CN-121991250-A - Recombinant protein, encoding gene and application thereof

Abstract

The invention discloses a recombinant protein, a coding gene and application thereof, and belongs to the technical field of bioengineering. The invention designs a recombinant protein, which consists of two subunits with different functions, and the secretory expression of the recombinant protein is promoted by design and transformation on the basis of a conserved sequence of an alpha herpesvirus protein with an immune escape function, so that the defects of inclusion body formation, excessive toxicity to a host and high purification difficulty of target protein are overcome. Experimental results show that the recombinant protein has remarkable effect of inhibiting the natural immunity of cells, and can effectively inhibit interferon produced by the natural immunity of cells, so that the titer of cultured viruses is improved.

Inventors

• XIANG JUN
• TIAN LIMEI
• XU JING
• KONG XUEYING
• QIU WENYING
• SHEN JIANQING

Assignees

• 华派生物技术(集团)股份有限公司

Dates

Publication Date

20260508

Application Date

20260206

Claims (8)

1. 1. The recombinant protein is characterized in that the amino acid sequence of the recombinant protein is shown as SEQ ID NO. 1.
2. 2. A gene encoding the recombinant protein of claim 1, wherein the nucleotide sequence of the gene is shown in SEQ ID No. 2.
3. 3. Use of the recombinant protein of claim 1 to increase viral culture titer.
4. 4. The use according to claim 3, wherein the virus is at least one of pseudorabies virus, porcine circovirus type 2 and porcine parvovirus.
5. 5. A method for increasing the culture titer of a virus, comprising adding the recombinant protein of claim 1 to a virus culture system during the culture of the virus.
6. 6. The method of claim 5, wherein the recombinant protein is present at a concentration of 20-200ng/mL.
7. 7. An agent for increasing the titer of a culture of a virus, comprising the recombinant protein of claim 1.
8. 8. The use of the recombinant protein of claim 1 for the preparation of a viral vaccine, wherein the virus is pseudorabies virus, porcine circovirus type 2 or porcine parvovirus.

Description

Recombinant protein, encoding gene and application thereof Technical Field The invention relates to the technical field of bioengineering, in particular to a recombinant protein and a coding gene and application thereof. Background Vaccination is a mode for effectively preventing virus infection, at present, vaccines are mainly inactivated vaccines and live vaccines, and the titer of the cultured viruses directly influences the production cost of the vaccines, so that the titer of the cultured viruses is improved to have important practical significance. However, the culture of viruses depends on the propagation of the virus in cells, which, after infection by the virus, produce a natural immune response against the virus, wherein the production of type I Interferon (IFN) is critical against the virus. Most proteins in alpha herpes viruses that inhibit the natural immune function of the cell are encoded by late genes, the protein products of which are produced mainly in the late stages of viral infection, and depend on replication and expression of the viral genome, while viral nucleic acids, once they enter the cell, trigger the antiviral response of the cell, thus preventing viral invasion and replication. If exogenous anti-natural immunity protein can be added simultaneously or in advance when virus infects cells, natural immune response of cells can be effectively inhibited. Therefore, the method for resisting the natural immunity of the cells by using a mature protein expression system such as an escherichia coli expression system, a baculovirus-insect cell expression system, a CHO cell expression system and the like to express the protein with the function of inhibiting the natural immunity and then adding the protein into a culture solution for culturing the cells is a method with broad prospect. However, most of these proteins are rich in basic amino acids, and exogenous expression of these proteins tends to form inclusion bodies to reduce the solubility of the proteins, and is toxic to the host to inhibit the growth of the host and even cause death, and has a strong positive charge to cause a high purification difficulty. Disclosure of Invention In order to solve the defects in the prior art, the invention aims to provide a recombinant protein, a coding gene and application thereof so as to improve the titer of virus culture. The technical scheme for solving the technical problems is as follows, and the invention provides a recombinant protein, the amino acid sequence of which is shown as SEQ ID NO. 1. The invention provides a gene for encoding the recombinant protein, and the nucleotide sequence of the gene is shown as SEQ ID NO. 2. The invention provides an application of the recombinant protein in improving virus culture titer. Further, the virus is at least one of pseudorabies virus, porcine circovirus type 2 and porcine parvovirus. The invention provides a method for improving virus culture titer, which is characterized in that the recombinant protein is added into a virus culture system to improve the virus culture titer in the process of culturing viruses. Further, the concentration of the recombinant protein is 20-200ng/mL. The invention provides a reagent for improving virus culture titer, which comprises the recombinant protein. The invention provides an application of the recombinant protein in preparing virus vaccines, wherein the virus is pseudorabies virus, porcine circovirus type 2 or porcine parvovirus. The invention has the following beneficial effects that the invention improves the alpha herpesvirus on the basis of the conserved sequence of the immune escape function protein, thereby promoting the secretion expression of the recombinant protein and overcoming the defects of inclusion body formation, excessive toxicity to hosts and high purification difficulty of target protein. Experimental results show that the recombinant protein has remarkable effect of inhibiting the natural immunity of cells, and can effectively inhibit interferon produced by the natural immunity of cells, so that the titer of cultured viruses is improved. Drawings FIG. 1 is a diagram showing the result of conservative sequence alignment of subunit 1 and subunit 2; FIG. 2 is a recombinant protein gene amplification map; FIG. 3 is a SDS-PAGE diagram of recombinant proteins containing a vector tag; FIG. 4 is a SDS-PAGE of recombinant proteins. Detailed Description The examples given below are only intended to illustrate the invention and are not intended to limit the scope thereof. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention. Example 1 plasmid construction and expression of recombinant proteins containing vector tags (1) The construction of plasmid, namely, selecting the conserved s