Author(s)

Wenping Zhu^*, Xinlu Wang, Liyan Dai, Weijie Yang, Yanxia Li, Ruilan Liu

College of Chemistry and Chemical Engineering, Zhoukou Normal University, Zhoukou, China.

Based on the principle that DNA ligation reaction mediates hydrolysis of the dumbbell probe (DP), a novel label-free strategy for sensitive detection of DNA ligase was developed. DNA ligase ligated the nick on DP to generate a fully closed dumbbell DNA structure, which could prevent the hydrolysis of exonuclease, after combining with SYBR Green I (SG I), a strong fluorescent signal was obtained. In the absence of DNA ligase, DP was hydrolyzed into single nucleotides by exonuclease, resulting in a rather weak signal. The linear range of this method for detecting T4 DNA ligase is 0.00004 - 0.004 U/μL, and the detection limit is 0.00003 U/μL. The proposed strategy is sensitive, inexpensive and easy to operate, which may offer an effective tool for further applications in new drug screening.

Dumbbell Probe, DNA Ligase, SYBR Green I, DNA Ligation Reaction, Exonuclease

Zhu, W.P., Wang, X.L., Dai, L.Y., Yang, W.J., Li, Y.X. and Liu, R.L. (2021) A Novel Label-Free Fluorescence Strategy Based on Dumbbell Probe for Sensitive Detection of DNA Ligase. Open Access Library Journal, 8, 1-9. doi: 10.4236/oalib.1107983.