C-Myc protein combinable DNA (Deoxyribonucleic Acid) segment and application thereof to c-Myc activity detection

A c-myc and protein technology, applied in the field of molecular biology, can solve problems such as changes and inability to reflect c-Myc activity

Active Publication Date: 2017-12-12
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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Problems solved by technology

[0004] At present, there are many methods for detecting c-Myc. Although methods such as western blot or qPCR have high sensitivity, they only detect the difference in the protein content of c-Myc, and cannot reflect the changes in the activity of c-Myc after binding to its specific target sequence.

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  • C-Myc protein combinable DNA (Deoxyribonucleic Acid) segment and application thereof to c-Myc activity detection
  • C-Myc protein combinable DNA (Deoxyribonucleic Acid) segment and application thereof to c-Myc activity detection
  • C-Myc protein combinable DNA (Deoxyribonucleic Acid) segment and application thereof to c-Myc activity detection

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Embodiment Construction

[0028] The principles and features of the present invention will be described below with reference to examples. The examples are only used to explain the present invention, but not to limit the scope of the present invention.

[0029] 1. Construction of DNA fragments that can be bound by c-Myc protein

[0030] The inventors conducted research and analysis on c-Myc and found that the DNA sequence bound by c-Myc protein was 5'-SCACGTGS-3' (S represents C or G). When synthesizing the DNA core sequence, the DNA core sequence was repeated three times. , separated by A / T bases with higher frequency near the core base, the sequence is shown in SEQ ID NO: 1, and the sequence is connected to the expression vector (pGL3.0-Basic) carrying firefly luciferase .

[0031] In order to ensure the successful construction of the vector, add the sticky end of the corresponding restriction endonuclease cleavage site at the end, in this example, add the sticky end (CATGG) of the Kpn I enzyme cleav...

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Abstract

The invention relates to a c-Myc protein combinable DNA (Deoxyribonucleic Acid) segment. The c-Myc protein combinable DNA segment comprises one or more c-Myc protein combining frames, wherein the sequence of each c-Myc protein combining frame is 5'-SCACGTGS-3' and S represents C or G; the invention further relates application of the DNA segment; the invention further relates to a method for detecting transcriptional regulation activity of c-Myc protein in cells. By utilizing the DNA segment and the method, provided by the invention, the transcriptional regulation activity of the c-Myc protein in the cells can be directly and specifically detected and the content of transcripts or the protein is also detected, so that the effect of the c-Myc protein, which is used as a transcription factor, in development of certain diseases can be more accurately analyzed.

Description

technical field [0001] The present invention relates to the field of molecular biology, more particularly, to a DNA fragment that can be combined with c-Myc protein and its application in detecting intracellular c-Myc transcription regulation activity. Background technique [0002] c-Myc is a member of the Myc family of cell cycle-related transcription factors, which can regulate apoptosis, cell cycle progression, and cellular metabolic reprogramming. The c-Myc gene is located on chromosome 8q24 and is mainly activated by amplification and chromosomal translocation and rearrangement, and is closely related to the occurrence, development and evolution of tumors. c-Myc gene is amplified and overexpressed in human myeloid leukemia, retinoblastoma, breast cancer, lung cancer, colon cancer, osteosarcoma, chondrosarcoma, chordoma, liposarcoma, rhabdomyosarcoma and other tumor tissues. early recurrence of tumors. [0003] Structurally, c-Myc protein can be divided into transcript...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/85C12Q1/66
CPCC12N15/11C12N15/85C12Q1/66
Inventor 童强松郑丽端洪梅王建群肖文晶叶霖李聃杨枫
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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