Preparation and Application of Tetrodotoxin DNA Aptamer

doi:10.7506/spkx1002-6630-201424039

Abstract

Abstract:

A 78-mer ssDNA library was synthesized in vitro. The tetrodotoxin (TTX) specific monoclonal DNA aptamer A3
was prepared using systematic evolution of ligands by exponential enrichment (SELEX) combined with mutagenic PCR by
screening, enrichment, cloning and sequencing. The secondary structure of the DNA aptamer A3 mainly contained stem ring
structure, and the affinity for tetrodotoxin was 1.254. The PBS buffer pH and fluorochrome-binding time were optimized.
The results showed that the optimum pH was 7.5 and the best binding time was 10 min. As a result, a DNA aptamerfluorochrome
method for rapidly screening and detecting tetrodotoxin was developed with a detection limit of 10-6 mo1/L.

Key words: tetrodotoxin, aptamer, fluorochrome, detection

CLC Number:

SHAO Bi-ying, CHEN Bin, CHEN Wen-bing, YANG Fang, MIAO Ting-yu, PENG Juan. Preparation and Application of Tetrodotoxin DNA Aptamer[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201424039.

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Preparation and Application of Tetrodotoxin DNA Aptamer

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