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animal fusion recombinant interferon

An interferon, animal technology, applied in the direction of interferon, fusion polypeptide, cytokine/lymphokine/interferon, etc., can solve problems such as ineffective effect and animal husbandry loss

Active Publication Date: 2015-08-26
维克香港贸易有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If there is no effective vaccine to prevent a certain animal virus, when the herd is infected by the virus, supportive therapy can only be used to deal with it, but the effect is often ineffective, resulting in heavy losses in animal husbandry; therefore, the development of animal interferon, An urgent and extremely important matter for the animal health industry

Method used

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  • animal fusion recombinant interferon
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Examples

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Embodiment 1

[0028] Embodiment 1 porcine interferon alpha 1 gene selection

[0029] Peripheral blood mononuclear cells (PBMC) were isolated from the blood of the three-strain hybrid pigs (L×Y-D), and total RNA was extracted by the guanidine thiocyanate (GTC) method. Then the extracted total RNA was subjected to reverse transcription polymerase chain reaction (reverse polymerase chain reaction, RT-PCR). 25mM dNTP, 1μl 3' end complementary primer, 11μl distilled water, 0.5μl RNasin and 0.5μl AMV reverse transcriptase reaction tubes were placed at 42°C for 30 minutes for cDNA synthesis; then the synthesized cDNA was subjected to polymerase chain reaction (polymerase chain reaction, PCR) to proliferate porcine interferon α1 gene (P IFNα1), add 10 μl cDNA, 5 μl 10x PCR reaction solution, 8 μl 1.25 mM dNTP, 1 μl 5’ end forward primer, 1 μl 3’ end reverse primer, 24 μl of distilled water and 1 μl of Taq polymerase were mixed evenly and placed in a PCR reactor (Applied Biosystems GeneAmp PCR syst...

Embodiment 2

[0037] Example 2 Selection and Cloning of Porcine Immunoglobulin IgG-Fc 4a Fragment Gene

[0038]Fresh pig spleen was taken, and total RNA was extracted by GTC method. Then RT-PCR was used to synthesize cDNA; take 20 μl total RNA, 8 μl 1.25 mM dNTP and 1 μl 3’-end complementary primer, put it on ice after reacting at 70 ° C for 5 minutes, add 1 μl sterilized water, 8 μl 5xAMV RT buffer, 1 μl RNasin and 1 μl AMV reverse transcriptase, and put it at 42°C for 1 hour to synthesize cDNA; then perform PCR reaction on the synthesized cDNA, add 1 μl cDNA, 5 μl 10x PCR reaction solution, 8 μl 1.25mM dNTP, 1 μl5 Forward primer at the ' end, 1 μl reverse primer at the 3' end, 33 μl sterilized water, and 1 μl Taq polymerase were mixed evenly and placed in a PCR reactor (Applied Biosystems GeneAmp PCR system2400). Denaturation followed by 30 cycles of 95°C for 1 minute, 55°C for 30 seconds, and 72°C for 30 seconds, and finally PCR reaction was completed at 72°C for 5 minutes. Wherein, th...

Embodiment 3

[0045] Example 3 Construction of porcine fusion recombinant interferon (P IFN-Fc) DNA sequence

[0046] In this example, the porcine interferon α1 gene (P IFNα1) (SEQ ID No: 1) obtained in Example 1 and the porcine immunoglobulin IgG-Fc 4a fragment gene (P IgG-Fc 4a ) obtained in Example 2 were ) (SEQ ID No: 3) is connected with the DNA sequence (SEQ ID No: 5) of the linker (linker) composed of glycine (Glycine, G) and serine (Serine, S), to construct pig fusion recombinant interferon ( DNA sequence of P IFNα1-Fc 4a) (SEQ ID No: 7).

[0047] First, the porcine interferon α1 gene (P IFNα1) (SEQ ID No: 1) and the porcine immunoglobulin IgG-Fc 4a fragment gene (P IgG-Fc 4a) (SEQ ID No: 3) were respectively amplified by PCR reaction, and The DNA sequence (SEQ ID No: 5) segment of the linker composed of glycine and serine was designed by using PCR primers with the porcine interferon α1 gene (P IFNα1) and the porcine immunoglobulin IgG-Fc 4a fragment gene (P IgG-Fc 4a) Amplify the...

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Abstract

Provided is a fusion protein containing an animal interferon and an animal immune globulin Fc section. The fusion protein further comprises a linker for linking the interferon and the immune globulin Fc section. Further provided are a polyribonucleotide for encoding the fusion protein, a method for preparing the fusion protein, and an application of the fusion protein in the preparation of an antiviral drug.

Description

technical field [0001] The present invention relates to an animal fusion recombinant interferon, in particular to a fusion recombinant interferon with anti-animal virus activity. Background technique [0002] Interferon was first discovered in 1957 by British scholars Alick Isaacs and Jean Lindenmann in experiments on influenza virus. When a cell is infected by a virus, it will immediately produce a cytokine to induce adjacent cells to produce antiviral proteins and interfere with the virus. copy. The cytokine was subsequently named interferon (Interferon, IFN). The antiviral effect of interferon is mainly responsible for the type I interferon (IFNα / β). In addition to its antiviral effect, interferon also has the functions of antitumor, promoting cell differentiation and immune regulation. [0003] Most of the interferon preparations currently on the market are developed and designed for humans, for example, they are used to treat viral diseases such as human hepatitis B ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12P21/02A61K38/21A61K47/48A61P31/12
CPCC07K2319/30C07K14/555A61K38/21A61P31/12A61K38/00A61K39/00A61K47/50C07K14/715C07K19/00C12N15/09C12N15/62C12N15/63
Inventor 郭村勇吴忠晋陈汉婷
Owner 维克香港贸易有限公司
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