Gold nanoparticle sensor based on pin locking deoxyribozyme probe and application of gold nanoparticle sensor in detecting MUC1

A deoxyribozyme probe and gold nanoparticle technology, applied in the field of biological analysis, can solve problems such as unsatisfactory sensitivity, and achieve the effects of improving sensitivity, simplifying the operation process, and good reproducibility

Active Publication Date: 2020-10-16
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the inventors of the present invention have found that the further application of these methods still has the problem of unsatisfactory sensitivity.

Method used

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  • Gold nanoparticle sensor based on pin locking deoxyribozyme probe and application of gold nanoparticle sensor in detecting MUC1
  • Gold nanoparticle sensor based on pin locking deoxyribozyme probe and application of gold nanoparticle sensor in detecting MUC1
  • Gold nanoparticle sensor based on pin locking deoxyribozyme probe and application of gold nanoparticle sensor in detecting MUC1

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Embodiment approach

[0027] A typical embodiment of the present invention provides a gold nanoparticle sensor based on a hairpin-locked DNAzyme probe, including a MUC1 aptamer probe, a hairpin-locked DNAzyme probe and gold nanoparticles;

[0028] The MUC1 aptamer probe includes an aptamer DNA sequence and a hairpin DNA sequence connected in sequence, and the aptamer DNA sequence and the hairpin DNA sequence form a hairpin structure, and the MUC1 aptamer probe can recognize MUC1 and Release the aptamer DNA sequence;

[0029] The hairpin-locked DNAzyme probe is a single-stranded DNA sequence, and the hairpin-locked DNAzyme probe includes a first connection sequence, a first binding sequence, a substrate sequence, and a second connection sequence from the 5' end to the 3' sequence, deoxyribozyme sequence and the second binding sequence, the first connecting sequence and the second connecting sequence can make the hairpin locking deoxyribozyme probe form a hairpin structure after the first connecting ...

Embodiment 1

[0050] Preparation of hairpin-locked DNAzyme-modified AuNPs: 1 mL of AuNPs solution (5.7×10 12 Particles / mL) was added 6nmol hairpin-locked DNAzyme probe (FAM-CAAAAAAGC ACA CCG GCT rAGT CTT TTT TTT TGATCCGAG CCG GAC GAA GCC CCA TAG GT-SH, the sequence is shown in SEQ ID NO.1), at room temperature Incubate for 16 h in the presence of 10 mM phosphate (NaH 2 PO 4 / Na 2 HPO 4 ) and 0.1M NaCl in PBS buffer (pH 7.0) at room temperature for 36 hours. Then, the mixture of AuNPs and DNAzyme was centrifuged three times at high speed (13000rpm, 30min) to remove the hairpin-locked DNAzyme probes not assembled on the surface of AuNPs, and the obtained precipitate was resuspended in 250 μL of PBS buffer, and finally stored at 4°C for use. In the hairpin-locked DNAzyme probe-AuNPs solution, the concentration of the hairpin-locked DNAzyme probe was about 18 μM.

[0051] Mg 2+ Dependent hairpin-locked DNAzyme probe self-cleavage cycle signal amplification: Synthetic DNA oligonucleotide s...

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Abstract

The invention discloses a gold nanoparticle sensor based on a pin locking deoxyribozyme probe and application of the gold nanoparticle sensor in detecting MUC1. The gold nanoparticle sensor comprisesan MUC1 aptamer probe, the pin locking deoxyribozyme probe and gold nanoparticles, wherein the MUC1 aptamer probe can recognize MUC1 and release an aptamer DNA sequence; the pin locking deoxyribozymeprobe sequentially comprises a first connecting sequence, a first binding sequence, a substrate sequence, a second connecting sequence, a deoxyribozyme sequence and a second binding sequence; the first connecting sequence and the second connecting sequence are complementary to enable the pin locking deoxyribozyme probe to form a pin structure; the substrate sequence comprises a cutting site; afterthe first binding sequence, the second binding sequence and the aptamer DNA sequence are complementary, the deoxyribozyme sequence forms an active secondary structure in the catalytic core, and the 5'end of the pin locking deoxyribozyme probe is connected with a fluorophore which can be quenched by the gold nanoparticles; and the 3'end of the pin locking deoxyribozyme probe is connected with goldnanoparticles.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and relates to a gold nanoparticle (AuNPs) sensor based on a hairpin-locked deoxyribozyme (DNAzyme) probe and its application for detecting MUC1. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Tumor-associated antigen mucin 1 (mucin 1, MUC1) is a transmembrane glycoprotein, which contains a hydrophobic transmembrane domain of 31 amino acids, a cytoplasmic domain of 69 amino acids, and a nearly identical repeat sequence (per An extracellular repeat region consisting of a stressed read sequence of 20 amino acids). MUC1 activates cytotoxic T lymphocytes (cytotoxic T-lymphocyte, CTL)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6825
CPCC12Q1/6825C12Q2525/205C12Q2525/301C12Q2521/345C12Q2563/137C12Q2563/107C12Q2565/607Y02A50/30
Inventor 张春阳张丹丹孟亚茹
Owner SHANDONG NORMAL UNIV
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