In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against Atrazine
| Autor: | Lisa A. Holland, Srikanth Gattu, Cassandra L. Crihfield, Ryan M. Williams, Letha J. Sooter |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Aptamer
DNA Single-Stranded 02 engineering and technology molecular recognition element (MRE) Biology In Vitro Techniques 01 natural sciences Catalysis Article Inorganic Chemistry lcsh:Chemistry chemistry.chemical_compound Molecular recognition Atrazine Physical and Theoretical Chemistry Molecular Biology lcsh:QH301-705.5 Spectroscopy SELEX 010401 analytical chemistry Organic Chemistry aptamer General Medicine Contamination 021001 nanoscience & nanotechnology Molecular biology In vitro 0104 chemical sciences Computer Science Applications Dissociation constant chemistry Biochemistry lcsh:Biology (General) lcsh:QD1-999 in vitro selection 0210 nano-technology atrazine DNA Systematic evolution of ligands by exponential enrichment |
| Zdroj: | International Journal of Molecular Sciences, Vol 15, Iss 8, Pp 14332-14347 (2014) International Journal of Molecular Sciences; Volume 15; Issue 8; Pages: 14332-14347 International Journal of Molecular Sciences |
| ISSN: | 1422-0067 |
| Popis: | Widespread use of the chlorotriazine herbicide, atrazine, has led to serious environmental and human health consequences. Current methods of detecting atrazine contamination are neither rapid nor cost-effective. In this work, atrazine-specific single-stranded DNA (ssDNA) molecular recognition elements (MRE) were isolated. We utilized a stringent Systematic Evolution of Ligands by Exponential Enrichment (SELEX) methodology that placed the greatest emphasis on what the MRE should not bind to. After twelve rounds of SELEX, an atrazine-specific MRE with high affinity was obtained. The equilibrium dissociation constant (Kd) of the ssDNA sequence is 0.62 ± 0.21 nM. It also has significant selectivity for atrazine over atrazine metabolites and other pesticides found in environmentally similar locations and concentrations. Furthermore, we have detected environmentally relevant atrazine concentrations in river water using this MRE. The strong affinity and selectivity of the selected atrazine-specific ssDNA validated the stringent SELEX methodology and identified a MRE that will be useful for rapid atrazine detection in environmental samples. |
| Databáze: | OpenAIRE |
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