Highly sensitive fluorescence assay of DNA methyltransferase activity by methylation-sensitive cleavage-based primer generation exponential isothermal amplification-induced G-quadruplex formation

文献类型： 外文期刊

第一作者： Xue, Qingwang

作者： Xue, Qingwang;Lv, Yanqin;Xu, Shuling;Zhang, Yuanfu;Wang, Lei;Li, Rui;Yue, Qiaoli;Li, Haibo;Liu, Jifeng;Gu, Xiaohong;Zhang, Shuqiu;Liu, Jifeng

作者机构：

关键词： Fluorescence assay;Methyltransferase;Label-free;EXPA;Supramolecular ZnPPIX/G-quadruplex

期刊名称：BIOSENSORS & BIOELECTRONICS （ 影响因子：10.618； 五年影响因子：9.323 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Site-specific identification of DNA methylation and assay of MTase activity are imperative for determining specific cancer types, provide insights into the mechanism of gene repression, and develop novel drugs to treat methylation-related diseases. Herein, we developed a highly sensitive fluorescence assay of DNA methyltransferase by methylation-sensitive cleavage-based primer generation exponential isothermal amplification (PG-EXPA) coupled with supramolecular fluorescent Zinc(II)-protoporphyrin IX (ZnPPIX)/G-quadruplex. In the presence of DNA adenine methylation (Dam) MTase, the methylation-responsive sequence of hairpin probe is methylated and cleaved by the methylation-sensitive restriction endonuclease Dpn I. The cleaved hairpin probe then functions as a signal primer to initiate the exponential isothermal amplification reaction (EXPAR) by hybridizing with a unimolecular DNA containing three functional domains as the amplification template, producing a large number of G-quadruplex nanostructures by utilizing polymerases and nicking enzymes as mechanical activators. The G-quadruplex nanostructures act as host for ZnPPIX that lead to supramolecular complexes ZnPPIX/G-quadruplex, which provides optical labels for amplified fluorescence detection of Dam MTase. While in the absence of Dam MTase, neither methylation/cleavage nor PG-EXPA reaction can be initiated and no fluorescence signal is observed. The proposed method exhibits a wide dynamic range from 0.0002 to 20 U/mL and an extremely low detection limit of 8.6 x 10(-5) U/mL, which is superior to most conventional approaches for the MTase assay. Owing to the specific site recognition of MTase toward its substrate, the proposed sensing system was able to readily discriminate Dam MTase from other MTase such as M.SssI and even detect the target in a complex biological matrix. Furthermore, the application of the proposed sensing strategy for screening Dam MTase inhibitors was also demonstrated with satisfactory results. This novel method not only provides a promising platform for monitoring activity and inhibition of DNA MTases, but also shows great potentials in biological process researches, drugs discovery and clinical diagnostics. (C) 2014 Elsevier B.V. All rights reserved.

分类号： Q

  • 相关文献

[1]A Comparative Analysis of DNA Methylation in Diploid and Tetraploid Apple (Malus x domestica Borkh.). He, Ping,Cheng, Lailiang,Li, Huifeng,Wang, Haibo,Li, Linguang. 2017

[2]Screening of strain-specific Actinobacillus pleuropneumoniae genes using a combination method. Lei, Liancheng,Wang, Jiaqi,Lei, Liancheng,Du, Chongtiao,Yang, Peng,Xie, Fang,Ou, Pingyang,Han, Wenyu.

[3]Establishment of an efficient transformation system for Pleurotus ostreatus. Lei, Min,Wu, Xiangli,Zhang, Jinxia,Huang, Chenyang,Lei, Min,Wang, Hexiang. 2017

[4]Development of a simple and efficient transformation system for the basidiomycetous medicinal fungus Ganoderma lucidum. Shi, Liang,Fang, Xing,Li, Mengjiao,Mu, Dashuai,Ren, Ang,Zhao, Mingwen,Tan, Qi. 2012

[5]Dual Reaction-Based Multimodal Assay for Dopamine with High Sensitivity and Selectivity Using Functionalized Gold Nanoparticles. Zeng, Zhanghua,Cui, Bo,Wang, Yan,Sun, Changjiao,Zhao, Xiang,Cui, Haixin. 2015

[6]Competitive fluorescence assay for specific recognition of atrazine by magnetic molecularly imprinted polymer based on Fe3O4-chitosan. Liu, Guangyang,Yang, Xin,Wang, Jing,Zhang, Min,Liu, Guangyang,Li, Tengfei,She, Yongxin,Wang, Miao,Wang, Jing,Wang, Shanshan,Jin, Fen,Jin, Maojun,Shao, Hua,Jiang, Zejun,Yu, Hailong.

[7]Aptamer based ultrasensitive determination of the beta-adrenergic agonist ractopamine using PicoGreen as a fluorescent DNA probe. Zhu, Chao,Zhang, Guilan,Huang, Yafei,Yan, Jiao,Chen, Ailiang,Zhu, Chao,Zhang, Guilan,Huang, Yafei,Yan, Jiao,Chen, Ailiang,Huang, Yafei,Yan, Jiao.

[8]Prohibitin-2 promotes hepatocellular carcinoma malignancy progression in hypoxia based on a label-free quantitative proteomics strategy. Cheng, Jun,Wu, Jian,Wu, Liming,Xie, Haiyang,Zheng, Shusen,Zhou, Lin,Cheng, Jun,Gao, Feng,Xing, Chunyang,Lv, Zhen,Wu, Liming,Ye, Sunyi,Xie, Haiyang,Zheng, Shusen,Zhou, Lin,Gao, Feng,Chen, Xiang,Xu, Wei,Xie, Qing.

[9]A simple and sensitive label-free fluorescent approach for protein detection based on a Perylene probe and aptamer. Lv, Zhenzhen,Liu, Jinchuan,Bai, Wenhui,Yang, Shuming,Chen, Ailiang,Lv, Zhenzhen,Liu, Jinchuan,Bai, Wenhui,Yang, Shuming,Chen, Ailiang,Lv, Zhenzhen.

[10]Comparative proteomics of matrix fractions between pimpled and normal chicken eggshells. Liu, Zhangguo,Song, Lingzi,Zhang, Xianfu,Lu, Lizhi,Zhang, Fuming,Linhardt, Robert J.,Zhang, Fuming,Linhardt, Robert J.,Zhang, Fuming,Linhardt, Robert J.,Zhang, Fuming,Linhardt, Robert J.,Wang, Kehua.

[11]A label-free SERS approach to quantitative and selective detection of mercury (II) based on DNA aptamer-modified SiO2@Au core/shell nanoparticles. Lu, Yilin,Zhong, Jie,Yao, Guohua,Huang, Qing,Zhong, Jie,Huang, Qing,Lu, Yilin. 2018

[12]Label-free quantitative proteomics analysis of cotton leaf response to nitric oxide. Yanyan Meng,Feng Liu,Chaoyou Pang,Shuli Fan,Meizhen Song,Dan Wang,Weihua Li,Shuxun Yu.

[13]Rapid Visual Detection of Aflatoxin B1 by Label-Free Aptasensor Using Unmodified Gold Nanoparticles. Luan, Yunxia,Chen, Jiayi,Lu, Anxiang,Li, Cheng,Fu, Hailong,Ma, Zhihong,Wang, Jihua,Chen, Zhengbo,Xie, Gang.

[14]Label-free and rapid detection of ATP based on structure switching of aptamers. Ji, Danyang,Ge, Jia,Zhang, Lin,Li, Jianjun,Bai, Dongmei,Chen, Juan,Li, Zhaohui,Wang, Hongqi,Li, Zhaohui.

[15]Label-free DNA Y junction for bisphenol A monitoring using exonuclease III-based signal protection strategy. Chen, Junhua,Zhou, Shungui.

[16]High sensitive rapid visual detection of sulfadimethoxine by label-free aptasensor. Chen, Ailiang,Jiang, Xiaoling,Zhang, Weiwei,Chen, Gang,Zhao, Yan,Tunio, Tarique Muhammad,Liu, Jinchuan,Lv, Zhenzhen,Yang, Shuming,Chen, Ailiang,Jiang, Xiaoling,Zhang, Weiwei,Chen, Gang,Zhao, Yan,Tunio, Tarique Muhammad,Liu, Jinchuan,Yang, Shuming,Lv, Zhenzhen,Li, Cheng.

作者其他论文 更多>>

微信小程序