期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 238, 期 -, 页码 626-632出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2016.07.087
关键词
Colorimetric; DNA methyltransferase; Gold nanoparticles; Strand displacement amplification
资金
- National Natural Science Foundation of China [21265012, 21365015]
- Program for New Century Excellent Talents in University [NCET-11-1002, NCET-13-0848]
- Department of Education (Jiangxi Province, China [GJJ13109]
In this paper, we present a colorimetric method for the assay of DNA methyltransferase (MTase) activity based on strand displacement amplification (SDA). In our study, a well-designed hairpin DNA I (HPI) containing the sequence of 5'-CCGG-3' is specifically recognized by CpG methyltransferase (M.SssI) and HpaII endonuclease. The methylated HPI is able to coexist with all the DNA and enzymes in the solution while the unmethylated HPI can be cleaved into single-stranded DNA (ssDNA) fragments. The amplification can be triggered by the HpaII digestion products hybridization with another hairpin structure DNA II (HPII) to form a duplex, which would be replaced by probe DNA, leading to the aggregation of gold nanoparticles (AuNPs). Simultaneously, ssDNA fragments released from the duplex, and triggered the cycle anew. Varying concentrations of M.SssI in the solution therefore would lead to differences of absorption and color changed from red to pale. A linear response was obtained when the M.SssI concentration ranging from 0.2 to 50 U mL(-1) with a detection limit of 0.08 U mL(-1). In addition, the developed assay in this study can also be applied to screen the inhibitors of M.SssI. (C) 2016 Elsevier B.V. All rights reserved.
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