Ma Changbei, Liu Haisheng, Li Wei, Chen Hanchun, Jin Shunxin, Wang Jing, Wang Jun
State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
Mol Cell Probes. 2016 Apr;30(2):118-21. doi: 10.1016/j.mcp.2016.02.001. Epub 2016 Feb 5.
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.
我们基于末端脱氧核苷酸转移酶（TdT），利用硫黄素T探针开发了一种用于DNA甲基转移酶（MTase）活性的荧光开启检测新方法。该方法具有高度的选择性和灵敏性。在0.1至8.0 U/mL范围内，荧光强度与Dam MTase浓度成正比，检测限为0.1 U/mL。并且由于无需用荧光团-猝灭剂对进行标记，所以该方法简单且成本低。我们设想，我们用于Dam MTase活性的新型荧光检测方法可作为生物医学研究中的一种有用工具。
