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Anal Biochem. 2013 Dec 15;443(2):166-8. doi: 10.1016/j.ab.2013.09.006. Epub 2013 Sep 10.

Simultaneous detection of kinase and phosphatase activities of polynucleotide kinase using molecular beacon probes.

Author information

1
School of Life Sciences, The State Key Labratory of Medical Genetics, Central South University, Changsha 410013, China. Electronic address: macb2012@csu.edu.cn.

Abstract

Phosphorylation and dephosphorylation of DNA by polynucleotide kinase (PNK) has an important role in DNA damage repair, replication, and recombination. Traditionally, it is assayed by denaturing gel electrophoresis and autoradiography, which are tedious and not sensitive. We report on the development of a sensitive and simple method for PNK assay based on DNA ligation using a molecular beacon. Enzyme activity of PNK is measured down to a limit of 0.002 unit/ml. The method not only provides a universal platform for simultaneous monitoring of kinase and phosphatase activities, but also shows great potential in biological research, drug discovery, and clinical diagnostics.

KEYWORDS:

Kinase and phosphatase activities; Molecular beacon; Polynucleotide kinase

PMID:
24036036
DOI:
10.1016/j.ab.2013.09.006
[Indexed for MEDLINE]
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