Recombinant strain of mink IFN-Gamma gene

A technology of recombinant strain and γ gene, applied in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., can solve problems such as unsatisfactory treatment effect, restricting the development of mink breeding industry, and economic loss.

Inactive Publication Date: 2009-09-16
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Abstract
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AI Technical Summary

Problems solved by technology

With the development of this industry, infectious diseases such as parvovirus enteritis, canine distemper, and Aliu, which seriously endanger the mink breeding industry, often occur, causing serious economic losses and limiting the development of the mink breeding industry; due to the onset of these viral infectious diseases The mortality rate is high, and the treatment effect of ordinary drugs is not ideal. Therefore, the research and development of genetically engineered mink recombinant interferon biological products with antiviral activity and immune regulation activity is of great significance in the prevention and treatment of mink viral diseases.

Method used

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  • Recombinant strain of mink IFN-Gamma gene
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  • Recombinant strain of mink IFN-Gamma gene

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Embodiment Construction

[0017] 1. Materials and methods

[0018] (1) Restriction endonucleases, Taq enzymes, and DNA molecular weight standards were all purchased from Treasure Bio (Dalian) Co., Ltd. Bacterial culture yeast extract and peptone were purchased from Oxford, UK. Inorganic salts are all domestic analytically pure.

[0019] (2) Strains and plasmids, Escherichia coli DH5α are preserved by the applicant.

[0020] Common methods such as plasmid preparation, competent preparation, DNA transformation and transformant screening, and restriction endonuclease hydrolysis can be found in (Molecular Cloning)

[0021] 2. In view of the similar genus relationship and close kinship between mink and dog in animal taxonomy, a pair of interferon gamma was designed and synthesized according to the gene sequence of canine γ-interferon published by GenBank, and the sequence accession number is: (NM 001003174). Specific primers, the primer sequence is

[0022] p1: ATGACCAGTAGATGCATC,

[0023] p2: TTCAGTTC...

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Abstract

The invention relates to a recombinant strain of a mink IFN-Gamma gene, which is characterized by being an IFN-Gamma gene sequence which is cloned from peripheral mink bloodlymph cells (PMBC) simulated by phytohemagglutinin (PHA) and being a recombinant plasmid vector containing a nucleotide sequence; i.e., using a plasmid vector pGM-T which is formed by the cutting of a cloning vector by taking an EcoR V enzyme as the cutting point and then adding a 'T' from the '3' end at both sides; and TA cloning is carried out on the nucleotide sequence of a coding Gamma-interferon which is amplified from the peripheral mink bloodlymph cells so as to construct the recombinant plasmid of the nucleotide sequence of the mink Gamma-interferon. In addition, Escherichia coli conversed from the plasmid vector is used as a host. The recombinant strain aims at laying down foundation for researching and developing genetically engineered mink recombinant interferons biological products with anti-viral activity and immunomodulatory activity.

Description

Technical field: [0001] The invention relates to a recombinant strain of mink IFN-γ gene. The γ-interferon gene was amplified from the isolated mink peripheral blood lymphocytes (PBMC), connected to the pGM-T vector, and the pT-MiIFN-γ recombinant plasmid was constructed, transformed into Escherichia coli JM109, and the mink IFN-γ gene was obtained. Gene recombinant strains. Background technique: [0002] Interferon (Interferon, IFN) is a cytokine with broad-spectrum anti-virus, anti-tumor and immune regulation activities. It is a kind of secreted glycoprotein produced by vertebrate cells, and it is the earliest discovered and most studied cytokine. It has broad-spectrum antiviral and immune response enhancing effects. It can non-specifically inhibit the proliferation of various viruses and plays a central role in the regulation of immune responses. Its antiviral and anti-tumorigenic properties lend it potential for immunoprophylaxis and therapy. [0003] According to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/23C12N15/63C12R1/19
Inventor 张海玲闫喜军赵春霏罗国良王凤雪柴秀丽易立
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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