Title
**In vitro** selection and characterization of DNA aptamers recognizing chloramphenicol **In vitro** selection and characterization of DNA aptamers recognizing chloramphenicol
Author
Faculty/Department
Faculty of Sciences. Biology
Faculty of Sciences. Chemistry
Publication type
article
Publication
Amsterdam ,
Subject
Chemistry
Biology
Source (journal)
Journal of biotechnology. - Amsterdam
Volume/pages
155(2011) :4 , p. 361-369
ISSN
0168-1656
ISI
000294488900001
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Chloramphenicol (Cam), although an effective antibiotic, has lost favour due to some fatal side effects. Thus there is an urgent need for rapid and sensitive methods to detect residues in food, feed and environment. We engineered DNA aptamers that recognize Cam as their target, by conducting in vitro selections. Aptamers are nucleic acid recognition elements that are highly specific and sensitive towards their targets and can be synthetically produced in an animal-friendly manner, making them ethical innovative alternatives to antibodies. None of the isolated aptamers in this study shared sequence homology or structural similarities with each other, indicating that specific Cam recognition could be achieved by various DNA sequences under the selection conditions used. Analyzing the binding affinities of the sequences, demonstrated that dissociation constants (Kd) in the extremely low micromolar range, which were lower than those previously reported for Cam-specific RNA aptamers, were achieved. The two best aptamers had G rich (>35%) nucleotide regions, an attribute distinguishing them from the rest and apparently responsible for their high selectivity and affinity (Kd ∼ 0.8 and 1 μM respectively). These aptamers open up possibilities to allow easy detection of Cam via aptamer-based biosensors.
E-info
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