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Porcine circovirus type Ⅱ genetic engineering subunit vaccine and its preparation method and application

A porcine circovirus and vaccine technology, applied in the direction of antiviral agents, virus antigen components, chemical instruments and methods, etc., can solve the problems of poor virus multiplication ability and difficulty, and achieve the effects of low cost, increased weight gain, and high purity

Active Publication Date: 2015-12-02
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because PCV2 does not produce cytopathic changes when cultured in vitro, and the ability of virus proliferation is poor, it is very difficult to prepare the virus vaccine by traditional methods.

Method used

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  • Porcine circovirus type Ⅱ genetic engineering subunit vaccine and its preparation method and application
  • Porcine circovirus type Ⅱ genetic engineering subunit vaccine and its preparation method and application
  • Porcine circovirus type Ⅱ genetic engineering subunit vaccine and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0053] 1. Construction of cloning vector of PCV2cap protein gene

[0054] 1Materials and methods

[0055] 1.1 Extraction of disease material DNA

[0056] Take 0.5 g of the inguinal lymph node sample of the dead pig after porcine circovirus infection, add 4.5 ml of sterile normal saline, and place it in a glass homogenizer to make a homogenate. Take 472μl of lymph node homogenate, add 25μl of 10% SDS and 2.5μl of proteinase K (25mg / ml), place in a 50℃ water bath for 2 hours, add 500μl of water-saturated phenol, vortex for 30 seconds, and centrifuge at 12000rpm for 10 minutes. Transfer the supernatant to another centrifuge tube, add 500 μl of phenol:chloroform:isoamyl alcohol (volume ratio 25:24:1) to mix and centrifuge at 12000rpm for 10 minutes. Take the supernatant, add 3mol / L sodium acetate (pH 5.3) at 1 / 10 volume, add 2.5 times the volume of absolute ethanol, and put it in the refrigerator at -20°C for 30 minutes. Centrifuge at 15000 rpm for 10 minutes, discard the supernatant ...

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Abstract

The invention discloses a porcine circovirus type II gene engineering subunit vaccine, and a preparation method and application thereof. The porcine circovirus type II vaccine comprises a soluble fusion protein composed of a protein obtained by expressing a porcine circovirus type II capsid protein gene by Escherichia coli; and the porcine circovirus type II capsid protein gene is subjected to positioning signal zone shear and site-directed mutagenesis, and the site-directed mutagenesis comprises mutation of codon AGA or AGG to CGC. A porcine circovirus type II vaccine prepared by the technical scheme provided by the invention has high antigen purity, good safety, strong immunogenicity, and no pathogenicity to animals like pig. Furthermore, the vaccine antigen in the invention is expressed by Escherichia coli, so a preparation process is relatively simple and low-cost.

Description

Technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a preparation method and application of a porcine circovirus type II genetic engineering subunit vaccine. Background technique [0002] In 1991, John Harding of Canada reported the postweaning multisystemic wasting syndrome (PMWS) of weaned piglets. Later studies confirmed that Porcinecircovirus 2 (PCV2) was the main pathogen of the syndrome. [0003] Existing studies have shown that Porcine circovirus disease (PCVD) is a clinical syndrome caused by PCV2 infection. Its main symptoms are progressive weight loss, shortness of breath, anemia, diarrhea, jaundice, and interstitial Pneumonia, lymphadenitis and nephritis, etc. Porcine circovirus type II mainly affects 5-12 weeks old weaned piglets, and is related to PMWS, porcine dermatitis and nephropathy syndrome (PDNS), porcine respiratory syndrome (porcinerespiratorydiseasecomplex, PRDC), A2 congenital Congenitaltremor (CT), p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/21A61K39/12A61P31/20
Inventor 荣俊杜元钊范根成韩建文蔡联燊
Owner YEBIO BIOENG OF QINGDAO
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