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

A protein complex, cell technology, applied in the field of bioengineering, can solve the problems of inability to identify the body components of human cells, and no human cells have been seen yet.

Inactive Publication Date: 2005-12-28
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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Experimental program
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Embodiment Construction

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

[0011] 1. Collect human cells at the end of mitosis

[0012] (1) After culturing human cultured cells in 5mM Thymidine (Thymidine) culture medium for 16 hours, the cells are synchronized in G1 / S phase;

[0013] (2) Wash off the drug, culture with fresh medium (DMEM solution containing 5-8% fetal bovine serum) for 8 hours to allow the cells to pass through the S phase synchronously.

[0014] (3) Add 50ng / ml Nocodazole and incubate for 4 hours to synchronize the cells in the pre-middle stage of mitosis;

[0015] (4) Wash away the drugs, and use fresh medium (DMEM containing 5-8% fetal bovine serum) to culture in a shaker at 37°C for 75 minutes to make the cells highly synchronized at the end of mitosis;

[0016] (5) Collect cells at the end of mitosis using conventional methods;

[0017] 2. Collect midbody complexes in human cells at the end stage

[0018] (6) Add 25 times the cell volume of...

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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 present invention relates to the field of bioengineering technology, in particular to the separation technology of human cells. Background technique [0002] Current research shows that the midbody of a cell is a dense complex formed by microtubules and some other protein complexes. Although the mechanism of the formation of the midbody is not yet fully understood, the important regulatory role of the midbody in the process of cell division has been recognized by many cell biologists in the world. The cell division cycle is a strictly regulated and highly ordered process. Mitosis only accounts for about 1 / 20 of the entire cell cycle. This short process involves the equal distribution and regulation of very important genetic material (such as chromosomes, organelles, etc.). The regulation of life involves a wide range. Among them, genetic information includes the holder of the gene chromosome and the main unit of cytoplasmic inheritance. The organelle is...

Claims

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Application Information

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