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Infectious bursal disease virus recombinant ubiquitinated DNA vaccine and preparation method thereof

A bursal disease, infectious technology, applied in the field of DNA vaccine, to achieve the effect of high cost, short preparation time and simple operation method

Inactive Publication Date: 2017-09-22
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Despite extensive use of traditional attenuated live vaccines and inactivated vaccines for the prevention and treatment of IBD, the disease is still prevalent across the country

Method used

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  • Infectious bursal disease virus recombinant ubiquitinated DNA vaccine and preparation method thereof
  • Infectious bursal disease virus recombinant ubiquitinated DNA vaccine and preparation method thereof
  • Infectious bursal disease virus recombinant ubiquitinated DNA vaccine and preparation method thereof

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Experimental program
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Effect test

experiment example

[0034] Step 1, amplification of chicken ubiquitin gene fragment Ub and infectious bursal disease virus structural protein VP2 gene

[0035] 1) Mix 200 μg chicken muscle tissue (or 200 μL infectious bursal disease virus liquid) with 1 mL Trizol solution, shake and let stand for 5 minutes, add 200 μL chloroform, shake for 30 seconds, then centrifuge at 12000 r / min for 30 minutes, take 400 μL of supernatant in Add 400 μL isopropanol to a new centrifuge tube, mix well, let stand at -20°C for 30 minutes, centrifuge at 12,000 r / min for 30 minutes, discard the supernatant, add 1 mL of 75% ethanol, centrifuge at 12,000 r / min for 10 minutes, discard the supernatant, and add 30 μL ddH 2 O Obtain chicken muscle tissue (or infectious bursal disease virus) RNA and store it at -20°C for later use.

[0036] 2) Add the RNA of chicken muscle tissue (or infectious bursal disease virus) obtained in step 1 to the following mixture: 4 μL of buffer solution (250 mM Tris-HCl, pH 8.3, 375 mM KCl, 15 ...

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Abstract

The invention discloses a recombinant plasmid for expressing infectious bursal disease virus structural protein VP2; the recombinant plasmid mainly comprises infectious bursal disease virus structural protein VP2 gene, avian Ub gene and eukaryotic expression vector pVAX1. Therefore, the inventor designs and establishes a corresponding infectious bursal disease virus recombinant ubiquitinated DNA vaccine and a preparation method thereof. Compared with the prior art, the vaccine has the advantages of good manufacture simplicity, low cost, high safety, and good immunization effect. Tests indicate that the vaccine is effective in avoiding and decreasing damage and chicken mortality of immune organs, such as bursa of Fabricius, due to infectious bursal disease.

Description

technical field [0001] The invention belongs to the technical field of DNA vaccines, in particular to an infectious bursal disease virus recombinant ubiquitinated DNA vaccine and a preparation method thereof. Background technique [0002] Infectious bursal disease (IBD) is a severe and highly contact immunosuppressive disease caused by infectious bursal disease virus that harms young chickens. It has been nearly 40 years since IBD was first discovered in China in 1979. Despite extensive use of traditional attenuated live vaccines and inactivated vaccines for the prevention and treatment of infectious bursal disease, the disease is still prevalent throughout the country. The prevalence of infectious bursal disease has seriously affected the healthy and stable development of the national poultry industry and caused huge economic losses to the poultry industry. [0003] The genome of infectious bursal disease virus encodes two major structural proteins, VP2 and VP3. Among th...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/66A61K39/12A61K48/00A61K9/00A61P31/14
CPCA61K9/0019A61K39/12A61K48/005A61K2039/53A61K2039/552C12N15/66C12N15/85C12N2720/10034
Inventor 陈果姬忠华焦鹏涛赵增志张誉文何秀苗磨美兰韦天超黄腾韦平
Owner GUANGXI UNIV
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