Method for preparing recombinant goose interferon I and II

A technology for interferon and purpose, applied in the field of preparation of goose recombinant I and II interferon

Inactive Publication Date: 2006-08-23
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no research reports on goose interferon at home and abroad. The preparation of goose genetically engineered interferon has laid a material foundation for the study of the physicochemical properties and biological activities of goose interferon.

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0061] The prokaryotic expression protein encoding the goose IFN-α mature protein (mIFN-α) and the IFN-γ mature protein (mIFN-γ) gene, the baculovirus / insect cell expression protein and the preparation method thereof include:

[0062] (1) Designing specific primers for amplifying the genes encoding goose IFN-α mature protein and encoding IFN-γ mature protein;

[0063] (2) Obtain the genes encoding goose IFN-α mature protein and IFN-γ mature protein by RT-PCR amplification;

[0064] (3) Cloning and sequencing identification of mIFN-α and mIFN-γ gene sequences respectively;

[0065] (4) Directional subcloning of mIFN-α and mIFN-γ genes into pET30a prokaryotic expression vector;

[0066] (5) Induced expression of mIFN-α and mIFN-γ genes in Escherichia coli respectively;

[0067] (6) Affinity chromatography purification of mIFN-α and mIFN-γ gene fusion expression protein, mainly including:

[0068] ① Affinity chromatography purification of mIFN-α gene fusion expression protein ...

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Abstract

The present invention provides the preparation process of recombinant type I and II goose interferon. The preparation process includes: RT-PCR amplifying the gene coding goose IFN-alpha mature protein and IFN-gamma mature protein; sequencing identification of gene sequences, directionally sub-cloning target genes to pET30a prokaryotic expression vactor separately; inducing expressing in colibacillus BL21(DE3)plysS separately, separately performing affinity chromatographic purification of the fusion expressed target protein, separately directionally sub-cloning to pBlueBacHis2A and pMel BacA baculovirus transfer vector, separately cotransfecting insect cell Sf9 with the transfer vector and linearized baculovirus DNA, recombining virus infected insect cell, and expressing goose mIFN-alpha and mIFN-gamma in the prokaryotic expression vactor. The products may be used as goose antiviral vaccine medicine and immunity adjuvant.

Description

(1) Technical field [0001] The invention relates to a preparation method of genetic engineering products, specifically a preparation method for expressing goose IFN-alpha mature protein and IFN-gamma mature protein by an Escherichia coli prokaryotic expression system and a baculovirus / insect cell eukaryotic expression system. (2) Background technology [0002] At present, the prevention and control of poultry diseases still adopts vaccine immunization, chemical medicine and antibiotics. However, the extensive use of antibiotics and chemical drugs is likely to cause environmental problems and endanger human health. Vaccine immunization prevention also has problems such as single serotype and the speed of vaccine development cannot catch up with the speed of strain mutation, which often leads to immune failure. Cytokines, as endogenous substances in the body, have high-efficiency immune regulation. IFN is the earliest in the body's anti-viral infection defense response, and h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N15/21C12N15/23C12N1/21C12N15/866C07K14/56C07K14/57A61K38/21
Inventor 王君伟李洪涛马波秘晶纬
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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