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A cux1 protein that can bind to DNA fragments and its application in the detection of cux1 activity

A protein and fragment technology, applied in the field of molecular biology

Active Publication Date: 2019-12-13
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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Problems solved by technology

However, the current detection of CUX1 endogenous activity still relies on methods such as Western Blot or qRCR. Although the sensitivity is high, what is detected is only the difference in the content of CUX1 protein and mRNA, which cannot directly reflect that it binds to the target sequence as a transcription factor to regulate transcription. activity changes

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  • A cux1 protein that can bind to DNA fragments and its application in the detection of cux1 activity
  • A cux1 protein that can bind to DNA fragments and its application in the detection of cux1 activity
  • A cux1 protein that can bind to DNA fragments and its application in the detection of cux1 activity

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Embodiment Construction

[0027] The principles and features of the present invention are described below in conjunction with examples, which are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0028] 1. Construction of DNA fragments that CUX1 protein can bind

[0029] The inventors researched and analyzed CUX1, and found that the DNA sequence bound by CUX1 protein is ATCRAT (R represents A, G). When synthesizing the DNA core sequence, this DNA core sequence was repeated four times, and the core base was adjacent to the base. spaced and connected to the expression vector (pGL3.0-Basic) carrying firefly luciferase. After successful construction, the luciferase reporter gene expression vector contains four repeats of the DNA core sequence bound by the CUX1 protein. The base adjacent to the base is spaced, and its sequence is shown in SEQ ID NO:1.

[0030] In order to ensure the success of the vector construction, the cohesive end of the corr...

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Abstract

The present invention relates to a DNA fragment that can bind to a CUX1 protein, which comprises one or more CUX1 protein binding boxes, and the sequence of the CUX1 protein binding box is 5'-ATCGAT-3' or 5'-ATCAAT-3'; It relates to the application of the DNA fragment; it also relates to a method for detecting the CUX1 transcription regulation activity in cells. By using the DNA fragments and methods of the present invention, the transcriptional regulatory activity of CUX1 in cells can be directly and specifically detected, rather than just detecting its transcript or protein content, thereby enabling more accurate analysis of CUX1 as a transcription factor in some diseases. role in the development of science.

Description

technical field [0001] The invention relates to the field of molecular biology, and more particularly relates to a DNA fragment to which a CUX1 protein can bind and its application in detecting the CUX1 transcription regulation activity in cells. Background technique [0002] CUX1 (CCAAT displacement protein), also known as CDP / Cux / Cut (CCAAT-displacement protein / cut homeobox), is a kind of transcription family protein that is widely distributed in multicellular species and plays a vital role in gene transcription regulation In addition, this molecule also participates in the regulation of gene post-transcriptional level and has various biological functions. A number of studies have shown that: CUX1, as a transcription factor, can bind to and regulate the expression of various proto-oncogene promoter regions (about -2000bp upstream of the transcription initiation site). [0003] Studies suggest that: CUX1 protein can combine with various proteins and DNA through its unique ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/66C12N15/85
CPCC12N15/11C12N15/85C12Q1/66
Inventor 童强松郑丽端李欢欢杨枫叶霖陈亚俊宋华杰李聃
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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