Serine protease inhibitor of Rana grahami, its gene and application

A serine protease and inhibitor technology, applied in the field of biomedicine, to achieve the effect of inhibiting tumor growth, simple structure, and obvious tumor inhibiting effect

Inactive Publication Date: 2006-12-27
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are few reports about the research on the active substances of the fingerless frog skin

Method used

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  • Serine protease inhibitor of Rana grahami, its gene and application

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Embodiment Construction

[0029] The substantive content of the present invention is further described below with embodiment, but content of the present invention is not limited thereto.

[0030] Cloning of the Serine Protease Inhibitor Gene of the Fingerless Frog:

[0031] I. Extraction of total RNA from the skin of the fingerless frog:

[0032] A. The live fingerless frog is cleaned with water, put into liquid nitrogen and quick-freeze for 4 hours, get the skin tissue, weigh, get 300mg skin tissue, add 10ml total RNA extraction buffer (Trizol solution, U.S. GIBCOBRL company product), Homogenize for 30 minutes in a 20ml glass homogenizer.

[0033] B. Add an equal volume of phenol / chloroform solution, mix vigorously, leave at room temperature for 10 minutes, centrifuge at 12,000 rpm for 10 minutes at 4°C, and discard the precipitate.

[0034]C. Add an equal volume of isopropanol to the supernatant, leave it at room temperature for 10 minutes, centrifuge at 12,000 rpm for 10 minutes at 4°C, wash the p...

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Abstract

Disclosed is an pharmacy application of both a serine protease inhibitor of Rana grahami Graham's frog and a gene thereof, pertaining to biomedicine field. Serine protease inhibitor of Rana grahami Graham`s frog is a cyclic polypeptide encoded by a gene of Rana grahami Graham's frog, which is a Chinese amphibian. Said serine protease inhibitor has a molecular weight of 1951.38 dolton, an isoelectric point of 9.51, and no enzymatic activity. The complete sequence thereof is NH2-AVNIPFKVHFRCKAAFC-COOH,in which Cys at 12th site and Cys at 17th site form an intramolecularly disulfide bond. The coding gene is composed of 301 nucleotides, and the mature coding part is nucleotides from 160th site to 213th site. Artificial serine protease inhibitor of Rana grahami Graham's frog has strong suppression activity for serine protease, and can be used in pharmacy for preparing medicine for tumor, gastritis, and pancreatitis. Said serine protease inhibitor has a simple sequence and can be synthesized with ease.

Description

Technical field: [0001] The invention provides a Rana grahami serine protease inhibitor and its gene and application, which belong to the technical field of biomedicine. Background technique: [0002] The most basic function of serine protease inhibitor (serpin) is to prevent proteolysis and regulate the hydrolysis balance of serine protease. Through the regulation of serine proteases, serine protease inhibitors have important effects on many important physiological and biochemical functions in organisms. The following 14 physiological and biochemical functions are regulated by serine protease inhibitors: blood coagulation, complement formation, fibrinolysis, protein folding, cell migration, cell differentiation, cell matrix remodeling, hormone formation and transport, intracellular proteolysis, blood pressure regulation, Tumor suppression and formation of viral or parasitic pathogenicity. Because so many physiological functions are regulated by it, serine protease inhibit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/57A61P35/00A61P1/04A61P1/18
Inventor 赖仞李建许韩耀平李东升梁建国徐学清杨海龙
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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