African swine fever B and T cell tandem epitope fusion vaccine
An African swine fever and fusion vaccine technology, applied in the field of vaccines, can solve the problems of increased antibody levels, lack, and inability to produce antibodies quickly, achieving good safety, good immune effect, and avoiding the risk of accelerating virus infection.
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Embodiment 1
[0032] Example 1. Obtaining neutralizing epitope peptides of p72 protein, p54 protein, p30 protein, CD2v protein, C-type lectin protein, PP62 protein, p17 protein or p12 protein:
[0033] Using http: / / tools.iedb.org / bcell / , http: / / crdd.osdd.net / raghava / abcpred / ABC_submission.html, SYFPEITHI and BIMAS, etc., combined with existing literature analysis and design, the obtained African swine fever virus The potential epitopes of neutralizing antibodies are as follows:
[0034] 1.1 p72 protein neutralizing epitope peptide:
[0035] SEQ ID NO. 1: MASGGAFCLIANDGKADKI;
[0036] SEQ ID NO. 2: NVNKSYGKPDPEPTLSQIEETHLVHFNAHFKPYVPVGFEYNKVRPHTGTPTLGNKLTFGIPQYGDFFHD;
[0037] SEQ ID NO. 3: HSSWQDAPIQGTSQMGAHGQLQTFPRNGYDWDNQTPLEGAVYTLVDPFGRPIVPGTKNAYRNLVYYCEYPGERL;
[0038] SEQ ID NO. 4: VSVEGTSGPLLCNIHDLHKPHQSKPILTDENDTQRTCSHTNPKFLSQHFPENSHNIQTAGKQDITPITDA;
[0039] SEQ ID NO.5: RRNIRFKPWFIPGVINEISLTNNELYINNLFVTPEIHNLFVKRVRFSLIRVHKTQ1.2p30 protein neutralizing epitope peptide:
[0040]...
Embodiment 2
[0070]Embodiment 2. The specific amino acid sequence of the African swine fever tandem epitope fusion protein of the present invention (only enumerating part as an example,
[0071] Not all possible amino acid sequences of the invention):
[0072] 2.1 When explaining the African swine fever tandem epitope fusion protein, it is characterized by the sequence number representing the amino acid sequence of the neutralizing epitope peptide, and the connecting arm is indicated in bold, NP 147-155 Amino acid sequence for (in italics) and the immunoactive peptide tuftsin TKPR (underlined) indicates that different constituents are connected by connecting arms and "-".
[0073] 2.2 The African swine fever tandem epitope fusion protein of the present invention includes p30, p54, p72 neutralizing epitope peptide and NP 147-155 , its amino acid sequence can be expressed as:
[0074] (1) Including 1 neutralizing antigenic epitope of p30 protein, 1 neutralizing antigenic epitope of p54...
Embodiment 3
[0081] Example 3 According to the amino acid sequence of the African swine fever tandem epitope fusion protein of the present invention, the fusion protein is expressed and purified by genetic engineering
[0082] 3.1 Expression and purification of fusion protein by prokaryotic expression system
[0083] 3.1.1 According to the amino acid sequence of the designed fusion protein (including other necessary fragments without antigenic activity such as purification tag His if necessary), the synthesis of the nucleic acid molecule encoding the fusion protein is carried out by Sangon Bioengineering (Shanghai) Co., Ltd. And the construction of carrier (carry out codon optimization according to the amino acid sequence of fusion protein and the expression system that the nucleic acid molecule of coding this fusion protein uses, synthesize this nucleic acid molecule), prokaryotic expression vector selects pET28a for use, inserts the synthetic nucleic acid between NocI and XhoI Molecules,...
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