Quencher-free hairpin probes for real-time detection of T4 polynucleotide kinase activity |
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| Authors: | Changbei Ma Haisheng Liu Junyan Du Hanchun Chen Hailun He Shunxin Jin Kemin Wang Jun Wang |
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| Affiliation: | 1. State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China;2. State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, China |
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| Abstract: | Traditional methods of assaying polynucleotide kinase (PNK) activity are discontinuous, time-consuming, and laborious. Here we report a new quencher-free approach to real-time monitoring of PNK activity using a 2-aminopurine probe. When the 2-aminopurine probe was 5′-phosphorylated by PNK, it could be efficiently degraded by lambda exonuclease to release free 2-aminopurine molecules and generate a fluorescence signal. This method not only provides a universal approach to real-time monitoring of PNK activity, but also shows great potential for screening suitable inhibitor drugs for PNK. |
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| Keywords: | 2-Aminopurine Quencher-free Polynucleotide kinase activity Inhibitor |
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