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Microbiology and Biotechnology Letters

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Microbial Biotechnology (MB)  |  Cell Culture and Biomedical Engineering

Microbiol. Biotechnol. Lett. 2021; 49(3): 316-319

https://doi.org/10.48022/mbl.2103.03001

Received: March 1, 2021; Revised: March 31, 2021; Accepted: May 26, 2021

Detection of AluI Endonuclease Activity by Using Double Stranded DNA-Templated Copper Nanoclusters

Jisu Yang and Jongback Gang*

Department of Chemistry, Gachon University, Sungnam 1342, Republic of Korea

Correspondence to :
Jongback Gang,      jbgang@gachon.ac.kr

Restriction endonucleases play an important role in molecular cloning, clinical diagnosis, and pharmacological drug studies. In this study, DNA-templated copper nanoclusters (DNA-CuNCs) were used to detect AluI endonuclease activity due to their high fluorescence emission and rapid synthesis of DNA-CuNCs under ambient conditions. Results showed that AluI activity was detected in a highly sensitive manner at low concentrations of AluI endonuclease by the fluorescence intensity of DNA-CuNCs. Additionally, its inhibition was monitored in the presence of daidzein under optimal conditions.

Keywords: Fluorescence assay, DNA-CuNCs, AluI endonuclease assay

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  1. Brueckner B, Lyko F. 2006. DNA methyltransferase inhibitors: old and new drugs for an epigenetic cancer therapy. Trends Pharmacol. Sci. 25: 551-554.
    Pubmed
  2. Shames DS, Minna JD, Gazdar AF. 2007. DNA methylation in health, disease, cander. Curr. Mol. Med. 7: 85-102.
    Pubmed
  3. Norberg P, Bergstrom T, Liljeqvist JA. 2006. Genotyping of clinical herpes simplex virus type 1 isolates by use of restriction enzymes. J. Clin. Microbiol. 44: 4511-4514.
    Pubmed KoreaMed
  4. Xu X, Han MS, Mirkin CA. 2007. A gold-nanoparticle-based realtime colorimetric screening method for endonuclease activity and inhibition. Angew. Chem. Int. Ed. 46: 3468-3470.
    Pubmed
  5. Harford SE, Goodall AJ. 1988. Mode of DNA cleavage by the EcoRV restriction endonuclease. Biochemistry 27: 1771-1777.
    Pubmed
  6. Jeltsch A, Fritz A, Alves A, Wolfes H, Pingoud A. 1993. A fast and accurate enzyme-linked immunosorbent assay for the determination of the DNA cleavage activity of restriction endonucleases. Anal. Biochem. 213: 234-240.
    Pubmed
  7. Sharp PA, Sugden B, Sambrock J. 1973. Detection of two restriction endonuclease activities in Haemophilus Parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry 12: 3055-3063.
    Pubmed
  8. Song G, Chen C, Ren J, Qu X. 2009. A simple, universal colorimetric assay for endonuclease/methyltransferase activity and inhibition based on an enzyme-responsive nanoparticle system. ACS Nano 3: 1183-1189.
    Pubmed
  9. Zhao H, Dong J, Zhou F, Li B. 2017. One facile fluorescence strategy for sensitive detection of endonuclease activity using DNA-templated copper nanoclusters as signal indicators. Sens. Actuators B Chem. 238: 828-833.
  10. Feng X, Duan X, Liu L, Feng F, Wang S, Li Y, et al. 2009. Fluorescence logic‐signal‐based multiplex detection of nucleases with the assembly of a cationic conjugated polymer and branched DNA. Angew. Chem. Int. Ed. 48: 5316-5321.
    Pubmed
  11. Deng J, Jin Y, Wang L, Chen G, Zhang C. 2012. Sensitive detection of endonuclease activity and inhibition using gold nanorods. Biosens. Bioelectron. 34: 144-150.
    Pubmed
  12. Lee C, Gang J. 2018. A Label-free detection of NdeI endonuclease activity by using DNA-templated silver nanoclusters. J. Nanosci. Nanotechnol. 18: 6339-6342.
    Pubmed
  13. Huang Y, Zhao S, Chen ZF, Liu YC, Liang H. 2011. Ultrasensitive endonuclease activity and inhibition detection using gold nanoparticle-enhanced fluorescence polarization. Chem. Commun. 47: 4763-4765.
    Pubmed
  14. Li W, Liu Z, Lin H, Nie Z, Chen J, Xu X, et al. 2010. Label-free colorimetric assay for methyltransferase activity based on a novel methylation-responsive DNAzyme strategy. Anal. Chem. 82:1935-1941.
    Pubmed
  15. Zhao Y, Chen F, Wu Y, Dong Y, Fan C. 2013. Highly sensitive fluorescence assay of DNA methyltransferase activity via methylationsensitive cleavage coupled with nicking enzyme-assisted signal amplification. Biosens. Bioelectron. 42: 56-61.
    Pubmed
  16. Bi S, Zhao T, Luo B, Zhu JJ. 2013. Hybridization chain reactionbased branched rolling circle amplification for chemiluminescence detection of DNA methylation. Chem. Commun. 49: 69066908.
    Pubmed
  17. Cao A, Zhang CY. 2012. Sensitive and Label-free DNA methylation detection by ligation-mediated hyperbranched rolling circle amplification. Anal. Chem. 84: 6199-6205.
    Pubmed
  18. Jia H, Li Z, Liu C, Cheng Y. 2010. Ultrasensitive detection of microRNAs by exponential isothermal amplification. Angew. Chem. Int. Ed. 49: 5498-5501.
    Pubmed
  19. Zhou Z, Du Y, Zhang L, Dong S. 2012. A Label-free, G-quadruplex DNAzyme-based fluorescent probe for signal-amplified DNA detection and turn-on assay of endonuclease. Biosens. Bioelectron. 34: 100-105.
    Pubmed
  20. Jiang XQ, Guo SM, Zhou M, Ye BC. 2014. DNA-hosted Hoechst dyes: application for label-free fluorescent monitoring of endonuclease activity and inhibition. Analyst 139: 5682-5685.
    Pubmed
  21. Zhao H, Dong J, Zho F, Li B. 2017. One facile fluorescence strategy for sensitive detection of endonuclease activity using DNA-templated copper nanoclusters as signal indicators. Sens. Acuators B Chem. 238: 828-833.
  22. Rotaru A, Dutta S, Jentzsch E, Gothelf K, Mokhir A. 2010. Selective dsDNA-templated formation of copper nanoparticles in solution. Angew. Chem. Int. Ed. 49: 5665-5667.
    Pubmed
  23. Maniotis AJ, Valyi-Nagy K, Karavitis J, Moses J, Boddipali V, Wang Y, et al. 2005. Chromatin organization measured by AluI restriction endonuclease changes with malignancy and is regulated by the extracellular matrix and the cytoskeleton. Am. J. Pathol. 166:1187-1203.
  24. Sandal T, Valyi-Nagy K, Spencer VA, Folberg R, Bissell MJ, Maniotis AJ. 2007. Epigenetic reversion of breast carcinoma phenotype is accompanied by changes in DNA sequestration as measured by AluI restriction enzyme. Am. J. Pathol. 170: 1739-1749.
    Pubmed KoreaMed

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