Real-time Monitoring of Nucleotide Excision of Molecular Beacon Catalyzed by Klenow Fragment  

作　　者：LIU Bin, YANG Xiao-hai, WANG Ke-min and TAN Wei-hong State Key Laboratory of Chemo/Biosensing and Chemometrics, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, College of Chemistry and Chemical Engineering, College of Biology, Hunan University, Changsha 410082, P. R. China 

出　　处：《Chemical Research in Chinese Universities》2012年第1期37-40,共4页高等学校化学研究（英文版）

基　　金：Supported by the Key Project of Natural Science Foundation of China(No.90606003);the Hunan Provincial Natural Science Foundation, China(No.08JJ1002);the National High-Tech Research and Development Program(No.2007AA022007);the Changjiang Scholars and Innovative Research Team in University, China

摘　　要：Klenow fragment（KF） uses the activity of a separate exonuclease to excise nucleotide, which is a crucial step in DNA replication and repair. Here is a novel sensitive and convenient method introduced for real-time monitoring nucleotide excision by KF with a molecular beacon as a detecting probe in a homogeneous solution. This method, which overcomes the drawbacks of traditional methods such as discontinuity, time consuming and low sensitivity, was used to assay KF activity and the detection limit reached up to 0.4 U/mL. In addition, the method was applied to investigating the effects of metal ions and chemical drugs on the reaction. The results demonstrate that it is a potential high-throughput assay for screening inhibitors and activity analysis of KF in vitro.Klenow fragment（KF） uses the activity of a separate exonuclease to excise nucleotide, which is a crucial step in DNA replication and repair. Here is a novel sensitive and convenient method introduced for real-time monitoring nucleotide excision by KF with a molecular beacon as a detecting probe in a homogeneous solution. This method, which overcomes the drawbacks of traditional methods such as discontinuity, time consuming and low sensitivity, was used to assay KF activity and the detection limit reached up to 0.4 U/mL. In addition, the method was applied to investigating the effects of metal ions and chemical drugs on the reaction. The results demonstrate that it is a potential high-throughput assay for screening inhibitors and activity analysis of KF in vitro.

关 键 词：Real-time monitoring Klenow fragment Molecular beacon 

分 类 号：O643.32[理学—物理化学]