New method for detection of T4 polynucleotide kinase phosphatase activity through isothermal EXPonential amplification reaction
New method for detection of T4 polynucleotide kinase phosphatase activity through isothermal EXPonential amplification reaction
复制标题
等温指数扩增反应检测T4多核苷酸激酶磷酸酶活性的新方法
DOI：
10.1039/c8an02368a
复制
发表时间：
2019-03-21
期刊：
影响因子：
4.2
通讯作者：
Zhang, Juan
中科院分区：
文献类型：
--
作者：
Chen, Huinan;Wang, Zihan;Zhang, Juan
作为一种双功能酶，T4 多核苷酸激酶磷酸酶 (T4 PNKP) 催化 5'-羟基的磷酸化，并去除末端 3'-磷酸基团。这与核酸的重组、复制和损伤修复密切相关。在本文中，我们描述了一种基于等温指数扩增反应（EXPAR）灵敏检测 T4 PNKP 活性的新方法。 T4 PNKP 可在 0.001 至 0.01 U mL(-1) 范围内进行线性检测，检测限为 7.9 x 10(-4) U mL(-1)。此外，该方法具有较高的特异性和灵敏度，可应用于复杂血清样品的酶分析。鉴于其简单性和温和的实验条件，该方法可能适合在商业试剂盒中用于 T4 PNKP 活性的分析。
As a bifunctional enzyme, T4 polynucleotide kinase phosphatase (T4 PNKP) catalyzes the phosphorylation of 5'-hydroxyl, and also removes the terminal 3'-phosphate group. This is closely related to the restructuring, replication, and damage repair of nucleic acid. In this paper, we describe a new method for the sensitive detection of T4 PNKP activity based on the isothermal EXPonential amplification reaction (EXPAR). T4 PNKP can be linearly assayed in the range from 0.001 to 0.01 U mL(-1) with a detection limit of 7.9 x 10(-4) U mL(-1). Moreover, the method exhibits high specificity and sensitivity and can be applied in the enzyme analysis of complex serum samples. In view of its simplicity and moderate experimental conditions, the method may suitable for use in a commercial kit for the analysis of T4 PNKP activity.