Production process of chicken beta interferon optimized gene and recombinant chicken beta interferon

A technology of beta interferon and production process, applied in genetic engineering, plant gene improvement, recombinant DNA technology, etc., can solve the problems of large loss of denatured protein, low antiviral activity, and low expression of chicken beta interferon

Inactive Publication Date: 2017-02-15
SOUTH CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects of the prior art of low expression of chicken interferon beta, large loss of renaturation protein, and low antiviral activity, and provide an optimized production process of recombinant chicken interferon beta

Method used

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  • Production process of chicken beta interferon optimized gene and recombinant chicken beta interferon
  • Production process of chicken beta interferon optimized gene and recombinant chicken beta interferon
  • Production process of chicken beta interferon optimized gene and recombinant chicken beta interferon

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Embodiment Construction

[0043] The preferred embodiments of the present invention will be described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

[0044] Such as Figure 1 to Figure 3 Shown, the production technology of a kind of chicken interferon beta optimization gene of the present invention and recombinant chicken interferon beta, it comprises the following steps:

[0045]S1. Referring to the chicken interferon beta gene sequence published in NCBI, the codon optimization of the interferon gene without the signal peptide gene sequence was performed according to the codon preference of Escherichia coli, and the chicken interferon beta gene was artificially synthesized; after optimization The fragments were sequenced and synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd.

[0046] S2. Design primers: ...

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Abstract

The invention discloses a production process of a chicken beta interferon optimized gene and a recombinant chicken beta interferon. The process includes the steps of: S1. conducting codon optimization on published chicken beta interferon genes in NCBI according to the preference of Escherichia coli codon and performing artificial synthesis of the optimized chicken beta interferon gene; S2. designing primers: according to the codon optimized interferon gene, designing a pair of primers; S3. constructing a recombinant expression plasmid; S4. transforming and identifying the recombinant expression plasmid; and S5. carrying out induced expression of the recombinant expression plasmid in Escherichia coli; and S6. extracting the expression product and conducting protein refolding and purification: S61. extraction and treatment of an inclusion body; S62. inclusion body denaturation; S63. denaturation fluid renaturation; and S64. nickel column affinity purification. The process provided by the invention expresses and purifies the recombinant chicken beta interferon protein by means of prokaryotic expression, and detects the interferon's activity of inhibiting vesicular stomatitis virus proliferation by means of cytopathic effect inhibition, and the activity unit reaches 2.73*10<6>UI / mg.

Description

technical field [0001] The invention relates to the technical field of gene recombination, in particular to a chicken interferon-beta optimized gene and a production process for recombinant chicken interferon-beta. Background technique [0002] In recent years, my country's poultry industry has developed rapidly and its scale has continued to expand. However, the prevalence of viral diseases such as avian influenza, Newcastle disease, and chicken infectious bronchitis has restricted the important factors for the development of the poultry industry. At present, methods such as vaccine immunization, chemical drugs and antibiotic treatment are still used in my country to prevent and control poultry diseases. However, due to the complexity of pathogenic serotypes and rapid mutation, the immune effect of vaccines is often poor. At the same time, the use of antibiotics and chemical drugs limits the scope of their use due to the problem of pathogenic drug resistance and drug resid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/22C07K14/565C12N15/70C07K1/36C07K1/34C07K1/22C12R1/19
CPCC07K14/565C12N15/70C12N2800/101C12N2800/22
Inventor 焦培荣廖明冯赛祥吴斯宇孙超孙俭波廖想
Owner SOUTH CHINA AGRI UNIV
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