Unnatural base dna aptamers targeting bacillus cereus 5 / b / 6 metallo-beta-lactamase and methods of screening the same

CN121825978BActive Publication Date: 2026-05-29SOUTH CHINA UNIV OF TECH
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Patent Information

Application Number
CN202610303447.9
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2026-03-12
Publication Date
2026-05-29
Estimated Expiration
2046-03-12

AI Technical Summary

Technical Problem

In existing technologies, the binding affinity of nucleic acid aptamers for Bacillus cereus 5/B/6 metallo-β-lactamases is insufficient, which cannot meet the practical application requirements of high-sensitivity detection and precise targeting.

Method used

Using non-natural base DNA aptamers Apt-BcN, Apt-BcC1, and Apt-BcC2, and employing the SELEX screening method, a non-natural base DNA library was constructed using two non-natural base pairs, dNaM-dTPT3 and dCNMO-dTPT3, to screen for aptamers with high binding activity.

Benefits of technology

It achieves high-affinity binding to Bacillus cereus 5/B/6 metallo-β-lactamases, overcoming the sequence diversity limitations of the natural base system and providing higher binding activity and sensitivity.

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Abstract

The application discloses a non-natural base DNA aptamer targeting Bacillus cereus 5 / B / 6 metal beta-lactamase and a screening method thereof. The screening method uses dNaMTP and dTPT3TP, or dCNMOTP and dTPT3TP as non-natural base nucleoside triphosphate substrates, uses OneTaq DNA polymerase to amplify a DNA library containing non-natural bases, and uses lamda exonuclease to degrade 5' end phosphorylated DNA chains in PCR products, so as to obtain two ssDNA libraries. The non-natural base DNA aptamer targeting Bacillus cereus 5 / B / 6 metal beta-lactamase is screened based on the libraries, and three non-natural base DNA aptamers with a length of 70 nt are obtained. The results of a biological membrane layer interference analysis show that the three non-natural base DNA aptamers have good binding affinity to Bacillus cereus 5 / B / 6 metal beta-lactamase.
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Citation Information

Patent Citations

  • Method for template-free synthesis of non-natural base-containing oligonucleotide chain by using terminal deoxyribonucleotide transferase and application of non-natural base-containing oligonucleotide chain

    CN114196714A

  • 2 '-fully modified XNA aptamer of targeted 5 / B / 6 metal beta-lactamase and obtaining method of 2'-fully modified XNA aptamer

    CN117210450A