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Novel human cell growth inhibiting gene THAP11 and application thereof

A technology of genes and preparations, applied in the field of new human cell growth regulation genes

Inactive Publication Date: 2010-05-26
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no research report on the function of THAP11

Method used

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  • Novel human cell growth inhibiting gene THAP11 and application thereof
  • Novel human cell growth inhibiting gene THAP11 and application thereof
  • Novel human cell growth inhibiting gene THAP11 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Obtaining of human THAP11 cDNA

[0035] Using the EDAG gene as bait, using the common yeast two-hybrid technique in the field to study its interacting proteins, a DNA sequence was obtained, and the DNA sequence was used for BLAST retrieval. The present invention found that it has a high homology with the speculated human gene THAP11 . The total RNA of human bone marrow cells was extracted, and reverse transcription was performed according to the instructions of the reverse transcription kit (Promega Company). Take 1μg of quantified RNA, add water to make up to 10.5μl, 70℃, 10min, add 2μl of 10×buffer, 2μl of MgCl 2 , 2 μl of dNTP, 1 μl of oligo dT, 1 μl of rRNasin, 1.5 μl of AMV (15U), the total reaction system is 20 μl; 42°C, 1h; 94°C, 5min; 4°C, 5min. RNA is reverse transcribed into cDNA. The target fragment was amplified by PCR with primers 5'-GGGGTACCATGCCTGGCTTTACGTGCTGCG-3'; 5'-CGGAATTCCACATTCCGTGCTTC TTGCGGATG-3'. The PCR conditions are:

[0036] C...

Embodiment 2

[0046] Example 2THAP11 is expressed in various tissues

[0047] Using 5'-GGGGTACCATGCCTGGCTTTACGTGCTGCG-3'; 5'-CGGAATTCCACATTCCGTGCTTCTTGCGGATG-3 as primers, PCR technology was used to amplify THAP11 using 8 adult tissue cDNAs as templates, and G3PDH was used as an internal reference. The results showed that THAP11 was expressed in the eight adult tissues tested, and the expression level was higher in the heart and skeletal muscle ( figure 1 ). It can be seen that THAP11 is widely distributed in adults without obvious specific tissue distribution.

Embodiment 3

[0048] Example 3 THAP11 is localized in the nucleus

[0049] To observe the subcellular localization of THAP11, we constructed THAP11 cDNA into the EcoRI / KpnI site of the pEGFP vector to form a THAP11-GFP fusion gene expression vector. The expression vector was transfected into COS7 cells, and laser scanning confocal microscope detection showed that the fluorescent signal was distributed as the nucleus ( figure 2 ), after mutating the predicted nuclear localization signal inside the THAP11 molecule, the cellular localization changes and becomes the whole cell distribution, indicating that the nuclear localization signal of THAP11 is necessary for its localization in the nucleus.

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PUM

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Abstract

The invention provides a novel human cell growth inhibiting gene and an application of an encoded protein. The gene and the encoded protein are related to the regulation and control of a cell cycle, can inhibit the growth of tumor cells by inhibiting the expression of a proto oncogene c-Myc, and are down-regulated in various tumor tissues, thereby having a close relation with the generation of tumors. The gene and the encoded protein and derivatives thereof can be used for the diagnosis, treatment and drug screening for tumors caused by disordered regulation and control of cell cycles and neurodegenerative diseases and the like.

Description

technical field [0001] The invention relates to a new human cell growth regulation gene, and also relates to the DNA constituting the gene and the protein coded by the DNA and its application in the diagnosis and treatment of tumors and neurodegenerative diseases. Background of the invention: [0002] Cell growth is complexly regulated by a variety of genes. As a proto-oncogene, c-Myc plays an important role in the signal regulation network regulating cell proliferation, growth, differentiation and apoptosis. Multiple gene studies have reported that down-regulation of c-Myc can trigger cell growth arrest (Roussel et al., 1991; He et al., 1998). As one of the most important transcription factors, c-Myc is widely expressed in various stages of embryonic development and in tissues with high proliferation ability. The expression level of c-Myc in the resting period of the cell is very low. Under the action of growth factors, the c-Myc protein is synthesized rapidly, and forms ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/11C12N15/86C07K14/47C07K16/18C12Q1/68G01N33/577G01N33/574A61K38/17A61P35/00
Inventor 杨晓明李长燕朱晨雁胡德庆许望翔詹轶群
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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