Escherichia coli recombinant chicken alpha-gamma divalent interferon and preparation method thereof

A technology of Escherichia coli and interferon, applied in interferon, biochemical equipment and methods, cytokines/lymphokines/interferon, etc., can solve problems such as clinical combination applications that have not been seen

Inactive Publication Date: 2011-06-29
QINGDAO VLAND BIOTECH INC +1
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Problems solved by technology

[0003] A recombinant chicken α-γ bivalent interferon related to the present invention has dual ef

Method used

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Embodiment Construction

[0008] Design a primer pair according to the chicken IFN-α gene in GenBank, use PCR technology to clone the IFN-α gene, design a pair of primers on both sides of the mature peptide gene, add appropriate restriction sites, and subclone it on the expression vector PET , after transforming BL21(DE3), the expression was induced by IPTG, and the expression form was analyzed by SDS-PAGE electrophoresis. The result was that the protein was expressed in the form of inclusion bodies. Oxidation method refolds the purified inclusion body denaturation solution by fractional dilution. The antiviral activity of the refolding liquid was determined by cytopathic inhibition method to obtain chicken α-interferon. Then, according to the chicken IFN-γ gene sequence in GenBank, the signal peptide region was removed, and the codon preference of E. coli was modified for the mature peptide region and the earphone structure at the 5' end of the mRNA was analyzed, and BgI II and Hand III were established

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Abstract

The invention relates to an escherichia coli recombinant chicken alpha-gamma divalent interferon and a preparation method thereof. The invention aims at providing the interferon which has good anti-tumor effect and can improve immunity of livestock and poultry and realize control over diseases within a short time. The interferon comprises the components such as an escherichia coli recombinant chicken alpha-interferon and an escherichia coli recombinant chicken gamma-interferon; and the preparation method comprises the steps of gene design, construction of an expression vector, induction of expression, purification of a target protein and activity detection. The medicine is an alternative medicine for treating viral diseases of the livestock and the poultry, and escherichia coli is utilized for expressing the chicken alpha-interferon and the chicken gamma-interferon, thereby having a broad-spectrum antivirus role. The divalent interferon can have immunoregulation activity and a certain antivirus activity after purifying the protein, and the two interferons have obvious synergy when being combined and applied clinically. The divalent interferon prepared by the invention is better than any one interferon which is applied separately in the effects of treating and preventing newcastle disease and bursal disease.

Description

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Claims

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Application Information

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Owner QINGDAO VLAND BIOTECH INC
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