用于实时检测T4多核苷酸激酶活性的无猝灭剂发夹探针。
Quencher-free hairpin probes for real-time detection of T4 polynucleotide kinase activity.
作者信息
Ma Changbei, Liu Haisheng, Du Junyan, Chen Hanchun, He Hailun, Jin Shunxin, Wang Kemin, Wang Jun
机构信息
State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China; State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, China.
State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
出版信息
Anal Biochem. 2016 Feb 1;494:1-3. doi: 10.1016/j.ab.2015.10.007. Epub 2015 Oct 28.
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.
传统的多核苷酸激酶（PNK）活性检测方法是不连续的，耗时且费力。在此，我们报告一种使用2-氨基嘌呤探针实时监测PNK活性的无淬灭剂新方法。当2-氨基嘌呤探针被PNK 5'-磷酸化时，它可被λ核酸外切酶有效降解，释放出游离的2-氨基嘌呤分子并产生荧光信号。该方法不仅提供了一种实时监测PNK活性的通用方法，而且在筛选适合PNK的抑制剂药物方面也显示出巨大潜力。
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