Production method of chicken gamma interferon protein

A technology of interferon gamma and interferon, which is applied in the production field of soluble chicken interferon gamma protein, can solve the problems of difficulty in achieving renaturation, lack of biological activity of inclusion body protein, and complicated purification.

Inactive Publication Date: 2015-03-25
YEBIO BIOENG OF QINGDAO
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  • Abstract
  • Description
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Problems solved by technology

The inclusion body protein has no biological activity, and the subsequent purification is very complicated, and it is difficult to achieve complete renaturation during the renaturation process, and it is difficult to achieve 50% renaturation

Method used

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  • Production method of chicken gamma interferon protein
  • Production method of chicken gamma interferon protein
  • Production method of chicken gamma interferon protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Transformation of chicken gamma interferon gene

[0026] 1. Design of the chicken interferon gamma gene Referring to the chicken interferon gamma gene published in Genbank, combined with the codon preference expressed in Escherichia coli, the chicken interferon gamma gene was designed and synthesized. Simultaneously design primers for amplification.

[0027] 2. Amplification and recovery of the chicken interferon gamma gene The synthesized gene was used as a template for PCR amplification. After the reaction, 10 μL of the PCR product was taken to analyze the amplification result by 1% agarose gel electrophoresis. see results figure 1 (Examples of chicken gamma interferon gene PCR amplification identification diagram of the embodiment of the present invention). Cut out the target band at about 500bp, recover and purify according to the instructions of the recovery kit (Shanghai Sangon Bioengineering Co., Ltd.).

[0028] 2. Construction of recombinant ex...

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Abstract

The invention provides a gene of the chicken gamma interferon protein, wherein after being mutated by nucleotide, the gene can be used for efficiently expressing soluble chicken gamma interferon protein in escherichia coli; a sequence of the gene is SEQ ID NO:1. By changing the chicken gamma interferon gene sequence, the soluble expression of the chicken gamma interferon protein in the escherichia coli is realized. The expression product is a soluble protein in cytosol, and has the biological activity of the natural chicken gamma interferon protein. The invention also provides an inducible expression process of the engineering bacteria. According to a cytopathic effect inhibition test, a chicken embryo virus inhibition test, an immunological enhancement test, and an animal attacking protection test, the expressed chicken gamma interferon protein is proven to have good antiviral activity and immune enhancement activity.

Description

technical field [0001] The invention belongs to the technical field of recombinant protein expression, and in particular relates to a production method of soluble chicken gamma interferon protein. Background technique [0002] Interferon is a kind of low-molecular-weight soluble multifunctional glycoprotein produced by T lymphocytes and NK cells, which can enhance the function of cytotoxic T cells, promote the differentiation and activation of B cells, and induce animal cells to produce various antiviral and antitumor activities. Cytokines have good immunoregulatory effects (HOU Y-D. Molecular virology. Beijing: Xueyuan Press, 1990.). [0003] Chicken gamma interferon was first discovered by Issaacs et al. in 1957 when he used chicken embryo chorioallantoic membrane to study the interference phenomenon of influenza virus. Thanks to the rapid development of biotechnology, the biological research of chicken gamma interferon has entered a more in-depth Extensive research phase...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/23C12N15/11C12N15/70C12N1/21C07K14/57
Inventor 刘新文胡潇邹敏郭伟伟宫晓
Owner YEBIO BIOENG OF QINGDAO
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