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Interferon-thymosin broad-spectrum antiviral pharmaceutical preparation for pigs and preparation method thereof

An antiviral drug, swine interferon technology, applied in the field of bioengineering, can solve the problems of no report, less application of interferon and thymosin, etc.

Inactive Publication Date: 2010-06-09
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, interferon and thymosin are rarely used clinically in domestic veterinary medicine, especially the expression and application of the two fusion proteins have not been reported so far

Method used

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  • Interferon-thymosin broad-spectrum antiviral pharmaceutical preparation for pigs and preparation method thereof
  • Interferon-thymosin broad-spectrum antiviral pharmaceutical preparation for pigs and preparation method thereof
  • Interferon-thymosin broad-spectrum antiviral pharmaceutical preparation for pigs and preparation method thereof

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Experimental program
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Embodiment 1

[0069] 1. Sequence amplification and cloning of porcine IFN-α1 gene.

[0070] 1.1 The target gene of porcine IFNα1 was amplified by PCR from the porcine genome.

[0071] The porcine blood genome was extracted using Whole Blood Genomic DNA Mini-prep Kit, and PCR amplification was performed with IF1 and IF2 primers to obtain the target gene of porcine IFN-α1.

[0072] Amplify the IFNα1 gene by PCR, add each component once according to Table 2-1, and perform PCR reaction. The reaction conditions are as follows: pre-denaturation at 94°C for 5 minutes, 30s at 94°C, 30s at 54°C, 30s at 72°C, and extension at 72°C after 30 cycles , 8min.

[0073] Table 1-1 PCR reaction system of porcine IFNα1 gene

[0074]

[0075] 1.5% agarose gel electrophoresis, recovery.

[0076] 1.2 Connection of porcine IFN-α1 gene and pMD18-T vector

[0077] The porcine IFN-α1 gene was obtained by extending the PCR method, and connected to the pMD18-T vector. See Table 2-2 for the connection reaction s...

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Abstract

The invention discloses an interferon-thymosin broad-spectrum antiviral pharmaceutical preparation for pigs and a preparation method thereof. According to the molecular structure characteristics of interferon alpha 1 (IFN-alpha 1) and thymosin alpha 1 (THY-alpha 1) and the preference of Escherichia coli codon, primers are designed; the PCR method is used for acquiring the fusion gene of the IFN-alpha 1 and the THY-alpha 1; the gene is cloned to a pGEX4T-2 prokaryotic expression vector to construct pGEX4T-IFN alpha 1-THY alpha 1 prokaryotic expression plasmids; and the recombinant plasmids are converted into the BL21 (DE3) recipient bacterium to acquire IFN alpha 1-THY alpha 1 fusion protein through IPTG induction, expression and purification. The biological activity of the protein is detected through cytopathy inhibiting experiment and E-rosette forming experiment, and the result shows that the IFN alpha 1-THY alpha 1 fusion protein has double biological activity.

Description

technical field [0001] The invention provides a novel broad-spectrum antiviral biological preparation for pigs, and also provides a synthesis method of the biological preparation, which belongs to the technical field of bioengineering. Background technique [0002] Interferon (IFN) is a small peptide secreted by cells, which has a wide range of biological activities such as anti-virus, anti-proliferation and immune regulation. Studies have shown that interferon is ubiquitous in animals. Not only cells of humans and higher animals can produce interferon, but even many lower animals and bacteria can also produce interferon. Interferon-inducing agents or viruses can induce animal bodies or animal cells to produce interferon. All mammals studied contain IFN-α, IFN-β (type I) and IFN-γ (type II) genes. Among them, IFN-β and IFN-γ have species specificity, while IFN-α has the antiviral activity of different animals. Normal cells generally do not spontaneously produce interferon...

Claims

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Application Information

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IPC IPC(8): A61K38/22A61K38/21A61P31/12C12N15/62C12N15/70C07K19/00
Inventor 刘松财王佳
Owner JILIN UNIV
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