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Process for detection of dna modifications and protein binding by single molecule manipulation

A protein and molecular technology, applied in the field of determining whether a protein binds to a specific DNA sequence, can solve problems such as not providing insights into genome-wide biological functions, and achieve the effects of improving statistics, reducing instrument drift, and improving reliability

Inactive Publication Date: 2015-12-02
CENT NAT DE LA RECHERCHE SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many of these principles appear to be valid when studying other genes, these principles often do not provide genome-wide insights into biological function

Method used

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  • Process for detection of dna modifications and protein binding by single molecule manipulation
  • Process for detection of dna modifications and protein binding by single molecule manipulation
  • Process for detection of dna modifications and protein binding by single molecule manipulation

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Embodiment Construction

[0194] Experimental Example

Background of the invention

[0195] Binding of proteins to DNA is an important phenomenon in biology; it is a very general process that controls many reactions. While the thermodynamic equilibrium nature of this mechanism is well known, measuring its kinetics is a more challenging problem. The use of single molecules not only provides the ability to measure the time it takes a protein to find its DNA target but also the exact location of a binding event. We describe here a new single-molecule assay that achieves these goals.

[0196] Although this assay is broad in scope, we first demonstrate its applicability to binding of specific oligonucleotides, as well as non-specific binding of helicases to ssDNA.

[0197] Finally, we discuss the specific binding of antibodies that recognize methylated sites in DNA.

[0198] Summary

[0199] The present invention relates to novel methods for detecting a wide variety of DNA modification and DNA protei...

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Abstract

The present invention relates to a method for determining whether a protein binds to a specific DNA sequence. This method is useful in particular for identifying modifications to the DNA sequence (e.g. methylations) via the binding of proteins that specifically recognize those modifications (e.g. antibodies), but also to identify the binding sequence on DNA of a variety of proteins.

Description

technical field [0001] The present invention relates to methods for determining whether a protein binds to a specific DNA sequence. This method is particularly useful for identifying DNA sequence modifications (eg, methylation) through the binding of proteins (eg, antibodies) that specifically recognize these modifications, and also for identifying binding sequences of various proteins on DNA. Background technique [0002] Binding of proteins to DNA is an important phenomenon in biology; it has fundamental roles in regulating cellular and viral functions. These include fundamental cellular processes such as DNA replication, transcription, DNA repair and DNA recombination, but also DNA modification or maintenance of chromosome structure. [0003] Several proteins bind to specific sites in the genome to regulate genome expression and maintenance. DNA-binding proteins constitute a large family of proteins with diverse and important biological functions. The DNA-binding prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/557C12Q1/68
CPCG01N33/557C12Q1/6874G01N33/53G01N33/577C12Q1/6883C12Q2600/154C12Q2600/156
Inventor D·邦西蒙V·克罗凯特H·古埃J-F·阿莱曼德丁方圆
Owner CENT NAT DE LA RECHERCHE SCI
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