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Recombined chicken alpha interferon gene and recombinant vector thereof

An alpha interferon and genome technology is applied in the field of recombinant chicken alpha interferon gene sequence and its recombinant vector, which can solve the problems of low yield of expressed protein, cumbersome processes, few sources, etc., and achieves strong antiviral activity and high antiviral activity. Activity, activity-enhancing effect

Inactive Publication Date: 2009-01-07
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0006] Chicken IFN-α induced by inducers is expensive due to many limitations such as few sources, high cost, and complicated purification process, which greatly limits clinical and scientific applications.
Prokaryotic expression has problems such as low yield of expressed protein, difficulty in purification, and unstable strains, which limit the transformation to large-scale production
Moreover, the chicken interferon expressed by Escherichia coli mainly exists in the form of inclusion bodies, which require high technical personnel and equipment in terms of denaturation and renaturation, and the procedures are cumbersome and cause a lot of loss, which increases the interference of recombinant chickens. The cost and difficulty of production of prime industrial products

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  • Recombined chicken alpha interferon gene and recombinant vector thereof
  • Recombined chicken alpha interferon gene and recombinant vector thereof
  • Recombined chicken alpha interferon gene and recombinant vector thereof

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Embodiment Construction

[0057] 1. Test materials

[0058] Escherichia coli DH5a, yeast X-33, and pPICZα-A vector were purchased from Nanjing Tiangen Biochemical Technology Company; Newcastle disease virus and avian influenza virus were purchased from the Veterinary Institute of Jiangsu Academy of Agricultural Sciences; SPF chicken embryos were purchased from Nanjing Qianyuanhao Biotechnology Co., Ltd. The company's Nanjing Biopharmaceutical Factory; the chemical reagents used were purchased from Takara Company; commercial chicken natural alpha interferon was purchased from a Shandong Biotechnology Co., Ltd.;

[0059] When designing the interferon, select the wild-type and α-interferon subtypes of varieties with strong disease resistance as templates; compare and analyze the collected interferon amino acid sequences with software such as DANMAN or Lasergene DNAstar, and use the principle of selective admission. Select the amino acid with the highest frequency to splice out an artificial interferon ami...

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Abstract

The invention relates to a novel gene sequence of a recombinant chicken Alpha interferon, constructs a recombinant expression vector thereof and belongs to a gene engineering biological product obtained by a molecular biology method. The gene sequence of a newly designed chicken Alpha interferon is recombined into a pPICZ Alpha-A vector and then is confirmed on the special position of a microzyme by an electric conversion mode. Besides, a pichia expression system is adopted to express a foreign gene, thus being beneficial to the commercial production of the chicken interferon. The protein expressed by a gene group after being diluted by 4365158.3 times can completely restrain the attraction of vesicular stomatitis virus of 100-1000TCID50. Compared with a natural chicken Alpha interferon, the novel gene sequence of a recombinant chicken Alpha interferon has a higher anti-virus effect; the anti-virus effect thereof is improved by 8 times. Test also detects that the protein expressed by the gene can restrain the proliferation of a newcastle disease virus and an avian influenza virus; besides, the effect of the interferon with high concentration is more remarkable.

Description

1. Technical field [0001] The invention discloses a gene sequence of recombinant chicken alpha interferon and its recombinant carrier, belonging to the product field of genetic engineering biological products. Compared with natural chicken alpha interferon, it has stronger antiviral activity, and can be used for the prevention and early treatment of various viral diseases in chickens. 2. Background technology [0002] Chicken α-interferon (Interferon alpha, IFN-α) is a class of anti-virus, anti-tumor, anti-proliferation and other functions (OritaniK, Medina KL, Tomiyama Y et al. Limitin: an interferon like cytokine that preferentially influences B2lymphocyte precursors) glycoprotein. In 1994, Sekellick et al. first cloned the IFN-α gene of chicken embryo fibroblasts, and carried out structural analysis. In 1996, Sick and Scultz cloned and expressed chicken alpha interferon gene, and studied the antiviral activity of the expressed product. Studies have shown that chicken i...

Claims

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Application Information

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IPC IPC(8): C12N15/21C12N15/81C12N1/19C07K14/56C12R1/84
Inventor 陈溥言侯凤香刘珂
Owner NANJING AGRICULTURAL UNIVERSITY
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