Electrochemical DNA Cleavage Sensing for EcoRV Activity and Inhibition with an ERGO Electrodeopen access
- Authors
- Oh, Da Eun; Kim, Hyun Beom; Kim, Tae Hyun
- Issue Date
- Feb-2024
- Publisher
- MDPI
- Keywords
- endonuclease; EcoRV; electrochemical assay; graphene
- Citation
- BIOSENSORS-BASEL, v.14, no.2
- Journal Title
- BIOSENSORS-BASEL
- Volume
- 14
- Number
- 2
- URI
- https://scholarworks.bwise.kr/sch/handle/2021.sw.sch/25926
- DOI
- 10.3390/bios14020073
- ISSN
- 2079-6374
2079-6374
- Abstract
- An electrochemically reduced graphene oxide (ERGO) electrode-based electrochemical assay was developed for rapid, sensitive, and straightforward analysis of both activity and inhibition of the endonuclease EcoRV. The procedure uses a DNA substrate designed for EcoRV, featuring a double-stranded DNA (dsDNA) region labeled with methylene blue (MB) and a single-stranded DNA (ssDNA) region immobilized on the ERGO surface. The ERGO electrode, immobilized with the DNA substrate, was subsequently exposed to a sample containing EcoRV. Upon enzymatic hydrolysis, the cleaved dsDNA fragments were detached from the ERGO surface, leading to a decrease in the MB concentration near the electrode. This diminished the electron transfer efficiency for MB reduction, resulting in a decreased reduction current. This assay demonstrates excellent specificity and high sensitivity, with a limit of detection (LOD) of 9.5 x 10-3 U mL-1. Importantly, it can also measure EcoRV activity in the presence of aurintricarboxylic acid, a known inhibitor, highlighting its potential for drug discovery and clinical diagnostic applications.
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Collections - College of Natural Sciences > Department of Chemistry > 1. Journal Articles
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