Method for preparing venom edema factor inhibition protein and application thereof

Abstract

The invention relates to method for preparing venom edema factor inhibition protein and the application thereof, belonging to the field of biotechnology and relating to a method for separating and purifying a protein having inhibition action on venom edema factors from the blood serum of Gloydius shedaoensis. The invention is characterized by the method adopted for separating and purifying the inhibition protein, and the method comprises the following steps: performing the ammonium sulphate precipitation on the crude blood serum of Gloydius shedaoensis, performing gel filtration twice, passing through an anion exchange column, an affinity chromatography column and a reversed phase HPLC to obtain a single component with the molecular weight of 25KDa, and identifying 70 amino acid sequencesof the N terminal and 20 amino acid sequences of the C terminal of the protein. The invention is also characterized by obtaining the gene sequence corresponding to the venom edema factor inhibition protein, the gene sequence is established on a TOPO expression vector for producing in vitro and expressing the protein. The invention has the advantages of providing a preparation method for producingin vitro venom edema factor inhibition protein, and the inhibition protein can be applied to the clinical treatment of snakebite and other diseases.

Description

technical field [0001] The invention relates to the preparation and application of a snake venom edema factor inhibitory protein, and belongs to the field of biotechnology. Background technique [0002] Snakes belong to lower animals. There are about 13 families, 400 genera, and more than 2,700 species in the world, of which 1 / 4 are poisonous snakes. There are about 8 families, 53 genera, and 173 species in China. It represents the corresponding changes of genes and functional proteins in various living environments, and is the most direct and true biological evolution material. [0003] The whole body of the snake is full of treasures, and it has a long history of medicinal use in my country. "Compendium of Materia Medica" contains: snake slough, snake skin, snake meat, snake blood, snake viscera, snake venom, snake gall, snake tongue, snake eggs, snake oil, etc. can be used as medicine, especially snake venom. Snake venom is the secretion of the specialized oral gland o...

AI Technical Summary

Problems solved by technology

There are many studies on the separation and purification of neurotoxins, muscle necrosis factors, edema factors, hemolytic factors, bleeding factors, etc. in snake venoms, but the development of snake venom inhibitory proteins is still limited to the neutralization and inhibition of snake venom mixtures. There is still very little research on the specific inhibition of a single component of snake venom components in the processing of mixed inhibitor protein resource products

In addition, most of the current snake venom inhibitors are obtained from animals (such as horses), which requires raising a large number of animals, and there are not many methods and means for obtaining snake venom inhibitors in vitro.

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