Detecting protein of transcriptional factor through probe of digestion FRET marke nucleic acid of nuclease

A technology of transcription factors and nucleic acid probes, applied in the field of nuclease digestion of FRET-labeled nucleic acid probes to detect transcription factor proteins, can solve the problems of high chip preparation and use costs

Inactive Publication Date: 2006-02-01
王进科 +1
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  • Detecting protein of transcriptional factor through probe of digestion FRET marke nucleic acid of nuclease
  • Detecting protein of transcriptional factor through probe of digestion FRET marke nucleic acid of nuclease
  • Detecting protein of transcriptional factor through probe of digestion FRET marke nucleic acid of nuclease

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Embodiment Construction

[0106] Here only FAM / Dabcyl labeled FRET nucleic acid probes were used to detect the transcription factor protein NF- κ The experiment of B is taken as an example to illustrate the specific implementation of the technology of the present invention.

[0107] 1. Design and chemical synthesis of FRET-labeled nucleic acid probes

[0108] We designed and synthesized by BIOASIA Biologic Technology Co.LTD., Shanghai, China, the following two oligonucleotides with reverse complementary base sequences for the preparation of FRET for detecting NF-κB Nucleic acid probe:

[0109] FAM-strand: 5′...AGTTGAG GGGACTTTCC CGAA TCG GGGACTTTCC CAGGC...3'( dT-FAM)

[0110] Dabcyl-strand: 3′...TCAACTC CCCTGAAAGG GCT AAGC CCCTGAAAGG GTCCG...5′( dT-Dabcyl)

[0111] The underlined sequence in the oligonucleotide is the NF-κB binding sequence.

[0112] Dissolve oligonucleotide FAM-strand and Dabcyl-strand in TEN buffer (10mM Tris, 1mM EDTA, 0.1M NaCl, pH8.0) at a concentration of 100...

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Abstract

A method for detecting transcription factor protein by nucleic acid probe with nuclease protection PRET label includes preparing fluorescence labeled nucleic acid probe, carrying out combination reaction of nucleic acid probe with transcription factor protein, adding nuclease in combination reaction system to carry out enzyme nick reaction and detecting reaction system by fluorescence to reflect expression and activation level of transcription factor protein.

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): G01N33/52G01N33/50G01N21/76C12Q1/68
Inventor 王进科
Owner 王进科
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