Selection and characterization of single stranded DNA aptamers recognizing fumonisin B1
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We present an improved method for the selection of single-stranded DNA aptamers that can recognize fumonisin B1 (FB1). FB1 is a carcinogenic mycotoxin mainly found in corn and corn-based food products worldwide, posing a global threat to feed and food safety. Selection was based on the mag-SELEX (magnetic bead systematic evolution of ligands by exponential enrichment) technology modified by adopting free analogs of targets rather than immobilized targets for counter selections. Firstly, aptamer candidates for FB1 were selected from an 80 nt random DNA library after 13 rounds of selection. Next, binding assays were performed for affinity evaluation, and circular dichroism spectroscopy was used to investigate their conformation. A high-affinity aptamer designated as F10 (with a dissociation constant of 62 ± 5 nM) was identified and tested for its specificity by competitive binding assays. The results demonstrate that this improved mag-SELEX technology facilitates aptamer screening because it avoids the tedious immobilization of counter-selection molecules on magnetic beads. The aptamers obtained by this technique open new possibilities for the detection of FB1 via aptasensors.
KeywordsFumonisin B1 Aptamer SELEX Magnetic beads Competition
This work was partly supported by NSFC (21375049), National S&T Support Program of China (2012BAK08B01), S&T Supporting Project of Jiangsu Province (BE2011621, BE2012614), Research Fund for the Doctoral Program of Higher Education (20110093110002), NCET-11-0663, JUSRP51309A and JUSRP21125.
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