Selection Of Dna Aptamers For Root Exudate L-Serine Using Multiple Selection Strategies
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY(2021)
摘要
Agricultural biosensing can aid decisions about crop health and maintenance, because crops release root exudates that can inform about their status. L-Serine has been found to be indicative of nitrogen uptake in wheat and canola. The development of a biosensor for L-serine could allow farmers to monitor crop nutrient demands more precisely. The development of robust L-serine-binding DNA aptamers is described. Because small molecules can be challenging targets for Systematic Evolution of Ligands by EXponential enrichment (SELEX), three separate DNA libraries were used for SELEX experiments. A L-homocysteine aptamer was randomized to create a starting library for a L-serine selection (randomized SELEX). The final selection rounds of the L-homocysteine selection were also used as a starting library for L-serine (redirected SELEX). Finally, an original DNA library was used (original SELEX). All three SELEX experiments produced L-serine-binding aptamers with micromolar affinity, with Red.1 aptamer having a K-d of 7.9 +/- 3.6 mu M. Truncation improved the binding affinity to 5.2 +/- 2.7 mu M, and from this sequence, a Spiegelmer with improved nuclease resistance was created with a K-d of 2.0 +/- 0.8 mu M. This L-serine-binding Spiegelmer has the affinity and stability to be incorporated into aptamer-based biosensors for agricultural applications.
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关键词
exudate, amino acids, aptamers, biosensing, precision agriculture, Spiegelmer
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