title: Label-free monitoring of DNA methyltransferase activity based on terminal deoxynucleotidyl transferase using a thioflavin T probe.
abstract: We have developed a new methodology for fluorescence turn-on detection of DNA methyltransferase (MTase) activity based on terminal deoxynucleotidyl transferase (TdT) using a thioflavin T probe. This method is highly selective and sensitive. The fluorescence intensity was direct proportion to Dam MTase concentration in the range from 0.1 to 8.0 U/mL with a detection limit of 0.1 U/mL. And because no labeling with a fluorophore-quencher pair was required, it is simple and low cost. We envision that our novel fluorescent detection method for Dam MTase activity could be applied as a useful tool in biomedical research.
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China. Electronic address: macb2012@csu.edu.cn.
authors: Changbei
authors: C
authors: Ma
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors: Haisheng
authors: H
authors: Liu
authors: School of Chemistry and Biological Engineering, Changsha University of Science and Technology, Changsha, 410004, China.
authors: Wei
authors: W
authors: Li
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors: Hanchun
authors: H
authors: Chen
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors: Shunxin
authors: S
authors: Jin
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors: Jing
authors: J
authors: Wang
authors: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China. Electronic address: junwang@csu.edu.cn.
authors: Jun
authors: J
authors: Wang
mesh_terms: D052160:Benzothiazoles; D015374:Biosensing Techniques / Q000379:methods*; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism; D015257:DNA-Cytosine Methylases / Q000378:metabolism*; D005453:Fluorescence; D013844:Thiazoles / Q000737:chemistry
chemical_list: D052160:Benzothiazoles; D013844:Thiazoles; C009462:thioflavin T; D015257:DNA-Cytosine Methylases; D004253:DNA Nucleotidylexotransferase
keywords: DNA methyltransferase; Label-free; Terminal deoxynucleotidyl transferase; Thioflavin T