One-step label-free chemiluminescent assay for determination of exonuclease III activity towards hairpin oligonucleotides
| Autor: | Oleg L. Bodulev, Anastasia V. Gribas, Ivan Yu. Sakharov, Konstantin M. Burkin |
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| Rok vydání: | 2019 |
| Předmět: |
Aptamer
Deoxyribozyme Oligonucleotides Bioengineering Applied Microbiology and Biotechnology Biochemistry Sensitivity and Specificity law.invention Luminol chemistry.chemical_compound law Chemiluminescence Detection limit Exonuclease III biology Chemistry Oligonucleotide Aptamers Nucleotide Combinatorial chemistry Exodeoxyribonucleases Luminescent Measurements biology.protein Hemin Nucleic Acid Conformation Biotechnology |
| Zdroj: | Enzyme and microbial technology. 131 |
| ISSN: | 1879-0909 |
| Popis: | Fast label-free chemiluminescent assay for determination of exonuclease III (ExoIII) activity measured towards hairpin oligonucleotide substrates was developed. The designed substrates consisted of EAD2 aptamer to hemin which was associated with DNA sequence complementary to 5’-terminus fragment of EAD2. In the presence of ExoIII the associated sequence of the hairpin stem was digested, producing EAD2 aptamer which reacted with hemin with the formation of peroxidase-mimicking DNAzyme (PMDNAzyme). The catalytic activity of the produced PMDNAzyme was measured towards luminol/H2O2. Under the optimized conditions the limit of detection and sensitivity of the one-step chemiluminescent assay of ExoIII were 7.3 nM and 1.7 × 108 M−1, respectively. The coefficient of variation (CV) was lower than 6%. |
| Databáze: | OpenAIRE |
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