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Gene synthesis of wild boar alpha-interferon, vector construction and method for producing outcome

A technology of gene synthesis and production method, which is applied in the fields of gene synthesis, carrier construction and product production of wild boar α-interferon, which can solve the problems of low expression level and no high-density fermentation process, and achieve the effect of increasing expression level

Inactive Publication Date: 2009-03-25
黑龙江省农业科学院畜牧研究中心 +1
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Problems solved by technology

Although relevant research has been carried out in China, there is still no complete downstream process for production. Domestic recombinant strains either have low expression levels, or the product is expressed in a fusion state, or the product is purified. labels, or there is no high-density fermentation process, there are many problems

Method used

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  • Gene synthesis of wild boar alpha-interferon, vector construction and method for producing outcome
  • Gene synthesis of wild boar alpha-interferon, vector construction and method for producing outcome
  • Gene synthesis of wild boar alpha-interferon, vector construction and method for producing outcome

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Embodiment 1

[0027]The gene synthesis of wild boar α-interferon, the construction of vector and the production method of the product, its composition includes: the wild boar α-interferon gene is synthesized by using Escherichia coli preferred codons and codon pairs, constructing high-efficiency expression vectors and transforming high-efficiency expression strains, High-density fermentation of engineering bacteria, isolation and purification of inclusion bodies, denaturation of target proteins, renaturation and purification, and biological activity assay methods of expressed products. The artificially synthesized gene encoding wild boar α-interferon contains the following nucleotide sequence ATG TGT GAT CTG CCG CAA ACC CATAGC CTG GCG CAC ACC CGT GCG CTG CGT CTG CTG GCG CAG ATG CGT CGC ATC AGCCCG TTT AGC TGC CTG GAT CAC CGT CGC GAT TTT GGT TTC CCG CAG GAA GCG CTGGGC GGC AAC CAG GTG CAG AAA GCG CAA GCC ATG GCG CTG GTG CAT GAG ATG CTGCAG CAG ACC TTC CAG CTG TTC AGC ACC GAA GGC AGC GCG GCG GCC ...

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Abstract

The invention discloses a gene synthesis of Alpha-interferon of wild boars and vector construction thereof as well as a production method of a product. A number of problems of low expression capacity, products expressed in a fusion state, products with purification tags or no fermentation technology with high density exist in the domestic recombinant strains. The invention includes a method that a codon and codon pairs, preferred by Escherichia coli, are used for synthesizing an Alpha-interferon gene of the wild boar, establishing a high-efficiency expression vector and transforming a high-efficiency expression strain as well as methods of high-density fermentation of engineering bacteria, separation and purification of inclusion bodies, the modification, the renaturation and the purification of target protein and the determination of biological activity of the expressed product. The gene synthesis, the vector construction and the production method pertain to the technical field of the production of polypeptide drugs by genetic engineering in biopharmaceuticals.

Description

Technical field: [0001] The invention relates to a method for gene synthesis, carrier construction and product production of wild boar α-interferon. Background technique: [0002] As the first virus defense system in organisms, interferon has always been one of the important cytokines in various organisms. The use of interferon to prevent and treat viral diseases in livestock is also a very effective means. Wild boar α-interferon is an induced protein, which is not produced by the animal body under normal circumstances. At present, in China, porcine white blood cells are cultured in vitro and induced to produce interferon, and then interferon is extracted from the culture supernatant. The interferon produced by this method has low biological activity and high cost, and pig blood is contaminated with various viruses, which will also bring problems to the safety of the product. Through genetic engineering technology, recombinant interferon protein can be produced in vitro o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/21C07K14/56A61K38/17A61P31/12
CPCY02A50/30
Inventor 刘娣王君伟李文辉
Owner 黑龙江省农业科学院畜牧研究中心
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