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A mzf1 protein can bind DNA fragments and its application in the detection of mzf1 activity

A protein and fragment technology, applied in the field of molecular biology, can solve problems such as changes in activity and differences in MZF1 protein content

Active Publication Date: 2019-12-20
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Claims
  • Application Information

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Problems solved by technology

However, the current detection of endogenous MZF1 activity in cells still relies on techniques such as Western Blot or qRCR. Although the sensitivity is high, only the difference in MZF1 protein content is detected, and it cannot directly reflect the change in activity after it binds to the target sequence.

Method used

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  • A mzf1 protein can bind DNA fragments and its application in the detection of mzf1 activity
  • A mzf1 protein can bind DNA fragments and its application in the detection of mzf1 activity
  • A mzf1 protein can bind DNA fragments and its application in the detection of mzf1 activity

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Embodiment Construction

[0028] 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.

[0029] 1. Construction of DNA fragments that MZF1 protein can bind

[0030] The inventors researched and analyzed MZF1, and found that the DNA sequence bound by the MZF1 protein is 5'-AGTGGGGA-3', and 5'-CGGGNGAGGGGGAA-3' (N represents A, T, C, G, such as SEQ ID NO: 1- 4), when synthesizing the DNA core sequence, in the embodiment of the present invention, when designing and synthesizing the DNA core sequence, the two DNA core sequences are repeated twice, separated by TA bases, to obtain a sequence such as SEQ ID NO : The DNA fragment shown in 5.

[0031] In order to ensure the success of the vector construction, the cohesive end of the corresponding restriction endonuclease site was added at the end. In this example, the cohesive end of the Kpn I ...

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Abstract

The present invention relates to a DNA fragment which can be combined with MZF1 protein, which contains multiple MZF1 protein binding boxes; also relates to the application of the DNA fragment; and also relates to a method for detecting intracellular MZF1 transcription regulation activity. By using the DNA fragments and methods of the present invention, the transcriptional regulatory activity of MZF1 in cells can be directly and specifically detected, rather than just detecting its transcript or protein content, thereby enabling more accurate analysis of MZF1 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 which can be combined with MZF1 protein and its application in detecting the activity of MZF1 transcription regulation in cells. Background technique [0002] Myeloid zinc finger protein 1 (MZF1) is a protein with a zinc finger structure encoded by the MZF1 gene of human chromosome 19. It belongs to the C2H2-like zinc finger protein family and exists in large quantities in human cells. MZF1 protein has a full length of 734 amino acids, about 82055 Da. After binding to the target gene, it combines with the DNA of the promoter region of the target gene through its unique zinc finger structure, and exerts gene activation or inhibition effect in the process of bone marrow hematopoiesis. [0003] Mutations in the MZF1 gene can cause certain regulation disorders and even diseases. However, the current detection of endogenous MZF1 activity in cells still ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115C12Q1/66
CPCC12N15/115C12N2310/16C12Q1/66
Inventor 童强松郑丽端王建群肖文晶洪梅叶霖李聃宋华杰
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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