Porcine circovirus type 3 virus-like particle as well as construction method and application thereof

A technology of porcine circovirus and type 3 virus, applied in the direction of viruses, viral peptides, antiviral agents, etc., can solve the problem of poor immunogenicity of Cap protein truncations, and achieve high immunogenicity and low preparation costs

Active Publication Date: 2022-05-27
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to overcome the formation of inclusion bodies, there is a case where a prokaryotic expression system is used to express a Cap protein truncation that removes nuclear localized amino acids. Although this method achieves soluble expression of the Cap protein, the prepared Cap protein truncation exists Poor immunogenicity and other disadvantages

Method used

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  • Porcine circovirus type 3 virus-like particle as well as construction method and application thereof
  • Porcine circovirus type 3 virus-like particle as well as construction method and application thereof
  • Porcine circovirus type 3 virus-like particle as well as construction method and application thereof

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Embodiment 1

[0047] 1. Porcine circovirus type 3 and structural protein gene Cap codon optimization and synthesis

[0048] Based on the published Capsid sequence of the structural protein gene of the porcine circovirus type 3 strain (GenBabnk accession number: NC_031753.1), and according to the codon bias of Escherichia coli, the porcine circovirus type 3 Cap gene was optimized and synthesized. The optimized porcine circovirus type 3 Cap gene sequence is shown in SEQ ID No.1.

[0049] 2. Construction of recombinant expression vector for porcine circovirus type 3 structural protein

[0050] (1) Enzyme cleavage of fragments and vectors

[0051] Referring to the endonuclease instructions, the synthesized Cap gene and pSMA vector (containing SUMO-His tag) were digested, and the reaction system was as follows: 20 μl of DNA fragment (Cap) or 30 μl of vector was added to 50 μl of the restriction enzyme digestion reaction system, containing the endonuclease Enzyme buffer and endonucleases BsmB I...

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Abstract

The invention relates to a porcine circovirus type 3 virus-like particle as well as a construction method and application thereof, and belongs to the technical field of preparation of virus-like particles. The invention provides a recombinant plasmid, the recombinant plasmid takes a pSMA plasmid as a basic carrier, an optimized Cap full-length gene is inserted, and the nucleotide sequence of the optimized Cap full-length gene is as shown in SEQ ID NO. 1. The defects of an existing eukaryotic expression system and a traditional prokaryotic expression system are overcome, the preparation method for soluble expression of complete Cap protein of the porcine circovirus type 3 and successful realization of the virus-like particles by using the prokaryotic expression system becomes possible, and the obtained virus-like particles are high in immunogenicity and low in cost.

Description

technical field [0001] The invention relates to the technical field of virus-like particle preparation, in particular to a porcine circovirus type 3 virus-like particle and a construction method and application thereof. Background technique [0002] Porcine circovirus (PCV) is a single-stranded circular DNA virus belonging to the genus Cyclovirus of the family Anelloviridae, and is one of the smallest animal viruses discovered so far. Porcine circovirus type 3 (PCV3), as one of them, is similar in shape, size and genome structure to PCV1 and PCV2. The virion has an icosahedral structure, no envelope, and a diameter of 17-20 nm. , is a single-stranded circular DNA with a total length of 2000 bp and contains 3 major open reading frames (ORFs), of which ORF2 is 645 bp in length and encodes the main antigenic capsid protein (Cap protein) of PCV type 3. Relevant studies have shown that the amino acid homology of PCV 3 and PCV2 Cap protein is only 30%, and the difference is very ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C12N1/21C07K14/01C12P21/06C07K1/34C07K1/22C07K1/36A61K39/12A61P31/20C07K16/08C12R1/19
CPCC12N15/70C07K14/005C12P21/06A61K39/12A61P31/20C07K16/081C12N2800/22C12N2750/10023C12N2750/10034A61K2039/5258A61K2039/552Y02A50/30
Inventor 孙世琪白满元郭慧琛裴辰辰董虎张韵丁耀忠尹双辉吴金恩郭建宏何继军靳野
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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