Electrochemical biosensor for DNA demethylase detection based on demethylation triggered endonuclease BstUI and Exonuclease III digestion

被引：22

作者：

Yin, Huanshun ^{[1
]}

Yang, Zhiqing ^{[1
]}

Li, Bingchen ^{[1
]}

Zhou, Yunlei ^{[1
]}

Ai, Shiyun ^{[1
]}

机构：

[1] Shandong Agr Univ, Coll Chem & Mat Sci, Tai An 271018, Shandong, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2015年 / 66卷

基金：

中国国家自然科学基金;

关键词：

DNA demethylase; Electrochemical biosensor; Endonuclease BstUI; Exonuclease III; BINDING DOMAIN PROTEIN; METHYLTRANSFERASE ACTIVITY; SIGNAL AMPLIFICATION; SENSITIVE DETECTION; ASSAY; METHYLATION; CANCER; GENE;

D O I：

10.1016/j.bios.2014.11.026

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

Herein, an electrochemical biosensor was fabricated for DNA demethylase detection based on DNA demethylation triggered endonuclease BstUl and Exonuclease III digestion. After the double-strand DNA was demethylated, it can be further digested by BstUI and formed a blunt end at the electrode surface. Then, the remained fragment of DNA-DNA duplex was further cleaved by exonuclease III and led to increased electrochemical signal. Based on this detection strategy, the biosensor showed high sensitivity with low detection limit of 0.15 ng/mL. Moreover, the developed method also presented high selectivity and acceptable reproducibility. This work provides a novel detection platform for DNA demethylase detection. (C) 2014 Elsevier B.V. All rights reserved.

引用

页码：266 / 270

页数：5

共 22 条

[1] A mammalian protein with specific demethylase activity for mCpG DNA [J].

Bhattacharya, SK ;

Ramchandani, S ;

Cervoni, N ;

Szyf, M .

NATURE, 1999, 397 (6720) :579-583

[2] Highly Sensitive Electrochemical Methyltransferase Activity Assay [J].

Deng, Huimin ;

Yang, Xinjian ;

Yeo, Stephanie Pei Xing ;

Gao, Zhiqiang .

ANALYTICAL CHEMISTRY, 2014, 86 (04) :2117-2123

[3] Exonuclease III-based and gold nanoparticle-assisted DNA detection with dual signal amplification [J].

Fan, Qi ;

Zhao, Jing ;

Li, Hao ;

Zhu, Li ;

Li, Genxi .

BIOSENSORS & BIOELECTRONICS, 2012, 33 (01) :211-215

[4] A sensitive signal-on electrochemical assay for MTase activity using AuNPs amplification [J].

He, Xiaoxiao ;

Su, Jing ;

Wang, Yonghong ;

Wang, Kemin ;

Ni, Xiaoqi ;

Chen, Zhifeng .

BIOSENSORS & BIOELECTRONICS, 2011, 28 (01) :298-303

[5] UNIDIRECTIONAL DIGESTION WITH EXONUCLEASE-III CREATES TARGETED BREAKPOINTS FOR DNA SEQUENCING [J].

HENIKOFF, S .

GENE, 1984, 28 (03) :351-359

[6] Structure and substrate recognition of the Escherichia coli DNA adenine methyltransferase [J].

Horton, JR ;

Liebert, K ;

Bekes, M ;

Jeltsch, A ;

Cheng, XD .

JOURNAL OF MOLECULAR BIOLOGY, 2006, 358 (02) :559-570

[7] Signal Amplification of Graphene Oxide Combining with Restriction Endonuclease for Site-Specific Determination of DNA Methylation and Assay of Methyltransferase Activity [J].

Li, Wen ;

Wu, Ping ;

Zhang, Hui ;

Cai, Chenxin .

ANALYTICAL CHEMISTRY, 2012, 84 (17) :7583-7590

[8] An electrochemical approach for detection of DNA methylation and assay of the methyltransferase activity [J].

Liu, Shuna ;

Wu, Ping ;

Li, Wen ;

Zhang, Hui ;

Cai, Chenxin .

CHEMICAL COMMUNICATIONS, 2011, 47 (10) :2844-2846

[9] A profile of methyl-CpG binding domain protein occupancy of hypermethylated promoter CpG islands of tumor suppressor genes in human cancer [J].

Lopez-Serra, Lidia ;

Ballestar, Esteban ;

Fraga, Mario F. ;

Alaminos, Miguel ;

Setien, Fernando ;

Esteller, Manel .

CANCER RESEARCH, 2006, 66 (17) :8342-8346

[10] A sensitive electrochemical DNA biosensor for specific detection of Enterobacteriaceae bacteria by Exonuclease III-assisted signal amplification [J].

Luo, Caihui ;

Tang, Hua ;

Cheng, Wei ;

Yan, Li ;

Zhang, Decai ;

Ju, Huangxian ;

Ding, Shijia .

BIOSENSORS & BIOELECTRONICS, 2013, 48 :132-137

← 1 2 3 →