Recombinant baculovirus for expressing dog serum albumin fused interferon gamma and application of recombinant baculovirus

A technology of recombinant baculovirus and serum albumin, which is applied in the field of mutation or genetic engineering, can solve the problem of large differences in the structure and function of natural proteins of veterinary interferon products, adverse reactions of animal immune tolerance, and the purification of veterinary interferon products Difficulties and other problems, to achieve the effect of fast and convenient fusion, simplified separation and purification work, and low cost

Inactive Publication Date: 2017-02-15
SOUTH CHINA AGRI UNIV
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Problems solved by technology

At present, the commonly used interferon for dog immune enhancement is conventional interferon. Repeated injections will cause immune tolerance and other adverse reactions in the animal body, which will affect its therapeutic effect.
[0008] 2. Most veterinary interferon products adopt prokaryotic expression, and the products are mostly in the form of inclusion bodies, and

Method used

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  • Recombinant baculovirus for expressing dog serum albumin fused interferon gamma and application of recombinant baculovirus
  • Recombinant baculovirus for expressing dog serum albumin fused interferon gamma and application of recombinant baculovirus
  • Recombinant baculovirus for expressing dog serum albumin fused interferon gamma and application of recombinant baculovirus

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Experimental program
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Embodiment

[0036] (1) Preparation of canine CSA-IFNγ fusion gene:

[0037] According to the canine serum albumin gene sequence (GenBank accession number: AB090854.1); the canine IFNγ gene sequence (GenBank accession number: EF095772.1) as the original sequence, according to the partial tropism of the insect cell expression system codon, optimize the synthesis of the new The nucleotide sequence of the canine interferon-γ mature peptide (the sequence was optimized and synthesized by Nanjing GenScript Biotechnology Co., Ltd.), and the synthetic primers were designed. The primer sequences are shown in Table 1.

[0038] Table 1 PCR primer sequence list

[0039]

[0040]

[0041]Step 1, the amplification method of canine serum albumin gene:

[0042] Take 0.5cm 3 Grind the canine liver sample from a cross grinder in an ice bath, collect the grinding solution in a 1.5mL centrifuge tube, freeze and thaw three times at -20°C, centrifuge at 12,000×g for 5min, collect the supernatant to extr...

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Abstract

The invention provides a recombinant baculovirus for expressing a dog serum albumin fused interferon gamma and application of the recombinant baculovirus. A dog serum albumin complete nucleotide sequence and a dog IFNgamma maturation protein nucleotide sequence are fused to construct a recombinant plasmid pF-CSA-IFNgamma for recombinant baculovirus screening; the recombinant baculovirus for stably expressing the dog serum albumin fused interferon gamma is screened. The recombinant baculovirus constructed through the method can stably secrete the novel dog gamma interferon with good expression activity and simple and convenient separation and purification, and lays a foundation for development of a novel dog long-acting interferon.

Description

technical field [0001] The invention relates to the technical field of mutation or genetic engineering, in particular to a recombinant baculovirus expressing canine serum albumin fusion interferon gamma and its application. [0002] technical background [0003] Interferon (Interferon, IFN) is a glycoprotein produced by viruses or other types of interferon inducers that stimulate the reticuloendothelial system, macrophages, lymphocytes and somatic cells. This protein has multiple biological activities, including anti-proliferative, immunomodulatory, antiviral and differentiation-inducing effects. Interferons are classified into type I and type II interferons based on specific receptors and sequence homology. IFN-γ is the only type II interferon, also known as immune interferon. Young (1996) and Frucht (2001) proposed that mainly by CD4 + Th1, CD8 + Secreted by cytotoxic T lymphocytes and NK cells, some B cells, NKT cells and APC cells can also secrete IFN-γ. Cantin (1999...

Claims

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Application Information

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IPC IPC(8): C12N7/01C07K19/00A61K38/21A61K47/64A61P37/04
CPCC07K14/57A61K38/217C07K2319/31
Inventor 罗满林李冰邹舒展张贺翟俊琼周霞
Owner SOUTH CHINA AGRI UNIV
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