Fusion protein of infectious bursal disease virus, preparation method, application, expression system of fusion protein and vaccine comprising fusion protein

A technology of fusion protein and bursal disease, applied in antiviral immunoglobulin, veterinary vaccines, biochemical equipment and methods, etc., can solve problems such as unstable efficacy and unsafe source of vaccine antigens, and achieve high protection rate , Reduce production cost, good safety effect

Inactive Publication Date: 2019-01-04
TECON BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] An object of the present invention is to provide a fusion protein of chicken infectious bursal disease virus, which expresses the VP2 and VP3 proteins of chicken infectious bursal disease virus in tandem, so as to alleviate the problems in the prior art. Unsafe source of vaccine antigen for chicken infectious bursal disease and technical problems of unstable potency

Method used

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  • Fusion protein of infectious bursal disease virus, preparation method, application, expression system of fusion protein and vaccine comprising fusion protein
  • Fusion protein of infectious bursal disease virus, preparation method, application, expression system of fusion protein and vaccine comprising fusion protein
  • Fusion protein of infectious bursal disease virus, preparation method, application, expression system of fusion protein and vaccine comprising fusion protein

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preparation example Construction

[0051] A method for preparing the above fusion protein, comprising expressing the gene encoding the fusion protein in a host. The host in the present invention can be, but not limited to, an Escherichia coli expression system, a yeast expression system, an insect expression system, a plant expression system or a mammalian expression system. Since the protein expressed by mammalian cells is translated and processed, its structure and biological properties are closer to natural proteins, so the present invention preferably uses a mammalian expression system to express the gene encoding the fusion protein.

[0052] In a preferred embodiment of the present invention, a mammalian expression system is used to express the gene encoding the fusion protein. Since the protein expressed by mammalian cells is translated and processed, its structure and biological properties are closer to natural proteins, so the present invention preferably uses a mammalian expression system to express th...

Embodiment 1

[0091] Example 1 VP2-VP3 Gene Design and Synthesis

[0092] VP2-VP3 gene design and synthesis: After optimization of the VP2 and VP3 sequences in GenBank accession number M97346.1, the length of VP2 is designed to be 1356bp, with the sequence shown in SEQ ID NO.1, and the length of VP3 is 870bp, with the sequence shown in SEQ ID NO. .2 the sequence shown. A Linker is used between VP2 and VP3, and the Linker sequence has the sequence shown in SEQ ID NO.3. The full length of the VP2-VP3 gene is 2256bp, and has the amino acid sequence shown in SEQ ID NO.4 and the nucleotide sequence shown in SEQ ID NO.5. VP2-VP3 gene synthesis was completed by Shanghai Sangon Biological Company.

Embodiment 2

[0093] Example 2 Construction of PEE6.4-VP2-VP3 recombinant plasmid

[0094]2.1 Add restriction sites: Add restriction sites respectively upstream and downstream of the VP2-VP3 gene sequence by PCR amplification: EcoR I, BamH I, PCR amplification upstream primers are shown in SEQ ID NO.6, PCR amplification The downstream primer is shown in SEQ ID NO.7. PEE6.4-VP2-VP3 plasmid map as figure 1 shown.

[0095] 2.2 VP2-VP3 gene and vector double enzyme digestion reaction

[0096] 2.2.1 Construct a 50 μL reaction system, mix the components according to the table below, and then bathe in water at 37°C for 2 hours.

[0097] 10×buffer

5μL

DNA sample

2μg

Eco RI

2.5μL

Bam HI

2.5μL

dd H 2 0

38μL

[0098] 2.2.2 Recovery of target DNA fragments: DNA gel recovery kit (purchased from Beijing Laibao Technology Co., Ltd.) was used to recover the enzyme-cut target fragments. The steps are as follows:

[0099] (1) After agarose ...

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Abstract

The invention relates to the technical field of biology, and particularly provides a fusion protein of an infectious bursal disease virus, a preparation method, application, an expression system of the fusion protein and a vaccine comprising the fusion protein. The fusion protein comprises a VP2 section and a VP3 section, wherein the VP2 section is expressed by a nucleotide sequence shown in SEQ ID NO.1, the VP3 is expressed by a nucleotide sequence shown in SEQ ID NO.2. The fusion protein is obtained by tandem expression of the gene sequences of the VP2 and the VP3 of the infectious bursal disease virus. By analyzing the gene sequences of the VP2 and the VP3 and selecting zones high in antigenicity and easy for high expression to link tandemly, the obtained fusion protein has the advantages of high antigenicity and high expression amount. By adopting the fusion protein as an antigen to prepare the vaccine, the prepared vaccine has the advantages of good safety, stable potency and no toxic and side effects. The invention further provides the preparation method and application of the fusion protein, and the vaccine prepared from the fusion protein.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fusion protein of chicken infectious bursal disease virus and its preparation method, application, expression system and vaccine. Background technique [0002] Chicken infectious bursal disease (Infectious bursal disease, IBD) is caused by chicken infectious bursal disease virus (Infectious bursal disease virus, IBDV) A kind of acute, highly Contact immunosuppressive infectious disease. [0003] IBDV is a non-enveloped virus with a double-segment, double-stranded RNA genome and an icosahedral symmetrical structure with a size of 60-70nm. Its nucleocapsid consists of dsRNA (double-stranded ribonucleic acid) and protein. In addition to complete virions, empty viral capsids without nucleic acid structures are common. Electron microscope observation of the three-dimensional structure of the negative staining virus showed that the IBDV virus particles were not spherical, but symmetri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C07K16/10C12N15/62C12N15/85G01N33/569A61K39/295A61K39/12A61P31/14
CPCA61K39/12A61K2039/552A61K2039/70A61P31/14C07K14/005C07K16/10C07K2319/00C12N15/85C12N2720/00022C12N2720/00034G01N33/56983
Inventor 贺笋任立松李延涛任郭子君
Owner TECON BIOLOGY CO LTD
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