Chimeric protein, virus-like particle and application thereof

A chimeric protein and chimeric virus technology, applied in the field of molecular biology, can solve the problems of large workload, low efficiency and high labor cost, and achieve the effects of low cost, simple preparation method and good immunogenicity

Active Publication Date: 2015-06-10
江苏省苏农科技术转移中心有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in order to prevent pigs from being infected with the above two diseases, it is necessary to inoculate the corresponding vaccines respectively, at least twice, the workload is large, the labor cost is high, and the efficiency is low

Method used

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  • Chimeric protein, virus-like particle and application thereof
  • Chimeric protein, virus-like particle and application thereof
  • Chimeric protein, virus-like particle and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1 Contains the chimeric protein and the virus-like particle of a main B cell epitope of FMDV VP1 protein

[0051] 1. Design of chimeric protein and construction of recombinant plasmid

[0052] The amino acid sequence of porcine circovirus type 2 Cap protein is shown in SEQ ID NO:9. The amino acid sequence of the main B cell epitope of FMDV VP1 protein is shown in SEQ ID NO:10. The gene encoding the Cap protein of porcine circovirus type 2 is shown in SEQ ID NO:1. The gene encoding the main B cell epitope of the porcine foot-and-mouth disease virus type O VP1 protein is shown in SEQ ID NO:2. The inventors used software to analyze the high-level structure of the PCV2 Cap protein, and designed the following four chimeric proteins and their coding genes:

[0053] (1) Chimeric protein Cap-loop1: The 58th to 66th amino acid residues of porcine circovirus type 2 Cap protein were replaced with a major B-cell epitope of FMDV VP1 protein. The sequence of the gene e...

Embodiment 2

[0089] Embodiment 2: the preparation of chimeric protein and virus-like particle containing 2 main B cell epitopes of FMDV VP1 protein

[0090] 1. Design of chimeric protein containing two main B cell epitopes of FMDV VP1 protein and construction of recombinant vector

[0091] The amino acid sequence of porcine circovirus type 2 Cap protein is shown in SEQ ID NO:9. The amino acid sequence of the B cell epitope of FMDV VP1 protein is shown in SEQ ID NO:10. On the basis of the research results in Example 1, the inventors used software to analyze the high-level structure of the PCV2 Cap protein, and designed a chimeric protein Cap-loop2-loop4: the 72nd to 94th positions of the porcine circovirus type 2 Cap protein The amino acid residues, the 162nd to 197th amino acid residues were respectively replaced by a main B cell epitope of FMDV VP1 protein. The sequence of the gene encoding the chimeric protein Cap-loop2-loop4 is shown in SEQ ID NO: 7, which was synthesized by Yingjun B...

Embodiment 3

[0106] Embodiment 3: the preparation of chimeric protein and virus-like particle containing 3 main B cell epitopes of FMDV VP1 protein

[0107] 1. The chimeric gene and identification of three main B-cell epitopes of FMDV VP1 protein

[0108] The amino acid sequence of porcine circovirus type 2 Cap protein is shown in SEQ ID NO:9. The amino acid sequence of the main B cell epitope of FMDV VP1 protein is shown in SEQ ID NO:10. On the basis of the results of Example 1 and Example 2, the inventors used software to analyze the high-level structure of the PCV2 Cap protein, designed a chimeric protein Cap-loop2-loop3-loop4, and incorporated the 72nd cap protein of porcine circovirus type 2 Amino acid residues from position 94, amino acid residues 122 to 147, and amino acid residues 162 to 197 were replaced by a major B-cell epitope of FMDV VP1 protein. The sequence of the gene encoding the chimeric protein Cap-loop2-loop3-loop4 is shown in SEQ ID NO: 8, which was synthesized by Yi...

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Abstract

The invention provides a chimeric protein, a virus-like particle and application thereof and belongs to the field of molecular biology. The chimeric protein is prepared by the following steps: replacing one, two or three of four sites in a porcine circovirus 2 type Cap protein by a main B cell epitope of a porcine O-shaped foot-and-mouth disease virus VP1 protein to obtain 58th-66th amino acid residues, 72nd-94th amino acid residues, 122nd-147th amino acid residues and 162nd-197th amino acid residues. The chimeric protein can form the chimeric virus-like particle after soluble expression, shows the main B cell epitope of the porcine O-shaped foot-and-mouth disease virus VP1 protein on the surface of the chimeric virus-like particle, has very good immunogenicity to both PCV2 and FMDV and can generate a high antibody to the PCV2 and the FMDV, by once immune.

Description

technical field [0001] The invention belongs to the field of molecular biology, and specifically relates to a chimeric protein, a virus-like particle and applications thereof. Background technique [0002] Porcine circovirus type 2 ( Porcine circovirus type 2 , PCV2) is the main pathogen that causes multisystemic wasting syndrome (PWMS) in weaned piglets. Porcine dermatitis nephritic syndrome (PDNS) is closely related to respiratory disease syndrome (PRDS). Since PMWS was first reported in Canada in 1991, the disease has spread to all parts of the world. It is very prevalent in my country's pig herds and has caused great economic losses to the pig industry all over the world. At the International Porcine Veterinary Congress (IPVS) in 2010, porcine circovirus type 2 infection became the disease with the highest attention, and it is one of the important infectious diseases recognized globally that endangers the pig industry. In recent years, the domestic porcine circovir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N1/21C12N7/04A61K39/295A61K39/135A61K39/12A61P31/20A61P31/14
Inventor 郑其升李图帅乔绪稳陈瑾赵晓云于晓明吴楠张元鹏李鹏成侯立婷侯继波
Owner 江苏省苏农科技术转移中心有限公司
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