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Porcine interferon alpha and application thereof

An alpha interferon and gene technology, applied in the field of protein engineering, can solve the problems of low codon expression and low antiviral activity of the original gene, and achieve the effect of good antiviral effect, good antiviral activity and strong market competitiveness.

Active Publication Date: 2017-07-11
SOUTH CHINA AGRI UNIV +1
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above technologies all have the defects of low expression of the original gene codon and low antiviral activity.

Method used

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  • Porcine interferon alpha and application thereof
  • Porcine interferon alpha and application thereof
  • Porcine interferon alpha and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 Gene modification and expression of porcine interferon alpha protein

[0031] 1. Using gene analysis software, referring to the mature peptide nucleotide sequence of porcine interferon α gene GenBank: AFK92985.1 published on NCBI GenBank, according to the partial tropism of codons in the expression system of Escherichia coli, select high-expression codons and redesign new ones The sequence, the nucleotide sequence of which is shown as SEQ ID NO.2, was sent to Biological Company to synthesize the sequence.

[0032] After the whole gene synthesis of the target gene, the target gene was cloned into pUC57 and transformed into Escherichia coli. The transformed clones were taken for PCR verification of the target gene and double-enzyme digestion identification. According to the experimental results, PCR and double enzyme digestion were all positive plasmids for sequencing the target gene. Sequencing results show that its nucleotide sequence is shown in SEQ ID N...

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Abstract

The invention provides a porcine interferon alpha and an application thereof. The porcine interferon alpha is synthesized through selecting high-expression codon design according to the codon bias of the of a Escherichia coli prokaryotic expression system, the amino acid sequence of the porcine interferon alpha is represented by SEQ ID NO. 1, and the nucleotide sequence for encoding the porcine interferon alpha is represented by SEQ ID NO. 2. A method for preparing the porcine interferon alpha has the advantages of simplicity, low cost, easiness in industrial production, high expression level of the porcine interferon alpha, thorough denaturation in the inclusion body treatment process, and high protein renaturation rate porcine. The porcine interferon alpha prepared in the invention has high titer, can inhibit 100TCID50 vesicular stomatitis virus, has a specific activity reaching up to 10<8>, has good bioactive functions, can be used to prepare medicines for preventing and treating the swine virus infection and enhancing the immune function, and has a broad market application prospect.

Description

technical field [0001] The invention relates to the field of protein engineering, in particular to a porcine alpha interferon and its application. Background technique [0002] In recent years, porcine viral infectious diseases have become increasingly rampant, which has seriously threatened the sustainable development of animal husbandry in my country. Immunosuppressive diseases and rapid mutation of viral antigens often lead to vaccine immunization failure; coupled with the continuous emergence of new viral diseases, there is no successful vaccination prevention yet. Vaccines, on the other hand, only prevent disease, and treatment also depends on the use of antibiotics. However, the emergence of some drug-resistant strains poses a great threat to human food and health. Some countries have explicitly banned the application of certain antibiotics and antibacterial agents in aquaculture production. Therefore, there is an urgent need for a new method that is both effective a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/56C12N15/21A61K38/21A61P31/12A61P35/00A61P37/04C12N15/70
CPCA61K38/00C07K14/56C12N15/70C12N2800/101
Inventor 廖明孙俭波焦培荣古明珠白大勇孙超廖想吴斯宇崔进冯赛祥徐成刚
Owner SOUTH CHINA AGRI UNIV
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