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Protein compound of human cell mesosome, its separation and use thereof

A protein complex and separation method technology, applied in the field of human cell separation, can solve the problems of human cells that have not been seen yet, and the inability to identify the body components in human cells.

Inactive Publication Date: 2007-12-26
UNIV OF SCI & TECH OF CHINA
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  • Abstract
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Problems solved by technology

Although researchers have sequenced the human genome in recent years, existing methods have not been able to identify the mesosome components of human cells
Therefore, there have been no reports on the study of body composition in human cells.

Method used

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Embodiment Construction

[0010] The present invention will be further described below through specific embodiments.

[0011] 1. Collection of Human Cells in Telophase

[0012] (1) Human cultured cells were cultured in 5mM Thymidine (Thymidine) culture solution for 16 hours, and the cells were synchronized in the G1 / S phase;

[0013] (2) The drug was washed away, and cultured with fresh medium (DMEM solution containing 5-8% fetal calf serum) for 8 hours, so that the cells passed through the S phase synchronously.

[0014] (3) Add 50 ng / ml Nocodazole (Nocodazole) and culture for 4 hours, so that the cells are synchronized in the prometaphase of mitosis;

[0015] (4) Wash away the drug, and culture it with fresh medium (DMEM containing 5-8% fetal bovine serum) in a shaker at 37° C. for 75 minutes, so that the cells are highly synchronized at the end of mitosis;

[0016] (5) collecting cells in the terminal phase of mitosis by conventional methods;

[0017] 2. Collection of Midbody Complexes in Telopha...

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PUM

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Abstract

Protein composites of human body cell mesosomem its separation and use are disclosed. The composites consist of at least 129 proteins: 30 cell skeleton proteins, 4 centrosome proteins, 6 vesicular transport related proteins, 6 chromosome regulatory proteins, 2 motor proteins, 3 check point proteins, 7 degradable regulatory proteins, 9 membrane proteins, 12 transcriptional and translational related proteins, 28 protein kinases, 5 mitochondria proteins, 9 molecular chaperone proteins and 7 missing proteins. The process is carried out by culturing, synchronizing and collecting for human body cell, adding hypotonic solution, re-suspending, incubating and expanding collected cell, breaking expanded cell, taking out supernatant, centrifugal separating from density gradient, extracting and purifying protein composite from decontaminate, and separation identifying mesosome protein type. It supplies a pattern system and target protein ezrin and target site.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the separation technology of human cells. Background technique [0002] Current research shows that the midbody of the cell is a dense complex formed by microtubules and some other protein complexes. Although the formation mechanism of midbody is not very clear at present, the important regulatory role of midbody in the process of cell division has been recognized by many cell biologists in the world. The cell division cycle is a tightly regulated, highly ordered process. The mitosis phase (mitosis) only accounts for about 1 / 20 of the entire cell cycle, and in this short process, it involves the equal distribution and regulation of quite important genetic materials (such as chromosomes, organelles, etc.). The regulation of life involves a wide range, among which genetic information includes the holder of genes, chromosomes and cytoplasmic inheritance, the main unit of or...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/30C07K1/14
Inventor 姚雪彪符传孩缪勇薛宇
Owner UNIV OF SCI & TECH OF CHINA
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