KR101578444B1 - Recombinant foot-and-mouth disease virus using Korean isolated strain of FMDV A serotype and the manufacturing method - Google Patents

Abstract

The present invention relates to a recombinant foot-and-mouth disease virus having protective immunity against a foot-and-mouth disease type A Asian region and a method for producing the same, including a plasmid in which a whole gene of foot-and-mouth disease type A isolate Korean isolate virus (A / KOR / Pocheon / 2010) will be.

Description

[0001] The present invention relates to recombinant foot-and-mouth disease virus (hereinafter referred to as " foot-and-mouth disease virus "

The present invention relates to a recombinant foot-and-mouth disease virus using a foot-and-mouth disease type A isolate in Korea, and more particularly, to a method for producing a foot-and-mouth disease virus including a plasmid in which a whole gene of foot-and-mouth disease type A isolate virus (A / KOR / Pocheon / 2010) , A foot-and-mouth disease type A Asian region, and a method for producing the recombinant foot-and-mouth disease virus.

Foot-and-mouth disease (FMD) is a virus-borne disease that infects animals with split hoofs and is characterized by rapid replication and rapid spreading. Due to its economic importance, this disease is classified as a very important animal disease that can be transmitted by the World Animal Health Organization (OIE). Due to these characteristics, the occurrence of livestock products in the international trade is the most important check factor .

The foot-and-mouth disease pathogen is a single-stranded bipolar RNA virus, which belongs to the genus Piconaviridae and Aphthovirus and has seven different serotypes (A, O, C, Asia, SAT 1, SAT 2 and SAT 3) Being classified.

Of the A-type foot-and-mouth disease viruses, there are three regional types: ME-SA (Middle East-South Asia), AFRICA (Africa), and ASIA (Asia). This type of vaccine is required to develop various types of vaccine against A virus because it is not protected against virus attack because the rate of change is high.

A type foot-and-mouth disease occurred in Korea for the first time in 2010. At that time, there was no vaccination against foot-and-mouth disease, but a type-A foot-and-mouth disease was occurring in Korea around 2009. However, the stockpile vaccine strain A22 / Iraq / 24/64 is widely used in the world, but it is presumed that the vaccine immunity will be relatively low even if the vaccine is inoculated because it has low antigenicity with viruses generated in Korea.

Since the outbreak of massive foot and mouth disease in November 2010, only the O-type vaccine has been inoculated. In the second half of 2011, vaccines for O, A and Asia type 1 trivalent vaccines have been started. Among them, A vaccine containing the A Malaysia 97 virus strain, which has a moderate correlation with sex, has been in use since the second half of 2011.

The virus isolated from Korea is an Asian type virus and belongs to the Sea-97 lineage genetically, and the virus is a virus that occurs in Asia and is still present in Asia, The development of a vaccine against such diseases is urgently required.

In order to adequately defend against foot-and-mouth disease type A Asian viruses, the inventors of the present invention have developed a vaccine based on a virus vector obtained by cloning a whole genome of a foot-and-mouth disease virus A (A / KOR / Pocheon / 2010) The inventors have found that a new foot-and-mouth disease recombinant virus having the same anti-circular form as the foot-and-mouth disease type A isolate of Korea can be provided.

Accordingly, it is an object of the present invention to provide a recombinant foot-and-mouth disease virus which has defense immunity against foot-and-mouth disease type A Asian region type and can be usefully used as a vaccine seedling.

Another object of the present invention is to provide a method for producing recombinant foot-and-mouth disease virus, which has defense immunity against foot-and-mouth disease type A type Asian region and can be usefully used as a vaccine seedling.

In order to achieve the above object, the present invention provides a plasmid in which a whole gene of a foot-and-mouth disease type A Korean isolate virus (A / KOR / Pocheon / 2010) having a nucleotide sequence of at least a gene bank number KC588943 is inserted, The present invention provides a recombinant foot-and-mouth disease virus having defense immunity against foot-and-mouth disease type A Asian type.

The present invention also relates to a method for producing a recombinant virus comprising the steps of: (1) inserting a whole gene of foot-and-mouth disease type A isolate virus having the nucleotide sequence of KC588943 in a plasmid; and (2) introducing the plasmid obtained in the above step And then proliferating the recombinant foot-and-mouth disease virus.

The recombinant foot-and-mouth disease virus according to the present invention can provide a foot-and-mouth disease vaccine capable of effectively defending the foot-and-mouth disease type A virus type Asian region occurring in Korea, such as East Asia, China or Taiwan.

In addition, the recombinant foot-and-mouth disease virus according to the present invention can effectively prevent the type of foot-and-mouth disease virus type Asian region occurring in the vicinity of Korea such as East Asia and China while the virus titer does not greatly differ from the original virus have.

FIG. 1 is a schematic diagram of a genome of a recombinant virus using a strain of foot-and-mouth disease type A Korean isolate.
Fig. 2 is a schematic diagram of a plasmid (pA-Kor-Poc) into which the whole gene of the foot-and-mouth disease type A isolate of Korea is inserted.
FIG. 3 shows that the protein expressed in the recombinant FMDV according to the present invention shows a FMD-positive reaction using a liver antigen kit.
4 is a photograph showing virus-infected cells (detection of fluorescent antibody method) and cytopathic effect in foot-and-mouth disease virus infected cells.
FIG. 5 is a graph showing that recombinant foot-and-mouth disease virus according to the present invention grows in a fetal tongue cell line (ZZ-R), a porcine kidney cell line (IBRS-2) and a hamster cell line (BHK-21) to form a virus titer.

The present invention includes a plasmid into which a whole gene of a foot-and-mouth disease type A isolate Korean isolate virus (A / KOR / Pocheon / 2010) having a nucleotide sequence of at least a genbank-buster session number KC588943 is inserted, The present invention relates to a recombinant foot-and-mouth disease virus having an immunological ability.

In the recombinant FMD virus of the present invention, the plasmid into which the recombinant gene is inserted is characterized by having the nucleotide sequence of SEQ ID NO: 1.

In the recombinant foot-and-mouth disease virus of the present invention, the foot-and-mouth disease type A Asian type is a Sea-97 genotype.

In the recombinant foot-and-mouth virus of the present invention, the recombinant virus is characterized by being a foot-and-mouth disease-backing virus.

The present invention also relates to a method for producing a recombinant virus comprising the steps of: (1) inserting a whole gene of foot-and-mouth disease type A isolate virus having the nucleotide sequence of KC588943 in a plasmid; and (2) introducing the plasmid obtained in the above step And a method for producing recombinant foot-and mouth disease virus.

The step of inserting the whole gene of foot-and-mouth disease type A isolate virus into the plasmid may be performed by inserting a gene into a plasmid, so that detailed description thereof will be omitted.

In the method for producing recombinant foot-and-mouth disease virus according to the present invention, the cells in the step (2) for injecting the recombinant plasmid constructed in the above step (1) into the cells are selected from the group consisting of fetal tongue cells ZZ- -2) and a hamster cell line (BHK-21) can be used.

In the method for producing recombinant foot-and-mouth virus according to the present invention, the recombinant virus is characterized by having defense immunity against the foot-and-mouth disease type A Asian region type.

The present invention also relates to a vaccine for the prevention of foot-and-mouth disease including the recombinant foot-and-mouth disease virus.

The vaccine of the present invention may be a vaccine live vaccine or an inactivated vaccine, and the vaccine may be manufactured using conventional vaccine production methods, so that detailed description thereof will be omitted.

Hereinafter, the content of the present invention will be described in detail through experimental examples. However, the following experimental examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited thereto.

Example 1 Preparation of Recombinant Plasmid pA-Kor-Poc

The gene of A / KOR / Pocheon / 2010 in the foot-and-mouth disease virus (A / KOR / Pocheon / 2010) was amplified by PCR and the gene of A / KOR / Pocheon / 2010 in the plasmid (pBluescript SK II) To construct a recombinant plasmid pA-Kor-Poc.

In the above, PCR amplification was carried out in the following manner.

Generally, a random primer used in cDNA synthesis is used to prepare cDNA for the Korean isolate virus, and then 5 '(5'-5' ') nucleotides based on specific primers (GenBank Accession No. KC588943) And 10 μl of 10 mM dNTPs (1 μl), Phusion enzyme (2 μl / μl, 0.5 μl each) was used for the PCR. ) And sterilized distilled water (35.5 占 퐇) at 98 占 폚 for 30 seconds, followed by 25 cycles at 98 占 폚 for 10 seconds, 65 占 폚 for 30 seconds, 72 占 폚 for 2 minutes and 30 seconds, and final 72 占 폚 for 10 minutes.

Fig. 1 is a schematic diagram of a plasmid (pA-Kor-Poc) into which the whole gene of foot-and-mouth disease type A isolate virus has been inserted. The recombinant plasmid pA-Kor-Poc has the nucleotide sequence of SEQ ID NO: 1.

≪ Example 2 > Recovery of recombinant foot-and-mouth disease virus

After recovering the recombinant foot-and-mouth disease virus, the recombinant plasmid (pA-Kor-Poc) was reacted with a restriction enzyme, and the virus was obtained from BHKT7-9 cells (cell line expressing T7 RNA polymerase) (IBRS-2) and BHK-21 (hamster cell line), and the cytopathic effect was confirmed in BHK-21 cells.

Fig. 2 is a schematic diagram of a plasmid (pA-Kor-Poc) into which the whole gene of the foot-and-mouth disease type A isolate of Korea is inserted.

FIG. 3 shows that the protein expressed in the recombinant FMDV according to the present invention shows a FMD-positive reaction using a liver antigen kit. 25 μl of the recovered viral culture supernatant can be confirmed with a positive antigen kit (PBM Co. Ltd.) for a positive response to the structural protein (SP) against foot-and-mouth disease.

4 is a photograph showing virus-infected cells (detection of fluorescent antibody method) and cytopathic effect in foot-and-mouth disease virus infected cells. Recovered viruses in the laboratory were infected with the cells, so that they could detect virus infection by fluorescence antibody method and showed the effect of denaturing the cells.

FIG. 5 is a graph showing that recombinant foot-and-mouth disease virus according to the present invention grows in a fetal tongue cell line (ZZ-R), a porcine kidney cell line (IBRS-2) and a hamster cell line (BHK-21) to form a virus titer. Referring to FIG. 5, the recombinant foot-and-mouth disease virus of the present invention shows a virus titer similar to that of an outdoor virus.

Although the present invention has been described and illustrated in detail, it should be understood by those skilled in the art that various changes and modifications may be made without departing from the scope of the present invention as defined by the appended claims. It will be understood that various modifications and changes may be made in the present invention.

Foot-and-mouth disease A Type Foot-and-mouth disease vaccine using the recombinant foot-and-mouth disease virus according to the present invention having defense immunity against Asian type can contribute to treatment and / or prevention of foot-and-mouth disease.

<110> Animal, Plant and Fisheries Quarantine and Inspection Agency <120> Recombinant foot-and-mouth disease virus using Korean isolated          strain of FMDV serotype and the manufacturing method <160> 1 <170> Kopatentin 1.71 <210> 1 <211> 11040 <212> DNA <213> Artificial Sequence <220> <223> Recombinant plasmid pA-Kor-Poc <400> 1 tatgtaatac gactcactat agggttgaaa gggggcgtta gggtcccacc cctagcatgc 60 caacgactgc tcctgcgttg cactccacac ttacgttcgt gtacgcggga accgatagac 120 taccgttcac ccacctgcag ctggactcat gacaccgcgt ggtcatttag ctggattgtg 180 cggacgaaca ccgcttgcgc acctcgcgtg accgattagt actctcacca ccttccgcct 240 acttggtcgt cagcgctgtc ctgggcaccc ctgttggggg ccgctcgatg ctctacggtt 300 tcccccgcag cgggctacgg tgatggggcc gcctcgtgcg ggttgatcgt ttggtgtgcc 360 tcggttgtga tccgaagctc acctttcatc cccccccccc cccctaagca ctaccgtcgt 420 tcccgacgtt aaagggatga aactacaagc ttagcgcctt cgtgtccgaa gtcaaaggac 480 tacaaccgca agtttgagcc gccttttccg gtgttaatgg aatgtaacca caagatggac 540 cttcttccgg aagtaaaacg gtaatctcac acagttttgc ccgttttcat gagaaatggg 600 acgtctgcgc acgaaacgcg ccgtcgcttg aggaggactt gtacaaacac gatctacgca 660 ggtctccaca actgacacag aacgtgcaac ttggaacccc gcctggtctt tccaggtcta 720 gaggggagac actttgtact gtgcctgact ccacgctcgg tccactggcg agtgttagta 780 gcggcactgt tgcttcgtag cggagcatga cggccgcggg aatccctcct tggtgacaag 840 gacccgcggg gccgaaagcc acgtcccacg gacccgtcat gtgtgcaacc ccagcacggc 900 aactttgcct cgaaacccac ttaaaggtga cactgatact ggtactcagt cactggtgac 960 aggctaagga tgcccttcag gtaccccgag gtaacaagcg acactcggga tctgagaagg 1020 ggactggggc ttctataaaa gcgcccagtt taaaaagctt ctatgcctgg ataggcgatc 1080 ggaggccggc gcctttcctt aattatcact acctttatga acacgactga ctgctttacc 1140 gctctgttgt acgccatcag agagattaag gcacgattct tttccaggac acaagagaaa 1200 atggaattca ctctttacaa cggtgagaag aaaacttttt actctaggcc caacaaccac 1260 gacactgtt ggttgaacac catcctccaa ctgtttaggt acgtcgacga accattcttc 1320 gactgggtct atgaatcacc tgagaacctc acttttgagg cgattaaaca attggaacag 1380 gtcactggtc ttgagctgcg cgagggtggc ccacccgccc tcgtaatttg gaacattaag 1440 cacttgctcc acaccggaat cggtaccgcc tcgcgaccta gtgaggtgtg tatggtagac 1500 gggacggaca tgtgtctggc tgatttccat gctggcattt tcctgaaagg gcaggaacac 1560 gcagtgtttg cctgtgttac ctccaacggg tggtacgcaa ttgacgacga ggacttttac 1620 ccctggacac cagacccgtc tgatgtcctg gtgtttgtcc cgtatgatca agaaccactc 1680 aacggagaat ggaaaacaaa ggttcagagg cgacttaaag gggcagggca atctagccct 1740 gccaccgggt cacagaacca gtcaggcaat accggcagca ttattaacaa ctactacatg 1800 cagcagtacc agaactccat ggacacacaa cttggtgaca acgccatcag cggaggatcc 1860 aacgaggggt ccacggacac aacctctacc cacacaacca acacccaaaa caatgactgg 1920 ttctcaaaac tggcaagttc cgcgttcacc ggtcttttcg gcgcactgct cgccgacaag 1980 aagaccgaag agacaactct tctggaggac cgtatcctca ccactcgtaa tggacacacc 2040 acttctacaa ctcagtcgag tgtgggggtc acctacgggt attcaactgg tgaggaccac 2100 gtttctggac ctaacacatc aggtttggag acgcgggtgg tacaagctga aaggttcttc 2160 aagaagcact tgtttgattg gacgacggac aaaccctttg gtcacgttga aaagctggaa 2220 cttcccactg atcacaaagg tgtctacgga cagctggtgg actcctttgc atacatgaga 2280 aatggctggg acgtggaggt gtctgctgtt ggcaaccagt tcaacggcgg gtgccttctc 2340 gtggccatgg tacctgagtt taaggagttc accacacgtg aaaagtacca gctcaccctg 2400 ttcccccacc agttcattag ccccagaacc aacatgaccg cgcacatcac ggtcccgtac 2460 cttggtgtga acaggtatga ccagtacaac aaacacaaac cctggacgtt ggtggtgatg 2520 gtggtttcac cacttaccac tagctccatt ggtgcatcac agattaaggt ctacaccaat 2580 atcgccccta cccacgttca cgtggctggc gagctcccgt cgaaagaggg gatcgtgccg 2640 gtcgcctgct cggacgggta cggtggcctg gtgacaacag accctaaaac agctgaccct 2700 gcttacggta tggtgtacaa cccacctagg accaactacc ccgggcggtt tacaaacttg 2760 ttggacgtgg cagaggcgtg tcccaccttc ctctgtttcg acgacgggaa accgtatgtt 2820 gtgacaagag cggatgagca gcgcctcttg gccaagtttg acctttccct tgctgcaaag 2880 cacatgtcaa acacctacct ttcagggata gcacagtact acgcacagta ctctggcacc 2940 atcaatttgc acttcatgtt tactggttcc actgactcaa aggcccgtta catggtggct 3000 tacgtcccgc ctggcgtgac aacgccaccg gacacgcctg agagagctgc gcactgcatc 3060 cacgcagaat gggacacggg cctaaactcc aaattcactt tttcaatccc gtacgtatct 3120 gctgcagatt acgcgtacac agcgtccgat gtggcagaca caacaaacgt acagggatgg 3180 gtttgcatct accaaatcac ccacgggaag gccgaacaag acactttggt cgtgtcggtc 3240 agcgccggca aagactttga gctgcgcctc cccattgacc cccgtgcgca aaccaccgcc 3300 accggggaat cagcagaccc cgtcacaacc accgtcgaga actacggtgg tgagacacaa 3360 gtgcagcgac gccaccacac cgacgtcagc tttataatgg acaggtttgt gcagatcaag 3420 cctgtgagcc ccacacatgt cattgacctc atgcaaacac accaacacgg gctggtaggc 3480 gctatgttgc gcgcggccac ctactacttt tctgatcttg agattgtggt gaaccacaca 3540 ggtcgcctaa cgtgggtacc caatggagca ccagaggcag cactggataa cacgagcaac 3600 cccactgctt accacaaagc accgttcaca aggcttgcac tcccttacac cgcgccacac 3660 cgcgtgttgg caaccgtgta caacgggact agcaggtact ctgcgcctgc aacacggcga 3720 ggtgacttgg ggtctctcgc ggcgaggctc gccgcacagc ttcctgcctc cttcaactac 3780 ggcgcggttc gagccacgga gatccaagaa ctcctcgtgc gcatgaagcg cgccgagctc 3840 tactgcccca ggccactgct ggcggtggag gtgacgtcac aagacagaca caagcagaaa 3900 atcattgccc ctgcaaagca actcctgaac cttgatctgc tcaagttggc gggagacgtt 3960 gagtccaacc ctgggccttt cttcttctcc gacgttaggt caaatttcac caaactggtt 4020 gaaaccatta accagatgca agaggacatg tcaacaaaac acgggcctga ctttagccgg 4080 ttggtgtccg cgtttgagga attggccgcc ggggtgaaag ccatcaggaa cggtctcgac 4140 gaggccaagc cctggtataa gctcatcaaa ctcctaagcc gcttgtcatg catggccgct 4200 gtagcagcac ggtccaagga cccagtcctt gtggccatca tgctggctga caccggcctt 4260 gagattctgg acagcacgtt tgtcgtgaag aagatctccg actcgctctc cagtctcttt 4320 cacgtgccgg cccccgtctt tagtttcgga gctccgatcc tattggctgg gttggtcaag 4380 gtcgcctcga gtttcttccg gtccacgccc gaagaccttg agagagcaga aaaacagctc 4440 aaagcacgtg acatcaatga catattcgcc attctcaaga acggcgagtg gctggtcaag 4500 ctgatcctag ctatccgcga ctggattaaa gcatggatcg cctcagaaga aaagtttgtc 4560 accatgacag acttggtgcc tggtatcctt gaaaagcagc gggatctcaa cgacccgagc 4620 aagtacaagg cagccaagga gtggcttgac aacgcgcgtc aagcgtgcct gaagagcggg 4680 aatgtccaca ttgccaacct ttgcaaagtg gtcgccccag cgccaagcaa gtcgagaccc 4740 gaacccgtgg tcgtttgcct ccgtggcaaa tccggccagg gcaaaagttt ccttgcgaac 4800 gtgctcgcac aagcaatttc aacccacttc actggcagaa tcgattcagt ttggtactgc 4860 ccacctgacc ctgaccactt cgacggttat aaccaacaaa ccgttgttgt gatggacgat 4920 ttgggccaga accccgacgg caaggacttt aagtacttcg cccagatggt ttcaaccacg 4980 gggttcatcc cgcccatggc ctcacttgag gacaagggga aacccttcaa cagcaaagtc 5040 atcattacca ccaccaacct ctatgcgggc tttaccccga gaaccatggt gtgtcctgac 5100 gcgctgaacc ggaggttcca ctttgacatt gatgtgagtg ccaaggacgg gtacaaaatt 5160 aacaacaaat tggacataat caaagcactt gaagacacac acaccaaccc ggtagcaatg 5220 ttccagtacg actgcgccct cctcaacggc atggcagtcg agatgaagag attgcaacag 5280 gatttgttca ctccccaacc acccatacag aatgtttacc aacttgttca agaagtgatt 5340 gatcgggtgg agctccacga gaaagtgtcg agccacccga ttttcaaaca gatctcaatt 5400 ccttcccaaa agtctgtgct gtactttctc attgagaaag gccaacacga ggcagcaatt 5460 gaattctttg aggggatggt gcatgactcc atcaaggagg agctccggcc cctcatccaa 5520 caaacctcat ttgtgaaacg cgctttcagg cgcctgaagg aaaactttga ggtcgttgcc 5580 ctgtgtttga ctctcttggc aaacatagtg atcatgatcc gcgaaactcg taagagacaa 5640 caaatggtgg atgatgcggt gaatgagtac attgagaagg caaacatcac cacagatgac 5700 aaaactcttg acgaggcgga aaagaaccct ctggagacta gcggtgccag cactgttggc 5760 ttcagagaga gatctctccc cggacataaa gtgagcgacg acgtgaactc cgagcccacc 5820 aaacccgcgg gaggacaacc acaagctgaa ggaccctgcg ccggaccgct tgagcgtcag 5880 aaacctctga aagtgaaagc caagctgcca caacaggagg gaccctacgc tggtccgatg 5940 gagagacaga aaccactgaa agtgaaagta aaagccccgg tcgtgaagga aggaccctac 6000 gaaggaccgg tgaagaagcc tgtcgctttg aaagtaaaag ctaagaacct gattgtcact 6060 gaaagtggtg cccccccgac cgacttgcaa aagatggtta tgggcaacac caaacccgtt 6120 gagcttatcc tcgatgggaa gacagtagcc atctgctgcg ctactggagt gttcggcact 6180 gcctacctcg tgcctcgtca tcttttcgct gagaagtatg acaagatcat gttggatggt 6240 agagccatga cagacagtga ctacagagtg tttgagtttg aaattaaggt aaaaggacag 6300 gacatgcttt cagacgctgc gctcatggtg ttgcaccgtg ggaatcgcgt gagagatatc 6360 acgaaacact ttcgtgacac agcaagaatg aagaaaggca cctccgtcgt cggcgtaata 6420 aacaacgctg atgttggtag actgattttc tctggtgagg cccttaccta caaagacatt 6480 gtagtgtgca tggatggaga caccatgccg ggcttgtttg cctacagagc cgccaccaag 6540 gctggttact gtggaggagc tgttcttgcc aaagacggag ctgacacatt catcgtcggc 6600 actcactccg caggtggtaa tggggttggg tactgctcgt gcgtatccag atccatgctt 6660 ttgaagatga aggcacacat tgaccctgag ccacaccatg aagggttgat tgttgacacc 6720 agggatgtgg aagagcgcgt gcacgtcatg cgtaaaacca agcttgcgcc caccgtggct 6780 cacggtgtgt tcaaccccga gtacggcccc gctgccttgt ccaacaagga tccgcggctg 6840 aatgagggag ttgtcctcga cgaggtcatc ttctccaaac ataagggaga cacaaggatg 6900 tcaccggagg ataaagcgct gttccgccgc tgtgctgccg actacgcgtc gcgtttgcac 6960 ggtgtgctgg gtacggcaaa tgccccattg agcgtctatg aggccatcaa gggtgtcgac 7020 ggacttgacg ccatggaacc tgacacagca cctggccttc cctgggccct ccaggggaag 7080 cgtcgtggcg cgctggttga ctttgagaac ggcacggtcg ggcccgaagt ccaggctgct 7140 ctagagctca tggagaaaag agaatacaag tttgcttgcc agaccttcct gaaggacgag 7200 attcgcccga tggaaaaagt acgtgccggc aagactcgta ttgtcgatgt cttgcctgtt 7260 gaacacattc tttacactag gatgatgatt ggcagatttt gtgcacaaat gcactcaaac 7320 aacggcccgc aaattggatc agcggtcggc tgtaatcctg atgttgattg gcaaagattc 7380 ggaacacact tcgcccaata caggaacgtg tgggatgtgg actattcggc cttcgatgct 7440 aaccactgca gtgatgcaat gaacatcatg tttgaggagg tgtttcgcac agactttggt 7500 ttccacccga acgctgagtg gattctgaag actcttgtga atacggaaca cgcctatgaa 7560 aacaaacgca ttactgttga aggcgggatg ccatcaggtt gttccgcgac aagcatcatt 7620 aacacaattt tgaacaacat ttacgtgctc tacgccctgc gtagacacta tgagggagtt 7680 gagctggaca cttacaccat gatctcctat ggggacgaca tcgtggtcgc aagtgattac 7740 gatctggact ttgaggctct caagccccac ttcaaatctc ttggtcaaac catcacccca 7800 gctgacaaaa gcgacaaagg ttttgttctt ggtcactcca tcaccgatgt cactttcctc 7860 aaaagacact ttcacatgga ctacggaact gggttttaca aacctgtgat ggcttcgaag 7920 accctcgaag ctatcctctc ctttgcacgc cgtgggacca tacaggagaa gttgatctcc 7980 gtggcaggac tcgccgtcca ctctggaccc gacgagtatc ggcgtctctt tgagcccttc 8040 cagggtctct ttgagattcc aagctacaga tcactttacc tgcgatgggt gaacgccgtg 8100 tgtggcgacg cataatccct cagaagtcac aattggcaga aagactctga ggcgcgcgac 8160 gccataggag tgaaaagccc gaaaaggctt ttcccgcttc ctatttcaaa aaaaaaaaaa 8220 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaacta gtaatcatgg tcatagctgt 8280 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 8340 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 8400 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 8460 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 8520 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 8580 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 8640 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 8700 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 8760 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 8820 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 8880 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 8940 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 9000 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 9060 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 9120 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 9180 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 9240 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 9300 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 9360 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 9420 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 9480 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 9540 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 9600 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 9660 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 9720 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 9780 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 9840 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 9900 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 9960 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 10020 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 10080 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 10140 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 10200 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 10260 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 10320 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 10380 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat 10440 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 10500 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 10560 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 10620 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 10680 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 10740 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 10800 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 10860 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc 10920 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 10980 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagca 11040                                                                        11040

Claims (9)

(A / KOR / Pocheon / 2010) having the nucleotide sequence of the genus Berk et al.'S session number KC588943, having a defense immunity against the foot-and-mouth disease type A Asian region, Wherein the plasmid has the nucleotide sequence of SEQ ID NO: 1. delete The recombinant foot-and-mouth disease virus according to claim 1, wherein the foot-and-mouth disease type A Asian type is Sea-97 genotype. The recombinant foot-and-mouth disease virus according to claim 1 or 3, wherein the recombinant virus is a foot-and-mouth disease vaccine virus. (One). Inserting the whole gene of foot-and-mouth disease type A isolate virus having the nucleotide sequence of KC588943 in a plasmid; And
(2). And injecting the plasmid obtained in the step (1) into a cell to proliferate the recombinant FMDV. The recombinant foot-and-mouth disease virus according to claim 5, wherein the cell is at least one selected from the group consisting of fetal goat fetal tongue (ZZ-R), porcine kidney cell line (IBRS-2) and hamster cell line (BHK-21) . 6. The method according to claim 5, wherein the recombinant virus has defense immunity against the foot-and-mouth disease type A Asian region type. The method for producing recombinant foot-and-mouth disease virus according to any one of claims 5 to 7, wherein the recombinant virus is a foot-and-mouth disease vaccine virus. A vaccine for the prevention of foot-and-mouth disease comprising the recombinant foot-and-mouth disease virus of claim 1 or 3.

Images