CN-122012511-A - Nucleic acid aptamer of bisphenol A, nucleic acid aptamer derivative and application thereof

Abstract

The invention relates to the technical field of biology, in particular to a bisphenol A special nucleic acid aptamer, a derivative thereof and application thereof. The nucleic acid aptamer has a DNA sequence shown as SEQ ID NO. 1, and covers variants with at least 90% identity with the sequence, and derivatives obtained through chemical modification or structural optimization. The aptamer can identify bisphenol A with high specificity and high affinity, is suitable for the fields of environmental monitoring, food safety detection, ecological risk assessment and the like, and is particularly suitable for on-site rapid screening of bisphenol A. The aptamer disclosed by the invention has the advantages of simplicity and convenience in preparation, good stability, low cost and the like, and provides a reliable identification element for constructing a high-sensitivity low-cost BPA rapid detection sensor.

Inventors

• XIE DANPING
• YIN HUA

Assignees

• 华南理工大学

Dates

Publication Date

20260512

Application Date

20260225

Claims (6)

1. 1. A nucleic acid aptamer of bisphenol A, characterized in that the nucleic acid aptamer has a DNA sequence shown in SEQ ID NO. 1.
2. 2. The nucleic acid aptamer of claim 1, wherein a position on the nucleotide sequence of the nucleic acid aptamer is phosphorylated, oxymethylated, methylated, aminated, sulfhydrylated, fluorinated, or isotopicized.
3. 3. The aptamer of claim 1, wherein the nucleotide sequence of the aptamer is labeled with biotin, digoxigenin, a fluorescent substance, a nanomaterial, polyethylene glycol, a peptide fragment, a protein, an enzyme, or folic acid.
4. 4. A nucleic acid aptamer derivative of bisphenol a, characterized in that the nucleic acid aptamer derivative is a phosphorothioate backbone derived from the backbone of the nucleotide sequence of the nucleic acid aptamer of claim 1,2 or 3, or is a corresponding locked nucleic acid or peptide nucleic acid modified from the nucleic acid aptamer of claim 1,2 or 3.
5. 5. A method of screening for a nucleic acid aptamer according to any one of claims 1 to 3, wherein the random library LibV and primer mixture are subjected to variegation using a PCR instrument.
6. 6. The method according to claim 5, wherein the aptamer or the aptamer derivative is selected by a magnetic bead capture method.

Description

Nucleic acid aptamer of bisphenol A, nucleic acid aptamer derivative and application thereof Technical Field The invention relates to a nucleic acid aptamer, a nucleic acid aptamer derivative and application thereof, in particular to a nucleic acid aptamer capable of combining bisphenol A, a nucleic acid aptamer derivative and application thereof. Background Bisphenol a (Bisphenol a, BPA), 2-bis (4-hydroxyphenyl propane), is a typical environmental endocrine disrupter (Endocrine disrupting chemicals, ECDS) and is mainly used for the production of polycarbonates, epoxy resins and other polymeric materials. When these materials are heated, the labile lipophilic compound BPA can migrate into water and food through the release of food contact materials, and can also enter the environment through the discharge of waste water from plastic manufacturing plants, severely compromising human health. Thus, sensitive detection of BPA is critical to the ecological environment and human health. Currently, bisphenol a (BPA) detection is primarily dependent on instrumental analysis methods, including High Performance Liquid Chromatography (HPLC), liquid chromatography-tandem mass spectrometry (LC-MS), and gas chromatography-mass spectrometry combined techniques (GC-MS). Although these methods have high detection accuracy and specificity, they are limited in practical application due to the reliance on expensive equipment, cumbersome pretreatment processes, and specialized technicians. In contrast, the biosensor can realize real-time quantitative analysis of the target object by converting the biological recognition process into a detectable signal, has the advantages of good selectivity, high sensitivity, simple and convenient operation, low cost, continuous monitoring and the like, and is widely focused in the fields of environment detection, food safety, medical diagnosis and the like. The aptamer is used as a key recognition element of the biosensor, has higher stability and lower preparation cost compared with the traditional antibody, and can realize high consistency among batches through chemical synthesis. In addition, the structure of the aptamer is easy to modify, a specific functional group can be introduced through molecular design, and a high-performance sensor is further constructed, so that the aptamer plays an important role in the fields of disease diagnosis, drug screening, environmental pollutant monitoring, food safety detection and the like. Up to now, studies on high affinity, high selectivity nucleic acid aptamer screening methods to obtain bisphenol a remain blank. If the specific aptamer aiming at bisphenol A can be successfully obtained, a series of rapid detection technology research and development including an aptamer biosensor, rapid detection test paper and the like can be promoted, so that a simple and economical analysis tool is provided for detecting environmental pollutants. Therefore, the screening method of the bisphenol A specific nucleic acid aptamer and the functionalized derivative thereof has remarkable practical significance and application prospect. Disclosure of Invention The invention aims to overcome the defects of the prior art and provide a bisphenol A aptamer, a nucleic acid aptamer derivative and application thereof, wherein the nucleic acid aptamer and the nucleic acid aptamer derivative can be combined with bisphenol A in high specificity and high affinity, can be chemically synthesized, have good biocompatibility, small molecular weight and stability and are easy to store. In order to solve the technical problems, the invention adopts the following technical scheme: A nucleic acid aptamer of bisphenol A, characterized in that the nucleic acid aptamer has a DNA sequence shown in SEQ ID NO. 1. Further, a position on the nucleotide sequence of the nucleic acid aptamer is phosphorylated, oxymethylated, methylated, aminated, sulfhydrylated, fluorinated or isotopically substituted. Further, the nucleotide sequence of the nucleic acid aptamer is combined with biotin, digoxin, fluorescent substances, nano materials, polyethylene glycol, peptide fragments, protein, enzyme or folic acid labels. Further, the nucleic acid aptamer derivative is a phosphorothioate backbone derived from the backbone of the nucleotide sequence of the nucleic acid aptamer, or a corresponding locked nucleic acid or peptide nucleic acid into which the nucleic acid aptamer is modified. A method for screening the aptamer comprises the step of performing variofying on a random library LibV and a primer mixture by using a PCR instrument. Further, the aptamer or aptamer derivative is screened by using a magnetic bead capture method. Compared with the prior art, the invention has the advantages that: (1) The nucleic acid aptamer provided by the invention has high specific recognition capability on bisphenol A molecules, has strong binding affinity and good selectivity, and can obviously improve the reliability o