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DNA (Deoxyribose Nucleic Acid) vaccine vector based on B2L gene of contagious pustular dermatitis virus and preparation method and application thereof

A technology of DNA vaccine and impetigo virus, which is applied in the field of DNA vaccine carrier and its preparation, DNA vaccine carrier for preventing sheep infectious pustule and its preparation, can solve the problem of complex production process of sheep oral ulcer vaccine, low protection efficiency, no Mouth vaccine and other issues

Inactive Publication Date: 2013-01-23
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, the strain used to produce aphthus vaccine was prepared 20 years ago, and as the virus mutates, the protection efficiency is not high
What's more serious is that due to the complex production process of the oral ulcer vaccine and the smaller market demand than other animal vaccines, many animal vaccine companies are unwilling to produce the oral oral ulcer vaccine, resulting in no oral ulcer vaccine supply on the market

Method used

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  • DNA (Deoxyribose Nucleic Acid) vaccine vector based on B2L gene of contagious pustular dermatitis virus and preparation method and application thereof
  • DNA (Deoxyribose Nucleic Acid) vaccine vector based on B2L gene of contagious pustular dermatitis virus and preparation method and application thereof
  • DNA (Deoxyribose Nucleic Acid) vaccine vector based on B2L gene of contagious pustular dermatitis virus and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation of a DNA vaccine vector based on the B2L gene of sheep infectious impetigo virus

[0031] 1. Cloning of B2L gene by genetic engineering technology

[0032] 1. Primer design:

[0033] A pair of primers were designed according to the complete coding region of the B2L gene, and the upstream and downstream primers P1 and P2 were respectively introduced into BamH Ⅰ ( GGATCC ) and Xho Ⅰ ( CTCGAG )Restriction sites.

[0034] Upstream primer P1: TTT GGATCC ATGTGGCCGTTTCCCTC

[0035] Downstream primer P2: TTT CTCGAG TTAATTTATTGGTTTGC

[0036] 2. PCR amplification of B2L gene

[0037] Primers P1 and P2 are used to amplify the B2L gene under the action of high-fidelity DNA polymerase and the DNA of sheep infectious impetigo virus as a template. The amplification conditions were pre-denatured at 95°C for 5 minutes and then cycled. The cycle parameters were 1 minute at 95°C, 1 minute at 57.5°C, 1 minute at 72°C, and 10 minutes at 72°C after 35 cycle...

Embodiment 2

[0077] Example 2 Immune Effect Verification of DNA Vaccine Vector

[0078] 1. Preparation of DNA vaccines for animal immunization

[0079] Dissolve a large amount of extracted DNA vaccine plasmid in phosphate buffered saline (PBS pH7.4) and mix it with an equal volume of conventional aluminum hydroxide vaccine adjuvant. 4 degrees refrigerator, shelf life of 12 months.

[0080] 2. Animal immunity test

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Abstract

The invention discloses a DNA (Deoxyribose Nucleic Acid) vaccine vector based on B2L gene of contagious pustular dermatitis virus and a preparation method of the DNA vaccine vector and application, and belongs to the field of biological medicine. The DNA vaccine vector based on B2L gene of contagious pustular dermatitis virus is characterized by being prepared by inserting the main immunogenicity gene B2L of the contagious pustular dermatitis virus into a eukaryotic expression vector pVAX1, and then implementing large-scale preparation to obtain the product. The DNA vaccine prepared by the preparation method disclosed by the invention is applied to animal immunization; as the results shown, animal with the DNA vaccine injected are able to effectively resist the infection of the contagious pustular dermatitis virus, and the immune protection efficiency is up to 90% by comparing with a control group, so that the DNA vaccine disclosed by the invention can be widely applied to prevention and treatment of contagious pustular dermatitis disease.

Description

technical field [0001] The invention relates to a DNA vaccine carrier and its preparation method and application, especially a DNA vaccine carrier which can be used to prevent sheep contagious pustules, its preparation method and application, and belongs to the field of biopharmaceuticals. Background technique [0002] Orf contagious impetigo (commonly known as "sheep aphthous ulcer") is a zoonotic infectious disease caused by Orf virus (ORFV) of the family Poxviridae and the genus Parapoxvirus. The disease mainly harms 3-6 month-old lambs and some adult sheep. The clinical symptoms are skin rashes, pustules, ulcers and scabs on the lips and mucous membranes of the mouth; Secondary or mixed infection with other pathogenic microorganisms has a higher mortality rate. The disease can be manifested as herpes and ulceration on the back of the hand, between the fingers and the forearm through wound infection. The disease occurs worldwide and is widespread. Therefore, the outbrea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/79C12N15/39C12N1/21C12N15/66A61K48/00A61P31/20C12R1/19
Inventor 刘湘涛张克山刘永杰孔汉金尚佑军田宏尹双辉逯忠新
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI