Novel Method to Detect DNA Methylation Using Gold Nanoparticles Coupled with Enzyme-Linkage Reactions

被引:139
作者
Liu, Tao [1 ,2 ]
Zhao, Jing [1 ,2 ]
Zhang, Dongmei [1 ,2 ]
Li, Genxi [1 ,2 ,3 ]
机构
[1] Nanjing Univ, Dept Biochem, Nanjing 210093, Peoples R China
[2] Nanjing Univ, Natl Key Lab Pharmaceut Biotechnol, Nanjing 210093, Peoples R China
[3] Shanghai Univ, Sch Life Sci, Lab Biosensing Technol, Shanghai 200444, Peoples R China
基金
中国国家自然科学基金;
关键词
ESCHERICHIA-COLI; NUCLEIC-ACIDS; BOUND DNA; ASSAY; MANIPULATION; CLEAVAGE; CANCER; NANOSTRUCTURES; ENDONUCLEASE; INHIBITORS;
D O I
10.1021/ac902198v
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
DNA methylation, catalyzed by methylases, plays a critical role in many biological processes, and methylases have been regarded as promising targets for antimicrobial drugs. In this paper, we propose a simple and sensitive colorimetric assay method to detect the activity of methylases so as to monitor DNA methylation using DNA-modified gold nanoparticles (AuNPs) coupled with enzyme-linkage reactions. The duplex DNA molecules modified on the surface of AuNPs are first methylated by DNA adenine methylation (Dam) methyltransferase (MTase) and then cut by methylation-sensitive restriction endonuclease Dpn I. Removal of duplex from the AuNP surfaces by the methylation/cleavage process will destabilize the nanoparticles, resulting in aggregation of AuNPs and a red-to-blue color change. Consequently, the enzyme activity of Dam MTase can be assayed and DNA methylation can be detected. Furthermore, this study may provide a sensitive platform to screen inhibitors for Dam MTase.
引用
收藏
页码:229 / 233
页数:5
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