Exonuclease III digesting FRET-dsDNA microarray chip for detecting transcription factor protein

A technology of microarray chips and exonucleases, which can be used in the determination/inspection of microorganisms, biological testing, material inspection products, etc., and can solve the problems of difficulty in designing DNA half-sites, difficulty in binding proteins, and impossibility

Inactive Publication Date: 2006-02-15
王进科 +1
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Problems solved by technology

The main advantage of method 1 is that only two DNA half-sites labeled by the donor and the acceptor are needed to detect the protein, the experiment is simple and efficient, and the specificity is good; however, there are difficulties in the design of the DNA half-site and receptor tags inserted into the protein binding site may prevent the normal binding of the protein or even prevent it from binding, and for those proteins whose DNA binding site is not long, it is more difficult o

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  • Exonuclease III digesting FRET-dsDNA microarray chip for detecting transcription factor protein
  • Exonuclease III digesting FRET-dsDNA microarray chip for detecting transcription factor protein
  • Exonuclease III digesting FRET-dsDNA microarray chip for detecting transcription factor protein

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[0093] Here, only the experiment of preparing FRET-dsDNA microarray chip and detecting transcription factor protein NF-κB is used to illustrate the specific implementation of the patent of the present invention.

[0094] 1. Design and chemical synthesis of FRET-dsDNA probe molecules

[0095] NF-κB (Nuclear Factor kappaB) is a sequence-specific transcription factor. When cells are stimulated by various substances such as inflammatory mediators, virus infection, and oxidative stress, NF-κB is activated in the cytoplasm, enters the nucleus and interacts with viruses. , cytokines, growth factors, cell adhesion molecules, acute phase response proteins, enzymes and other gene enhancer sequences combined to enhance gene transcription; thus play a role in the pathogenesis of a series of diseases involving cytokines, inflammatory mediators and proteases important role. A large number of studies have shown that NF-κB hyperactivation is associated with inflammation, vascular diseases, t...

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Abstract

Disclosed is a transcription factor protein detection method through exonuclease III digestion FRET-dsDNA micro array chips, wherein the expression and activation levels of the transcription factor are analyzed through the following steps: (1) preparing FRET-dsDNA micro array chips, (2) reacting the transcription factor with FRET-dsDNA micro array chips, (2) reacting the exonuclease III with FRET-dsDNA micro array chips, (1) proceeding detection and analysis to the FRET-dsDNA micro array chips.

Description

1. Technical field: [0001] Transcription factor (transcription factor) is a kind of important protein responsible for gene expression regulation in proteomics. ) and proteome research; many diseases are closely related to the abnormal expression and activation of transcription factors, and have become important targets for transcriptional therapy and drug research (drug target). The detection and analysis of transcription factor expression and activation degree is the main means to study its function. This patent proposes a new method for detecting the expression and activation level of transcription factors, which will be used in the research of transcription factors in the fields of molecular biology, functional genomics, proteomics and biomedicine Provide a new detection and analysis technology that can promote scientific research on transcription factors in these fields, and provide a diagnostic technology for disease-related transcription factors in the biomedical field,...

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68
Inventor 王进科
Owner 王进科
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