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A kind of molecular mimetic peptide of O-type foot-and-mouth disease virus epitope and its application
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A foot-and-mouth disease virus and antigen epitope technology, applied in the field of molecular immunology, to achieve low production costs, good economic and social benefits
Inactive Publication Date: 2011-12-21
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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But, the report of efficient genetically engineered FMDV vaccine is seldom, if can screen out on the basis of FMDV neutrality epitope, develop its genetically engineered vaccine, will help to develop more effective genetically engineered vaccine
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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 △(lacZ)M15proA+B+fhuA2supE thi△(1ac-proAB)△(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, HRP-conjugated Rabbit
[0077] anti-mouse IgG was purchased from Sigma Company.
[0078] PCR primers: Synthesized by Shanghai Jikang Co., Ltd.
[0079] Commonly used enzymes: restriction endonucleases Sac I, Bbe I, Nco I, and XhoI are all products of Takara Company. RNase is a product of Huashun Company.
[0080] Molecular biology kits: pGEM-T kit is the product of Huashun Company; PCR kit and T4 ligase kit are product...
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Abstract
The present invention relates to the field of molecular immunology. Although conventional vaccines such as attenuated foot-and-mouth disease vaccines and inactivated vaccines currently used have good immunogenicity, there are problems such as strong virus virulence, incomplete virus inactivation, and live virus escape from the preparation factory. Unsafe factors have prompted people to seek a safer and more effective FMD vaccine. The present invention utilizes phage surface display technology to screen effective FMDV epitopes, and provides a molecular mimic peptide of O-type foot-and-mouth disease virus antigen epitope, and the present invention also provides that the mimic peptide can be used in the preparation of immunogen and porcine foot-and-mouth disease epitope peptide vaccine Applications. The FMDV epitope peptide vaccine of the present invention has the antigenicity of the target molecule without toxicity, is very safe, and has low cost in large-scale production, and has good economic and social benefits.
Description
technical field [0001] The invention relates to the field of molecular immunology, in particular to a short peptide capable of simulating the antigenic epitope of O-type foot-and-mouth disease virus, and the application of the short peptide in preparing immunogen and porcine foot-and-mouth disease epitope vaccine. Background technique [0002] Foot and Mouth Disease (FMD) is an acute, febrile and highly contagious infectious disease of cloven-hoofed animals. The disease has many transmission routes and fast transmission speed, and it has become a pandemic in the world many times. Although the mortality rate of the disease is not high (except for young animals), it causes a decline in animal production performance, and culling animals costs a lot of manpower, material and financial resources, affects international trade, and causes heavy economic losses. For example, the FMD that broke out in my country's Taiwan in 1997, Japan and South Korea in 2000, and the United Kingdom ...
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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08A61K39/135A61P31/14
Inventor 孙树汉郭瀛军王越张毅周奇
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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