KR20190082081A - Vaccine composition for foot and mouth disease virus a serotype - Google Patents

Abstract

The present invention relates to a vaccine composition for foot-and-mouth disease virus type A and a method for generating immune responses to foot-and-mouth disease virus type A in a subject. According to the present invention, a vaccine composition and an immune response generation method are useful for effective defense against the Asian topotype of foot-and-mouth disease virus type A occurring in Korea and neighboring East Asian countries such as China or Taiwan.

Description

VACCINE COMPOSITION FOR FOOT AND MOUTH DISEASE VIRUS A SEROTYPE "

The present invention relates to polypeptides, polynucleotides, plasmids, and vaccine compositions containing them that are involved in the production of an immune response against foot-and-mouth disease virus type A. The present invention also relates to a method for producing an immune response against the FMD type A virus in an individual.

Foot and Mouth Disease (FMD) is a viral disease that occurs in hoofed chicks, including livestock (cattle, pigs, sheep, goats, deer) and various wild animals. It is characterized by the formation of blisters. FMD is highly contagious and has a mortality rate of over 40%, which is a socioeconomically significant disease. For this reason, the International Bureau of OIE (OIE) classifies and manages FMD as a Class A disease, and in most countries it has been designated as a Class 1 livestock infectious disease. In addition, the onset of the FMD population, which has caused tremendous economic damage, has occurred over almost every continent over the past decade.

Foot and Mouth Disease Virus (FMDV) is a single-stranded positive RNA virus belonging to the genus Adovidovirus, which is a picornavirus. The incubation period varies from animal to animal, but is about 6 to 10 days. In addition, since the virus is excreted in vitro before clinical symptoms, propagation can easily occur. FMDV is transmitted through respiratory, digestive, and reproductive systems, especially viruses that are released into the respiratory tract up to 250 km. FMDV is classified into seven serotypes (O, A, C, SAT1, SAT2, SAT3, and Asia 1) according to the coding region sequence of VP1. . In the meantime, most of the foot-and-mouth disease problems in Korea, including Asia and Europe, were O-type, so Korea has so far been inoculated with O-type vaccine. However, it has been confirmed that FMD occurring in some parts of the country recently caused additional infection of A virus. Type A FMDV has three regional types: ME-SA (Middle East-South Asia), AFRICA (Africa), and ASIA (Asia). Because of the high rate of change in this type of area, it is often not possible to protect against virus attacks. Therefore, it is necessary to develop various vaccines against type A virus. In particular, the recent FMD in Korea is due to the virus belonging to the Sea-97 lineage (Sea-97 lineage). Presently, there is a vaccine against FMDV type A, but it is presumed that the protective immunity is relatively low even if the vaccine is inoculated because the antigenicity of the virus is low. Therefore, there is a strong demand for a vaccine against viruses belonging to the A-type FMDV, particularly the Sea-97 lineage, in order to prevent effective prevention of FMD and economic damage caused by infection.

It is an object of the present invention to provide a polypeptide capable of generating an immune response against the FMD virus type A and a polynucleotide encoding the polypeptide.

It is another object of the present invention to provide a plasmid capable of generating an immune response against foot-and-mouth disease virus type A.

It is another object of the present invention to provide a vaccine composition for foot-and-mouth disease virus type A.

It is yet another object of the present invention to provide a method for producing an immune response against the FMD virus type A in an individual.

In order to achieve the above object, one aspect of the present invention provides a polypeptide comprising an amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having 90% or more homology thereto.

The amino acid sequence of SEQ ID NO: 1 or the amino acid sequence having 90% or more of the sequence homology thereto may be a sequence corresponding to the VP1 region of FMDV serotype.

As used herein, the term "homology" means the degree to which a given polypeptide sequence or polynucleotide sequence corresponds and may be expressed as a percentage. In the present specification, its homologous sequence having the same or similar activity as a given polypeptide sequence is indicated as "% homology ". The homology may be determined using standard software, e.g., BLAST 2.0, which computes parameters such as score, identity and similarity, or using hybridization techniques, such as BLAST 2.0, By comparing the sequences by experiment, and the appropriate hybridization conditions to be defined can be determined by methods well known to those skilled in the art.

The amino acid sequence of SEQ ID NO: 1 is preferably derived from the foot-and-mouth disease virus type A Asian type, and more preferably from the foot-and-mouth disease virus type ASIA Sea-97 type. Most preferably the amino acid sequence of SEQ ID NO: 1 is GDMM / CHA / 2013-S (KF450794), CAM / 2/2008 (HQ116294), MAY / 4/2012 (KT223560), A / VIT / 3/2008 (HQ116371) , MAY / 97 or VN / T11D / 2013 strains. In one embodiment, the common sequences of the VP1 region sequences of GDMM / CHA / 2013-S, CAM / 2/2008, MAY / 4/2012, and A / VIT / 3/2008 viruses are examined, Sequences important for neutralizing antibody production were recombined based on VP1 of VN / T11D / 2013 virus to generate the amino acid sequence of SEQ ID NO: 1. The term "recombinant" means the semisynthetic or synthetic origin of a polynucleotide or polypeptide linked to another polynucleotide or polypeptide in an arrangement not found in nature or found in nature.

The amino acid sequence having 90% or more sequence homology with the amino acid sequence of SEQ ID NO: 1 may preferably be the amino acid sequence of SEQ ID NO: 2. In one embodiment, the common sequences of the VP1 region sequences of the GDMM / CHA / 2013-S, CAM / 2/2008, MAY / 4/2012 and A / VIT / 3/2008 viruses were examined and then mismatched portions and neutralization The sequence important for antibody production was recombined based on VP1 of the MAY / 97 virus to generate the amino acid sequence of SEQ ID NO: 2.

The amino acid sequence of SEQ ID NO: 1 and the amino acid sequence of SEQ ID NO: 2 are highly homologous to the foot-and-mouth disease virus of the A ASIA Sea-97 strain recently developed in Korea and the MAY / 97 or VN / T11D / 2013 virus There is a high degree of homology. Particularly, the amino acid sequence of SEQ ID NO: 1 is more homologous with the foot-and-mouth disease virus of the A ASIA Sea-97 strain recently developed in Korea. Therefore, the polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or the amino acid sequence of SEQ ID NO: 2 can effectively function as an antigen capable of generating an immune response against the FMD type A virus.

Another aspect of the present invention provides a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having 90% or more homology thereto, and the amino acid sequence of VP2-4 protein of foot-and-mouth disease virus.

In the polypeptide, the amino acid sequence having the amino acid sequence of SEQ ID NO: 1 or having at least 90% homology thereto may be sequentially connected to the amino acid sequence corresponding to VP2, VP3 and VP4 proteins derived from foot-and-mouth disease virus. For example, VP4 region amino acid sequence-VP2 region amino acid sequence-VP3 region amino acid sequence-Sequence of amino acid sequence of SEQ ID NO: 1 (or amino acid sequence of SEQ ID NO: 2). Additional amino acid sequences may be included between each amino acid sequence. For example, the amino acid sequence of the furin cleavage site may be included between sequences of each VP region. The polypeptide may further comprise an IgE leader sequence and / or a 2A amino acid sequence. For example, the polypeptide comprises an amino acid sequence of IgE leader sequence - VP 4 region amino acid sequence - VP 2 region amino acid sequence - VP 3 region amino acid sequence - amino acid sequence of SEQ ID NO: 1 (or amino acid sequence of SEQ ID NO: 2) (Preferably in the above order). Also in this case, preferably, the amino acid sequence of the purine cleavage site may be further included between the sequences of the respective regions.

The amino acid sequence of the VP 2 to VP 4 protein may preferably be derived from the type A foot-and-mouth disease virus, more preferably from the A24 / Cruzeiro virus. The amino acid sequences of the VP 2 to VP 4 proteins may each preferably be the amino acid sequence of the corresponding region included in the amino acid sequence of SEQ ID NO: 3. Also, the amino acid sequence to which the amino acid sequence of the VP 2 to VP 4 protein is linked may preferably be the amino acid sequence of SEQ ID NO: 3. The amino acid sequence of SEQ ID NO: 3 contains the VP2, VP3 and VP4 protein amino acid sequences, and further includes the amino acid sequence of the purine cleavage site between each VP protein.

Another aspect of the present invention provides a polynucleotide encoding the amino acid sequence of SEQ ID NO: 1.

The polynucleotide encoding the amino acid sequence of SEQ ID NO: 1 may preferably consist of the nucleotide sequence of SEQ ID NO: Further, the polynucleotide encoding the amino acid sequence of SEQ ID NO: 1 may more preferably be a sequence having the nucleotide sequence of SEQ ID NO: 4 changed to the codon optimized for the animal in which the immune response is to be generated. The animal may be any animal that can develop foot-and-mouth disease, such as, for example, pigs, cows, sheep, and the like, preferably pigs. The polynucleotide encoding the amino acid sequence of SEQ ID NO: 1 may preferably consist of the nucleotide sequence of SEQ ID NO:

The present invention also provides a polynucleotide encoding an amino acid sequence having 90% homology with the sequence of SEQ ID NO: 1. The amino acid sequence having 90% homology with the sequence of SEQ ID NO: 1 may preferably be the amino acid sequence of SEQ ID NO: The polynucleotide encoding the amino acid sequence having 90% homology with the sequence of SEQ ID NO: 1 may preferably consist of the nucleotide sequence of SEQ ID NO: 8.

Another aspect of the present invention provides a plasmid comprising a polynucleotide encoding the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having 90% homology with the sequence of SEQ ID NO:

The term "plasmid" means a DNA construct containing a DNA sequence operably linked to a suitable regulatory sequence capable of expressing DNA within an appropriate host. The plasmid may be a vector, phage particle, or simply a potential genome insert. Plasmids herein can be used interchangeably with vectors (or viral vectors).

Polynucleotides included in the plasmid of the present invention are the same as those mentioned above unless otherwise stated. The polynucleotide involved may preferably be the nucleotide sequence of SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 8, and more preferably the nucleotide sequence of SEQ ID NO: 4 or SEQ ID NO:

The plasmid may further comprise a nucleotide sequence encoding the VP2-4 protein of foot-and-mouth disease virus. In this case, the nucleotide sequence of SEQ ID NO: 4 or SEQ ID NO: 5 in the plasmid may be linked to a nucleotide sequence encoding VP2-4 protein of foot-and-mouth disease virus, preferably VP2, VP3 and VP 4 < / RTI > protein. For example, the plasmid comprises a nucleotide sequence of the VP 4 region - a nucleotide sequence of the VP 2 region - a nucleotide sequence of the VP 3 region - a sequence of the nucleotide sequence of SEQ ID NO: 2 or 3 (based on 5 '-> 3' Nucleotides < / RTI > Additional nucleotide sequences may be included between the nucleotide sequences in each region. The additional nucleotide sequence may be a nucleotide sequence corresponding to a purine cleavage site region.

The nucleotide sequence coding for the VP 2 to VP 4 protein may be preferably derived from the type A foot-and-mouth disease virus, more preferably from the A24 / Cruzeiro virus. The nucleotide sequence encoding the VP 2 to VP 4 protein may each preferably be the nucleotide sequence of the corresponding region contained in the nucleotide sequence of SEQ ID NO: 6. Also, the polynucleotide sequence to which the nucleotide sequence of the VP 2, VP 3 and VP 4 regions are linked may preferably be the nucleotide sequence of SEQ ID NO: 6.

The plasmid may most preferably comprise the nucleotide sequence of SEQ ID NO: 7. The nucleotide sequence of SEQ ID NO: 7 may comprise the nucleotide sequence of the VP2, VP3 and VP4 regions derived from SEQ ID NO: 4 or 5, and the A24 / Cruzeiro virus. Also, the nucleotide sequence of SEQ ID NO: 7 may comprise a Kozak sequence.

Another aspect of the present invention provides a foot-and-mouth vaccine composition comprising a polypeptide of the present invention, a polynucleotide of the present invention, or a plasmid of the present invention.

In the foot and mouth vaccine composition, the polypeptide, the polynucleotide, and the plasmid are as described above unless otherwise stated.

The term "vaccine" refers to a biological agent containing an antigen that immunizes an individual. The term " vaccine " refers to an immunogenic or antigenic substance that causes immunization to a living body by injection or oral administration to a human or animal to prevent infection.

The vaccine may be a DNA vaccine. The term "DNA vaccine" refers to a vaccine that causes an immune response by artificially replicating some of the genes, such as pathogens or viruses, and administering them. These DNA vaccines have a variety of advantages over existing protein vaccines, i) it is possible to synthesize only genetic information of pure target pathogen antigens, so there is no need to handle dangerous pathogens directly, ii) genes necessary for toxicity induction And iii) it is possible to develop a vaccine by rapidly responding to various infectious diseases that occur suddenly due to the simplicity of plasmid DNA alone, and the like. do.

The foot-and-mouth disease vaccine composition may preferably have defense immunity against the foot-and-mouth disease type A Asian region type, and more preferably belong to the foot-and-mouth disease virus A Asia Sea-97 strain.

The vaccine composition of the present invention may comprise a veterinarily acceptable carrier. The term "veterinarily acceptable carrier" includes any and all solvents, dispersion media, coatings, adjuvants, stabilizers, diluents, preservatives, antibacterial and antifungal agents, isotonic agents, Examples of carriers, excipients and diluents that can be included in the vaccine composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, maltitol, starch, glycerin, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, , Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, the vaccine composition of the present invention can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like in the form of oral preparations and sterilized injection solutions according to a conventional method. In the case of formulation, it may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like which are usually used.

The vaccine composition can be injected into a subject in a variety of forms. "Infusion" may be carried out by any one of the methods selected from the group consisting of subcutaneous injection, intramuscular injection, subcutaneous injection, intraperitoneal injection, nasal administration, oral administration, transdermal administration and oral administration.

The vaccine composition may include one or more adjuvants and the like to improve or enhance the immune response. Suitable adjuvants include compositions consisting of peptides, mineral hydroxides, aluminum phosphates, aluminum oxides and mineral oils or vegetable oils such as Marcol 52, and one or more emulsifiers, or surface active substances such as ricosecetin, polyvalent cations, polyvalent anions, .

Another aspect of the present invention provides a method of producing an immune response to the FMD virus type A in an individual, comprising administering the vaccine composition of the present invention.

In the immunological reaction production method, the vaccine composition is as described above unless otherwise stated.

The term "immune response" refers to the activation of the immune system of a host, e. G., The immune system of a mammal, in response to the introduction of an antigen. The immune response may be in the form of a cellular response or a body fluid response, or both.

In the above immunological reaction production method, the vaccine composition of the present invention may contain an effective amount of an effective ingredient, i.e., an entire recombinant polypeptide, a recombinant polynucleotide or a plasmid containing it, together with a pharmaceutically acceptable carrier and adjuvant. The individual may be an animal, preferably an animal belonging to the milk family, for example pig, cow or sheep. The term "effective amount" means that the component of the vaccine is in an amount sufficient to induce a specific immune response against the FMD type A virus in the vaccinated animal. Effective amounts can be readily determined by those skilled in the art and can be determined, for example, through routine experimentation in animals.

The administration can be by any route suitable for administration of a DNA vaccine, and can be administered, for example, by subcutaneous, intramuscular, intraperitoneal, or intravenous injection.

The foot-and-mouth disease antiviral composition according to the present invention can provide a DNA vaccine composition exhibiting high immunogenicity against foot-and-mouth disease virus belonging to the A type, particularly the A type Asian Sea-97 strain. Accordingly, the vaccine composition of the present invention is very useful for effectively preventing A-type FMD virus infection in Asia, such as East Asia, China or Taiwan, in Korea and surrounding countries.

FIG. 1 shows the results of analysis of VP1 amino acid sequences of four types of FMV (GeneBank Accession Nos. HQ116371, HQ116294, KF450794 and KT223560) in Asia.
FIG. 2 is a graph showing the positions (epitopes) important for the generation of neutralizing antibodies in the mismatched portions of the VP1 amino acid sequences of four types of FMDV (GeneBank Accession Nos. HQ116371, HQ116294, KF450794 and KT223560) and A24 / Cruzeiro virus .
FIG. 3 shows results of aligning the two types of A viruses (MAY / 97 and VN / T11D / 2013) possessed by the quarantine source and the five foot-and-mouth disease viruses together.
Figure 4 shows the amino acid sequence (A-1 type) of a new form of VP1 derived based on the MAY / 97 virus.
Figure 5 shows the amino acid sequence (A-2 type) of a new form of VP1 derived on the basis of the VN / T11D / 2013 virus.
6 is a schematic diagram showing a strategy for replacing the VP1 region of pGX9008 with a novel synthetic gene VP1.
Figure 7 shows the result of aligning the sequence identified by sequencing with the VP1 sequence of the newly synthesized gene.
FIG. 8 shows the results of analyzing the effect of the vaccine administration of the present invention on mouse antibody formation by SPC ELISA.
FIG. 9 shows the result of analyzing the antibody-forming effect of vaccine administration of the present invention in a mini-pig by SPC ELISA.
FIG. 10 shows the effect of forming a neutralizing antibody upon administration of the vaccine of the present invention in a mini-pig.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited by these examples.

Example 1: Analysis of homology between pGX9008 and A-type foot-and-mouth disease virus

In February 2017, the foot-and-mouth disease that occurred in Yeoncheon, Korea was identified as serotype A virus and the gene was identified as A ASIA Sea-97 through genetic analysis. In addition, the results of the analysis of the A type virus and the homology analysis in Asia (633 nucleotide sequence comparisons of VP1 region), 99.8% with Vietnam (cattle and pig) virus (VIT / 14/2016) and 2016 Myanmar (99.7%) and 2013 (99.4%) of the Guangdong province (pig) virus (GDMM / CHA / 2013-S) in the same year.

Sequence analysis of GDMM / CHA / 2013-S (99.5% identical to the A type) available in the GB database (NCBI) was used for homology analysis between the foot-and-mouth disease virus in Yeoncheon and the South American FMD vaccine pGX9008 GDMM / CHA / 2013-S (99.5% identical to the A type) and about three nucleotide sequences. The amino acid sequence of the VP1 site, which is part of the P1 structural protein of GDMM / CHA / 2013-S and pGX9008, was 178/211 (84%) homologous.

Therefore, it is confirmed that the vaccine for domestic or Asian region needs to be developed since the current South American pandemic virus vaccine pGX9008 does not have a vaccine effect against the FMD-A serotype occurring in Southeast Asia including Korea.

Example 2. Investigation of A-type FMDV sequence occurring in Asia for recombination of VP1 region of pGX9008

From the WRLFMD website, recent reports of an ME report on VP1 of type A foot-and mouth disease virus in Asia have been made. The survey covers 17 countries including Vietnam, Taiwan, Korea, Russia, Laos, Myanmar, Kazakhstan, Cambodia, Malaysia, Mongolia, the Philippines, Thailand, Nepal, Japan, Hong Kong, Bhutan and Bangladesh. Of these, the Philippines, Taiwan, Nepal, Japan, Hong Kong, Bhutan and Bangladesh have recently been found to have never developed foot-and-mouth disease virus. From the ME report results, four strains (GeneBank Accession numbered) were selected which can identify the nucleotide sequence information in NCBI GeneBank as a strain for the recombination of VP1 region of pGX9008 (Table 1).

Country Related virus Accession no. Nucleotide% Id Vietnam (VIT) GDMM / CHA / 2013-S KF450794 99.5 to 99.8% South Korea GDMM / CHA / 2013-S KF450794 99.5% Russia GDMM / CHA / 2013-S KF450794 98.11 ~ 99.69% Myanmar GDMM / CHA / 2013-S KF450794 99.84% Laos GDMM / CHA / 2013-S KF450794 99.53% Kazakhstan GDMM / CHA / 2013-S KF450794 99.53% Mongolia GDMM / CHA / 2013-S KF450794 98.4% Thailand GDMM / CHA / 2013-S KF450794 98.58 to 98.74% Cambodia CAM / 2/2008 HQ116294 96.86% Malaysia MAY / 4/2012 KT223560 94.97% Malaysia A / VIT / 3/2008 HQ116371 96.52%

Four strains (GDMM / CHA / 2013-S (KF450794), CAM / 2/2008 (HQ116294), MAY / 4/2012 (KT223560) and A / VIT / 3/2008 (HQ116371) The nucleotide sequence was converted into an amino acid sequence. The amino acid sequences were analyzed by Clustal / muscle and 22 amino acid mismatches were confirmed. Figure 1 shows the data obtained by arranging the alignment results through Jalview.

Example 3. Investigation of an epitope region important for neutralizing antibody production

Of the mismatched sequences identified in Example 2, the site (epitope) important for neutralizing antibody generation is described in Res Vet Sci. 2017 Dec; 115: 374-381, Arch Virol. 2016 Oct; 161 (10): 2705- , Virus Res. 2014 Mar 6; 181: 72-6., And Vet Microbiol. 2011 Apr 21; 149 (1-2): 242-7). Figure 2 shows the alignment results of epitopes associated with neutralizing antibody production.

In addition, two strains of the A type foot-and-mouth disease virus (MAY / 97, VN / T11D / 2013) possessed by the quarantine personnel were aligned and the results are shown in FIG.

Example 4. Synthesis of new type A foot-and-mouth disease virus VP1 region

4.1. New VP1 region sequence determination

The sequence of the epitope region was determined according to MAY / 97 and VN / T11D / 2013 to be inoculated with GDMM / CHA / 2013-S as a comparative virus, and two kinds of VP1 regions The amino acid sequence was selected as a candidate for recombination of VP1 region of pGX9008. The VP1 amino acid sequence based on the MAY / 97 virus was named 'New type A-1' and the VP1 amino acid sequence based on VN / T11D / 2013 was named 'New type A-2'.

As a result of analysis of homology between the sequence of New type A-1 and New type A-2 and the conventional A-type virus and sequence, the following results were obtained.

In case of recombination using New type A-2, homology to MAY / 97 and VN / T11D / 2013 to be inoculated by attack is high, Accordingly, a novel DNA vaccine was generated by recombining the VP1 region of pGX9008 using New type A-2, and its efficacy was confirmed.

4.2. Recombination of new VP1 region

The FMDV VP1 base sequence (SEQ ID NO: 4) to be constructed with the new A serotype was optimized with the pig codon. The optimized gene sequence (SEQ ID NO: 5) with this pig codon was used for VP1 sequence replacement of pGX9008. A strategy for replacing the VP1 site of pGC9008 with a novel porcine codon-optimized synthetic gene A serotype is shown in Fig.

Specifically, inserts (774 bp) obtained by treating with the restriction enzyme BstBI / XhoI in pUC57-FMD new type A-2 plasmid (synthesized by Genscript), in which the synthetic gene VP1 site optimized for porcine codon A was cloned, and pGX9008 The vector backbone obtained by treating BstBI / XhoI with the plasmid was ligated to obtain multiple clones.

The sequenced clones finally confirmed the clone containing the intended pGX-FMD new type A-2 sequence. Sequence-confirmed sequences and FMD new type A-2 porcine codon optimization sequences were aligned and confirmed to be 100% identical (FIG. 7). The FMD New type A-2 porcine codon optimization sequence was used for the preparation of the FMD DNA plasmid vaccine in the following.

Example 5. Confirmation of Antibody Formation Effect of New DNA Vaccine

5.1. Production of FMD DNA plasmid vaccine

E. coli harboring the FMD DNA plasmid (containing the FMD new type A-2 porcine codon optimization sequence) was inoculated into 2.5 L of LB medium and incubated overnight at 37 ° C. Plasmids were then extracted from the cultured E. coli using EndoFree Plasmid Giga kit (QIAGEN, Cat # 12391). The extracted plasmid was dissolved in endotoxin-free water and stored at -70 ° C. The stored plasmid was then used in an experiment to determine the effect of antibody formation in an animal (confirmation of vaccination effect). Hereinafter, the novel DNA vaccine thus prepared is referred to as 'PLS-A'.

5.2. Identification of Antibody Formation Effect of Vaccine in Mouse

5.2.1. DNA vaccination

Mouse C57BL / 6 (6-week-old, female) were divided into three groups of 5 mice per group, and the vaccine, the existing vaccine and the mock plasmid of the present invention were inoculated three times at intervals of two weeks as follows Respectively. The cells were inoculated using an electroporation apparatus (cellectra 2000), and the DNA vaccine was administered at a concentration of 30 μg / ml.

Group 1 (control) - Mock plasmid DNA

· Group 2 (comparison group) - FMD DNA type A (South American-A24 cruzeiro, conventional vaccine)

Group 3 (experimental group) - FMD DNA type A (PLS-A, Asian type, vaccine of the present invention)

Blood sampling was performed using the orbital blood collection method before (0), after the second (3) and after the third (5) weeks. Serum was separated from the collected blood and stored at -80 ° C. The stored serum was dissolved and SPC (solid phase competition) ELISA analysis was performed.

5.2.2. Antibody formation assay - SPC ELISA assay

The SPC ELISA was performed using PrioCHECK FMDV Type A ELISA KIT (Prionics lelystad B.V / Holland) and the experimental protocol was performed according to the experimental protocol provided by the kit manufacturer.

The results are shown in Fig. As can be seen from FIG. 8, the vaccine PLS-A of the present invention was significantly less time-consuming to form antibody than the conventional vaccine (South American-A24), and had a higher reaction inhibition (PI value) Respectively. Here, the higher the degree of reaction inhibition (PI value), the more specific antibodies to foot-and mouth serotype A were produced.

As described above, foot-and-mouth disease is very contagious and has a characteristic that not only the animal-to-animal propagation is easily caused by excretion of foot-and-mouth disease virus before the clinical symptoms, but also the latency period is very short. Therefore, in order to exhibit an effective function as a vaccine against foot-and-mouth disease, not only should the effect of suppressing foot-and-mouth disease virus be excellent, but also the rate of antibody production against foot-and-mouth disease virus should be fast. The vaccine according to the present invention is remarkably superior in inhibiting the foot-and-mouth disease virus, and since the antibody production rate is significantly faster than that of the conventional vaccine, the foot-and-mouth disease can be suppressed early, It is very effective.

5.3. Identification of Antibody Formation Effect of Vaccine in Mini-pig

5.3.1. DNA vaccination

9 SPF-Göttingen mini-pigs (4 months, male) were divided into 3 groups of 3 mice per group, and the mice were vaccinated 3 times at 3-week intervals as follows. The vaccine was administered to the hindlimb muscles at a concentration of 0.5 mg / mouse using an electroporation device (cellectra 2000), and the inactivated vaccine was administered to the muscle behind 2 ml of the neck. At week 9, each vaccine was administered at a concentration of 3 mg / mouse.

Group 1 (negative control) - Mock plasmid DNA

· Group 2 (positive control) - inactivated vaccine (comipam, serotype O / A 2 vaccine)

Group 3 (experimental group) - FMD DNA type A (PLS-A, type A, vaccine of the present invention)

Blood samples were collected before (0), after (3), after (6) and after (4). Plasma was separated from the collected blood, and the separated plasma was stored at -80 ° C. The stored plasma was dissolved and SPC (solid phase competition) ELISA analysis was performed.

5.3.2. Antibody formation assay - SPC ELISA assay

The SPC ELISA was performed using PrioCHECK FMDV Type A ELISA KIT (Prionics lelystad B.V / Holland) and the experimental protocol was performed according to the experimental protocol provided by the kit manufacturer.

The results are shown in Fig. As can be seen from FIG. 9, the vaccine PLS-A of the present invention significantly increased the response inhibition (PI value) at the 12th week after the fourth administration, and especially higher than that of the inactivated vaccine strain of positive control .

5.4. Identification of neutralizing antibody formation by vaccination in mini-pig

5.4.1. DNA vaccination

9 SPF-Göttingen mini-pigs (4 months, female) were divided into 3 groups of 3 mice per group, and were vaccinated 3 times at 3-week intervals. The concentration of DNA vaccine administered was 6 mg / mouse, and the vaccine was administered to the muscles inside the hindlimb using an electric perforator (cellectra 2000). The inactivated vaccine was administered to the muscle 2 ml after the neck.

Group 1 (negative control) - Mock plasmid DNA

· Group 2 (positive control) - inactivated vaccine (comipam, serotype O / A 2 vaccine)

· Group 3 (experimental group) - FMD DNA type A (PLS-A, type A) + O type + Asia type 1

Groups 3 and 3 purchased vaccines of type O and Asia1, which were used to make vaccines. Plasma was separated from the collected blood after the administration (week 9), and the separated plasma was stored at -80 ° C. The SPN (solid phase competition) ELISA assay and the neutralization test were carried out to dissolve the separated plasma and to test the neutralization ability of VNT.

5.4.2. Virus neutralization test (VNT-neutralization test)

The neutralization test was carried out at the Vaccine Research Center for Foot and Mouth Disease of the Ministry of Agriculture, Forestry and Livestock.

The VNT for identification of neutral region was A type MAY / 97 foot-and-mouth disease virus. A / MAY / 97 100 TCID50, serum, positive serum and negative serum were diluted at an appropriate dilution ratio and reacted for 1 hour. LFBK (fetal porcine kidney cell line) cell line was divided into two groups and reacted for 2 ~ 3 days, and cytopathic effect was observed. The neutralization antibody was found to be positive 16 times (log value 1.20) and the domestic standard was 32 times (log value 1.51).

The results are shown in Fig. As can be seen from FIG. 10, the trivalent DNA vaccine containing PLS-A at 9 weeks after administration showed more than 200-fold neutralizing antibody titers in all individuals and was found to be positive.

FIG. 9 compares the inhibition (PI) values of the vaccine according to the present invention and the divalent vaccine positive control, while FIG. 10 shows the results of the comparison between the divalent vaccine positive control and the vaccine according to the present invention, (PI value) of the vaccine. It can be seen that the vaccine according to the present invention not only exerts excellent effects on A-type FMDV even when acting alone, but also exerts excellent effects on A-type FMDV even when it acts in the form of a trivalent vaccine together with other types. This means that the vaccine according to the present invention is both highly useful as a unit price vaccine and a multivalent vaccine.

<110> Plumbline Life Sciences, Inc. <120> VACCINE COMPOSITION FOR FOOT AND MOUTH DISEASE VIRUS A SEROTYPE <130> DP17052KR <160> 8 <170> KoPatentin 3.0 <210> 1 <211> 212 <212> PRT <213> Artificial Sequence <220> <223> New type A-2 VP1 <400> 1 Thr Ala Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu   1 5 10 15 Asn Tyr Gly Gly Glu Thr Gln Ala Gln Arg Arg His His Thr Asp Val              20 25 30 Gly Phe Leu Met Asp Arg Phe Val Gln Ile Lys Pro Val Ser Pro Thr          35 40 45 His Val Ile Asp Leu Met Gln Thr His Gln His Gly Leu Val Gly Ala      50 55 60 Met Leu Arg Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile Val Val  65 70 75 80 Asn His Thr Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ala                  85 90 95 Ala Leu Asn Asn Thr Ser Asn Pro Thr Ala Tyr His Lys Ala Pro Phe             100 105 110 Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala Thr         115 120 125 Val Tyr Ser Gly Thr Ser Lys Tyr Ser Thr Pro Gln Thr Arg Arg Gly     130 135 140 Asp Leu Gly Pro Leu Ala Ala Arg Leu Ala Ala Gln Leu Pro Ala Ser 145 150 155 160 Phe Asn Phe Gly Ala Ile Arg Ala Thr Glu Ile Gln Glu Leu Leu Val                 165 170 175 Arg Met Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Val             180 185 190 Glu Val Ser Ser Gln Asp Arg His Lys Gln Lys Ile Ile Ala Pro Ala         195 200 205 Lys Gln Leu Leu     210 <210> 2 <211> 212 <212> PRT <213> Artificial Sequence <220> <223> New type A-1 VP1 <400> 2 Thr Ala Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val Glu   1 5 10 15 Asn Tyr Gly Gly Glu Thr Gln Ala Gln Arg Arg His His Thr Asp Val              20 25 30 Ser Phe Ile Met Asp Arg Phe Val Arg Ile Lys Pro Val Ser Pro Thr          35 40 45 His Val Ile Asp Leu Met Gln Thr His Gln His Gly Leu Val Gly Ala      50 55 60 Met Leu Arg Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile Val Val  65 70 75 80 Asn His Thr Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu Ala                  85 90 95 Ala Leu Asp Asn Thr Ser Asn Pro Thr Ala Tyr His Lys Ala Pro Phe             100 105 110 Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala Thr         115 120 125 Val Tyr Asn Gly Thr Ser Lys Tyr Ser Thr Pro Gly Ala Arg Arg Gly     130 135 140 Asp Leu Gly Ser Leu Ala Ala Arg Leu Ala Ala Gln Leu Pro Ala Ser 145 150 155 160 Phe Asn Phe Gly Ala Ile Arg Ala Thr Glu Ile Gln Glu Leu Leu Val                 165 170 175 Arg Met Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala Val             180 185 190 Glu Val Leu Ser Gln Asp Arg His Lys Gln Lys Ile Ile Ala Pro Ala         195 200 205 Lys Gln Leu Leu     210 <210> 3 <211> 446 <212> PRT <213> Artificial Sequence <220> <223> PGX9008 VP4-VP2-VP3 <400> 3 Asp Lys Lys Thr Glu Glu Thr Thr Leu Leu Glu Asp Arg Ile Leu Thr   1 5 10 15 Thr Arg Asn Gly His Thr Thr Ser Thr Thr Gln Ser Ser Val Gly Val              20 25 30 Thr His Gly Tyr Ser Thr Glu Glu Asp His Val Ala Gly Pro Asn Thr          35 40 45 Ser Gly Leu Glu Thr Arg Val Val Gln Ala Glu Arg Phe Tyr Lys Lys      50 55 60 Tyr Leu Phe Asp Trp Thr Thr Asp Lys Ala Phe Gly His Leu Glu Lys  65 70 75 80 Leu Glu Leu Pro Ser Asp His His Gly Val Phe Gly His Leu Val Asp                  85 90 95 Ser Tyr Ala Tyr Met Arg Asn Gly Trp Asp Val Glu Val Ser Ala Val             100 105 110 Gly Asn Gln Phe Asn Gly Gly Cys Leu Leu Val Ala Met Val Pro Glu         115 120 125 Trp Lys Glu Phe Asp Thr Arg Glu Lys Tyr Gln Leu Thr Leu Phe Pro     130 135 140 His Gln Phe Ile Ser Pro Arg Thr Asn Met Thr Ala His Ile Thr Val 145 150 155 160 Pro Tyr Leu Gly Val Asn Arg Tyr Asp Gln Tyr Lys Lys His Lys Pro                 165 170 175 Trp Thr Leu Val Val Met Val Val Ser Pro Leu Thr Val Asn Asn Thr             180 185 190 Ser Ala Ala Gln Ile Lys Val Tyr Ala Asn Ile Ala Pro Thr Tyr Val         195 200 205 His Val Ala Gly Glu Leu Pro Ser Lys Glu Arg Gly Arg Lys Arg Arg     210 215 220 Ser Gly Ile Phe Pro Val Ala Cys Ala Asp Gly Tyr Gly Gly Leu Val 225 230 235 240 Thr Thr Asp Pro Lys Thr Ala Asp Pro Ala Tyr Gly Lys Val Tyr Asn                 245 250 255 Pro Pro Arg Thr Asn Tyr Pro Gly Arg Phe Thr Asn Leu Leu Asp Val             260 265 270 Ala Glu Ala Cys Pro Thr Phe Leu Cys Phe Asp Asp Gly Lys Pro Tyr         275 280 285 Val Thr Thr Arg Thr Asp Asp Thr Arg Leu Leu Ala Lys Phe Asp Leu     290 295 300 Ser Leu Ala Ala Lys His Met Ser Asn Thr Tyr Leu Ser Gly Ile Ala 305 310 315 320 Gln Tyr Tyr Thr Gln Tyr Ser Gly Thr Ile Asn Leu His Phe Met Phe                 325 330 335 Thr Gly Ser Thr Asp Ser Lys Ala Arg Tyr Met Val Ala Tyr Ile Pro             340 345 350 Pro Gly Val Glu Thr Pro Pro Asp Thr Pro Glu Arg Ala Ala His Cys         355 360 365 Ile His Ala Glu Trp Asp Thr Gly Leu Asn Ser Lys Phe Thr Phe Ser     370 375 380 Ile Pro Tyr Val Ser Ala Asp Tyr Ala Tyr Thr Ala Ser Asp Thr 385 390 395 400 Ala Glu Thr Ile Asn Val Gln Gly Trp Val Cys Ile Tyr Gln Ile Thr                 405 410 415 His Gly Lys Ala Glu Asn Asp Thr Leu Val Val Ser Val Ser Ala Gly             420 425 430 Lys Asp Phe Glu Leu Arg Leu Pro Ile Asp Pro Arg Gln Gln         435 440 445 <210> 4 <211> 636 <212> DNA <213> Artificial Sequence <220> <223> Nex type A-2 VP1 DNA <400> 4 accaccgcca ccggggaatc agcagaccct gtcacaacca ccgttgagaa ctacggtggc 60 gagacacaag cacagcggcg tcaccacacc gacgtcggct tcttaatgga caggttcgtg 120 cagatcaagc ctgtgagccc cacacatgtc attgacctca tgcagacaca ccaacacggg 180 ctggtgggcg ccatgttgcg cgcggccacc tactactttt ctgatcttga gattgtggtg 240 aaccacacgg gtaacctaac gtgggtaccc aatggagcac ccgaggcagc actgaacaac 300 acgagcaacc ccactgctta ccacaaagcg ccgttcacga ggcttgcgct cccctacacc 360 gcgccacacc gcgtgctggc aactgtgtac agcgggacga gcaagtactc cacacctcaa 420 acacggcgag gtgacctggg tcctctcgcg gcgaggctcg ctgcacagct ccctgcctcc 480 ttcaacttcg gtgcaattcg ggccacggag atccaagaac tccttgtgcg catgaagcgc 540 gccgagctct actgccccag gccactgttg gcggtggagg tgtcgtcgca agacagacac 600 aagcagaaaa tcattgcccc tgcaaaacaa ctcctg 636 <210> 5 <211> 636 <212> DNA <213> Artificial Sequence <220> <223> Nex type A-2 VP1 DNA porcine codon optimization <400> 5 acaacagcca caggagaatc cgccgaccca gtgacaacca ccgtggagaa ctacggagga 60 gagacacagg ctcagcgccg gcaccacacc gacgtgggct tcctgatgga caggttcgtg 120 cagatcaagc ccgtgagccc aacccacgtg atcgacctga tgcagaccca ccagcacggc 180 ctggtgggag ctatgctgag ggctgccacc tactacttct ccgacctgga gatcgtggtg 240 aaccacaccg gaaacctgac ctgggtgcca aacggagctc cagaggccgc cctgaacaac 300 accagcaacc ccaccgccta ccacaaggct ccattcacca ggctggccct gccatacacc 360 gctccacacc gggtgctggc taccgtgtac tccggcacca gcaagtactc caccccacag 420 accaggagag gcgacctggg accactggct gctaggctgg ctgctcagct gccagcctcc 480 ttcaacttcg gagctatcag ggctaccgag atccaggagc tgctggtgag gatgaagaga 540 gccgagctgt actgccccag gccactgctg gctgtggagg tgtccagcca ggacagacac 600 aagcagaaga tcatcgcccc agccaagcag ctgctg 636 <210> 6 <211> 1338 <212> DNA <213> Artificial Sequence <220> <223> pGX9008 VP4-VP2-VP3 DNA <400> 6 gataagaaaa cagaggaaac caccctgctg gaggacagaa tcctgaccac aagaaacggg 60 cacactacca gcacaactca gtcttcagtg ggcgtcacac acggatactc aactgaggaa 120 gaccatgtgg ccgggccaaa taccagtggc ctggagacac gagtggtcca ggctgaaagg 180 ttctacaaga aatatctgtt tgactggacc acagataagg ccttcggcca cctggagaaa 240 ctggaactcc cctcagacca ccacggcgtg ttcggccatc tggtcgatag ctacgcctat 300 atgagaaacg gatgggacgt ggaggtctcc gctgtgggca accagttcaa tggcggatgc 360 ctgctcgtgg ctatggtgcc cgagtggaag gaatttgata ccagggaaaa ataccagctg 420 acactcttcc cacaccagtt tatctctcct agaactaaca tgaccgccca tattaccgtg 480 ccttatctgg gcgtcaatcg gtacgaccag tataagaaac acaaaccttg gaccctggtg 540 gtcatggtgg tcagtcccct cacagtgaac aatactagcg ccgctcagat caaggtctac 600 gccaacattg ctccaaccta tgtgcacgtc gcaggagagc tgccttccaa ggaacgggga 660 cgcaaacggc gctctgggat cttcccagtg gcatgtgctg acggatacgg agggctggtc 720 actaccgacc ctaagaccgc agatcccgcc tacggaaaag tgtataaccc acccaggact 780 aattacccag ggcggttcac caacctgctc gatgtggcag aggcctgccc caccttcctg 840 tgctttgacg atggcaagcc atacgtgaca actcggacag acgatactcg cctgctcgcc 900 aagtttgacc tgagcctcgc agccaaacac atgtcaaaca cctacctgag tggaatcgcc 960 cagtactata ctcagtattc cgggaccatt aatctgcatt tcatgtttac cggctctaca 1020 gactcaaagg ctcgctacat ggtggcatat atccctcccg gcgtcgagac cccacctgat 1080 acacctgaaa gggctgcaca ctgcatccat gccgagtggg acacaggact gaacagcaag 1140 ttcacttttt ccattcccta cgtgtctgcc gctgactacg cttataccgc atccgatact 1200 gccgaaacca ttaacgtgca gggatgggtc tgtatctacc agattactca cgggaaagcc 1260 gagaatgaca ccctggtggt ctccgtgtct gctggcaagg acttcgaact gcgcctccct 1320 atcgatcccc gacagcag 1338 <210> 7 <211> 2400 <212> DNA <213> Artificial Sequence <220> <223> new type VP for FMDV 2 serotype vaccine <400> 7 gccaccatgg attggacatg gattctgttc ctggtggctg ctgctactag agtgcattca 60 ggggccggac agtcttcacc cgcaaccgga tcacagaacc agagtggaaa taccgggagc 120 atcattaaca attactatat gcagcagtac cagaacagca tggacacaca gctgggggat 180 aacgccatca gcggcggcag caatgagggc tccacagata ccacatctac tcacactacc 240 aatacccaga acaatgactg gttctctaaa ctggcaagct ccgccttcac cggcctcttt 300 ggagctctgc tcgcaagggg aagaaagagg agaagcgata agaaaacaga ggaaaccacc 360 ctgctggagg acagaatcct gaccacaaga aacgggcaca ctaccagcac aactcagtct 420 tcagtgggcg tcacacacgg atactcaact gaggaagacc atgtggccgg gccaaatacc 480 agtggcctgg agacacgagt ggtccaggct gaaaggttct acaagaaata tctgtttgac 540 tggaccacag ataaggcctt cggccacctg gagaaactgg aactcccctc agaccaccac 600 ggcgtgttcg gccatctggt cgatagctac gcctatatga gaaacggatg ggacgtggag 660 gtctccgctg tgggcaacca gttcaatggc ggatgcctgc tcgtggctat ggtgcccgag 720 tggaaggaat ttgataccag ggaaaaatac cagctgacac tcttcccaca ccagtttatc 780 tctcctagaa ctaacatgac cgcccatatt accgtgcctt atctgggcgt caatcggtac 840 gaccagtata agaaacacaa accttggacc ctggtggtca tggtggtcag tcccctcaca 900 gtgaacaata ctagcgccgc tcagatcaag gtctacgcca acattgctcc aacctatgtg 960 cacgtcgcag gagagctgcc ttccaaggaa cggggacgca aacggcgctc tgggatcttc 1020 ccagtggcat gtgctgacgg atacggaggg ctggtcacta ccgaccctaa gaccgcagat 1080 cccgcctacg gaaaagtgta taacccaccc aggactaatt acccagggcg gttcaccaac 1140 ctgctcgatg tggcagaggc ctgccccacc ttcctgtgct ttgacgatgg caagccatac 1200 gtgacaactc ggacagacga tactcgcctg ctcgccaagt ttgacctgag cctcgcagcc 1260 aaacacatgt caaacaccta cctgagtgga atcgcccagt actatactca gtattccggg 1320 accattaatc tgcatttcat gtttaccggc tctacagact caaaggctcg ctacatggtg 1380 gcatatatcc ctcccggcgt cgagacccca cctgatacac ctgaaagggc tgcacactgc 1440 atccatgccg agtgggacac aggactgaac agcaagttca ctttttccat tccctacgtg 1500 tctgccgctg actacgctta taccgcatcc gatactgccg aaaccattaa cgtgcaggga 1560 tgggtctgta tctaccagat tactcacggg aaagccgaga atgacaccct ggtggtctcc 1620 gtgtctgctg gcaaggactt cgaactcaga ctccccattg acccaagaca gcagagaggc 1680 agaaaaagac gcagcacaac agccacagga gaatccgccg acccagtgac aaccaccgtg 1740 gagaactacg gaggagagac acaggctcag cgccggcacc acaccgacgt gggcttcctg 1800 atggacaggt tcgtgcagat caagcccgtg agcccaaccc acgtgatcga cctgatgcag 1860 acccaccagc acggcctggt gggagctatg ctgagggctg ccacctacta cttctccgac 1920 ctggagatcg tggtgaacca caccggaaac ctgacctggg tgccaaacgg agctccagag 1980 gccgccctga acaacaccag caaccccacc gcctaccaca aggctccatt caccaggctg 2040 gccctgccat acaccgctcc acaccgggtg ctggctaccg tgtactccgg caccagcaag 2100 tactccaccc cacagaccag gagaggcgac ctgggaccac tggctgctag gctggctgct 2160 cagctgccag cctccttcaa cttcggagct atcagggcta ccgagatcca ggagctgctg 2220 gtgaggatga agagagccga gctgtactgc cccaggccac tgctggctgt ggaggtgtcc 2280 agccaggaca gacacaagca gaagatcatc gccccagcca agcagctgct gcgcggccgg 2340 aaacgacgct ctaattttga cctcctgaaa ctggctggcg atgtggaatc taaccctggc 2400                                                                         2400 <210> 8 <211> 636 <212> DNA <213> Artificial Sequence <220> <223> new type A-1 DNA <400> 8 accaccgcca ccggggaatc agcagaccct gtcacaacca ccgttgagaa ctacggtggc 60 gagacacaag cacagcggcg tcaccacacc gacgtcagct tcataatgga caggttcgtg 120 cggatcaagc ctgtgagccc cacacatgtc attgacctca tgcagacaca ccaacacggg 180 ctggtgggcg ccatgttgcg cgcggccacc tactactttt ctgatcttga gattgtggtg 240 aaccacacgg gtaacctaac gtgggtaccc aatggagcac ccgaggcagc actggataac 300 acgagcaacc ccactgctta ccacaaagcg ccgttcacga ggcttgcgct cccctacacc 360 gcgccacacc gcgtgctggc aactgtgtac aatgggacga gcaagtactc cacacctgga 420 gcccggcgag gtgacctggg ttctctcgcg gcgagactcg ctgcacagct ccctgcctcc 480 ttcaacttcg gtgcaattcg ggccacggag atccaggaac tccttgtgcg catgaagcgc 540 gccgagctct actgccccag gccactgttg gcggtggagg tgctgtcgca agacagacac 600 aagcagaaaa tcattgcccc tgcaaaacaa ctcctg 636

Claims (13)

A polypeptide comprising an amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having 90% or more homology thereto. A polypeptide comprising an amino acid sequence having the amino acid sequence of SEQ ID NO: 1 or 90% or more homology thereto and an amino acid sequence of VP2 to VP4 protein of foot-and-mouth disease virus. 3. The method of claim 2,
Wherein the VP 2 to VP 4 proteins of foot-and-mouth disease virus are derived from type A foot-and-mouth disease virus. 1. A polynucleotide which encodes an amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having 90% or more homology thereto. A plasmid comprising the polynucleotide of claim 4. 6. The method of claim 5,
Wherein the plasmid further comprises a nucleotide sequence encoding the VP 2 to VP 4 protein of foot-and-mouth disease virus. The method according to claim 6,
Wherein the VP 2 to VP 4 proteins of foot-and-mouth disease virus are derived from type A foot-and-mouth disease virus. 9. The method of claim 8,
Wherein the plasmid comprises the nucleotide sequence of SEQ ID &lt; RTI ID = 0.0 &gt; 7. &Lt; / RTI &gt; A foot-and-mouth vaccine composition comprising the polypeptide of any one of claims 1 to 3, the polynucleotide of claim 4, or the plasmid of any one of claims 5 to 8. 10. The method of claim 9,
Wherein the foot-and-mouth disease vaccine composition has protective immunity against Type-I foot-and-mouth disease virus. 11. The method of claim 10,
Wherein the type A foot-and-mouth disease virus is an Asian-type foot-and-mouth disease. 10. The method of claim 9,
Wherein the vaccine is a unit price or a polyvalent vaccine. 11. A method of producing an immune response against an A type FMD virus in an individual, comprising administering the vaccine composition of claim 9 to the individual.

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