KR20200053825A - Recombinant adenovirus expressing three kinds of antigen for anti-foot-and-mouth disease virus vaccination - Google Patents

Abstract

The present invention relates to adenovirus for simultaneously expressing three serotypes of antigen for anti-foot-and-mouth disease virus vaccination, and more particularly, to a recombinant vector wherein VP1 gene of food-and-mouth disease virus type A, VP1 gene of food-and-mouth disease virus type Asia1 and VP1 gene of food-and-mouth disease virus type O are inserted into adenovirus vector, as well as recombinant adenovirus which simultaneously expresses each VP1 protein of three serotypes as an antigen for foot-and-mouth disease virus, including type A, type Asia1 and type O prepared by the recombinant vector. The recombinant adenovirus of the present invention provides each VP1 protein of three serotypes for foot-and-mouth virus including type A, type Asia1 and type O as an antigen, thereby showing a vaccination effect on the three serotypes, which mainly occur in South Korea, with just one administration. Also, the recombinant adenovirus of the present invention may also continuously provide an antigen to an animal dosed therewith, thereby showing a continuous vaccination effect.

Description

Recombinant adenovirus expressing three kinds of antigen for anti-foot-and-mouth disease virus vaccination}

The present invention relates to an adenovirus for a foot and mouth vaccine that simultaneously expresses three serotype foot and mouth virus antigens, specifically, the VP1 gene of the foot and mouth A virus, the VP1 gene of the foot and mouth disease Asia1 virus and the VP1 gene of the foot and mouth O virus A recombinant vector characterized by being inserted into an adenoviral vector and a recombinant adenovirus expressed by the VP1 protein of each of the three serotype foot and mouth disease viruses of type A, Asia1 type, and type O by the recombinant vector at the same time as an antigen It is about.

Foot-and-mouth disease (FMD) is a viral vesicular disease that infects hoofed animals such as pigs, cows, sheep, goats, deer, and wild ruminants. It is classified as a very important animal disease because it has a high mortality rate and there is no special treatment, so foot and mouth disease can cause a huge obstacle to the livestock industry.

The pathogen causing this foot and mouth disease is foot-and-mouth disease virus (FMDV), which belongs to the genus Picornaviridae and Aphthovirus and is a single-strand bipolar RNA virus. Currently, it is classified into 7 different serotypes (A, O, C, Asia1, SAT1, SAT2, and SAT3), but it is difficult to cope with infection due to high variability within these serotypes.

In Korea, since foot-and-mouth disease occurred in 2010, O-type, A-type, and Asia-type vaccines have been tried to prevent them, but these vaccines have low antigenic association with domestically occurring viruses, and the development of foot-and-mouth disease vaccine suitable for Korea is urgently required. Is becoming.

In addition, since most of the existing vaccines are vaccines for one serotype, there is a hassle of applying the vaccine for each serotype separately, and it is applied in the form of administering an inactivated vaccine or an antigen to expect a continuous vaccine effect. It is a difficult situation.

In order to solve the above problems, the present inventors tried to develop a method suitable for foot-and-mouth disease occurring in Korea and preparing for major foot-and-mouth disease serotypes in Korea with one administration, and at the same time, exhibiting a continuous vaccine effect.

Republic of Korea Registered Patent No. 10-1578445

Therefore, the main object of the present invention is to provide a new vaccine that is suitable for foot and mouth disease occurring in Korea and can prepare for major foot and mouth disease serotypes in Korea with one administration, and at the same time, exhibits a continuous vaccine effect.

Another object of the present invention is to provide an effective method for preventing foot and mouth disease in Korea, particularly when using the vaccine.

According to one aspect of the present invention, the present invention provides a recombinant vector characterized in that the VP1 gene of foot and mouth disease A virus, the VP1 gene of foot and mouth disease Asia1 virus and the VP1 gene of foot and mouth disease type O virus are inserted into an adenovirus vector. do.

In the recombinant vector of the present invention, the VP1 gene of the foot-and-mouth disease type A virus encodes the amino acid sequence of SEQ ID NO: 1, and the VP1 gene of the foot-and-mouth disease Asia1 type virus encodes the amino acid sequence of SEQ ID NO: 2 and the foot-and-mouth disease type O It is preferable that the VP1 gene of the virus encodes the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4.

In the recombinant vector of the present invention, the VP1 gene of the foot-and-mouth disease type A virus consists of the nucleotide sequence of SEQ ID NO: 5, and the VP1 gene of the foot-and-mouth disease Asia1 type virus consists of the base sequence of SEQ ID NO: 6, the foot and mouth disease O The VP1 gene of the type virus is preferably composed of the nucleotide sequence of SEQ ID NO: 7 or SEQ ID NO: 8.

According to another aspect of the present invention, the present invention provides a recombinant adenovirus produced by the recombinant vector.

According to another aspect of the present invention, the present invention provides a composition for a foot and mouth vaccine containing the recombinant adenovirus as an active ingredient.

According to another aspect of the present invention, the present invention provides a method for preventing foot and mouth disease characterized in that the recombinant adenovirus is administered to animals other than humans.

Recombinant adenovirus of the present invention shows vaccine effects against major three serotypes occurring in Korea with one administration by providing VP1 protein of each of three serotype foot and mouth disease virus types A, Asia1 and O In addition, it is possible to continuously provide an antigen to the administered animal, thereby exhibiting a continuous vaccine effect.

1 is a diagram illustrating the process of introducing the VP1 gene of foot and mouth disease virus into the shuttle vector during the process of manufacturing a recombinant vector according to an embodiment of the present invention.
FIG. 2 is a schematic diagram showing the process of introducing a VP1 gene expression construct into an adenovirus vector using a shuttle vector into which a VP1 gene has been introduced during the manufacturing process of a recombinant vector according to an embodiment of the present invention.
Figure 3 shows the titer measurement results of the recombinant adenovirus according to an embodiment of the present invention.

The recombinant vector of the present invention is a vector capable of generating the recombinant adenovirus of the present invention in animal cells, the VP1 gene of foot and mouth disease type A virus, the VP1 gene of foot and mouth disease Asia1 type virus and the VP1 gene of foot and mouth type O virus adenovirus It is characterized by being inserted into a vector.

In the present invention, the adenovirus vector system is used as a platform, and this adenovirus vector is one of virus-derived vectors and is used in various fields. Up to 10kb of gene can be inserted, and excluding elements necessary for expression of the inserted gene, it is possible to deliver an external gene of about 2 to 3kb.

In the present invention, a vector of an adenovirus type 5 (Ad5) family can be used as an adenovirus vector, for example, a pAdEasy-1 vector, which is a vector of the Ad5 family, can be used.

In the present invention, a base sequence encoding the amino acid sequence of SEQ ID NO: 1 may be used as the VP1 gene of the foot-and-mouth disease virus. The amino acid sequence of SEQ ID NO: 1 is an amino acid sequence derived from the VP1 protein of A / Pocheon / KOR / 2010, a foot-and-mouth disease virus. As the base sequence encoding the amino acid sequence of SEQ ID NO: 1, base sequences such as SEQ ID NO: 9, SEQ ID NO: 5 are possible, and in the present invention, it is preferable to use the base sequence of SEQ ID NO: 5.

In the present invention, as the VP1 gene of the foot-and-mouth disease Asia1 virus, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 2 may be used. The amino acid sequence of SEQ ID NO: 2 is the amino acid sequence derived from the VP1 protein of foot-and-mouth disease Asia1 type virus NKR / 2/2007. As the base sequence encoding the amino acid sequence of SEQ ID NO: 2, base sequences such as SEQ ID NO: 10, SEQ ID NO: 6 are possible, and in the present invention, it is preferable to use the base sequence of SEQ ID NO: 6.

In the present invention, as the VP1 gene of the foot-and-mouth disease O virus, a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4 may be used. SEQ ID NO: 3 is an amino acid sequence derived from the VP1 protein of foot-and-mouth disease O virus, O / SKR / 2002, and SEQ ID NO: 4 is an amino acid sequence derived from the VP1 protein of foot-and-mouth disease O virus SKR / 5/2010. As the base sequence encoding the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4, base sequences such as SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 7, SEQ ID NO: 8 are possible, and in the present invention, SEQ ID NO: 7 or SEQ ID NO: 8 It is preferable to use the base sequence of.

The recombinant vector of the present invention is preferably made such that the transcription of the VP1 gene is regulated by a T7 promoter and a T7 terminator. The T7 promoter and terminator are one of various types of promoters and terminators used in the field of gene expression, and various vector systems using them are known. Therefore, by cloning the VP1 gene in these vectors, construct a construct in which the T7 promoter is located 5 'in the front of the VP1 gene and the T7 terminator is located in the 3' rear, and this is used using a recombinant method or homologous recombination using restriction enzymes. Recombinant methods can be used to prepare such recombinant vectors.

The recombinant adenovirus of the present invention can be produced by a method of transfecting the recombinant vector of the present invention into animal cells to express the gene present in the recombinant vector. In this case, 293A cells may be used as animal cells.

Recombinant adenovirus of the present invention can express three VP1 proteins of foot and mouth disease type A, Asia1 type, and O type due to the composition of the recombinant vector of the present invention, thereby providing an antigen for the production of immune antibodies when administered to animals. It will continue to provide. Accordingly, the composition for the foot-and-mouth disease vaccine of the present invention containing the recombinant adenovirus of the present invention as an active ingredient and the foot-and-mouth disease prevention method of the present invention for administering the recombinant adenovirus of the present invention are more efficient for the animal to control the foot-and-mouth disease of the three serotypes. Can have immunity.

The foot-and-mouth disease vaccine composition of the present invention may further include additives such as excipients commonly used in viral vaccines in addition to the recombinant adenovirus of the present invention.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Recombinant vector preparation

VP1 gene of foot-and-mouth disease A virus (SEQ ID NO: 5), foot-and-mouth disease Asia1 type VP1 gene (SEQ ID NO: 6), foot-and-mouth disease type O (type O1 or O2) virus VP1 gene using MCS site in pShuttle-CMV vector (SEQ ID NO: 7 or SEQ ID NO: 8) or A, Asia1 type and O type (optionally selected from O1 type and O2 type) VP1 gene is inserted into the form of the linked form, the 5 'direction of the VP1 gene of foot and mouth disease virus A recombinant shuttle vector in the form of a T7 promoter in the front and a T7 terminator in the 3 'rear is prepared (see FIG. 1).

The recombinant shuttle vector is introduced into BJ5183 cells together with the pAdEasy-1 vector to prepare a recombinant adenovirus vector in the form of inserting the VP1 gene through homologous recombination (see FIG. 2).

Recombinant adenovirus production

The recombinant adenovirus vector is transfected into 293A cells to produce a recombinant adenovirus (see FIG. 2).

Concentration and purification of recombinant adenovirus

After inoculating the 293A cells with the recombinant adenovirus, after 3 days, when the cytopathic effect (CPE) appears, the cells are collected (used while storing at -80 ° C), dissolved at 37 ° C, and then dissolved in a sonicator. The cells are crushed and centrifuged at 12K rpm for 10 minutes to recover the supernatant.

Virus is recovered by ultracentrifuge for 2 hours at 25K rpm with a gradient of CsCl concentration of 4M to 2.2M, and the virus is purified by dialysis overnight in phosphate buffered saline (PBS) containing 10% glycerol. .

Measure the titer of recombinant adenovirus

Since the genome of the recombinant adenovirus contains CMV-derived DNA, the titer of the recombinant adenovirus is measured by real-time PCR using a CMV-specific primer.

After converting the structural DNA of the control adenovirus (adenovirus derived from the non-recombined pAdEasy-1 vector) into a copy number, a standard curve (p = 0.9991) was prepared through CT values from 10 ⁹ to 10 ⁵ , and then from the recombinant adenovirus. The titer is measured against the standard curve based on the concentration of the extracted gDNA.

The adenovirus derived from the recombinant vector into which the VP1 gene (SEQ ID NO: 5) of the foot-and-mouth disease A virus was introduced showed a titer of about 1.24x10 ¹³ per ml (see FIG. 3).

<110> PharmaenTech, Inc. <120> Recombinant adenovirus expressing three kinds of antigen for          anti-foot-and-mouth disease virus vaccination <130> PA-D18119 <160> 12 <170> KoPatentIn 3.0 <210> 1 <211> 213 <212> PRT <213> Foot-and-mouth disease virus <400> 1 Met Thr Thr Ala Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val   1 5 10 15 Glu Asn Tyr Gly Gly Glu Thr Gln Val Gln Arg Arg His His Thr Asp              20 25 30 Val Ser Phe Ile Met Asp Arg Phe Val Gln Ile Lys Pro Val Ser Pro          35 40 45 Thr His Val Ile Asp Leu Met Gln Thr His Gln His Gly Leu Val Gly      50 55 60 Ala Met Leu Arg Ala Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Ile Val  65 70 75 80 Val Asn His Thr Gly Arg Leu Thr Trp Val Pro Asn Gly Ala Pro Glu                  85 90 95 Ala Ala Leu Asp Asn Thr Ser Asn Pro Thr Ala Tyr His Lys Ala Pro             100 105 110 Phe Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala         115 120 125 Thr Val Tyr Asn Gly Thr Ser Arg Tyr Ser Ala Pro Ala Thr Arg Arg     130 135 140 Gly Asp Leu Gly Ser Leu Ala Ala Arg Leu Ala Ala Gln Leu Pro Ala 145 150 155 160 Ser Phe Asn Tyr Gly Ala Val Arg Ala Thr Glu Ile Gln Glu Leu Leu                 165 170 175 Val Arg Met Lys Arg Ala Glu Leu Tyr Cys Pro Arg Pro Leu Leu Ala             180 185 190 Val Glu Val Thr Ser Gln Asp Arg His Lys Gln Lys Ile Ile Ala Pro         195 200 205 Ala Lys Gln Leu Leu     210 <210> 2 <211> 212 <212> PRT <213> Foot-and-mouth disease virus <400> 2 Met Thr Thr Thr Thr Gly Glu Ser Ala Asp Pro Val Thr Thr Thr Val   1 5 10 15 Glu Asn Tyr Gly Gly Glu Thr Gln Thr Ala Arg Arg Leu His Thr Asp              20 25 30 Val Ala Phe Val Leu Asp Arg Phe Val Lys Leu Thr Gln Pro Lys Ser          35 40 45 Thr Gln Thr Leu Asp Leu Met Gln Ile Pro Ser His Thr Leu Val Gly      50 55 60 Ala Leu Leu Arg Ser Ala Thr Tyr Tyr Phe Ser Asp Leu Glu Val Ala  65 70 75 80 Leu Val His Thr Gly Pro Val Thr Trp Val Pro Asn Gly Ala Pro Lys                  85 90 95 Thr Ala Leu Asp Asn His Thr Asn Pro Thr Ala Tyr Gln Lys Gln Pro             100 105 110 Ile Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ser         115 120 125 Thr Val Tyr Asn Gly Lys Thr Thr Tyr Gly Gly Glu Pro Pro Arg Arg     130 135 140 Gly Asp Leu Ala Ala Leu Ala Arg Arg Val Ser Asn Arg Leu Pro Thr 145 150 155 160 Ser Phe Asn Tyr Gly Ala Val Lys Ala Asp Thr Ile Thr Glu Leu Leu                 165 170 175 Ile Arg Met Lys Arg Ala Glu Thr Tyr Cys Pro Arg Pro Leu Leu Ala             180 185 190 Leu Asp Thr Thr Gln Asp Arg Arg Lys Gln Glu Ile Ile Ala Pro Glu         195 200 205 Lys Gln Thr Leu     210 <210> 3 <211> 214 <212> PRT <213> Foot-and-mouth disease virus <400> 3 Met Thr Thr Ser Thr Gly Glu Ser Ala Asp Pro Val Thr Ala Thr Val   1 5 10 15 Glu Asn Tyr Gly Gly Glu Thr Gln Val Gln Arg Arg Gln His Thr Asp              20 25 30 Val Ser Phe Ile Leu Asp Arg Phe Val Lys Val Thr Pro Lys Asp Gln          35 40 45 Ile Asn Val Leu Asp Leu Met Gln Ile Pro Ala His Thr Leu Val Gly      50 55 60 Ala Leu Leu Arg Thr Ala Thr Tyr Tyr Phe Ala Asp Leu Glu Val Ala  65 70 75 80 Val Lys His Glu Gly Asn Leu Thr Trp Val Pro Asn Gly Ala Pro Glu                  85 90 95 Ala Ala Leu Asp Asn Thr Thr Asn Pro Thr Ala Tyr His Lys Ala Pro             100 105 110 Leu Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala         115 120 125 Thr Val Tyr Asn Gly Asn Cys Lys Tyr Gly Glu Ser Pro Val Thr Asn     130 135 140 Leu Arg Gly Asp Leu Gln Val Leu Thr Gln Lys Ala Ala Arg Thr Leu 145 150 155 160 Pro Thr Ser Phe Asn Tyr Gly Ala Ile Lys Ala Thr Arg Val Thr Glu                 165 170 175 Leu Leu Tyr Arg Met Lys Arg Ala Glu Thr Tyr Cys Pro Arg Pro Leu             180 185 190 Leu Ala Ile His Pro Ser Glu Ala Arg His Lys Gln Lys Ile Val Ala         195 200 205 Pro Val Lys Gln Leu Leu     210 <210> 4 <211> 214 <212> PRT <213> Foot-and-mouth disease virus <400> 4 Met Thr Thr Ser Thr Gly Glu Ser Ala Asp Pro Val Thr Ala Thr Val   1 5 10 15 Glu Asn Tyr Gly Gly Glu Thr Gln Val Gln Arg Arg His His Thr Asp              20 25 30 Val Ser Phe Ile Leu Asp Arg Phe Val Lys Val Thr Pro Lys Asp Ser          35 40 45 Ile Asn Val Leu Asp Leu Met Gln Thr Pro Ser His Thr Leu Val Gly      50 55 60 Ala Leu Leu Arg Thr Ala Thr Tyr Tyr Phe Ala Asp Leu Glu Val Ala  65 70 75 80 Val Lys His Glu Gly Asp Leu Thr Trp Val Pro Asn Gly Ala Pro Glu                  85 90 95 Ala Ala Leu Asp Asn Thr Thr Asn Pro Thr Ala Tyr His Lys Ala Pro             100 105 110 Leu Thr Arg Leu Ala Leu Pro Tyr Thr Ala Pro His Arg Val Leu Ala         115 120 125 Thr Val Tyr Asn Gly Asn Cys Lys Tyr Ala Gly Gly Ser Leu Pro Asn     130 135 140 Val Arg Gly Asp Leu Gln Val Leu Ala Gln Lys Ala Ala Arg Pro Leu 145 150 155 160 Pro Thr Ser Phe Asn Tyr Gly Ala Ile Lys Ala Thr Arg Val Thr Glu                 165 170 175 Leu Leu Tyr Arg Met Lys Arg Ala Glu Thr Tyr Cys Pro Arg Pro Leu             180 185 190 Leu Ala Val His Pro Ser Ala Ala Arg His Lys Gln Lys Ile Val Ala         195 200 205 Pro Val Lys Gln Ser Leu     210 <210> 5 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> Optimized VP1 sequence of foot-and-mouth disease virus serotype A <400> 5 atgaccaccg cgaccggtga atctgctgat ccggttacta ccactgttga aaactatggt 60 ggtgaaaccc aggttcagcg tcgtcatcac actgatgttt ctttcatcat ggatcgtttc 120 gttcagatta aaccagtttc accaactcac gtaattgatc tgatgcagac tcatcagcat 180 ggtttagttg gcgcgatgtt acgtgctgct acttattatt tctctgattt agaaattgtt 240 gttaatcata ccggtcgttt aacctgggtt ccgaacggcg caccggaagc ggcgctggat 300 aacacttcta acccgaccgc gtaccacaaa gctccgttta ctcgtttagc tctgccatat 360 actgcgccgc accgtgttct ggcaaccgtg tataacggta cctctcgtta ttctgcgccg 420 gctactcgcc gcggcgatct gggtagcctg gcagcacgtc tggctgctca gctgccagct 480 tcttttaact acggcgctgt tcgtgcaacc gaaatccagg aattattagt gcgtatgaaa 540 cgtgcagaac tgtattgccc gcgtccgctg ctggctgttg aagtgacctc tcaggatcgt 600 cacaaacaaa aaattatcgc accggctaaa cagctgctgt aa 642 <210> 6 <211> 639 <212> DNA <213> Artificial Sequence <220> <223> Optimized VP1 sequence of foot-and-mouth disease virus serotype          Asia1 <400> 6 atgaccacca ccaccggtga gtccgcggac ccggtcacca ccaccgttga aaactacggc 60 ggtgaaaccc agaccgcccg tcgtctgcac accgacgttg cattcgttct ggaccgcttc 120 gttaaactga cccagccaaa aagcacccag accctggacc tgatgcagat cccgagccac 180 accctggttg gtgctctgct gcgttctgca acttattact tttctgattt agaagttgcg 240 ctggttcaca ctggtccggt tacttgggtt cctaatggtg ctccgaaaac tgcactggat 300 aatcacacca acccgaccgc ttaccaaaaa cagccgatta cccgtctggc actgccgtat 360 actgcaccgc atcgtgttct gagcactgtt tataacggta aaaccactta tggtggtgaa 420 ccgccgcgtc gtggtgatct ggcagcgtta gcacgtcgtg ttagcaaccg tctgccgacc 480 tctttcaact atggtgcggt taaagctgat actattaccg aattactgat tcgtatgaaa 540 cgtgctgaaa cctattgtcc gcgtccgctg ctggcactgg ataccaccca ggatcgtcgt 600 aaacaggaaa tcatcgcgcc ggaaaaacag accctgtaa 639 <210> 7 <211> 645 <212> DNA <213> Artificial Sequence <220> <223> Optimized VP1 sequence of foot-and-mouth disease virus serotype O <400> 7 atgaccacct ctaccggtga aagcgctgat ccggttaccg ctaccgttga aaactatggt 60 ggtgaaaccc aggttcagcg tcgtcagcac accgatgttt ctttcattct ggatcgtttt 120 gttaaagtta ccccgaaaga tcagattaac gttctggatt taatgcagat cccggctcat 180 accctggttg gtgctctgct gcgtaccgcg acctactact tcgctgatct ggaagttgct 240 gttaaacacg aaggtaactt aacctgggtt ccgaacggtg caccggaagc agcactggat 300 aacaccacta acccgaccgc ttatcataaa gcgccgctga ctcgtctggc tctgccgtat 360 accgcgccgc atcgtgtttt agctactgtt tataacggta attgtaaata cggtgaatct 420 ccggttacca acctgcgtgg tgatctgcag gttctgactc agaaagcggc tcgtaccctg 480 ccaacctctt ttaattatgg cgcgattaaa gcaacccgtg ttactgaact gctgtatcgt 540 atgaaacgtg ctgaaaccta ttgcccgcgt ccgctgctgg ctatccaccc gtctgaagcg 600 cgtcataaac agaaaatcgt tgctccggtt aaacagctgc tgtaa 645 <210> 8 <211> 645 <212> DNA <213> Artificial Sequence <220> <223> Optimized VP1 sequence of foot-and-mouth disease virus serotype O <400> 8 atgaccacca gcaccggtga atctgcggat ccggttaccg cgaccgttga aaactacggt 60 ggtgaaaccc aggttcagcg tcgtcaccat accgatgtta gctttatcct ggatcgtttc 120 gttaaagtta ctccaaaaga ttctattaac gttctggatc tgatgcagac tccgtctcat 180 accttagttg gtgctctgct gcgtaccgca acttactatt tcgctgattt agaagttgca 240 gttaaacatg aaggtgatct gacctgggtt ccgaacggtg ctccggaagc agctttagat 300 aacaccacta acccgaccgc atatcacaaa gcaccgctga cccgtttagc tctgccgtat 360 accgctccgc atcgtgttct ggctaccgtt tataacggca actgtaaata cgcaggtggt 420 agcctgccga acgttcgtgg tgatctgcag gtgctggctc agaaagctgc tcgtccgctg 480 ccgacctctt ttaactatgg cgctattaaa gctacccgtg ttaccgaact gctgtatcgt 540 atgaaacgtg ctgaaaccta ttgcccgcgt ccgctgttag ctgttcatcc gtctgctgca 600 cgtcataaac agaaaatcgt tgcgccggtt aaacagtctc tgtaa 645 <210> 9 <211> 642 <212> DNA <213> Foot-and-mouth disease virus <400> 9 atgaccaccg ccaccgggga atcagcagac cccgtcacaa ccaccgtcga gaactacggt 60 ggtgagacac aagtgcagcg acgccaccac accgacgtca gctttataat ggacaggttt 120 gtgcagatca agcctgtgag ccccacacat gtcattgacc tcatgcaaac acaccaacac 180 gggctggtag gcgctatgtt gcgcgcggcc acctactact tttctgatct tgagattgtg 240 gtgaaccaca caggtcgcct aacgtgggta cccaatggag caccagaggc agcactggat 300 aacacgagca accccactgc ttaccacaaa gcaccgttca caaggcttgc actcccttac 360 accgcgccac accgcgtgtt ggcaaccgtg tacaacggga ctagcaggta ctctgcgcct 420 gcaacacggc gaggtgactt ggggtctctc gcggcgaggc tcgccgcaca gcttcctgcc 480 tccttcaact acggcgcggt tcgagccacg gagatccaag aactcctcgt gcgcatgaag 540 cgcgccgagc tctactgccc caggccactg ctggcggtgg aggtgacgtc acaagacaga 600 cacaagcaga aaatcattgc ccctgcaaag caactcctgt ga 642 <210> 10 <211> 639 <212> DNA <213> Foot-and-mouth disease virus <400> 10 atgactacca ccactggcga gtccgcggac ccagtcacca ccacggttga gaactacgga 60 ggagagaccc agacggcccg acggcttcac actgatgtcg ctttcgttct cgacaggttc 120 gtgaaactca cccagcccaa gagcacccag acccttgatc tcatgcagat cccctcacac 180 acactggtcg gggcgcttct ccggtctgcg acgtactact tttcagacct ggaggttgcg 240 ctcgtccaca caggaccggt cacgtgggtg cccaacggtg cgcctaagac cgccttggac 300 aaccacacca acccgactgc ctaccagaag caacctatca cccgcttggc actcccctac 360 accgctcccc accgtgtgct gtcaacagtg tacaacggga agacaacgta cggaggagaa 420 cccccgcggc gcggtgatct tgccgccctc gcacgcagag tgagcaaccg gctgcccact 480 tccttcaact acggcgctgt gaaggccgac accatcacgg agctgttgat ccgcatgaag 540 cgtgcggaaa catactgccc caggcccttg ctggctcttg acaccacaca agaccgccgt 600 aaacaggaga tcattgcacc tgagaaacag actttgtga 639 <210> 11 <211> 645 <212> DNA <213> Foot-and-mouth disease virus <400> 11 atgaccacct ccacaggtga gtcggctgac cccgtgactg ccaccgttga gaactacggt 60 ggtgagacac aggtccagag acgccaacac acggatgtct cgttcatact agacagattt 120 gtgaaagtaa caccaaaaga ccaaattaat gtgttggacc tgatgcaaat ccctgcacac 180 actttggtag gcgcgctcct ccgtactgcc acctactact tcgcagatct ggaagtggca 240 gtgaaacacg aggggaacct cacctgggtc ccgaacgggg cgcccgaggc agcgttggac 300 aacaccacca atccaacggc ctatcacaag gcgccgctca cccggcttgc actgccttac 360 acggcaccac accgtgtctt ggctactgtt tacaacggga actgcaagta tggcgagagc 420 cccgtgacca atctgagagg tgacctgcaa gtgttgaccc agaaggcggc aagaacgctg 480 cctacctcct tcaattacgg tgccatcaaa gccactcggg tgactgaact gctttaccgc 540 atgaagaggg ccgaaacata ctgcccccgg cctcttttgg ctattcaccc gagcgaagct 600 agacacaaac aaaagattgt ggcgcctgtg aaacagctgt tgtga 645 <210> 12 <211> 645 <212> DNA <213> Foot-and-mouth disease virus <400> 12 atgaccactt cgacaggcga gtcggctgac cccgtgactg ccaccgttga gaattacggc 60 ggcgagacac aggtccagag gcgccaccac acagacgtct cattcatatt ggacagattt 120 gtgaaagtca caccaaaaga ctcaataaat gtattggacc tgatgcagac cccctcccac 180 accctagtag gggcgctcct ccgcactgct acttactatt tcgctgattt agaggtggca 240 gtgaaacacg agggggacct tacctgggtg ccaaatggag cacctgaagc agccttggac 300 aacaccacca acccaacggc gtaccataag gcgccgctta cccggctcgc attgccctac 360 acggcaccac accgtgtttt ggccaccgtt tacaacggga actgcaaata cgccgggggc 420 tcactgccca acgtgagagg cgatctccaa gtgctggctc agaaggcagc gaggccgctg 480 cctacttctt tcaactacgg tgccatcaaa gccactcggg tgacagaact gctgtaccgc 540 atgaagaggg ccgagacgta ctgtcctcgg cccctcttgg ctgttcaccc gagtgcggcc 600 agacacaaac agaaaatagt ggcacctgta aagcagtcct tatga 645

Claims (6)

A recombinant vector comprising the VP1 gene of the foot and mouth A virus, the VP1 gene of the foot and mouth Asia1 virus, and the VP1 gene of the foot and mouth O virus, inserted into the adenovirus vector. According to claim 1,
The VP1 gene of the foot and mouth disease A virus encodes the amino acid sequence of SEQ ID NO: 1,
The VP1 gene of the foot-and-mouth disease Asia1 virus encodes the amino acid sequence of SEQ ID NO: 2,
Recombinant vector, characterized in that the VP1 gene of the foot-and-mouth disease virus encodes the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4. According to claim 2,
The foot-and-mouth disease type A virus VP1 gene consists of the nucleotide sequence of SEQ ID NO: 5,
The foot-and-mouth disease Asia1 virus VP1 gene consists of the nucleotide sequence of SEQ ID NO: 6,
Recombinant vector, characterized in that the foot-and-mouth disease virus VP1 gene consists of the nucleotide sequence of SEQ ID NO: 7 or SEQ ID NO: 8. Recombinant adenovirus produced by the recombinant vector of any one of claims 1 to 3. A composition for a foot and mouth vaccine containing the recombinant adenovirus of claim 4 as an active ingredient. Method for preventing foot and mouth disease, characterized in that the recombinant adenovirus of claim 4 is administered to animals other than humans.

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