CN-119751703-B - Recombinant protein, nucleic acid molecule, recombinant phage and application thereof

Abstract

The invention relates to the field of biological medicine, in particular to recombinant protein, nucleic acid molecules, recombinant phage and application thereof. The invention provides a recombinant protein with an amino acid sequence shown as SEQ ID NO.6 and a recombinant protein with an amino acid sequence shown as SEQ ID NO.10 and application thereof in preparing vaccines. The vaccine prepared by injecting the recombinant protein can effectively prevent coronavirus infection. According to the invention, the PEDV S protein is selected as a candidate antigen for epitope design, two targeting peptides connected with specific S protein epitopes are selected, M13KE phage is selected as a delivery carrier of the PEDV epitope, and animal experiments are utilized to evaluate the effect of the PEDV vaccine. The results show that lymphocytes of the vaccinated mice show a significant proliferative response to PEDV antigen, and that IFN-gamma levels in serum of the vaccinated mice are significantly increased.

Inventors

• SUN JIANHE
• WANG ZHAOFEI
• YAN YAXIAN
• ZHAO GUOQING

Assignees

• 上海交通大学

Dates

Publication Date

20260508

Application Date

20241231

Claims (8)

1. 1. A recombinant protein is characterized in that the recombinant protein is BXT1-PEYC1 with an amino acid sequence shown as SEQ ID NO.6 or BXT2-PEYC with an amino acid sequence shown as SEQ ID NO. 10.
2. 2. A nucleic acid molecule encoding the recombinant protein of claim 1.
3. 3. A biological material comprising the nucleic acid molecule of claim 2.
4. 4. A biomaterial according to claim 3, wherein the biomaterial comprises recombinant phage.
5. 5. Use of the recombinant protein of claim 1, the nucleic acid molecule of claim 2 or the biological material of claim 3 or 4 for the preparation of a porcine epidemic diarrhea virus vaccine.
6. 6. A vaccine comprising the recombinant protein of claim 1, the nucleic acid molecule of claim 2, or the biological material of claim 3 or 4.
7. 7. The vaccine of claim 6, further comprising a pharmaceutically acceptable adjuvant.
8. 8. The vaccine of claim 6, wherein the vaccine comprises a porcine epidemic diarrhea virus vaccine.

Description

Recombinant protein, nucleic acid molecule, recombinant phage and application thereof Technical Field The invention relates to the field of biological medicine, in particular to recombinant protein, nucleic acid molecules, recombinant phage and application thereof. Background Porcine Epidemic Diarrhea Virus (PEDV), belonging to the family Coronaviridae, alpha coronaviridae, is a enveloped single-stranded positive-strand RNA virus, and has a polymorphic virion with a diameter of about 95-190nm. The virus genome is about 28kb in length, and encodes 4 structural proteins such as spike glycoprotein (S protein), envelope protein (E protein), membrane protein (M protein) and nucleocapsid protein (N protein), two non-structural proteins of multimeric protein complexes pp1a and pp1ab, and 1 ORF3 auxiliary protein. PEDV is the major causative agent of Porcine epidemic diarrhea (Porcine EPIDEMIC DIARRHEA, PED), which is mainly responsible for Porcine intestinal contagious disease, characterized by vomiting, diarrhea, dehydration, etc. In 1971, it was found for the first time in the pig farm for fattening pigs in the uk that the effect on weaned pigs of 4-5 weeks of age was great. Subsequently, europe and asian countries have begun to prevail. In north america 2013, outbreaks began, PEDV became a globally prevalent virus and severely compromised the global farming industry. For PEDV control, vaccination is currently the primary means of controlling PEDV infection in addition to biological control. The commercialized vaccine mainly comprises whole virus inactivated vaccine and attenuated vaccine, but the protection effect on the current epidemic strains is not ideal, and the reason is probably that the inactivated vaccine cannot generate effective mucosal immunity after intramuscular injection, the stability and proliferation capacity of the attenuated vaccine in intestinal canal are reduced after the attenuated vaccine is weakened, and the generated protection capability is limited. In addition, some farms are protected by means of counter feeding, and although the farms have a protective effect to a certain extent, other risks of viral infection are also present. It is therefore extremely important to develop a vaccine that can effectively control PEDV. Disclosure of Invention The invention aims to provide a recombinant protein, a nucleic acid molecule, a recombinant phage and application thereof, so as to solve the problems in the commercialized vaccine or the vaccine development process. The vaccine prepared by injecting the recombinant protein, the nucleic acid molecule and the recombinant phage provided by the invention induces a large amount of antigen specificity PEDVIgG antibodies and neutralizing antibodies, thereby achieving the effect of preventing and controlling PEDV. In order to achieve the above object, the present invention provides the following solutions: The invention provides a recombinant protein, which comprises BXT1-PEYC1 with an amino acid sequence shown as SEQ ID NO.6 and/or BXT2-PEYC with an amino acid sequence shown as SEQ ID NO. 10. The present invention provides a nucleic acid molecule encoding the recombinant protein described above. The invention provides a biological material containing the nucleic acid molecule. Preferably, the biological material comprises recombinant phage. Further preferred, the recombinant phage vector comprises a base phage that is an M13KE phage. The invention provides the application of the recombinant protein, the nucleic acid molecule or the biological material in preparing vaccines. Preferably, the vaccine comprises a porcine coronavirus recombinant phage vaccine. Preferably, the porcine coronavirus comprises porcine epidemic diarrhea virus. The present invention provides a vaccine comprising the recombinant protein described above, the nucleic acid molecule described above or the biological material described above. Preferably, the vaccine further comprises pharmaceutically acceptable excipients. Preferably, the vaccine comprises a porcine coronavirus recombinant phage vaccine. Further preferably, the porcine coronavirus comprises porcine epidemic diarrhea virus. The invention discloses the following technical effects: The invention provides a recombinant protein, which comprises BXT1-PEYC1 with an amino acid sequence shown as SEQ ID NO.6 and BXT2-PEYC with an amino acid sequence shown as SEQ ID NO. 10. The vaccine prepared by injecting the recombinant protein provided by the invention induces antigen specificity PEDVIgG antibody, PEDV specificity T cells and neutralizing antibodies, and the neutralizing antibodies can be combined with antigens on the surface of the PEDV, so that the PEDV is prevented from adhering to target cell receptors, the PEDV is prevented from invading cells, and the coronavirus infection can be effectively prevented. In order to study vaccine development and antiviral treatment strategies of PEDV, in a specific embodiment of the pres