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Method for detecting if interaction between nucleic acid conjugated protein -target protein exist based on biological chip

A technology for binding proteins and target proteins, which is applied in the field of detecting whether there is an interaction between nucleic acid binding proteins and target proteins based on biochips, and can solve the problem that there is no high-throughput detection method for the interaction between nucleic acid binding proteins and target proteins. Achieve the effect of high sensitivity, strong specificity and broad application prospects

Active Publication Date: 2008-08-06
CAPITALBIO CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] To date, there is no high-throughput method for detecting the presence or absence of nucleic acid-binding protein-target protein interactions

Method used

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  • Method for detecting if interaction between nucleic acid conjugated protein -target protein exist based on biological chip
  • Method for detecting if interaction between nucleic acid conjugated protein -target protein exist based on biological chip

Examples

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Embodiment 1

[0038] Example 1. Detection of the specificity of the method for detecting whether there is an interaction between the nucleic acid binding protein-target protein based on the biochip of the present invention

[0039] Taking the transcription factor NFkB as the target protein, using pure NFkB, to detect the specificity of the method of the present invention based on biochip detection of whether there is an interaction between the nucleic acid binding protein-target protein, since this embodiment is to detect the specificity of the method, Among them, there is only one type of protein, NFkB, so there is no nucleic acid binding protein interacting with it. The specific detection method includes the following steps:

[0040] A. Preparation of DNA probes: the DNA chip-immobilized probes in each group were first dissolved in water and prepared in a 50% dimethyl sulfoxide (DMSO) solution with a final concentration of 10 μM. The nucleic acid-binding protein capture probes were all d...

Embodiment 2

[0047] Embodiment 2, detect the nucleic acid binding protein that interacts with protein STAT1 by the method of the present invention

[0048] Use the method of the present invention to detect whether there is an interaction between the nucleic acid binding protein-target protein (this embodiment is the transcription factor STAT1, GenBank number: NM_007315) based on the biochip to detect whether there is an interaction between the nuclear protein extracted after IFNγ stimulates the mouse 3T3L1 cell line and For nucleic acid binding proteins interacting with STAT1, the mixture of nucleic acid capture probes was added to the protein extract (the nuclear protein extracted after IFNγ stimulated the mouse 3T3L1 cell line in this example), and then passed through the antibody of the target protein (this example For example, the antibody of STAT1) separates the target protein and the nucleic acid binding protein interacting with it, and by detecting the nucleic acid sequence bound to ...

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Abstract

The invention discloses a method of detecting whether an interaction exists between nucleic acid protein and target protein based on biochip. The method comprises the following steps: (1)adding a plurality of groups of nucleic acid acquisition probes into a biological sample system containing the target protein to form a target-protein-nucleic-acid-conjugated-protein-nucleic-acid-acquisition-probe compound, and the nucleic acid acquisition probes containing at least one section of sequence capable of integrating with the nucleic acid conjugated protein; (2) separating the target-protein-nucleic-acid-conjugated-protein-nucleic-acid-acquisition-probe compound through molecules capable of specifically integrating with the target protein, and then recovering the nucleic acid acquisition probes; (3) carrying out hybridization of the nucleic acid acquisition probes recovered in the step 2 and a plurality of single chain immobilized probes which are fixed on the substrate of the biochip and are corresponding to the nucleic acid acquisition probes, and the nucleic acid sequence of the immobilized probes complementing the corresponding nucleic acid acquisition probes or one chain of the nucleic acid acquisition probes; (4) detecting the hybridization results to see whether the interaction exists between the nucleic acid conjugated protein and the target protein.

Description

technical field [0001] The invention relates to a method for detecting whether there is an interaction between protein-proteins, in particular to a method for detecting whether there is an interaction between a nucleic acid binding protein and a target protein based on a biochip. Background technique [0002] Nucleic acid-binding proteins include double-stranded DNA-binding proteins, single-stranded DNA-binding proteins, RNA-binding proteins, and the like. [0003] Double-stranded DNA-binding protein is a kind of protein molecule or protein molecular complex that can specifically bind to a specific sequence of double-stranded DNA. Double-stranded DNA binding proteins mainly include repressors and operator proteins in prokaryotes and transcription factors in eukaryotes. [0004] The end of the Human Genome Project (HGP) and the completion of the sequencing of dozens of model organisms marked a gradual shift in the focus of biological research from the era of genomes to the e...

Claims

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Application Information

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IPC IPC(8): G01N33/68C12Q1/68
Inventor 孙义民张亮曾令琴任爱辉魏丽程京
Owner CAPITALBIO CORP
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