Recombinant baculovirus expressing porcine circovirus type 3 Cap protein and construction method and primers thereof

A technology of recombinant baculovirus and porcine circovirus, applied in the direction of virus, virus peptide, virus/phage, etc., to achieve the effect of good immunogenicity and good biological activity

Active Publication Date: 2019-01-15
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on PCV3 vaccines, and the expression of PCV3 Cap protein by constructing recombinant baculovirus can lay the foundation for the development of PCV3 subunit vaccines

Method used

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  • Recombinant baculovirus expressing porcine circovirus type 3 Cap protein and construction method and primers thereof
  • Recombinant baculovirus expressing porcine circovirus type 3 Cap protein and construction method and primers thereof
  • Recombinant baculovirus expressing porcine circovirus type 3 Cap protein and construction method and primers thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0039] 1. Construction of recombinant baculovirus rBac-Cap expressing porcine circovirus type 3 Cap gene:

[0040] The primer sequences designed for amplification are as follows:

[0041] Table 1 Required primers and enzyme cutting sites

[0042]

[0043]

[0044] Note: The underlined part is each enzyme cutting site

[0045] 1. Construct pSK-sPCV3 containing the whole PCV3 genome:

[0046] Using the extracted PCV3 DNA as a template, use the primer Xho I-3F and the primer EcoR I-3R to amplify the whole genome of PCV3; recover the PCV3 whole genome gel and use the pBluescript SK (+) vector stored in the laboratory at the same time with Xho I and EcoRI double enzyme digestion, the two enzyme digestion products were purified and recovered, then ligated at a molar ratio of 3:1, and finally the recombinant plasmid pSK-sPCV3 was obtained. The recombinant plasmid was identified by double digestion with Xho I and EcoR I, and the digested product was electrophoresed on a 0.7% ...

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Abstract

The invention belongs to the technical field of vaccine manufacturing. The invention discloses a recombinant baculovirus expressing a porcine circovirus type 3 Cap protein. The invention also discloses primers for constructing a recombinant baculovirus expressing a porcine circovirus type 3 Cap protein. A method for constructing the recombinant baculovirus-expressing porcine circovirus type 3 Capprotein is also disclosed and comprises that follow steps of: 1, constructing a pSK-SPCV3 inluding a PCV3 whole genome; 2. constructing a transfer vector; 3. Constructing a shuttle plasmid; 4, obtaining the recombinant baculovirus. Sf9 cells are transfected with positive shuttle plasmid Bacmid-Cap to obtain the recombinant baculovirus rBac-Cap expressing porcine circovirus type 3 Cap gene. The Capprotein expressed by the invention has good biological activity and is found to have good immunogenicity by immunizing mice.

Description

technical field [0001] The invention relates to a construction method of a recombinant baculovirus expressing porcine circovirus type 3 (PCV3) Cap gene, which is used for making vaccines. Background technique [0002] Porcine circovirus disease (PCVD) is one of the important diseases that endanger the swine industry at present, and PCV2 is the main pathogenic factor leading to this kind of disease. In 2016, Phan and other scholars reported a new type of porcine circovirus PCV3, which can cause porcine dermatitis and nephrotic syndrome (PDNS), porcine reproductive disorders respiratory diseases and digestive system inflammation [Phan TG, Giannitti F, Rossow S , et al. Detection of a novel circovirus PCV3 in pigs with cardiac and multi-systemic inflammation [J]. Virology Journal, 2016, 13(1): 184.]. Through the analysis of PCV3 genome-wide genetic evolution, it was found that PCV3 is in a different evolutionary branch from the other two genotypes of PCV1 and PCV2, and there a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/11C12N15/866C12N15/34C12R1/93
CPCC12N7/00C12N15/86C07K14/005C12N2710/14143C12N2710/14121C12N2800/105C12N2750/10022
Inventor 高崧程清如高清清王小波郇长超刘秀梵
Owner YANGZHOU UNIV
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