Novel process for preparing recombinant chicken interferon alpha with strong activity and high expression quantity

An alpha interferon, high expression technology, applied in the field of interferon, can solve problems such as unfavorable purification, low protein expression, and achieve the effect of improving protein expression and high expression

Pending Publication Date: 2021-01-05
SHIJIAZHUANG SHIMU ANIMAL HUSBANDRY PHARMA CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention proposes a new process for preparing recombinant chicken interferon alpha with strong activity and high expression, which solves the problem of low protein expression of recombinant chicken interferon alpha gene in the prior art and is unfavorable for purification

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  • Novel process for preparing recombinant chicken interferon alpha with strong activity and high expression quantity
  • Novel process for preparing recombinant chicken interferon alpha with strong activity and high expression quantity
  • Novel process for preparing recombinant chicken interferon alpha with strong activity and high expression quantity

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Embodiment 1

[0087] A new process for preparing recombinant chicken interferon alpha with strong activity and high expression, comprising the following steps:

[0088] S1. Construction of a cloning vector for recombinant chicken interferon-α

[0089] S11. Extract RNA from chicken lymphocytes, reverse transcribe the RNA to obtain chicken α-interferon cDNA, use ChIFNα primers to perform PCR amplification on cDNA, obtain chicken α-interferon gene PCR product, connect with plasmid pJET, and transform , to obtain the recombinant plasmid pJET-ChIFNα, which specifically includes the following steps:

[0090] S111. Extract RNA from chicken lymphocytes: aseptically collect 5 mL of chicken whole blood intravenously, add 5 mL of 1% sterile sodium heparin 1640 culture solution to obtain whole blood containing heparin sodium, and pre- Add 5 mL of lymphocyte separation medium, slowly add 5 mL of whole blood containing heparin sodium on top of the separation liquid, centrifuge at 1500 rpm for 20 minutes a...

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Abstract

The invention belongs to the technical field of interferon, and provides a novel process for preparing recombinant chicken interferon alpha with strong activity and high expression quantity. The process comprises the following steps of S1, constructing a cloning vector of the recombinant chicken interferon alpha; S2, constructing a pichia pastoris expression vector of the recombinant chicken interferon alpha; and S3, performing protein expression of the recombinant chicken interferon alpha. Through the technical scheme, the problems that in the prior art, the protein expression quantity of a recombinant chicken interferon alpha gene is low, and purification is not facilitated are solved.

Description

technical field [0001] The invention belongs to the technical field of interferon, and relates to a new process for preparing recombinant chicken alpha interferon with strong activity and high expression. Background technique [0002] Chicken alpha interferon (IFN-α) is a kind of protein with broad-spectrum anti-virus, anti-tumor and immunomodulatory effects induced by viruses and lipopolysaccharides. It was first discovered by Issacs and Lindeman in 1957. It is a Multifunctional cytokines, after binding to cell receptors, can induce the body to produce a variety of specific proteins and enzymes, mainly by inhibiting viral gene transcription and degrading viral RNA to inhibit the growth and reproduction of viruses and exert anti-tumor activities . It is now known that chicken α-interferon can selectively act on virus-infected cells in vivo, exert broad-spectrum and high-efficiency antiviral effects by inhibiting the biosynthesis of viral proteins in infected cells, but has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/56C12N15/81
CPCC07K14/56C12N15/815
Inventor 陈巨清赵海成
Owner SHIJIAZHUANG SHIMU ANIMAL HUSBANDRY PHARMA CO LTD
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