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Porcine circovirus type 2 Cap gene modified recombinant antigen and application thereof

A technology of porcine circovirus and gene modification, applied in the direction of virus antigen components, plant gene improvement, application, etc., can solve the problems of unsatisfactory immune protection effect, high titer virus difficulty, high price, etc., and achieve good application prospects , easy storage and application, high antibody level effect

Active Publication Date: 2016-05-04
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although commercialized PCV2 vaccines have been widely used, and production practices have shown that these vaccines can effectively reduce the clinical symptoms of PCVAD, but these vaccines cannot completely prevent the infection and spread of PCV2
Based on the fact that the current commercialized subunit vaccine is expensive and the circovirus is weak, it is difficult to obtain high-titer virus by culturing the whole virus, the immune protection effect is not ideal, and the current situation of the domestic use rate is not high. Necessary for safe, efficient, broad-spectrum, and inexpensive vaccines

Method used

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  • Porcine circovirus type 2 Cap gene modified recombinant antigen and application thereof
  • Porcine circovirus type 2 Cap gene modified recombinant antigen and application thereof
  • Porcine circovirus type 2 Cap gene modified recombinant antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of porcine circovirus type 2 modified recombinant antigen

[0033] 1. Screening of a gene with significant immune enhancement effect:

[0034] Porcine circovirus type 2 Cap protein is the main structural protein of the virus, which constitutes the nucleocapsid of the virus, and the encoded protein can bind to the host cell receptor. It is the main immunogenic protein of the virus and can stimulate the body to produce antibodies , significantly reducing the incidence of immunized pigs, but in practical applications, both prokaryotic expression and eukaryotic expression of Cap protein have defects such as high cost, limited expression yield, high requirements for protein purification, and poor immunogenicity. In order to improve the immunogenicity of Cap protein, this study used DNAStar biological software to analyze a gene with significant immune enhancement effect, and use this gene to modify the Cap gene in order to achieve the purpose of enhancin...

Embodiment 2

[0039] Example 2 Preparation of Porcine Circovirus Type 2 Cap Gene Modified Subunit Vaccine and Its Immunological Efficacy Analysis

[0040] 1. Vaccine preparation:

[0041] The endotoxin-removed protein (amino acid sequence shown in SEQIDNO: 2 and SEQIDNO: 4, respectively) was quantified by the BCA protein quantitative analysis kit and diluted to an appropriate concentration, and the modified protein was emulsified by adding an equal volume of Freund's adjuvant (Sigma) into vaccine preparations.

[0042] The amino acid sequence of the porcine circovirus type 2 LG strain Cap protein after removal of the nuclear localization signal is shown in SEQ ID NO:4.

[0043] Said SEQ ID NO: 4 is as follows:

[0044] NGIFNTRLSCTFGYTVKATTVRTPSWAVDMMRFNINDFVPPGGGTNEISIPFEYYRIRKVKVEFWPCSPITQGDRGVGSTAVILDDNFVTRATALTYGPYVNYSSRHTIPQPFSYHSRYFTPKPVLDSTIDYFQPNNKRNQLWLRLQTSANVDHVGLGIAFENSTYDQDYNIRVTMYKDPLKEFPL.

[0045] The nucleotide sequence of the porcine circovirus type 2 LG strain Cap prote...

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Abstract

The invention discloses a porcine circovirus type 2 Cap gene modified recombinant antigen and an application thereof, and belongs to the field of veterinary vaccine researches. The modified recombinant antigen is obtained by modification and construction of a Cap gene after a nuclear localization sequence of a porcine circovirus type 2 LG strain widely prevalent in swinery in China at present is deleted. A vaccine prepared from the modified recombinant antigen can stimulate organisms to produce high-level neutralizing antibodies, is superior to commercially available inactivated influenza virus vaccines and separate Cap protein immunization groups after nuclear localization sequences are deleted, has a highest antibody level, is safe and harmless to immunized animals after inoculation, has the advantages of low price, safety, stability, high yield and easiness in preservation and application, and has great significance to related diseases caused by porcine circovirus type 2 infection in China, so that the porcine circovirus type 2 Cap gene modified recombinant antigen has a good application prospect.

Description

technical field [0001] The invention belongs to the field of veterinary vaccine research and relates to a porcine circovirus type 2 antigen and its application, in particular to a porcine circovirus type 2 Cap gene modified recombined antigen and its application. Background technique [0002] Porcine circovirus type 2 (porcine circovirus type 2, PCV2) is the main viral pathogen of porcine circovirus-associated disease (porcine circovirus-associated disease, PCVAD), mainly infects the immune organs of pigs, leads to immunosuppression of pigs, and causes various viruses or bacteria Concurrent or secondary infection of the disease, leading to growth inhibition or death of pigs. Common mixed infection virus diseases such as: PRDC (chronic respiratory disease syndrome of fattening pigs), PRRSV, PPV, etc., common bacterial mixed infection pathogens such as: Pasteurella multocida, Mycoplasma pneumoniae, etc. PCVAD mainly includes porcine multisystemic failure syndrome (PMWS), porc...

Claims

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Application Information

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IPC IPC(8): C07K14/01C12N15/34C12N15/63A61K39/12A61K48/00A61P31/20
CPCA61K39/12A61K48/0016C07K14/005C12N2750/00034C12N2750/10022
Inventor 陈金顶吴云燕孙秀漆月朱俊灵王利军李鹏赵明秋
Owner SOUTH CHINA AGRI UNIV
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