Recombinant baculovirus expressing porcine circovirus type 3 truncated cap protein and its construction method and primers

A technology of recombinant baculovirus and porcine circovirus, applied in the direction of virus, viral peptide, virus/bacteriophage, etc., to achieve improved expression and good expression effect

Active Publication Date: 2022-05-27
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are few reports on PCV3 vaccines, and the expression of PCV3Cap 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 truncated cap protein and its construction method and primers
  • Recombinant baculovirus expressing porcine circovirus type 3 truncated cap protein and its construction method and primers
  • Recombinant baculovirus expressing porcine circovirus type 3 truncated cap protein and its construction method and primers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040]I. Construction of recombinant baculovirus rBac-ΔCap expressing the truncated Cap gene of porcine circovirus type 3:

[0041] The primer sequences designed for amplification are as follows:

[0042] Table 2 Required primers and digestion sites

[0043]

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

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

[0046] Taking the extracted PCV3DNA as the template, the PCV3 whole genome was amplified with primers Xho I-3F and primer EcoR I-3R; pcV3 whole genome gum was recovered and simultaneously cut with Xho I and EcoR I bienzymes with pBluescript SK(+) vectors, and the two digestion products were purified and recovered and connected according to a molar ratio of 3:1, and finally the recombinant plasmid pSK-sPCV3 was obtained. Recombinant plasmids were identified by XhoI and EcoR I dual digestion, and the digestion products were electrophoretic with a 0.7% agarose gel to obtain bands of about 2958 ...

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Abstract

The invention belongs to the technical field of vaccine manufacture. The invention discloses a recombinant baculovirus rBac-ΔCap expressing porcine circovirus type 3 truncated Cap gene, and the preservation number is CGMCC No.15691. The invention also discloses a primer for constructing recombinant baculovirus rBac-ΔCap expressing porcine circovirus type 3 truncated Cap gene. The present invention also discloses a method for constructing a recombinant baculovirus rBac-ΔCap expressing a porcine circovirus type 3 truncated Cap gene, comprising the following steps: 1. Constructing pSK-sPCV3 containing the entire PCV3 genome; 2. Construction of a transfer vector 3. Construction of shuttle plasmid; 4. Obtaining of recombinant baculovirus: transfect Sf9 cells with positive shuttle plasmid Bacmid-ΔCap, and obtain recombinant baculovirus rBac-ΔCap expressing porcine circovirus type 3 truncated Cap gene. The invention can conveniently and quickly construct the recombinant baculovirus expressing the PCV3 truncated Cap protein, and can confirm that the expressed truncated Cap protein has good biological activity, and the immunized mice are found to have good immunogenicity.

Description

Technical field [0001] The present invention relates to a method for constructing a recombinant baculovirus expressing porcine circovirus type 3 (PCV3) truncated Cap gene for the manufacture of a vaccine. Background [0002] Swine circovirus disease (PCVD) is one of the most important diseases currently endangering the swine industry, of which PCV2 is the main causative agent of this type of disease. In 2016, Phan et al. reported on the novel porcine circavirus PCV3, which can cause diseases such as 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 PCV3in pigs withcardiac and multi-systemic inflammation[J]. Virology Journal,2016,13(1):184.]。 Through the analysis of PCV3 genome-wide genetic evolution, it is found that PCV3 and the viruses of the other two genotypes of PCV1 and PCV2 are in different evolutionary branches, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/866C12N15/66
CPCC12N15/86C07K14/005C12N2710/14043C12N2750/10022
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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