KR20160025360A - Novel Atractylodes mottle virus gene from Atractylodes japonica plant - Google Patents

Abstract

The present invention relates to a novel Atractylodes mottle virus gene found in Atractylodes ovata (Thunb.) DC. More specifically, the present invention relates to an Atractylodes mottle virus gene comprising a base sequence represented by sequence number 1, a recombinant vector comprising the Atractylodes mottle virus gene, and a host cell transformed by the recombinant vector. According to the present invention, the gene of the present invention can contribute to distribution of disease-free host plants including Atractylodes ovata (Thunb.) DC. and also to quality improvement.

Description

(Novel Atractylodes mottle virus gene from Atractylodes japonica plant)

More particularly, the present invention relates to a gene for an extract of Atractylodes mottle virus comprising the nucleotide sequence of SEQ ID NO: 1 derived from a ssp. Plant, And a host cell transformed with the recombinant vector.

The emergence and risk of emergent viruses and new viral diseases are increasing gradually due to ecosystem changes due to climate warming, evolution and variation of plants and viruses, and increased trade in agricultural products. About 1,000 plant viruses have been reported so far worldwide, and new viruses are still reported in various plants. Currently, about 100 viruses have been reported in various plants in Korea. However, detailed investigations of the viruses have been carried out only in some crops, and most crops are only fragmented. Thus, it is not yet clear which viral diseases are occurring in many major crops.

Plant diseases are abnormalities of the host cells and tissues that result from the continued influences of pathogens or environmental factors on the plant host, including abnormalities in plant morphology, physiology, etc., resulting in a reduction in the resulting economic value It is also said. Causes of these plant diseases include fungi, bacteria, Nematode, Mycoplasma, protozoa and viruses. However, the virus is a pathogen having a size of less than 1 占 퐉 and its substance can not be observed even with an optical microscope, and since it requires an electron microscope, it is difficult to diagnose a virus that has developed in a plant with the naked eye. Accordingly, various methods for diagnosing viruses have been developed and used. In particular, it is necessary to study viruses that cause diseases in sow plants.

Korean Patent No. 10-0961156 discloses a probe set for detecting plant virus. Korean Patent No. 10-1153458 discloses a duplex primer for simultaneous diagnosis of INSV and TSWV and its use, As in the case of the present invention, there has been no report on a novel transgenic mosquito virus gene derived from a ssp. Plant.

The present invention has been made in view of the above-mentioned needs. The present inventors prepared a cDNA library of 138 plant samples suspected of being infected with a plant virus, and obtained PCR results of the primer set based on the contaminants thereof , Confirming that the gene is a novel Atractylodes mottle virus gene of about 8.9 kb which eventually causes disease in the shoot. Thus, the present invention has been completed.

In order to achieve the object of the present invention, the present invention provides an Atractylodes mottle virus gene comprising the nucleotide sequence of SEQ ID NO: 1 derived from a ssp. Plant.

The present invention also provides a recombinant vector comprising the Atractylodes mottle virus gene.

The present invention also provides a host cell transformed with the recombinant vector.

The present invention relates to the isolation of a novel scrapie mutant virus gene from a scion plant whose pathology is due to the virus but whose causative virus has yet to be revealed. Since the viral infection can be diagnosed using the viral genome of the present invention, the viral disease occurrence pattern in domestic crops can be used for monitoring or investigation to be used for the disease occurrence observation. Especially, And can contribute to improving the quality of the host plant and improving the quality of the host plant.

In order to achieve the object of the present invention, the present invention provides an Atractylodes mottle virus gene derived from a ssp. Plant.

The sister mouse virus gene derived from the ssp. Plant may be composed of the nucleotide sequence of SEQ ID NO: 1, but is not limited thereto. In addition, homologues of the nucleotide sequences are included within the scope of the present invention. Specifically, the gene has a nucleotide sequence having a sequence homology of 70% or more, more preferably 80% or more, still more preferably 90% or more, and most preferably 95% or more, with the nucleotide sequence of SEQ ID NO: 1 . "% Of sequence homology to polynucleotides" is ascertained by comparing the comparison region with two optimally aligned sequences, and a portion of the polynucleotide sequence in the comparison region is the reference sequence for the optimal alignment of the two sequences (I. E., A gap) relative to the < / RTI >

In addition, the present invention provides a recombinant vector comprising the S. cerevisiae virus gene.

The term "recombinant" refers to a cell in which a cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a protein encoded by a peptide, heterologous peptide or heterologous nucleic acid. The recombinant cell can express a gene or a gene fragment that is not found in the natural form of the cell in one of the sense or antisense form. In addition, the recombinant cell can express a gene found in a cell in its natural state, but the gene has been modified and reintroduced intracellularly by an artificial means.

In the present invention, the S. cerevisiae virus gene sequence can be inserted into a recombinant vector. The term "recombinant vector" means a bacterial plasmid, a phage, a yeast plasmid, a plant cell virus, a mammalian cell virus, or other vector. The recombinant vector is preferably a plant expression vector.

In principle, any plasmid and vector can be used if it can replicate and stabilize within the host. An important characteristic of the expression vector is that it has a replication origin, a promoter, a marker gene and a translation control element.

Expression vectors containing the viral vector gene sequences and suitable transcription / translation control signals from the vet plant can be constructed by methods known to those skilled in the art. Such methods include in vitro recombinant DNA technology, DNA synthesis techniques, and in vivo recombination techniques. The DNA sequence can be effectively linked to appropriate promoters in the expression vector to drive mRNA synthesis. The expression vector may also include a ribosome binding site and a transcription terminator as a translation initiation site.

The expression vector will preferably comprise one or more selectable markers. The marker is typically a nucleic acid sequence having a property that can be selected by a chemical method, and includes all genes capable of distinguishing a transformed cell from a non-transformed cell. Examples include herbicide resistance genes such as glyphosate or phosphinothricin, antibiotics such as kanamycin, G418, Bleomycin, hygromycin, chloramphenicol, Resistant gene, aadA gene, and the like, but are not limited thereto.

In the plant expression vector according to the present invention, the promoter may be CaMV 35S, actin, ubiquitin, pEMU, MAS or histone promoter, but is not limited thereto. The term "promoter " refers to a region of DNA upstream from a structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. A "constitutive promoter" is a promoter that is active under most environmental conditions and developmental conditions or cell differentiation. Constructive promoters may be preferred in the present invention because the choice of transformants can be made by various tissues at various stages. Thus, constitutive promoters do not limit selectivity.

In the plant expression vector according to the present invention, the terminator can be a conventional terminator such as nopaline synthase (NOS), rice alpha amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaci And the terminator of the Octopine gene of Agrobacterium tumefaciens , but is not limited thereto.

The present invention also provides a host cell transformed with the recombinant vector.

Any host cell known in the art may be used as the host cell capable of continuously cloning and expressing the vector of the present invention in a stable and prokaryotic cell, for example, E. coli JM109, E. coli BL21, E. coli RR1 , E. coli LE392, E. coli B, E. coli X 1776, E. coli W3110, Bacillus subtilis, Bacillus tulifene, and Salmonella typhimurium, Serratia marcesensus and various Pseudomonas species And enterobacteria and strains such as the species.

When the vector of the present invention is transformed into eukaryotic cells, yeast (Saccharomyce cerevisiae), insect cells, human cells (e.g., Chinese hamster ovary, W138, BHK, COS-7, 293 , HepG2, 3T3, RIN and MDCK cell lines) and plant cells. The host cell is preferably a plant cell.

The method of delivering the vector of the present invention into a host cell can be carried out by the CaCl ₂ method, one method (Hanahan, D., J. MoI. Biol., 166: 557-580 (1983)) when the host cell is a prokaryotic cell, And an electric drilling method or the like. When the host cell is a eukaryotic cell, the vector is injected into the host cell by microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, DEAE-dextran treatment, gene bombardment or the like .

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Experimental Method

One. Sample collection and total RNA  extraction

138 plant samples suspected of being infected with plant viruses were collected and all samples were each ground using liquid nitrogen. All samples were stored at cryogenic temperatures. Total RNA was extracted after mixing the same amount of each sample in the same amount. Total RNA extraction was performed with Tri Reagent (MRC).

2. Ribosome RNA  Removal, double strand cDNA  Synthesis, library creation and sequencing

Ribosomal RNA was removed from the extracted total RNA using Ribo-Zero ™ Magnetic Kit (plant leaf) (Epicenter). The prepared RNA was disrupted with a short fragment and then synthesized with double-stranded cDNA. The synthesized double-stranded cDNA was end-repaired and an adapter was attached for the cloning required for library production. This series of procedures was performed using the TrueqRNA sample prep kit (Illumina). Inserted DNA (insert DNA) of appropriate size purified using BluePippin ™ 2% agarose gel cassette (Sage Science) was selected for PCR amplification. Finally, the size and quantity / quality of the inserted DNA suitable for library production were measured using an Agilent 2100 BioAnalyzer 2100 (Agilent Technologies). The library production was completed, and the size of the library segment was confirmed to be approximately 350-450bp. The library was sequenced using Illumina HiSeq2500 and raw data was generated at about 40 Gbp using a paired-end sequencing method.

3. Sequenced low  data( raw data ) And plant virus-related contig ( contig ) Production

After selecting high-quality data from low-level data, we obtained plant virus-related readings and created contig. This process was carried out using de novo transcriptome analysis (SG-de novo assembler). Approximately 7,000 contigs with plant virus-related sequences were obtained. When the length of the conti was too short (<500 bp), data with low coverage was removed when compared with the virus database (DataBase), and about 700 contigs were finally obtained.

4. primer  Fabrication and identification of whole genome

When compared with the virus-related database (DataBase), about 150 to 90 contigues were obtained by removing the contigus that showed about 85-90% or more homology. We confirmed the most homologous virus through each database, They were grouped into two groups, each of which had high homology to the same virus. The contigs belonging to each group aligned their plant viral genome as a reference sequence and confirmed their position.

Based on the nucleotide sequences of these contigs, primers were prepared and PCR was performed on 138 plant samples using the primers. The PCR results were confirmed by agarose gel electrophoresis or by sequencing of the banded PCR products. Based on the nucleotide sequence obtained from the PCR result of the primer set based on the contig, a primer was further prepared to obtain an additional base sequence.

Example  1. Identification of the nucleotide sequence of a new plant virus

(SEQ ID NO: 1), the genome information was confirmed to be a single of about 8.9 kb (SEQ ID NO: 1), and the nucleotide sequence of the primer set Stranded RNA genome, and the name of the virus was named as Atractylodes mottle virus. The maximum nucleotide sequence identity to existing viral genes was 69%.

<110> Korea Research Institute of Bioscience and Biotechnology <120> Novel Atractylodes mottle virus gene from Atractylodes japonica          plant <130> PN14233 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 8881 <212> DNA <213> Atractylodes mottle virus <400> 1 ggataaacaa cattctcatg ccatatattg aacaatagaa atatcgttta atatacgcac 60 aatgtctcta acttaccgga gcccattgga agagaatctt ggagcatatg attcaaccgt 120 gcaagcagct attgccagca catctgcaaa ccactaccgg gaggctgaag cggctaattt 180 ccagtacttc aatttccatt tgcgaccaga agcaaagaaa cacctaattg aagcagggat 240 ctaccttagc cccttctccg ctgtgcctca ctcccatcca gcttgtaaaa cattggaaaa 300 ccatttactg tacaatgttt tacccaaatt aatagataat agattttatt ttgtaggaat 360 taagaatcat aagttagatc tactaaaaac gcgcaattca agtttaagta ctataagtca 420 gataaacagg tatgtcacca gtgctgacaa agcacgttac ggctcagacc tggtccgagt 480 atcaagtgat gaacatgtct gcattgctag gcacaagttt gaattgtcta aaccaacttt 540 gcaggagttg gtaccgcgtt tgcaattagt caaccatgac tacctctttc tgcacgacga 600 gctgcattac tggagtccta aagctttaat aactttccta gaggtgttga aacccaaagt 660 aatgtatgca acgttggttt accctccaga actgctggct gggtcaaaat actccctgaa 720 taaatggtgt tatactttta aggtcttcag gcgagactta ctgttttatc ccgatggcgt 780 catgtgcgag ggttatcaac aaccgttaaa gggtggttat ttgttgcgtg ccaataagat 840 cattttgaat aacggtgatg tctacatggt tgatgttatc catagcaaat tcgctcatca 900 tcttgtgtcc atcacgcggg gtgagttagc tgcgaacact gtgcgatcat ttggaccttt 960 tgaagcaacc agcacacgtg ggcttgaacc cctactcagg aacgtgcaac attgtttccc 1020 cgtatcatac tctgtggtct caaagattta caggtatctc aacacgctca agaaacctga 1080 tgcacagtcg gctatggcca aattgagtca actcctagaa gagcctagtg gtttagaaat 1140 gaaatttcta aaagactttg caaatctcgt cattaacact agtgggatta acactatgat 1200 caagcctgaa cgcttgcgta tgttttttgg gaagtggctt aaagagttac caactatcat 1260 ttcttcccaa ttatcacttg tgcaaacagt gagtctagat gattttgtga catacttagc 1320 cccatttacc ttcgaggtac aattgggtga agtccattat gatcacaaag agatactctc 1380 catgtttgat atggccgagt gcgcagagga aatggacgta cctgatttga tggaccgctt 1440 caccctcggg aaatccgcca tgctcgcaga tagagtgcca caacctgtca gtgctgatct 1500 attcaatgtg ttcagaaaaa caagcacttt gctaaaggta cctcttccca tctttgcacg 1560 ttacctagta tgtgaaataa catacatgct ggatccgctc tgtcgggttg tgagcctcag 1620 tgtcatcagg gaagctattg cacttcgtct gattcgcggt agcccgatga tcctcccgct 1680 gcagcaattg ctcaatctta tacgttctgg gccattattg cgggattgtg tcaataaggt 1740 cattagtcac tatgccaggc tactaactgt gttttggcat gaagattggt ttaactggtg 1800 cattgaccca aatcgccaca atctccgctt cttaactgcg cagccagatc atgtaagtac 1860 cttcaggtca atcactcaac cattccgttt tgtggtggat gaagtgtcga atttttgcaa 1920 gagttcgctt cggaaacggc gcatgcctgc tccatactgc tttgcagaat tagcaggaac 1980 agacaggcaa agggggtcgg tgattgagcg caaagaaaat gattacggag ggacgcaaga 2040 agcttctggg aaacatacat atgttggctc aataccgatt tcaatcgggc acgtaccagc 2100 aacaatacca gtcaatgaca caatcgaatg cgaattacct actttgcaga ttgtgcgtca 2160 ggtaggagct ttttcctgcc cgtgctctct tgacattcca atcatgacga tgccgttgcc 2220 tgacaatcat aatttcaaaa gcccagatct tctcaaaggg cgtaggggtg gctggtatac 2280 caaacgaggt aatgtaccat atacatatca tggtggcaaa caccaggatc taggctggcc 2340 acgttggttg cctgactgga tgctttgtaa tggtattgat ttggactatt atgattgtgt 2400 cttgtaccaa aaatatgaga aacatgggcg cattggtttg cacagtgatg atgaagcaat 2460 ttttgatccc aatggtcgcg tgcatacttg caatctttcc ggctcggttg agtttaccat 2520 cacttgtggt aagggtttta cttctgcatc tcttgagcct gggacacaat tcacaatgcc 2580 agctggtatg cagcaaactc acaagcatgc cgtcatgggg gcaagtgctg ggcgagaatc 2640 tctgaccttc cgcaagttgg ctgtaaaggt ggaatcgcca cctgaaatac ctactgttga 2700 tgtgaggagt ggaggggagc cagaagctac gcatggacct gaggtttctg tccctgaaca 2760 acaaaaagaa ccccttgagc gagctggtga cttcgatatt gccggcgggc gattgagcat 2820 agttgctgtg gatgctgtcc ttgagcagat taattatcgc tgccatgctg tgcctgggga 2880 tgggaactgt ttttggtata gcatcgaaaa agccttgggg tttgaggcca tgcagtttaa 2940 gagcaaatgc agccaggtta tcttcaacca agatggtgga gccatcaaaa aattgtcaca 3000 acaaatgcaa cctcaggcct ttgctgagga tgaagctata atggctgctg ctatatgctc 3060 caaacactgc atccgcatat tgtgccctga tgagaaattg agttacacgt ataccccaaa 3120 aactgatata attgggtgtg tgcacatgcg tttaagtggt gagcattttg agcatattga 3180 acctgtaaat aaatgcctcg tcaccgctat tgctgcggca cttggtcgca gtgaaactgc 3240 aattatgaat gtcttggagc aacaagaaca gggagcttat caggcaaaca tttgggccgg 3300 agatgggata gacatccagg atcttgagtt ctattttacg ctcttcgata tccaggcagt 3360 gatttttgtt ggggaccaaa caatcaaata caatgaaggt ggtcacgtcc cagcctgttt 3420 caacttggat gatgggcata tttcattttg caggaaaaat tgccctgtca atgttgaggt 3480 gctcaaagga gaagcaagta cactaagcgt ttctgataaa agtgtgttcg aattggcaac 3540 tttgtgtact caaattagtt ataccgcgag tcacgctcgg gctaaggtgc tggctgatag 3600 tctcgagagt ggttgcacag ggatcacact gtcaaaattg ttcaacgatg cgcacaacct 3660 catgcctgag cacgaacagg gggaggtcac agctacttta aattgccttt ttggcacttt 3720 cggttgtggc aagagtacac tattcacgaa attcacgcag aaaaatccag gtaaaggcgt 3780 gttctacgtc tcacctagga aagcccttgc agtggaacac gagcgcaaat gtattggcgg 3840 acaacgaaca gatgttaatg atgcagcagg tcgcaagaag aatagaagac ctcctcatgg 3900 taaaaattgg cacacactta ctttcgagaa gtttcttaag caggtgcacc tagtcaaacc 3960 caggatggcg ttgatcattg atgagataca gttgtaccct ccaggatatc tcgatcttgc 4020 actattgctc attgcaaaag gggtgcacat ctttatcgga ggagatccct gtcagagcga 4080 ttatgataat gaaaaagatc gtgcatggtt gggcaccatg gggagcgatg ttgatcgctt 4140 gctaggcgac caaacgtata agttcaacac atgcagtcaa cgttttaaaa actccaactt 4200 tgtgggtaga ttgccatgtg ctctatttga aaacaaaaac cccgagcccc aaagtgagga 4260 agctcatctg ttgttcaccg gctgcgacga attggtgcaa cttgaaccag agtactgtga 4320 gt; ggcgaacaaa gctatcttga ctttcgggga aagtactggg cttaattaca aaaaaggaac 4440 aattttgatc acgaatgtgt ccagctacac taatgagagg cgatgggtca ccgcactgag 4500 ccgtttcagt agtaatgtgt gcttggtgaa tctagttggt gtggattgga atggcattgc 4560 tatggcgtat cgtggacgtg tactagggcg gtttctagct cgcactgcaa agaccctgga 4620 tctgtgtgag ttacttcctg gaaaacctga atttgtcatg ggattccaga gccacgtggg 4680 taaggatgag ggcgttcgtg aggccaagtt ggagggcgac ccctggctaa aatgcatgat 4740 tgatctgggt caaaccgagg atgtcgaaga ggttgaagac ttgcaagagg tcatgcagga 4800 agaatggttc aaaacacact taccgcaagc ggagttggaa agtgtgcgtg cacgttgggt 4860 gcataggttt ttagctaaag agaaacgtga ggtacgcatg ggctctatgg tttctgagca 4920 gttcacagat gaatatgcaa aggaaggggg tctcatcttg accaatgctg cagaacgttt 4980 tgaagctatc tacccaaggc atcgggcgaa cgataccgtc accttcatca tggcagttaa 5040 gaaacgcttg cgcttttcca ggcctgcagt tgagcaagcc aaattgaatg aagcagagct 5100 ttatggtgag ttcctactcc gggaattttg caagaaagtg ccattaaatg gaatgaagga 5160 tgaacgtatg atggaacagg caaagcgagc ttttgaagag aaaaaggtta gcaaaagtgc 5220 tgcaataatt gaaaatcatg caggacgatc gtgtagggat tggctgcttg acattgggca 5280 gattttttcc aagagtcaac tgtgcaccaa atttgataac cggttccgcg ttgctaaagc 5340 tgcgcagagt atagtgtgtt tccaacacgc tgtattgtgc aggtttgccc cttacataag 5400 gtatattgaa gccaagttga atgaggccct accaaaaaat ttctatatcc actccggcaa 5460 aggcttagaa gaactacaag agtgggttat aaagggtaaa ttctcaggaa tctgcacgga 5520 gtctgattat gaggcttttg atgcctcgca agatcagtac atcgttgctt tcgaggtggc 5580 tgtgatgcgg aaattgggat taccggagag tctgatcagc gattacaagt tcatcaagac 5640 gcatcttggt tctaaacttg gaaatttcgc gattatgcgg ttctcagggg aggccagcac 5700 ttttcttttc aatactatgg ccaatatgct attcacattc ctgaggtatg acttgaaggg 5760 acgagagtac atttgcttcg ctggggatga tatgtgcgca tctgaacggc tgccacttag 5820 gcacgagcac gatggtttct taaagaaatt gaagttgaag gccaaagttt tcatggttga 5880 gaaaccaact ttttgcggct ggcacctctg tcccgatgga atttacaaaa aacctcagct 5940 ggtactcgaa agaatgctca ttgcaaaaga gaaaaacaac cttgcaaatt gccttgataa 6000 ttatgccata gaggtggctt atgcctacaa gttgggggaa cgggctgtga accgcatgga 6060 cgctgaggaa ctggaagcgg cctacaattg tgttcgtata ataatcaaga ataagaaact 6120 gctgaaatct gacatactac atttctactc caatttggag tcaaaaatgt agttttatct 6180 tgatgaggcg ttcagttgcc tgcataaggg ttctgtggtt accactgcac tctataggca 6240 ggttgttgtg ttagcttagg ttgtagctgt gtggattgta tatggatgtg ttactcagag 6300 tcgtaggtaa atataggttc actaggttaa atagtagttt gcgccaaccc atcgttttcc 6360 actgtgtccc cggagctggt aaaagcactt gtattcgcga gctaatcaat ctagatagtc 6420 ggttttcagc atatacgctc ggtatcccag atcgtccaaa tttgcaggga attggaatca 6480 aacaatttga aggacagtgc gagcagggta aattgtgcat tttggacgag tatactctag 6540 gtccattcga gcctaatttg ttctttgccg tatttgggga tccgatccaa cataaaccaa 6600 gcttgtcact tagggctgac ttcatatgct gtgaaagccg tcgttttggt cgttgcacag 6660 cacaattact gaggaccctg ggttttgaaa ttcaagcaga gggcgatgat gtggtgaatc 6720 ttggtaaagt gtatggtagt gatttaacgt ccgtaattct atactacgaa cccgaggttg 6780 gatgtatcct cagagctcat aatttgcgtg ctttcaatcc cgaggaggta gtgggccaaa 6840 catttgagtg cgtgactttc atcaccggta agactgtgct acccatagaa gagcgagaat 6900 tggcatacca gtgccttact agacactcca aagagttgca tgtgttgaca ccagatgcaa 6960 cttactccgc cacctgacca ttcaaagacc tacctgggtc tggcgattgg tgcaggtctt 7020 gctttggtaa tttacacgta ctctagaagc actctgccgc acgttggaga taatttacat 7080 catctgccac acggaggttg ttaccaggac ggcaccaaat caattttcta cggtcagccg 7140 cacaagctta attctctcga gaggccaact caattgctgc agcagccctg ggcttatgtt 7200 ataatccttg gtgcgatcat cttcttattg aacatatggg agtcaagaac ttgccgttgt 7260 gggcgttgtg ttagatgagg gatttccagg tgttaatgct agtattagcc ttattggtgc 7320 taggttttgt gtatgttaat attaatcctt attccccgtg tgtcgtagta attacaggtg 7380 agtcagttag gatagtatct tgtgagttca cgcccgagtt tgtcgcttta gctagggatt 7440 taagacccgc aggttcgtgt tgagctctta ggtttgtgag cattgaattg atatactgta 7500 ccatttgacg atgccgccca agcctgatcc agacgctgaa agctctaacg cccaaaggcc 7560 accagtgccg cctccacgcc ccccagctgc taatgctgag gaagcgcgtc aacgcttagc 7620 tgagatggag agggagcgga atgagaactt gaatgaacgt cctccagtta ctgagaacga 7680 ggaggaccta cagatgataa gccgtttggg gcgcttagct gacatgttgc gtcatgagag 7740 atcagctatt gttgtgacca atgctgctct ggagactggt agaccaaccc tgcagcctgc 7800 agagaatatg cgtggggatc cgactaatat atatagtcgc gtgtccactg aattgctgtg 7860 gcggattaag cccaagagag tgtcaaacaa catcgcaact gctgaagaga tgataaggat 7920 acagatcgct cttgaagggc atggtatacc aactgagaat gtggctgagg ttatactcca 7980 gatggccctt atatgtgcca acacaagcag ctcgtccttt caggatcctc agggcaccat 8040 tgaatgggaa gggggtggaa ccattattgt tgacgatgtg gttggtacca taaatgaaat 8100 cagtacttta cgcaaggtct gccgtttgta tgctgccata gtttggaacc acatgcatat 8160 tcatcactcc ccgccagctg attgggctgc tctgggtttt caccacagta caaggtttgc 8220 agcgtttgac ttctttgact acgttgagaa taacgcttct ataaagccag ctgggggtgt 8280 ggtgccgagg ccaactcgag cagagtatga cgcgtatttc acttacaagc atttggcgtt 8340 gaataaggcg aatgtcaatg acactttcgc caactttgat gcccagataa caggagggag 8400 gcagggtcca gcgatcagta ccaatttcaa taatgcgaat aaccggagtc ttcagtaatg 8460 aattcaagcg atgctactag gataaagatg ctcataatag gggctttcgc acaacactca 8520 tcccaagtga gtgttcctat ctgtataaat atatataggc gtgcttttgt taaagttgtt 8580 ggacaagggc gtagtactta cgctcgcaaa cgtcgtgcgt tgtccattgg tcggtgccac 8640 cggtgctacc gagtttatcc tcctttaccg ttttccaaga agtgcgataa ccgcacatgt 8700 gtgcctggaa tttcgtataa tgtgaaggta gctaattata tcctatgggg agtaaccgag 8760 gtgatacccc atcctggtta taatttctaa aagctgcata taaaacttaa ataatatatg 8820 tgtgtgtagc tataaaaaga gtgaggtttt aagatatttt tcccttaaaa aaaaaaaaaa 8880 a 8881

Claims (3)

The Atractylodes mottle virus gene comprising the nucleotide sequence of SEQ ID NO: 1 derived from a ssp. Plant. 1. A recombinant vector comprising the gene of the extract vector of claim 1, which is derived from an extractor plant mottle virus. A host cell transformed with the recombinant vector of claim 2.