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Targeted genetic engineering vaccine for African swine fever immune system

An African swine fever, targeted technology, applied in the field of vaccines, can solve problems such as poor immune effect, achieve good immune effect, prevent antibody dependence (ADE) effect, and enhance the effect of cellular immune response

Active Publication Date: 2020-02-07
河南省生物工程技术研究中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The invention provides an African swine fever fusion protein. When the African swine fever fusion protein is used to make a vaccine, the immune effect is better, so as to solve the problem of poor immune effect of subunit vaccines in the prior art

Method used

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  • Targeted genetic engineering vaccine for African swine fever immune system
  • Targeted genetic engineering vaccine for African swine fever immune system
  • Targeted genetic engineering vaccine for African swine fever immune system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Obtaining the epitopes of p72 protein, p54 protein and p30 protein

[0027] 1.1 Screen suitable neutralizing epitopes from the amino acid sequences of p72 protein, p54 protein and p30 protein, and then design the amino acid sequence of the African swine fever fusion protein of the present invention.

[0028] The amino acid sequence of p72 protein is: MASGGAFCLIANDGKADKIILAQDLLNSRISNIKNVNKSYGKPDPEPTLSQIEETHLVHFNAHFKPYVPVGFEYNKVRPHTGTPTLGNKLTFGIPQYGDFFHDMVGHHILGACHSSWQDAPIQGTSQMGAHGQLQTFPRNGYDWDNQTPLEGAVYTLVDPFGRPIVPGTKNAYRNLVYYCEYPGERLYENVRFDVNGNSLDEYSSDVTTLVRKFCIPGDKMTGYKHLVGQEVSVEGTSGPLLCNIHDLHKPHQSKPILTDENDTQRTCSHTNPKFLSQHFPENSHNIQTAGKQDITPITDATYLDIRRNVHYSCNGPQTPKYYQPPLALWIKLRFWFNENVNLAIPSVSIPFGERFITIKLASQKDLVNEFPGLFVRQSRFIAGRPS RRNIRFKPWFIPGVINEISLTNNELYINNLFVTPEIHNL FVKRVRFSLIRVHKTQ VTHTNNNHHDEKLMSALKWPIEYMFIGLKPTWNISDQNPHQHRDWHKFGHVVNAIMQPTHHAEISFQDRDTALPDACSSISDISPVTYPITLPIIKNISVTAHGINLIDKFPSKFCSSYIPFHYGGNAIKTPDDPGAMVSAAINMITFALKPREEYQPSGHINDYVSASLLMITFALKPREE...

Embodiment 2

[0074] Example 2 Prokaryotic expression system was used to express and purify the fusion protein according to the amino acid sequence of the designed fusion protein

[0075] (1) Construction of a prokaryotic expression plasmid: the amino acid sequence of the African swine fever fusion protein of the present invention was handed over to Shenggong Bioengineering (Shanghai) Co., Ltd., and the company synthesized the corresponding code for the above-mentioned African pig according to the provided amino acid sequence The nucleotide sequence of the plague fusion protein, and clone the synthesized nucleotide into a suitable plasmid vector.

[0076] (2) Expression of African swine fever fusion protein: The synthetic plasmid containing the African swine fever fusion protein was transferred into BL21(DE3), and the expression engineering bacteria were constructed, and inoculated to 100μg / mL Kan at an inoculum of 1%. (Kanamycin) 100mL LB liquid culture medium, 37℃, 200r / min, when the A600 is 0...

Embodiment 3

[0079] 3.1 Preparation of African swine fever vaccine: take the African swine fever fusion protein solution prepared in Example 2 and dilute it to 0.2 mg / ml, mix it with adjuvant 201 (Sebik, France) in equal volume, and emulsify it with an emulsifier for 25 minutes.

[0080] 3.2 Verify the immune effect of the prepared African swine fever vaccine

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Abstract

The invention belongs to the technical field of vaccines, and particularly relates to a targeted genetic engineering vaccine for an African swine fever immune system. The main raw material of the African swine fever immune system targeted genetic engineering vaccine is African swine fever fusion protein. The African swine fever fusion protein comprises a fragment selected from p72 protein, a fragment selected from p54 protein and a fragment selected from p30 protein. Wherein the fragment selected from the p72 protein at least comprises a sequence as shown in SEQ ID NO.1, the fragment selectedfrom the p54 protein at least comprises a sequence as shown in SEQ ID NO.2, and the fragment selected from the p30 protein at least comprises a sequence as shown in SEQ ID NO.3. The African swine fever fusion protein has the advantages of good immune effect and the like.

Description

Technical field [0001] The invention belongs to the technical field of vaccines, and specifically relates to a targeted genetic engineering vaccine for the immune system of African swine fever. Background technique [0002] African Swine Fever (ASF) was introduced into my country in August 2018, and it has severely affected the development of animal husbandry and people's lives. It has been nearly a hundred years since the disease was discovered in 1921, but there is currently no vaccine or specific medicine approved globally. With the development of modern biology, the research and development of ASF vaccine has made great progress. In view of the existing research, it seems unlikely to develop an effective inactivated ASF vaccine. The current research and development directions mainly focus on attenuated vaccines, live virus vector vaccines, subunit vaccines and nucleic acid vaccines. [0003] Live attenuated vaccines have the advantage of being able to effectively activate th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C07K16/08G01N33/569C12N15/70A61K39/187A61P31/20C12R1/19
CPCC07K14/005C07K16/081G01N33/56983C12N15/70A61K39/12A61P31/20C12N2710/12022C07K2319/00C07K2319/21C07K2319/33G01N2333/01C12N2710/12034A61K2039/552Y02A50/30
Inventor 毕胜利王云龙殷宏常慧芸范雪亭李玉林伊瑶张怡青王继创宋长绪王国强
Owner 河南省生物工程技术研究中心
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