Unlock instant, AI-driven research and patent intelligence for your innovation.

Nucleic acid aptamer and application thereof in specific recognition of p-hydroxybenzyl hydrogen sulfite

A technology of p-hydroxybenzyl bisulfite and nucleic acid aptamer, which is applied in the field of biomedicine, can solve the problems of being unable to be widely used, and achieve the effect of high specificity and high affinity

Active Publication Date: 2022-02-15
KUNMING UNIV OF SCI & TECH
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this detection method requires specific equipment and certain operating skills, and cannot be widely used.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Nucleic acid aptamer and application thereof in specific recognition of p-hydroxybenzyl hydrogen sulfite
  • Nucleic acid aptamer and application thereof in specific recognition of p-hydroxybenzyl hydrogen sulfite
  • Nucleic acid aptamer and application thereof in specific recognition of p-hydroxybenzyl hydrogen sulfite

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Screening and secondary structure analysis of p-hydroxybenzyl bisulfite nucleic acid aptamers

[0025] 1. Using the magnetic bead-SELEX method to screen, immobilize the target p-hydroxybenzyl bisulfite on magnetic beads (DynabeadsMyOne Streptavidin C1 magnetic beads), and then incubate the target-magnetic bead complex with the oligonucleotide library, which can bind to the target The bound oligonucleotide will bind to the target-magnetic bead complex to form an oligonucleotide-target-magnetic bead complex, and finally the oligonucleotide-target-magnetic bead complex can be combined by the magnetic attraction of the magnetic frame The ssDNA enriched library with affinity and specificity for p-hydroxybenzyl bisulfite was obtained after multiple rounds of screening;

[0026] Each round adopts the method of SA magnetic bead immobilized library screening. After the library is enriched, the affinity of the secondary library is first verified by Q-PCR. If the affini...

Embodiment 2

[0039] Example 2: Using graphene oxide (GO) fluorescence method to detect the specificity of p-hydroxybenzyl bisulfite nucleic acid aptamer SEQ16

[0040] In this experiment, compounds p-hydroxybenzyl alcohol, p-hydroxybenzenesulfonic acid, gastrodin, p-hydroxybenzaldehyde, and barisenoside A, which are similar in structure to p-hydroxybenzyl bisulfite, were selected to verify the nucleic acid aptamer SEQ16 p-hydroxybenzylsulfite Hydrogen Specificity.

[0041] Add an appropriate amount of 100 μmol / L FAM-modified nucleic acid aptamer SEQ16 (synthesized by Shanghai Sangon Bioengineering Co., Ltd.) to 200 μL binding buffer (Tris-HCl buffer with pH=7.2) to make a final concentration of 200 nmol / L The nucleic acid aptamer solution was pre-denatured in a boiling water bath at 95°C for 10 minutes, then ice-bathed for 10 minutes, and placed at room temperature for 10 minutes; then 200 μL of 100 μg / mL graphene oxide aqueous solution was added and incubated at room temperature for 2 hou...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a nucleic acid aptamer. The nucleotide sequence of the nucleic acid aptamer is shown as SEQ ID NO: 1, the nucleic acid aptamer is single-stranded DNA and is composed of 76 nucleotides, the secondary structure of the nucleic acid aptamer contains protruding rings and stems, and the Gibbs free energy DG of SEQ 16 is-16.65; based on the evaluation of the specificity, affinity and the like of the aptamer by a graphene oxide fluorescence method, the result shows that the dissociation constant Kd of the aptamer SEQ16 is equal to 32.12 + / -2.91, and the aptamer can be specifically combined with p-hydroxybenzyl hydrogen sulfite; the nucleic acid aptamer SEQ16 has the characteristics of high specificity and high affinity, can be applied to preparation of a reagent for detecting sulfur fumigated gastrodia elata or preparation of a kit for detecting sulfur fumigated gastrodia elata, and provides an effective method for detection of sulfur fumigated gastrodia elata.

Description

technical field [0001] The invention discloses a nucleic acid aptamer capable of specifically binding to a marker p-hydroxybenzyl bisulfite produced by gastrodia elata after sulfur fumigation and its application, belonging to the technical field of biomedicine. Background technique [0002] Oligonucleotide aptamers are single-stranded DNA or RNA molecules with a length of 10–100 nucleotides that are traditionally screened using the technique of systematic evolution of ligands by exponential enrichment (SELEX). Nucleic acid aptamers have a wide range of targets, including small molecular substances such as amino acids, metal ions, mycotoxins, and antibiotics. At the same time, macromolecular substances such as peptides, nucleic acids, and proteins, and complete individuals such as bacteria, cells, and viruses can also be used as nucleic acid aptamers. target. Aptamers bind to targets through hydrogen bonds, hydrophobic interactions, van der Waals forces, and other non-covale...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115G01N33/53
CPCC12N15/115G01N33/5308C12N2310/16C12N2310/531Y02A50/30
Inventor 杨野万芬崔秀明王承潇曲媛刘源杨晓艳
Owner KUNMING UNIV OF SCI & TECH