O-type foot and mouth disease virus antigen epitope molecular mimic peptide and application thereof
A foot-and-mouth disease virus and antigen epitope technology, applied in the field of molecular immunology, to achieve good economic and social benefits, and low production costs
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Embodiment 1
[0033] Embodiment 1: Screening and sequence analysis of porcine foot-and-mouth disease virus VP1 antigen mimotope
[0034] 1. Experimental materials
[0035] 1. Phage Random Peptide Library Kit
[0036] The linear 12-peptide library kit for phage surface capsid protein III display was purchased from NewEngland Biolabs, USA. The titer of the peptide library was 4×10 12 pfu / ml, random diversity 1.9×10 9 ; The genotype of the recipient strain E.coli ER2537 is F'lac q Δ(1acZ)M15proA+B+fhuA2supE thiΔ(1ac-proAB)Δ(hsdMS-mcrB)5(rk - mk - McrBC).
[0037] 2. Chemical reagents
[0038] BSA was purchased from Roche Company; IPTG and X-gal were purchased from Bebco Company; PEG8000, PEG20000, ABTS and Tween20 were purchased from Sigma Company; foot-and-mouth disease ELISA detection kit was produced by Lanzhou Institute of Veterinary Medicine, Chinese Academy of Agricultural Sciences; HRP-labeled anti-M13 monoclonal Antibodies and DEAE sephadex A50 were purchased from Pharmacia; porc...
Embodiment 2
[0071] Example 2: Application of Mimotope Peptides in Vaccines
[0072] 1. Experimental materials:
[0073] 1. Experimental animals C57BL / 6 (H-2kb) mice and Balb / c mice, 6-8 weeks old, were purchased from the Experimental Animal Center of Second Military Medical University.
[0074]2. Bacterial Strains and Genetic Engineering Vectors Host bacteria for genetic engineering: Escherichia coli DH5α, Top10′; pVAX1 vector, pBAD / gIII A vector (purchased from Invitrogen Life Science).
[0075] 3. Common reagents
[0076] Anti-HBcAg mAb was purchased from Biodesign Company, and HRP-conjugated Rabbitanti-mouse IgG was purchased from Sigma Company.
[0077] PCR primers: Synthesized by Shanghai Jikang Co., Ltd.
[0078] Commonly used enzymes: restriction endonucleases SacI, Bbe I, Nco I, Xho I, all of which are products of Takara Company. RNase is a product of Huashun Company.
[0079] Molecular biology kits: pGEM-T kit is the product of Huashun Company; PCR kit and T4 ligase kit are ...
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