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Swine alpha-interferon gene synthesis, expression vector establishment and product preparing method

An expression vector and interferon technology, applied in the production field of porcine alpha-interferon, can solve the problems of low expression level, porcine blood virus contamination, low activity titer, etc., and achieve the effect of high-efficiency expression

Inactive Publication Date: 2005-05-04
BAOYITE BIOTECH INST QINGDAO
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Problems solved by technology

At the same time, there are also problems of low activity titer per unit volume and potential virus contamination of pig blood
And genetically engineered pig α-interferon currently has the problem of high production cost, the main reason is: the expression level is on the low side, the interferon protein expression only accounts for 4-7% of the bacterial protein; the acquisition of the interferon finished product needs to go through multiple processes Complicated technological process, the recovery rate is only 10% to 20%

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  • Swine alpha-interferon gene synthesis, expression vector establishment and product preparing method
  • Swine alpha-interferon gene synthesis, expression vector establishment and product preparing method
  • Swine alpha-interferon gene synthesis, expression vector establishment and product preparing method

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Embodiment Construction

[0032] The following describes the embodiments of the present invention in detail with reference to the drawings:

[0033] In the embodiment of the present invention, the preferred codons of Escherichia coli are selected, and the translation initiation region and the minimum free energy of generation of the 5'end secondary structure of the porcine interferon gene are simulated and calculated by computer software. Synthesize porcine α-interferon gene, add EcoRI and SalI restriction sites to 5'and 3'respectively, and clone into pGEM-T plasmid to construct pGEM-T-poIFN-α. The designed and synthesized porcine alpha-interferon gene sequence, its comparison with natural porcine alpha-interferon and the sequencing results are shown in the following table and Figure 1 and Figure 2 respectively.

[0034] With reference to Figure 4, the embodiment of the present invention uses EcoRI and SalI to cut out the poIFN-α gene from pGEM-T-poIFN-α and clone it into pRLC to obtain the expression plas...

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Abstract

One producing method of gene composition of pig a-interferon, expression vector structuring and its offspring includes synthesing pig a-interferon consisted of bacilluscoli preference codon and constructs highly active expression vector with this gene, and realizes the highly active expression in bacilluscoli of pig-a interferon, with 26% expression of the whole mycoprotein. At the same time, it provides a purely chemical engineering system of technique of induced expression, inclusion bodies separating and cracking, and complexity of albumen. Pig a-interferon has the functions of highly active antivirus and immunoregulation, and can be applied in preventing and treatment of pig contagion. This invention is able to produce efficient, safe, and cheap pig a-interferon which has no potential virus pollution. This lays a solid foundation for mass-production of pig a-interferon in our country.

Description

Technical field [0001] The invention belongs to the technical field of gene engineering production of polypeptide medicines in biopharmaceuticals, and particularly relates to a method for producing pig alpha-interferon. Background technique [0002] Interferon (Interferor, IFN) is a type of cytokine produced by the animal body with anti-virus, anti-tumor and immune regulation functions. According to the different physical and chemical properties of the source cell and its own, it is mainly divided into α, β and γ-interferon. Alpha-interferon is widely used in the prevention and treatment of human viral diseases due to its significant antiviral function. With the urgent needs of animal health care and the continuous development of bioengineering technology, various methods of producing interferon for animals have become a research focus in recent years. [0003] Porcine interferon alpha (PoIFNα) can be divided into two subtypes, with at least 17 gene copies,...

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Application Information

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IPC IPC(8): A61K38/21A61P31/12C12N1/20C12N15/20C12N15/63
Inventor 陈溥言曹瑞兵蒋贻海张存
Owner BAOYITE BIOTECH INST QINGDAO
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