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GATA2 protein binding DNA fragment and application in GATA2 activity detection

A protein and fragment technology, applied in the field of molecular biology, can solve the problems of low sensitivity and complicated operation

Active Publication Date: 2017-08-11
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the current detection of endogenous GATA2 still relies on Western Blot technology. Although this type of method has high specificity, it is low in sensitivity and complicated to operate. What is detected is only the difference in GATA2 protein content, and cannot directly reflect the change in its activity. , and the activity of GATA2 is closely related to the occurrence and development of tumors. What we need to detect is not only the transcript or protein content of GATA2, but also the activity of GATA2 binding to effective sites to affect transcription.

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  • GATA2 protein binding DNA fragment and application in GATA2 activity detection
  • GATA2 protein binding DNA fragment and application in GATA2 activity detection
  • GATA2 protein binding DNA fragment and application in GATA2 activity detection

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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 GATA2 protein can bind

[0030] The inventor researched and analyzed GATA2 and found that the DNA sequence bound by the GATA2 protein is 5'-T / A(GATA)A / G-3'. When synthesizing the DNA core sequence, this DNA core sequence was repeated four times to obtain AG, AC or AA bases are 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 GATA2 protein The combined DNA core sequence is separated by AG, AC or AA bases, and its sequence is shown in SEQ ID NO:1.

[0031] In order to ensure the success of the vector construction, the cohesive end of the corresponding restriction en...

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Abstract

The invention relates to GATA2 protein binding DNA fragment, which comprises one or more GATA2 protein binding cassettes; the invention also relates to an application of the DNA fragment; and the invention also relates to a method for detecting GATA2 transcription regulating activity in cells. With the application of the DNA fragment and the method provided by the invention, the transcription regulating activity of GATA2 in the cells, instead of merely a transcript or a protein content, can be directly detected in a targeted mode, so that effects of the GATA2, as a transcription factor, in some developmental biology and pathological development can be analyzed more accurately.

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 GATA2 protein and its application in detecting the activity of GATA2 transcription regulation in cells. Background technique [0002] GATA-binding protein 2 (GATA2), encoded by the GATA2 gene, is a member of the GATA family of transcription factors. Current studies have found that: GATA2 plays an important role in transcriptional regulation of genes involved in the proliferation and differentiation of hematopoietic and endocrine system tumor cells. Through its C-terminal zinc finger structure, GATA2 binds to the target gene promoter, upstream of the transcription start site or close to the promoter region to exert its biological function. [0003] GATA2 protein has attracted much attention as a transcriptional regulator. Studies have shown that in solid tumors, high expression of GATA2 is closely related to poor prognosi...

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Application Information

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IPC IPC(8): C12N15/115C12Q1/66
CPCC12N15/115C12N2310/16C12Q1/66
Inventor 童强松郑丽端陈亚俊杨枫李欢欢叶霖李聃宋华杰
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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