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Protein interaction detection method based on nucleotide sequence

A protein and DNA sequence technology, applied in the field of protein research, can solve the problems of limited fluorescent protein fluorescence intensity, protein space structure, simultaneous detection of protein interaction network, insufficient dynamic detection and quantitative detection capabilities, etc., to achieve the goal of improving throughput Effect

Inactive Publication Date: 2014-10-15
UNIV OF SCI & TECH OF CHINA
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Problems solved by technology

For example, although the yeast two-hybrid technology (Y2H) is simple to implement, its dynamic detection and quantitative detection capabilities are obviously insufficient. Although the fluorescence resonance energy transfer technology (FRET) has good spatiotemporal resolution and quantitative capabilities, it is limited. The fluorescence intensity of fluorescent protein and the protein's spatial structure and other properties
Moreover, these traditional detection techniques can only detect a single protein interaction pair, and cannot simultaneously detect a protein interaction network containing multiple pairs of interactions in the same cell

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  • Protein interaction detection method based on nucleotide sequence
  • Protein interaction detection method based on nucleotide sequence
  • Protein interaction detection method based on nucleotide sequence

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Embodiment Construction

[0031] The technical solution of the present invention will be described in further detail below. It should be noted that the various embodiments of the present invention can be combined in any way as desired.

[0032] The first aspect of the present invention provides a protein dimerization footprint method (PDf) for obtaining information on protein-protein interactions. The influence of the binding kinetics between the sequences realizes that the interaction between the target proteins is expressed by the corresponding specific DNA sequence, and the interaction strength between the target proteins is given by the copy number of the specific DNA sequence.

[0033] In one embodiment, the method comprises the steps of:

[0034] (1) Expressing a target protein fused with a DNA-binding domain in a host cell, the affinity of the DNA-binding domain to the corresponding specific DNA is much higher than that of a monomer when dimerized;

[0035] (2) Treating with a cross-linking ag...

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Abstract

The invention relates to a protein interaction detection method, namely, a PDf (protein dimerization footprinting) method based on a nucleotide sequence, a kit and an application of the kit. The basic principle is that based on the influence of the interaction of target protein on the combined structural domain of DNAs fused by the protein and combination dynamics of a corresponding special DNA (deoxyribonucleic acid) sequence, the interaction of target protein can be represented by the corresponding special DNA sequence, and the target protein interaction strength is obtained by a copy number of the special DNA sequence.

Description

technical field [0001] The invention belongs to the technical field of protein research, and specifically relates to the detection of protein interaction, in particular to an in vivo detection method for protein interaction using DNA-encoded protein interaction information, a kit and application thereof. Background technique [0002] Although traditional in vivo protein-protein interaction detection techniques have been widely used in biological research, there are many shortcomings. For example, although the yeast two-hybrid technology (Y2H) is simple to implement, its dynamic detection and quantitative detection capabilities are obviously insufficient. Although the fluorescence resonance energy transfer technology (FRET) has good spatiotemporal resolution and quantitative capabilities, it is limited. Fluorescence intensity of fluorescent protein and properties such as protein spatial structure. Moreover, these traditional detection techniques can only detect a single prot...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6806C12Q2522/101C12Q2521/301
Inventor 吴家睿梁治骆斯伟
Owner UNIV OF SCI & TECH OF CHINA
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