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Angiogenin mutant as agonist, and its preparing method

An angiogenin, mutant technology, applied in the direction of angiogenin, angiogenesis factors, biochemical equipment and methods, etc., can solve problems such as the inability to meet structural molecular biology research and other problems

Inactive Publication Date: 2006-10-25
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
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Problems solved by technology

However, due to the limitations of various techniques, relying solely on experimental techniques to solve the problem of protein stereostructure is obviously far from meeting the needs of structural molecular biology research.

Method used

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  • Angiogenin mutant as agonist, and its preparing method
  • Angiogenin mutant as agonist, and its preparing method
  • Angiogenin mutant as agonist, and its preparing method

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Experimental program
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Effect test

Embodiment Construction

[0043] The invention provides an angiopoietin mutant as an antagonist and a preparation method thereof.

[0044] 1. Experimental materials and equipment

[0045] 1.1 Strains and expression vectors

[0046] The strain E.coliDH5α was preserved (Institute of Basic Medical Sciences, Academy of Military Medical Sciences), the pT-Ang plasmid (both Institute of Basic Medical Sciences, Academy of Military Medical Sciences) was preserved, and the expression vector pGEX-4T-2 was purchased from Amershampharmacia.

[0047] 1.2 Enzymes and biochemical reagents

[0048] Restriction enzymes BamH I, EcoR I, Sal I, Nco I, Taq DNA polymerase and T4 DNA ligase were purchased from Promega Company; nucleic acid molecular weight standard DL2000 was purchased from TaKaRa Company; protein molecular weight standard was purchased from Jingke Bioengineering Company; Affinity chromatography column GSTrap FF and thrombin (Thrombin) were purchased from Amersham pharmacia; reduced glutathione (GSH) and M4...

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Abstract

The invention uses biology information to reconstruct Ang crystal structure, and construct the 3D structure, and construct the lacunae mutant that the N end lack 5 amino acid and the C end lack 12 amino acid, the reconstructed Ang mutant would be constructed by gene reconstructing and expression technology. The invention supplies a new method for curing tumor.

Description

technical field [0001] The invention belongs to the technical fields of structural molecular biology and biological anti-cancer, and particularly relates to an angiopoietin mutant as an antagonist and a preparation method thereof. Background technique [0002] In 1985, Fett JW et al. isolated and identified a protein factor for the first time from a serum-free cultured human adenocarcinoma cell line (HT-29), which has a significant function of promoting angiogenesis, and named it angiopoietin (Angiogenin, Ang). It has been found that many kinds of cells can produce this protein factor, and its expression level is increased to varying degrees in almost all tumor patients. Angiogenin induces a second messenger response in endothelial cells, enhancing the adhesion of these cells to the surface of the dish. Studies have found that angiogenin is involved in morphological changes and angiogenesis in the ovary. In addition, it has some non-angiogenic functions, including: inhibi...

Claims

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Application Information

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IPC IPC(8): C07K14/515A61P35/00C12N15/12C12P21/02
Inventor 徐东刚邹民吉冯健男王颖徐涛王嘉玺
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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