL (As2+), and 1.80 mu g/mL (Cu2+). Finally, the new method was applied to detect DNase levels in complicated tumor tissue and cell samples and the results showed that DNase levels increased in tumor tissues compared with that of adjacent tissue. From the above results, we conclude that the method can be widely used for high - throughput assay of DNase I in biological samples as well as drug screening in vitro.

• 出版日期2016-5
• 单位湖南大学