Long-noded pit viper poison dissolving fiber protein No.2 gene and use thereof

A gene and genetic engineering technology, applied in the preparation of fibrin-dissolving drugs and the field of embolism diseases, can solve the problems of low market share, inexplicability, toxic and side effects, etc., achieve huge social and economic benefits, and good thrombolytic effect , the effect of less bleeding reaction

Inactive Publication Date: 2006-06-07
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, there are regional and seasonal variations in the composition and biological activity of natural-sourced five-step snake venom, and it is actually difficult to obtain stable clinical curative effect; in addition, due to the complex composition of crude snake venom, it is still difficult to purify it. Obtaining a truly single chemical composition makes clinical application imply certain toxic and side effects; the production of natural snake venom is limited, the cost is high, and the market share is low; its molecular structure and the relationship between structure and activity cannot be clarified by protein technology

Method used

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  • Long-noded pit viper poison dissolving fiber protein No.2 gene and use thereof
  • Long-noded pit viper poison dissolving fiber protein No.2 gene and use thereof
  • Long-noded pit viper poison dissolving fiber protein No.2 gene and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0038] 1. Separation of fibrinolytic factor FII from snake venom in five steps

[0039] 1. DEAE-SephadexA-50 ion exchange chromatography

[0040] Method: DEAE-SephadexA-50 ion-exchange gel was rinsed repeatedly in 0.05M ammonium acetate buffer solution with pH 8.0, soaked at 25°C for about 24 hours, and then packed into a column (gel column diameter 2.6cm, height 100cm). When the pH value of the effluent is close to 8.0, the balance is completed. 3g of five-step snake venom crude venom is dissolved in 10ml of 0.05M ammonium acetate buffer solution with pH 8.0, centrifuged at 1500rpm for 15 minutes to remove sediment, and then put on the column. Use 750ml each of 0.05M pH 8.0 ammonium acetate buffer and 1M pH 5.0 ammonium acetate buffer for linear gradient elution, and the eluent collection rate is 4tube / h, 3ml / tube. Light absorption values ​​were measured at a wavelength of 280 nm. The fibrinolytic activity of each peak was measured, and the components with fibrinolytic acti...

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Abstract

The invention relates to the technical field of genetic engineering, in particular to the No. 2 gene of Agkistrodon akistrodon (pentapod venom) dissolving fibrin (original), a carrier containing the gene, a host cell genetically engineered by using the carrier and the The gene is used to prepare medicines for dissolving fibrin (ogen) to resist embolism diseases caused by thrombus formation. Isolate the fibrinolytic factor FII from the crude venom of the five-step snake; complete the screening and cloning of the fibrinolytic factor FII gene; express the fibrinolytic factor FII in yeast cells; purify the expressed product and measure the fibrinolytic activity. The cellolytic factor expressed by the genetically engineered yeast system in the present invention has high activity, large output, stable quality and low cost, and can be a new type of powerful thrombolytic drug with huge social and economic benefits.

Description

[technical field] [0001] The invention relates to the technical field of genetic engineering, in particular to the No. 2 gene of Agkistrodon akistrodon (pentapod venom) dissolving fibrin (original), a carrier containing the gene, a host cell genetically engineered by using the carrier and the The gene is used to prepare medicines for dissolving fibrin (ogen) to resist embolism diseases caused by thrombus formation. [Background technique] [0002] Embolism diseases caused by intravascular thrombosis, especially myocardial infarction and cerebral apoplexy, are currently the most harmful diseases to human health in the world, and their morbidity and fatality rates rank first among all diseases. In my country, with the control of most infectious diseases, the significant improvement of people's living standards, the general extension of population life expectancy and the development of population aging, the incidence of heart and cerebrovascular thrombosis has risen to The first...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/63C07K14/75C07H21/04A61K38/17A61K38/36A61P9/00
Inventor 颜光美陈家树邱鹏新单鸿
Owner SUN YAT SEN UNIV
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