Chicken alpha interferon/interleukin 2 chimeric gene

A technology for interleukin and alpha interferon, which is applied in the fields of chicken alpha interferon and interleukin 2 chimeric gene and its construction, fusion expression and purification, to achieve good antiviral activity, simplified operation process, significant anti-NDV and Effects of IBDV activity and immune-enhancing activity

Active Publication Date: 2011-05-04
HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION
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Problems solved by technology

Since both IFN-α and IL-2 have broad-spectrum antiviral activity, the combined application of the two can exert a st

Method used

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  • Chicken alpha interferon/interleukin 2 chimeric gene
  • Chicken alpha interferon/interleukin 2 chimeric gene
  • Chicken alpha interferon/interleukin 2 chimeric gene

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Embodiment Construction

[0045] 1 method

[0046] 1.1 Design and synthesis of primers

[0047] The gene linker is a flexible linker rich in glycine (G) and serine (S) (G 4 S) 3 , a total of 15 amino acids. According to the ChIFN-α gene sequence in rpQE-30 / ChIFN-α (GenBank accession number: AB255630) and the ChIL-2 gene sequence (AB194099) in GenBank that we have sequenced, we designed a total of 4 primers for SOE using Oligo6.0 software -PCR amplification, the sequence is as follows: P1: 5'-TCAGCATGCG CCTGCAACCA CCTTCGCC-3' (containing Sph I restriction site); P2: 5'-GCCACCGCCA GAGCCACCTCCGCCTGAACCGCCTCCACCAGTGCGCGTGTTGCCTGTG-3', for the amplification of ChIFN-α-linker- The downstream primer used for the ChIFN-α-linker part of the ChIL-2 chimeric gene; P3: 5'-GGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGCATCTCTATCATCAGCAAA-3', the upstream primer used to amplify the linker+ChIL2 part of the chimeric gene; P4: 5'-TACAAGCTTTTTTTGCAGATATCTCAAAG-3 ', (containing Hind III restriction site). The above prime...

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Abstract

The invention belongs to the technical field of biologic genetic engineering, and discloses a chicken alpha interferon (ChIFN-alpha)/interleukin 2(ChIL-2) chimeric gene, the technology comprises using an overlap extension PRC technique to constitute a mature peptide gene of the ChIFN-alpha and the ChIL-2 as a ChIFN-alpha-linker-ChIL-2 chimeric gene through a gene flexible linker (linker)(G4S), and cloning the constituted gene into a pGEM-TEasy carrier, subcloning the chimeric gene in a pQE-30 expression carrier to perform prokaryotic expression and perform rapid renaturation and purification to the expressed recombinant fusion protein. The activities of the purified rChIFN-alpha-liner-ChIL-2 fusion protein on the cell and in vitro and in vivo of the chicken are all provided with the superposition of the protein activity of the ChIFN-alpha and the ChIL-2, and the antiviral activity thereof is superior to the rChIFN-alpha protein. The chicken alpha interferon (ChIFN-alpha)interleukin 2(ChIL-2) chimeric gene is used as the main component of the antiviral preparation for preventing and treating the viral disease of the chicken, which is efficient, broad-spectrum, safe and low in cost.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and relates to a chimeric gene of chicken alpha interferon and interleukin 2 and its construction, fusion expression and purification technology. Background technique [0002] my country is a big chicken raising country in the world, and there are many kinds of chicken diseases, especially the highest morbidity and mortality caused by viral infectious diseases, which cause huge economic losses to the chicken industry every year, and some viral diseases (such as highly pathogenic Sexual avian influenza, etc.) is also related to the health of human beings themselves. Therefore, the treatment and prevention of these viral diseases become the primary task of the poultry industry. [0003] At present, there is no specific drug to treat viral diseases. Although some chemically synthesized antiviral drugs such as amantadine hydrochloride and ribavirin have certain effects on a small numbe...

Claims

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Application Information

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IPC IPC(8): C12N15/62C12N15/11C12N15/70C07K19/00A61K38/21A61P31/12A61K38/20
Inventor 吴志明闫若潜张志凌张健谢彩华刘光辉张盼盼盛敏刘梅芬荆汝顶王东方钱勇程俊贞李桂喜刘先敏安春霞孙清莲
Owner HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION
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