CN101864435A - Turbot reovirus whole genome and application thereof - Google Patents
Turbot reovirus whole genome and application thereof Download PDFInfo
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Abstract
The invention discloses a turbot reovirus whole genome as well as a preparation method and application thereof. The method comprises the following steps of: A. extraction of turbot reovirus nucleic acid: extracting turbot reovirus dsRNA (double-stranded Ribonucleic Acid) by using a Trizol Reagent; B. artificial joint connection: carrying out agarose gel electrophoresis on purified virus nucleic acid and recycling all sections of dsRNA by using a gel recovery kit; C. reverse transcription: carrying out reverse transcription on the dsRNA connected with an artificial joint by using a primer; D. PCR (Polymerase Chain Reaction) amplification and sequence detection: carrying out PCR amplification on 2 microliters of cDNA, cutting off an amplified belt, connecting the amplified belt to a carrier after recycled and selecting a positive clone for sequence detection; and E. expression of an NS22 gene in a fish culture cell and confirmation of an initiation codon: designing and constructing a nucleotide sequence of a 1-613 basic group in the S7 section in a carrier into an eukaryotic expression plasmid. The invention also provides application of a turbot reovirus gene sequence and the coded protein thereof in preparing a turbot reovirus diagnostic reagent or being used as a cell fusion inductive agent.
Description
Technical field
The present invention relates to hydrocoles virus, be specifically related to a kind of turbot reovirus genome, also relate to the application of a kind of turbot reovirus genome in virus detects, particularly from the function and usage of turbot reovirus S7 sections clone's an inducing cell fusion rotein.
Background technology
Reovirus is to have the genomic virus of segmented double-stranded RNA, extensively infected person, other vertebratess, insect and plant.The early diagnosis of virus disease and vaccine immunity play keying action to the prevention of virus disease, and virus genomic nucleotide sequence can be used as the specific molecular marker of the inside and outside virus of detection bodies, also is the target of development antiviral and the basic material of virus vaccines.Therefore, separate, identify that aquatic reovirus and genome thereof are the preconditions of the hydrocoles virus disease being carried out molecular diagnosis and development virus vaccines.
Aquatic reovirus is the reovirus that infects hydrocoles.Be separated to reovirus from multiple ill hydrocoles at present, as be separated to the GCRV cause of disease from the grass carp that suffers from hemorrhagic disease, this viroid cause of disease causes serious harm to each department, the Asia grass carp aquaculture that comprises China.According to RNA-RNA blot hybridization analysis and genome sequence similarity, Aquareovirus virus has been divided into six hypotypes (aquatic reovirus A-F).So far, the aquatic reovirus that has obtained whole genome sequence information has: grass carp hemorrhage virus (GCHV) 873 strains (Grass carp reovirus 873, GCRV 873), America gold body bream reovirus (Golden shiner reoviru, GSRV), U.S.'s GCRV (American grass carp reovirus, AGCRV).Other has some virus strain partial sequences to measure (Attoui, H et al, (2002), general virology magazine (Journal of General Virology), 83:1941-1951; Mohd Jaafar, F et al, (2008), virusology (Virology), 373:310-321).The acquisition of genome sequence column information is for further relation between each virus strain of research and viral functional gene provide solid basis.
The symptom of virus infection and its virulence have closely related property.During existing known aquatic reovirus cells infected, but inducing cell merge, cause producing synplasm, caused typical cytopathic when this also is the reovirus cells infected.Turbot reovirus (Scophthalmus maximus reovirus, SMReV) be by our the reovirus cause of disease of isolation identification from ill turbot first, belong to Reoviridae (reoviridae) Aquareovirus (aquareovirus) member, the important economic fish turbot of main infection, this viral genome sequence column information and functional gene yet there are no report.
Summary of the invention
The objective of the invention is to be to provide a kind of turbot reovirus full genome, it forms length overall 24042bp by 11 segmented double-stranded RNAs.Each sections size is respectively: L1,3947bp; L2,3866bp; L3,3687bp; M4,2640bp; M5,2241bp; M6,2057bp; S7,1399bp; S8,1317bp; S9,1118bp; S10,986bp; S11,784bp.Wherein, encode the respectively structural protein of virus of L1, L2, L3, M5, M6, S8, S10 sections, and the Nonstructural Protein of other sections coding virus.The genome that utilization the present invention obtains can be made into available preparation or product by existing technology, is applied to aspects such as the detection of turbot reovirus and immunoprophylaxis.
Another object of the present invention is to be to provide the application of the full gene of a kind of turbot reovirus in virus detects.
A further object of the present invention is to be to provide the inducing cell fusion rotein of a kind of turbot reovirus coding to be used for the application of cytogamy inductor as preparation.
In order to realize above-mentioned task, the present invention has adopted following technical measures:
A kind of preparation method of turbot reovirus genome sequence the steps include:
The extraction of A, turbot reovirus nucleic acid
Use Trizol Reagent reagent (Invitrogen) to extract turbot reovirus dsRNA nucleic acid, see the reagent service manual for details.
B, connection manual splice
The viral nucleic acid of purifying, uses gel to reclaim test kit (Fermentas) and reclaims each sections dsRNA the band cutting-out of each sections correspondence in 1% (W/V) agarose gel electrophoresis.
The nucleic acid that reclaims adopts the tailing method to measure sequence.Use T4RNA ligase (Takara) the oligonucleotide joint P1 of synthetic to be connected to the 3 ' end of each sections dsRNA.Connect product with isopyknic phenol/chloroform extracting and purifying, be dissolved in 20 μ lTE solution.
C, reverse transcription
The dsRNA that connects manual splice uses primer P2 (5 ' AGTCTAAGTGAGGATGGCGGG3 ') reverse transcription.RNA composition final concentration in the reverse transcription product is that the NaOH hydrolysis of 0.1M is removed, and uses the HCl neutralization reaction.Add Tris-HCl damping fluid (PH7.5) to final concentration 20mM.CDNA was 68 ℃ of annealing 20 hours, and phenol/chloroform extracting and purifying is dissolved in 10 μ lTE solution.
D, pcr amplification and sequencing
Get 2 μ l cDNA pcr amplifications.Extend 2min prior to 72 ℃ in advance before the PCR circulation.The PCR product downcuts amplified band in 1% (W/V) agarose gel electrophoresis, is connected to pMD18-T carrier (Takara) after the recovery, selects the positive colony order-checking.Obtain the sequence information of each sections.
E, NS22 gene expression and the determining of initiator codon in the fish culturing cell
The nucleotides sequence of S7 sections 1-613 base is listed in pcDNA3.1 carrier (Invitrogen) design construction become eukaryon expression plasmid.The plasmid that successfully constructs uses Lipofectamine2000 transfection reagent (Invitrogen) transfection in fish cell system, is undertaken by the reagent operational manual.24h after the transfection uses nucleic acid dye Hoechst33342 to dye nuclear, observes down in fluorescent microscope.
As stated above, respectively with S7 sections 11-613,14-613,15-613, the nucleotide sequence of 18-613 base inserts the pcDNA3.1 carrier, transfection fish culturing cell system, observe phenomena.Follow the initiator codon of determining the NS22 gene according to the appearance of cytogamy phenomenon.The result shows that the NS22 gene originates in non-classical initiator codon CUG, the fusion that encoded protein can inducing cell.
The full genome of a kind of isolating turbot reovirus, its sequence is the nucleotide sequence shown in the SEQ ID NO.1, wherein 1-3947 is a L1 sections sequence, 3948-7813 is a L2 sections sequence, 7814-11500 is a L3 sections sequence, 11501-14140 is a M4 sections sequence, 14141-16381 is a M5 sections sequence, 16382-18438 is a M6 sections sequence, 18439-19837 is a S7 sections sequence, and 19838-21154 is a S8 sections sequence, and 21155-22272 is a S9 sections sequence, 22273-23258 is a S10 sections sequence, and 23259-24042 is a S11 sections sequence.In step D, obtain, form length overall 24042bp by 11 segmented double-stranded RNAs.Each sections size is respectively: L1,3947bp; L2,3866bp; L3,3687bp; M4,2640bp; M5,2241bp; M6,2057bp; S7,1399bp; S8,1317bp; S9,1118bp; S10,986bp; S11,784bp.Wherein, encode the respectively structural protein of virus of L1, L2, L3, M5, M6, S8, S10 sections, and the Nonstructural Protein of other sections coding virus.This genome is acquisition first, and its sequence and encoded protein can be used for the specific detection of virus.
A kind of isolating protein, its sequence are the aminoacid sequence shown in the SEQ ID NO.2, obtain in step e, and the gene NS22 that is comprised by turbot reovirus S7 sections encodes.Obtain the NS22 full length gene by pcr amplification, and determined the initiator codon of this gene by deletion mutantion.This gene cDNA total length 597bp, 198 amino acid of encoding originate in a non-classical initiator codon CTG, contain one and stride the film district.The albumen of this genes encoding can be applicable to the preparation of cytogamy inductor.
A kind of turbot reovirus gene order and proteins encoded thereof the purposes in preparation turbot reovirus diagnostic reagent.
The genome sequence of virus and albumen thereof can be used for the existence of specific this virus of detection, with S11 sections proteins encoded is that example has been illustrated the application of virus gene sequence in the viral detection of drugs of preparation, by S11 sections encoding gene is cloned among the prokaryotic expression carrier pET32a (Novagen), vivoexpression, purifying protein, with the protein immune animal of purifying, the preparation antiserum(antisera) has been investigated the application of serum in virus diagnose reagent of antiviral protein.The result shows: the antiserum(antisera) of antiviral protein can be used as the existence that a kind of pharmaceutical agent detects virus.
A kind of inducing cell fusion rotein of turbot reovirus coding is as the purposes in a kind of cytogamy inductor.
By the NS22 gene clone is arrived among the carrier for expression of eukaryon pcDNA3.1 (Invitrogen), in the fish cell of vitro culture, express, investigated of the influence of this expression of gene to fish cell.The result shows: this expression of gene causes that tangible fusion phenomenon appears in the fish culturing cell, and synplasm appears in cell.Illustrate that this gene can induce the fusion of fish cell.Owing to when the albumen of NS22 genes encoding is expressed, can cause tangible cytogamy phenomenon in fish cell, so this gene can be used as a kind of cytogamy inductor, plays an important role in the fusion of inducing various kinds of cell.
Advantage of the present invention:
1. obtained the genomic full sequence structural information of turbot reovirus first, the sequence information that utilization obtains, detection turbot reovirus disease that can be special can be used widely in agriculture production; The preparation of viral nucleic acid material programmable is produced in batches.
2. the purposes of utilization turbot reovirus genome sequence and protein Preparation virus-specific diagnostic reagent thereof is provided, set up special, responsive, the localized diagnostic method of turbot reovirus disease, but standard operation, quick diagnosis turbot reovirus disease.
3.S7 the evaluation of sections coding inducing cell fusion rotein provides a kind of purposes of development of novel cell fusion-inducing agent.Expression explanation in multiple fish cell, it can induce the fusion between the multiple fish cell, thereby can be used as the fusion that novel cytogamy inductor is widely used in inducing other cell.
Description of drawings
Fig. 1 is a turbot reovirus genome electrophoretogram, and virogene is organized structural representation.
Fig. 2 is that the prokaryotic expression and the turbot reovirus western of S11 sections detects.
Figure A is the prokaryotic expression of S11 sections:
M, marker; 1, contrast; 2, abduction delivering.
Figure B is that western detects virus:
1 is contrast, and 2 is the cell of virus infection.
Fig. 3 expresses NS22 gene and deletion fragment thereof in fish cell.
Figure a, b, c are that phase microscope is observed;
Figure d, e, f observes for the nucleus fluorescent dye;
Figure g, h, i differ and the overlapping of Fluirescence observation picture.
Microscopic examination shows that the NS22 gene can induce the fusion of fish cell, and sequence 15-613 and sequence 18-613 all can not merge by inducing cell, according to S7 sections 14-19 bit sequence AUCCUG and eukaryotic initiation codon conserved sequence
A/
GThe NNAUG comparative analysis illustrates that the NS22 gene uses the initial albumen of non-classical initiator codon CUG synthetic.
Embodiment
The invention will be further described below in conjunction with drawings and Examples, but not as the restriction to interest field of the present invention.
Embodiment 1:
The preparation method of the full gene of a kind of turbot reovirus the steps include:
The extraction of A, turbot reovirus nucleic acid:
Use Trizol Reagent reagent (Invitrogen) to extract turbot reovirus dsRNA nucleic acid, see the reagent service manual for details.100 μ l viral suspensions are added 900 μ l Trizol reagent, put upside down mixing 5-10 time, 15-30 ℃ leaves standstill 5min, thoroughly lytic virus albumen.Add the 0.2ml chloroform, firmly shake Eppdorf pipe 15s, 15-30 ℃ leaves standstill 2-3min; 2-8 ℃ to be no more than 12, the centrifugal 15min of 000g; The careful upper strata water that takes out adds isopyknic Virahol mixing in another clean Eppdorf pipe, 15-30 ℃ leaves standstill 10min; 2-8 ℃ to be no more than 12, the centrifugal 10min of 000g; The precipitation with 1ml 75% (V/V) washing with alcohol, shake up the back at 2-8 ℃ to be no more than 7, the centrifugal 5min of 500g; Abandon ethanol, air drying 10min; Add 20 μ l DEPC treating water ,-80 ℃ of preservations are standby.
B, connection manual splice:
The viral nucleic acid of purifying, uses gel to reclaim test kit (Fermentas) and reclaims each sections dsRNA the band cutting-out of each sections correspondence in 1% (W/V) agarose gel electrophoresis.
The nucleic acid that reclaims adopts the tailing method to measure sequence.Use the oligonucleotide joint P1 (5 ' PO of T4RNA ligase (Takara) with synthetic
4-CCCGCCATCCTCACTTAGACT-NH
23 ') be connected to the 3 ' end of each sections dsRNA.Undertaken by kit method, 20 μ l linked systems comprise 50mM Tris-HCl (PH7.8), 100ng dsRNA, 10mM MgCl
2, 10mM DTT, 10mM ATP, 50pM P1,40%DMSO, 10%PEG6000,20U T4RNA ligase enzyme, 15 ℃ of connections of spending the night.Connect product with isopyknic phenol/chloroform extracting and purifying, be dissolved in 20 μ lTE solution.
C, reverse transcription:
The dsRNA that connects manual splice uses primer P2 (5 ' AGTCTAAGTGAGGATGGCGGG 3 ') reverse transcription.DsRNA 10 μ l add P2 primer 1 μ l, dimethyl sulfoxide (DMSO) 4 μ l, 94 ℃ of sex change 5min behind the mixing.Be to add reaction buffer 5 μ l, RNA enzyme inhibitors 1 μ l, dNTP1 μ l, MMLV reversed transcriptive enzyme 1 μ l, H in the system of 25 μ l in final volume then
2 O 2 μ l.Reverse transcription was carried out in 42 ℃ of insulations in 1 hour.RNA composition final concentration in the reverse transcription product is that the NaOH hydrolysis of 0.1M is removed, and uses the HCl neutralization reaction.Add Tris-HCl damping fluid (PH7.5) to final concentration 20mM.CDNA was 68 ℃ of annealing 20 hours, and phenol/chloroform extracting and purifying is dissolved in 10 μ lTE solution.
D, pcr amplification and sequencing:
Get 2 μ l cDNA and add 10xPCR damping fluid 5 μ l, dNTP 2 μ l, primer P21 μ l, Ex Taq enzyme (Takara) 1 μ l carries out pcr amplification in 50 μ l reaction systems.Extend 2min prior to 72 ℃ in advance before the PCR circulation.The PCR circulation is as follows: behind 94 ℃ of sex change 4min, and with 94 ℃ of 30s, 58 ℃ of 30s, 32 circulations of 72 ℃ of 2min reactions, last 72 ℃ are extended 10min.The PCR product downcuts amplified band (about 1.4kb) in 1% (W/V) agarose gel electrophoresis, is connected to pMD18-T carrier (Takara) after the recovery, selects the positive colony order-checking.
E, NS22 gene expression and the determining of initiator codon in the fish culturing cell:
The nucleotides sequence of S7 sections 1-613 base is listed in the pcDNA3.1 carrier design be built into eukaryon expression plasmid: use primer P3/P4 (P3:5 ' ACTGGTACCGTTTTAGTCAATCATCCTG 3 '; P4:5 ' ATTCTCGAGTAAAGTC AACGGAAG 3 ') amplification, amplified production and pcDNA3.1 carrier be double digestion simultaneously, uses restriction endonuclease Kpn I and Xho I.Enzyme is cut product and is used the T4DNA ligase enzyme to connect, and transformed into escherichia coli is selected positive colony then.
The plasmid that successfully constructs uses Lipofectamine2000 transfection reagent (Invitrogen) transfection in fish cell system, is undertaken by the reagent operational manual.24h after the transfection uses nucleic acid dye Hoechst33342 to dye nuclear, observes down in fluorescent microscope.
As stated above, respectively with S7 sections 11-613,14-613,15-613, the nucleotide sequence of 18-613 base inserts the pcDNA3.1 carrier, transfection fish culturing cell system, observe phenomena is to determine the initiator codon of NS22 gene.The result shows that the NS22 gene originates in non-classical initiator codon CUG, the fusion that encoded protein can inducing cell.
F, experimental result:
From 100ul turbot reovirus suspension, obtained highly purified viral nucleic acid material, as shown in Figure 1, in 1% agarose gel electrophoresis, show 10 genome bands clearly, and obtained the turbot reovirus genomic information first, as shown in table 1.
Table 1 turbot reovirus genomic information
Embodiment 2: turbot reovirus gene order and proteins encoded thereof are applied to the preparation of turbot reovirus diagnostic reagent.
Be that example explanation turbot reovirus genome sequence is listed in the application in the viral detection of drugs with S11 sections proteins encoded below:
The vivoexpression of A, S11 sections:
The nucleotides sequence of S11 sections encoding gene is listed in the pET32a carrier design be built into prokaryotic expression plasmid: use primer P5/P6 (P5:5 ' GAAGAATTCGGAGCAGGAATG 3 '; P6:5 ' GCCAAGCTTACTCAAGACTCTACTC 3 ') amplification, amplified production and pET32a carrier be double digestion simultaneously, uses restriction endonuclease EcoR I and Hind III.Enzyme is cut product and is used the T4DNA ligase enzyme to connect, and Transformed E .coli DE3 selects positive colony.
The positive colony bacterium is cultured to logarithmic phase, and the adding final concentration is that the IPTG of 1mmol/L induces 4-8h at 37 ℃, and expressed proteins can be used affinitive layer purification.
B, Antiserum Preparation:
The albumen of purifying adopts the intraperitoneal injection immune mouse, and the per injection amount is 50ug.First immunisation is used Freund's complete adjuvant, back 4 use Freunds.Mouse blood was got in the 5th immunity in back 3 days.Blood is put 4 ℃ spend the night behind 37 ℃ of 1h, behind the centrifugal 10min of 4000g, draw the upper strata polyvalent antibody ,-80 ℃ of preservations are standby.
C, immunological method detect turbot reovirus:
Immunological method detects can be divided into ELISA, and Western etc. are that example is set forth the application of viral protein polyvalent antibody in viral detection of drugs below with Western:
The protein sample that need are detected separates through 12% (W/V) SDS-PAGE glue; Electrophoresis finishes the back and changes film Sandwich according to the assembling of MiniTrans-Blot electrophoretic transfer system operation guide, and the electrotransfer system is installed, and changes film 40min with 70V voltage ice bath, and albumen is transferred on the pvdf membrane that the aperture is 0.45 μ m; The PVDF behind the film is changeed in ponceau dyeing, mark Marker after, wash film three times with TBST solution, at every turn 10min; 5% (W/V) skim-milk (being dissolved among the TBST) room temperature sealing 1h; With 1: 500 Dilution ratio (according to the antibody titer adjustment) antiserum(antisera) is diluted among the TBST incubated at room 2h; TBST washes film 3 times, each 10min; With the goat anti-rabbit igg (or sheep anti mouse) of 1: 1000 Dilution ratio adding alkali phosphatase enzyme mark, incubated at room 1h; TBST washes film 3 times, each 10min; Adopt NBT/BCIP colouring reagents box to develop the color, observe the colour developing situation at any time, purpose is taken now out of and is washed termination reaction with flowing water; After film dries, deposit figure with the scanning of gel scanning analysis system.
D, detected result
Use S11 sections encoded protein successfully to prepare the serum of antiviral protein as antigen, shown in Fig. 2 A, the prokaryotic expression bacterial strain of structure has obtained a large amount of expressing protein (Fig. 2 A swimming lane 2) after inducing 6h.Still can the special existence that effectively detects viral protein after using this antiserum(antisera) with dilution in 1: 500, shown in Fig. 2 B swimming lane 2, obtained special band after the colour developing.The about 10ug of total protein that detects, thereby the precision of detection viral protein can reach Gamma Magnitude, has very high susceptibility, overview such as table 2.
The anti-S11 albumen of table 2 serum detects viral protein
| Serum dilution | Normal control | Virus infected cell |
| ??1∶100 | ??- | ??+++++ |
| ??1∶200 | ??- | ??+++++ |
| ??1∶300 | ??- | ??+++++ |
| ??1∶400 | ??- | ??++++ |
| ??1∶500 | ??- | ??+++ |
| Serum dilution | Normal control | Virus infected cell |
| ??1∶1000 | ??+ |
"-" expression does not have the positive signal of detection, and the quantitaes of "+" detects the power of positive signal.
Embodiment 3:NS22 gene is as artificial cytogamy inductor
A, the nucleotides sequence of S7 sections 1-613 base listed in the pcDNA3.1 carrier design be built into eukaryon expression plasmid:
Use primer P3/P4 (P3:5 ' ACTGGTACCGTTTTAGTCAATCATCCTG 3 '; P4:5 ' ATTCTCGAGTAAAGTC AACGGAAG 3 ') carries out pcr amplification;
B, amplified production and pcDNA3.1 carrier be double digestion simultaneously, uses restriction endonuclease Kpn I and Xho I.Enzyme is cut product and is used the T4DNA ligase enzyme to connect, and transformed into escherichia coli DH5 α selects positive colony then;
C, the clone that successfully constructs are in the substratum that contains the ammonia benzyl, and 37 degree are cultivated 12h, make to spend endotoxic plasmid extraction kit (OMEGA) and extract plasmid, are undertaken by operational manual;
D, the plasmid that successfully constructs use Lipofectamine2000 transfection reagent (Invitrogen) transfection in fish cell system, are undertaken by the reagent operational manual.24h after the transfection uses nucleic acid dye Hoechst33342 to dye nuclear, observes down in fluorescent microscope.
After E, the about 2ug of use contained the plasmid transfection cell of NS22 gene, in 24h, tangible fusion phenomenon appearred in cell, and as Fig. 3 g, shown in the h, the cytolemma that shows as flanking cell is fused to together, and nucleus is assembled the formation synplasm.And in the cell of the different cells of fish of the same race and different fishes, all can cause the fusion of cell.
Result (Fig. 3) according to the observation, the protokaryon of NS22 gene or eukaryotic expression product can be used as a kind of cytogamy inductor of artificial design, are used to induce the fusion of other fish cell or zooblast.Also can be used to promote the fusion of allogenic material and cytolemma, and then help allogenic material and enter cell.
The protein induced different cytogamy overviews of table 3NS22
| Cell | Inducing cell merges situation | The cytogamy time of occurrence |
| Grass carp fin ray cell | Good | ??16h |
| The grass carp kidney cell | Good | ??24h |
| The grass carp gonad cell | Good | ??24h |
| A fertile carp cell | Good | ??16h |
Sequence table (SEQUENCE LISTING)
＜110〉Inst. of Hydrobiology, Chinese Academy of Sciences
＜120〉full genome of turbot reovirus and application thereof
<130>2
<160>2
<170>PatentIn?version?3.1
<210>1
<211>24042
<212>RNA
＜213〉turbot reovirus
<400>1
guuuuaucac?gacauggcga?ccguauacgg?cauucaacuc?acugaccgac?ugaacaccgc????60
aacuguccga?cgcccccuua?gacuucgacg?cuacgacuca?uacauuacca?cuuuuaccac???120
uccuaauggc?aucucccaac?uguaccgcgc?ucuggauuuc?caaccaacuc?aguuuagcgc???180
uucuguuuua?caaacuuucc?cuccacuuaa?cgccuggagu?cccgcuccuc?aauuccuucc???240
cgaugaucua?gaccuauccc?aguggaaaga?auggauuacu?gaucgaaugc?gaucccuugc???300
caccguacuu?caacgugcuu?acccacuagu?agccaacuca?cgucgggagg?ugaaccccau???360
cguuaucggu?cuggucacau?cagcguuccu?caaccaacgc?ccaauugagc?gauuucuucc???420
auugcuauuc?cuucgaccag?gaaaccgcga?uccuaucgcg?cccuugguaa?cgguggaugc???480
cacuuucucu?gacgauaccu?acgucucaaa?ccguguucug?uacacgccug?ccggucucaa???540
guaucuaacc?uuaaggucgu?augauuccac?gaaaucaucc?gcuaucugua?ccuucgguaa???600
acacguucca?uucuacgcca?ucgcugcuuu?cuaucccgac?gagcacgccc?gacugaccau???660
ccuccaucgu?uacaacggcg?gaccaccucu?aauagagcau?uuugaucaac?caacguacgg???720
accccacgua?cucauucccg?cacuuggcuc?cccugagggu?uaugauacgg?ccaagccgac???780
guucgccgau?cuccucuuga?cugagggacu?ucucgauuca?uuccgccuaa?acgccucagc???840
cggccccuca?acugccgugg?cucguaucga?ccagacuuau?cauauuguca?ugaauggcaa???900
cccaagugau?cacacccagu?uggccacucg?ccuguccaau?cugucacuac?ucgcugucca???960
agguugucag?augacgguuc?agguuguuga?ccauccaucc?augucggacg?ucuguggcuu??1020
ucuagugcgc?cuucucggcc?ccggugaccc?ucaacgguug?cucaacuauc?aaaccgauca??1080
gauucuaauc?uggcaggcua?gccccuuccc?cuuugguaac?aacccucguu?auaucagacg??1140
ucaaggucgc?guaccguuca?ccgucgguaa?cacgacguac?gugccagaca?ccaagacccc??1200
acuaccguuc?cuuccccagu?aucggcgugc?uguugugaau?aagaacaacg?cucuggcguc??1260
auaccaagag?aaucucuugc?cuaguuuacc?aaucuaccac?gguuucgcuc?ucaccggugg??1320
ugccuucuuc?caaucucgug?acauuacugg?ugaucccgcu?aauguauggc?cggugaauac??1380
uaugccuacu?cugccucaag?acuauuuuag?cauccguucc?aggcagcguc?gcgagcuacu??1440
cucccggcug?cgaucaccgu?cagaucguuc?guacaucaag?gaccuucaua?acaucucguu??1500
cgcugcuacc?guucucaacc?ccgugaauaa?ucagaucguc?cucucugaag?guuucuccau??1560
ggcuuaccuu?ggcgccgccu?ccacccacgg?ugcaucggau?caaccacuca?ucaucgacgc??1620
gcucaagaau?ggaacugucc?caggaguccc?cgugccaucc?aaaguggcuc?aauuugguua??1680
ugacgucgcc?aaugguagca?ucauggacgc?gacucucucg?ccaccaacug?gaacguucuc??1740
cuuuguuuac?ucagacgucg?accaagugga?ggacgcuggc?cuauccaucg?uugccgcuaa??1800
ccgggcugca?gcugcuguga?cgaccgucgc?auuauccaug?acuacggcag?guggucuaac??1860
gaucgugaag?auuaacuucc?cgacucccgc?uuucuggacu?cacuuguucc?ggcagcacgc??1920
caugcaugcg?cgcgcgaugu?acauccuaaa?accguugauc?gugaacucag?uggaggucuu??1980
ccuccuguuc?gucagccgcg?cuuccgacgg?uaaccugguc?uccucccccg?ccuugcggca????2040
auuccuaguc?cagcuuuuug?acaggucagc?guaucuaacu?gacuugaugg?cucacguccc????2100
uuugcucggu?gauguagaug?auggcgcgac?cucccucggc?uuuaacgcuu?gucgacugua????2160
uagcccugac?cuccccucua?cuaacgucac?gcccgagaua?cagacucuug?ccuaccaacu????2220
ggcuacgauc?gugccgucca?ccucguucau?cgcccgugag?gauuaugaug?gggcuacagc????2280
cgucacguuc?uacgguaaac?guacauuccu?gucccaaaac?cgccuugacc?guuugguaga????2340
cgucccaguu?cccgcaacca?augccaucaa?ucaucagacg?cgcuucacug?gcuccccggu????2400
guaccagcug?uuuccugcca?aucccgcucc?ugucacccaa?cuucucgcug?gcgcguauaa????2460
ccguuuaguu?cacucucagc?ucgcucgugu?ccagccccaa?gcucuagugg?acuguggaac????2520
ggguccggaa?ugccgcgucc?ucucgcucau?accucccacg?acugcuauca?ccaugauuga????2580
cgcgcggccc?ccggcugaau?ugcuugcagc?cuucaauccg?gcuaugaacc?aguacauuga????2640
ggaugacuuc?cuuaacccgg?cucuuugggu?cgcgcauccg?uaugacgccu?uaaccgcuau????2700
cuucucuuua?ggagcugccu?ucgcaggugc?caacuuagau?cuugucgucg?gucugacagc????2760
cuuccugcgc?cuuguucaac?cagguaaccu?ccagcaccuu?uggcugcagc?uuaacacacc????2820
ccucacaucc?aacacuucuc?uccccggccu?auuggagauu?gacaccagga?cuaaucaaua????2880
caucuucaac?ggaggacagc?gcaccgagcc?cuacgcuuua?cccaaugaca?uuuugacugc????2940
agugcgucag?guauuccccg?cggccaccac?cuccuggcug?acugcuuccc?caucgaugga????3000
uugggcugag?uacguuaucg?cucugggcuc?cucaaugucg?uuagaugacg?ucaacacgau????3060
gaucacuuac?ucaggccugg?cucccauucu?ucacauugau?caaacucagc?ggcccaugga????3120
cguuccugua?ccuuugguug?uuggagucca?agcuaauauc?caugucgcug?cccccgucca????3180
acagaccacc?guaauuggca?cgaucgccgg?cguucaaguc?uuuaccgcug?auggcgugac????3240
cgcccccucg?accuugggcc?ccuuagcugc?cguuugggac?gcugcgcuau?cccguuggaa????3300
ccuuacucuc?acgcccaauc?aggccggagu?cuuggacguu?guuguggacc?augcuggugu????3360
uaugcucaac?cguggcucaa?ccacuaucgc?auuaccaccc?gcuaccauca?augucaccuu????3420
cccucaagcg?gcuaaccgag?acuacacuaa?cgcugguaac?gacgcggcca?ucguaugcga????3480
cccuuuuuau?cgucucgggg?ucuucguuag?ugugaacggu?acuuuccagc?ccguuaaccc????3540
agaacgugcc?gccaucauua?cugcugccaa?cgucagaucu?cuccacuaug?uauaugaccu????3600
cucugacaau?cacguucuga?uguacauaug?ugacaucacu?gacaacaacg?ugggccgcaa????3660
cauugccuuc?ccgcucgccg?acaucuucca?aacuguguuc?ccaaacaaua?cuccacuccu????3720
cgccuccccc?cccuaccccu?cugccucggg?ucgacuacug?cugaauggcc?agccguuccu????3780
agaccuggac?ccccucccac?cgguauuacc?uccagguguu?cagauccagg?ccuuaucuac????3840
cgcggucgaa?ccagcgcgcc?agacgguuga?ggugccagcg?ggugucuacg?uguauguggu????3900
uguuuagucu?uggugagcgc?cccgcgguuc?gcgucgugcu?auucaucguu?uuauccacua????3960
uguccgcguu?guucaacgcg?cugccacccg?agcuccagca?acuuucacuc?gcacuaucug????4020
guucccaacc?acucaccgac?aaaaccuuua?ccgcugccgc?ugaugccugg?cauauccgcc????4080
cccgauccca?agccuaccac?cuacucgaua?cccucacauu?ucgauccucc?gugguuaucc????4140
cuaacucuau?cuucgucggu?cgagucuggu?cggauuauug?ggccuuacag?gacaacaucg????4200
ucauccgaau?cagcccggaa?ggugcgaaag?acgccgacua?uugccacaau?ucgaacaucg????4260
caccuguccu?gucgccucuu?aagaagauuc?cggaauaugg?uacgcuccau?ccgacgauug????4320
acaaggaugc?aucugaacgc?gguuacccuu?cagcccgcau?ggcgucuucc?uucuuuaagc????4380
ucgccuccuc?ccaagccaga?caaguaaaga?ucgauccaac?uagguuucuu?gaguuccuuc????4440
ucgucgucaa?cgccaccacg?cgagucccuu?cgggugucga?cucagaccag?ccuaauccau????4500
gguuacccga?auccaguccc?gcgcuucaag?cgauuuggca?gauuaugcaa?cgcuacaagg????4560
uagacuccaa?guacuacgcu?cccgcccucg?uugugaacac?cggagccguu?ugguggauuc????4620
cgcccccagg?ucgcaccaau?ugcguaacug?ugcaguuccu?caucacugau?cugauuaauc????4680
uagccgugaa?cgcuuucgcc?acucgguuau?ccccugagcu?ugagaugugu?gcuguuagag????4740
ucuaccuugc?cgcggcggcu?acgcccaauu?acgcucacgc?ccuccucgau?cugaaggcaa????4800
ucuuccccaa?ucucagccuc?cacagcaugg?accgcagugg?ugaauucggu?gguaagugcc????4860
cucgcauuga?auggacugaa?ccgcguucau?cguaucgauu?caaauggggu?ggugucacgc????4920
aacuucacga?gggacuacgc?ccccugaccc?cgucacgcga?cgagaaagcu?auggaaaaga????4980
ugcgcgcgua?uggcuuaagc?gacgucgccc?aagugaucau?ccgaaugcgu?caaucccacc????5040
cccgucacaa?cgccgauuca?gugcgguucg?uccgugaugu?acucagccuc?acaaguggca????5100
uguaccuugu?ucgaccucca?accauguccg?uccuccgaga?guacucccaa?accccccaga????5160
uugaagaacc?cauccccccu?gacuggugga?cuggagcugu?ugguucacuu?gcuuacuuca????5220
acgaacgcgc?caaaggacca?cucucucauc?ucuacuccgu?guggcuggaa?gcugcccgua????5280
aggucguuau?ggaucccucg?acgcaugacc?cucugacaca?agccaucuac?aagacucaau????5340
uuguuacccc?ucgcggaggc?uccagcgccg?cccucaagca?agcccuagcc?gagagcaaag????5400
uugaguugcc?cgacuucacc?aguacuggcg?ucaaacgauc?cucuaagauu?uaccagaccg????5460
cccaacuugc?ccaucucagc?uuccaagcuc?ucauuccagc?cauuaugggu?caagucacuc????5520
ucggaauuag?gaaucaagug?caacgucgcg?cccgguccau?caugccuaug?aguaaccccc????5580
agcaaacugu?cucgguucca?cacacuuugg?uggcuaauua?caucaacaaa?cacaugaacc????5640
gaucaaccac?uucugguagc?gccguucagg?auaaagucau?cccucuccuc?uuguacgcuu????5700
cgaccccacc?gaggacuguc?aucaaugucg?acaucaaggc?cugcgacgcg?ucgaucacau????5760
acgcggccuu?ccucgccccg?aucuguggcg?ccaugcauca?aggcuucgac?cucggugacc????5820
cauccgcucc?guucaugaac?guucccucgu?ccacccagua?cgaccgucgc?aauccugagg????5880
ccccauacaa?ucguccuguu?uccggucucc?agacaaugac?ccagcaccuc?gcgaaacuuu????5940
accaagcugg?cuucuccuac?aaggucgaug?accccuucuc?caguggcaac?agcuucgucu????6000
ucccuaccac?caccuucccc?ucagguucca?cugcuacauc?uaccgagcau?acugcaaaca????6060
auggagcuau?ggccgacuac?uuucuccgcg?aguacguucc?gcaacacgcc?accuccagca????6120
cucugaaauu?caucguuaaa?gacaugacca?uucagaacaa?cuaugucugc?caaggugaug????6180
ucggaauguu?gauccuucca?gaucucggca?cuaagagagu?cuccccugau?gaccuggccg????6240
aguugauguc?auuguuagag?aaguacggac?gcgguuucgg?cuggguuuac?gacauugaca????6300
guuccgauuc?agcugaauau?cugaaguugu?acgcccuguu?cggugcacgu?auccccaaca????6360
ucagucgcca?uccccccguu?ggcaaagagu?acgcuucccc?ugaaacuggu?gagaucuggc????6420
caucccucgu?gaacaucgcc?augggcuccu?ucuacaacgg?cgucacugac?ugccucgaau????6480
ggcgugauug?guuaagguuc?aguugggcau?ucgcauguuu?cgccucccgu?gguuccuucc????6540
auccgaagau?ugguccucgu?gucgacgcgc?aguaccccgu?gugguccuuc?auuuacaugg????6600
gcuuaccucc?aauccuccuu?ccuggccaga?ccccguuccu?uaccucggua?uacaugccug????6660
cgggagacca?agguauuuuu?gccaucuuac?accaguggcg?cgacuauuua?accgcccggg????6720
ccacugccga?auauccacca?cucacgcgcc?gccaccccgu?cuggcaguug?gccgacguuc????6780
ccucucuucu?agcugaccuu?ggcguuuacc?guggcuacug?ggcugcucag?gucucccgcc????6840
gacccgaacc?cucaccugau?gacgccgacc?cgaccaacgu?ggaagcgaug?agugcagcuc????6900
ucuccacuua?ccuucucaaa?gauccagugc?uccgcgaccg?aguuguucgu?gguacuaacg????6960
cauggcggcg?ucucaccgau?ucccaacccg?gucgcuuacc?uucucgcgug?cccucccuac????7020
uugauguccc?cacucguugg?auuaaagcug?gacgugacgc?ugaaaaaccc?agacccucgg????7080
cuguugccau?gaugaugaag?gacauucaac?gcgcugcuag?cucuucacgc?aaggacuucu????7140
cucgucuucu?ugaacuguac?cugcacgucc?auguccaccu?uggaccaccc?guaccccucg????7200
cuguugaucc?cgaaguaccu?cauguugcug?gugcugacau?ucuuaacgac?gaccacuggu????7260
auaaagugac?cucccucggu?cccaucgcuc?aauccaccaa?gaaguguuuc?gacgcuaccc????7320
uuuucgucgg?aaagacuguu?ucaggacuag?acguugaagc?cgucgacgca?accuuacucc????7380
gucucucuau?ucuuggcgcu?gaaccugaag?aguaccacgc?uuucuuagcu?ggcauuggca????7440
ugucugacgc?ugaagcucau?cgcauugcua?gcgccaucuc?ucuugcagac?gcucagauug????7500
uccaacucgc?ccgaaccguu?aaucuggccg?uccccucauc?cuggauguca?cuagauuucg????7560
acacccuaau?cagaucacac?ucauacccac?gucagcccgg?uaucagcgac?uccucuacuc????7620
ucgucaggga?acgcgcgucc?uggauaaauu?cgguccuucg?acucuuaugc?gccacugucg????7680
ccaugacucg?aguuggaccu?guaugccaag?cgacuguugc?aaguguugau?ggugguguga????7740
accaaaucgu?cggguguuug?cgugccugga?ugcgggaugu?gugagccgug?cggcgggcag????7800
ugguacaauc?aucguuuuau?caucacgcau?caugccacgc?accucgcgua?acguacgcgc????7860
caccgagguu?gcuacuacug?ccauuccucc?uaccaaugcc?gccacugacu?ccaccgucga????7920
cacuacuacu?gcuccaacua?uugcuagugc?agaugcagcu?caacaugcuu?cacacaucac????7980
aucagcucag?cuuggagcag?cuggaauagc?agacaaaguc?cccucauccg?ucgucacuaa????8040
cgauggagac?auuuccguca?ccccuauauc?cgagaacgca?gcucagcuag?cgucuuccaa????8100
ccaacccgcc?ucagucaucu?caaacccagc?aggagccgcc?ucugugucca?uugucaaccc????8160
auccgcuuua?cgcugccaac?aauguggugc?cgaauucuca?uccaugacuc?agcuggcuga????8220
acauguccgu?acugaacauc?gaacuggugc?agguucacuu?gucacaucgc?cagccaucaa????8280
ccaagcgauc?gaguccuggc?uccucaccug?ggaagaucuc?cgucuucuag?cacccaccau????8340
cgccaccgau?gcccucaaca?aguacauggg?ugaaucuguc?gccaaggcgc?cccuucugau????8400
cauugaggac?ucaggucucu?gcacauccuu?ccucgcuuca?gacaccgucu?caaucgcugg????8460
ucugcaacgu?gaacucgucg?gauucacaug?guucauggag?acccuccaaa?ugauccccgc????8520
guuacccgag?ggugccguua?aucaucuuau?cugucacacc?gguugggcau?caaaagacuc????8580
cgcaucccgc?aaccuaaacg?uuaggcuauc?accacccacg?cacggugccg?ucucagcaua????8640
caccacagug?cuguccaaag?gcuacgugaa?agauaugcag?uuuaaucucc?auaccuucag????8700
agcuaacgcc?cucaugcuua?gccucaaguu?cgugcugucc?aaccucaaga?uuaacaaguc????8760
cacuccccua?acccaagauc?agacuccacg?uucccgaggc?cgcuacauca?gauuccauga????8820
cgacaaggaa?uugcucgcug?uugcauaccc?cggucgugaa?gugcucaugg?aggccaaccg????8880
uaacgcccuu?uuccucgaug?aagccauccc?agaucgcguc?ggucguauug?gucgugcuca????8940
gaacguuucc?ggugauguca?gugcucacau?cgacaccuac?gagcuuugug?acgaccuaac????9000
ccuggccauu?cgggagaugu?aucacaacau?gcuguucagc?augcaccugg?auccugcguc????9060
ggugauggaa?auuguucagg?augugucuca?gcaacugguc?gccgccucca?uuccauucgc????9120
ucaaacugac?accauucugu?gccccugggc?ugcaucaucc?ccuacucucc?agcuuggaca????9180
gguucugaac?cugcugaaug?uggcuaacaa?uacaucagcc?gcucuucccc?uuaucgaagc????9240
cgccgcuacg?cucaucaugg?gcaucacccc?acuccggaug?gaaccuagga?uuuugucaga????9300
agccaucaaa?cguguaccag?agaccacuac?caucgugccc?ucucccacug?gugagcucac????9360
acgucuacua?aaaccguugg?guaaugacua?uucugccuuc?uaccgaugca?uugcuggcug????9420
gcuguauagu?gguaucgucc?agacuuucau?cuccgcggac?ucguacccag?aucccacgca????9480
guccaucacc?agcauuccag?ccauuuggaa?gucucucauu?gugacguugg?ccgcgccgau????9540
gacaacagac?ccccacgcgg?cugugaaggc?cuucaugucc?auggcaaauc?ugcuggccca????9600
accagagccg?aucauuaucc?ccgcuccugg?caugacucag?uccacgcccg?ccguucaguu????9660
uggacacccc?gaaguguggc?caccagguuu?uauugauccc?accaccuugg?aucggaaccg????9720
gaccccccug?cuucacgcuc?ucgccaccau?gauccaugcg?cauuggccac?aaccuggagu????9780
uaucccguac?ggcagugcuc?gccuugguuc?cgccaaccug?uuccuccccg?ccaaucagcu????9840
ggcuuauccu?uggcccacac?aaccccuucc?ccgcaucacg?guagguccua?ccuaugacuc????9900
cgccauguca?cgauggauug?acaacguuuu?cggcuuuuac?aucaacgucg?uuaauucccg?????9960
auacgucgca?acgaucguag?gugacacuac?ucgucggacc?cucauuggcc?ugauguccgc????10020
cuuacgacag?gugaagacga?ugacuccauu?cuacaucgag?cguaugugcc?ccacugagau????10080
cgccguugug?gguggaguua?cagucgugcc?cccguuucaa?gugccauucu?cacgguuaga????10140
ccccgaucag?gucaucacca?acgucauggu?uucgcgcguu?gacccccaac?uucgugcuga????10200
cgucgcaguc?gaccccaucg?ucaccaugcc?cacccuggcu?aauucccuuc?caguggaccc????10260
agcggccauu?gucguugcca?ugcugugcgg?ccagacagac?gcaacucucg?ucccuucgua????10320
ccacuaugga?ucggccauca?cgccaauguu?ccuguccgaa?ggaaucuuca?cacgaaacca????10380
acgcgccguc?aucgcaagug?aagccuucgu?gugcgcucga?ucaaucaucg?cccagugcau????10440
ccccgauggc?uuucagguac?cccggccucu?gcaagccuuc?aaccaguaua?acgcgucugg????10500
gagcaccgcc?gccgaucucc?ucaaagcagu?ggacgauaug?uucaaaaccg?cguucgaccu????10560
ugacggcucg?uugaucgagg?gcauuggauu?auauggugac?ccacgugucg?cagaucuauc????10620
gguugcauac?auacgucaaa?acggugcugu?ugaacggguc?cacacagcuc?cugacucauc????10680
cuucauucac?gaagcgaugc?aagugacuuc?ucagguuaug?aucaacgaac?cgaaccugug????10740
ggcaaucgcg?agaggugacg?ucauucuggc?ccagaacgcg?accaacaaca?auugggaccc????10800
caugaacccg?gucggccuuc?cccuuaucgu?uagaggcgcu?ccuggcguac?gugucgucgg????10860
ucagcauggc?augaucaucc?cucaaccugg?uggcuucuca?cccaugauua?gagacgaaac????10920
uggcaacccc?cagcccaucg?acggugauug?gaucuauccc?auuagcguuc?uccaaguuuc????10980
gguagccaac?uuccgugauc?acgucuggcc?caugauccaa?acuggccgca?cgcgcguacg????11040
caucgagaug?ggccacuuuc?ucuauuccau?ccacuaccac?gaacccuucg?gucagaucac????11100
ugaagccccc?gcuuuggaug?ccuggcuugc?uggcauuucc?ccgaccggug?ucccaccuuu????11160
cccguucagc?gcaccuaucc?cccagaucaa?cauucccauu?acagcgcgac?gcguguacuu????11220
cgguuacugu?acuaugaaca?acaacggagc?aaccuucucc?acucugggag?ccgccauuca????11280
guccgccugg?gguacugaug?uaaccaucca?acgcaaucga?uggccagcgc?ugauugaucc????11340
cgccuacauc?ccgggccauu?cccaacuucc?cgcucguauc?caacuguaca?aucccuugcg????11400
ccgcuacaac?uaccgcuacc?ccgugcugaa?gggcauguug?uauauccccg?guguugagua????11460
agcgugcgug?ccgcgcgccu?ggcuaguggu?gauuuucauc?guuuuaagug?aacugcucga????11520
gaagauggcc?gcccgcauca?accuccgaau?gcucagugcc?gacucuaguc?uuacuacaaa????11580
caaaccgucu?accccuuccc?auccuccuuc?cgacaaccca?uccaccuccg?ccgcagccgg????11640
cuuccagucc?cugccuaacc?uguccuuccg?uuccccaccc?accugguccc?ucaccuacaa????11700
agguguugcu?uuccauggcg?ucugugaccc?uccuugcgag?ccuuucaucc?ccauugcugg????11760
cuaccuuagu?cagaugauau?cgcacaugac?cccuuccucc?gaaccccauc?gcaucgagga????11820
ugucaacaac?cugguugccg?uuggucuuuc?ccagcucggu?guaacaccca?gcaugucucu????11880
cggcgaagug?gaaguccuuc?ugaaugaucg?uguggcucgu?guccaugacg?gaucuggccc????11940
uuucuucgac?ccuguccccg?cuccgguucc?ugcuccucuc?uccccuguuc?cagccgcuac????12000
ugucucuccu?cccccacccg?caccucccgc?aucucuggcu?ccggccguca?ccucccuuaa????12060
cccggcucuc?gccgcagccc?ccacccucaa?gcccuccucu?gucccgacug?gcacccagac????12120
ccuccucaug?ccacuccaac?ugaccgcugc?caccucuacc?gcuacaccga?cugccucccc????12180
uacugugacc?gacgcucucu?ccaccacccg?ugcugacugu?ggcccgacau?ccgugaacga????12240
acccuccacc?cugugguccg?aaucugcucu?ggaugaauuu?gauggcccug?cucacuguuc????12300
ucucgcuacc?aauccaccuc?ugauugaugg?ccgccuguac?ucccaagucg?uucagccccc????12360
cgucaagccc?gucgaccagu?ucaucgugug?ugacucacgu?cgugugucca?uugguuccac????12420
cuccaaggcu?uccucugucg?cuagcgagac?cccggcauug?auggagcuua?augucacccc????12480
agcuucgacu?gguccccguc?guccuccucg?cccugccccc?ggccccuucc?caccucggac????12540
ugguagacgu?agggcccacc?auaucgcuga?cgacaacacg?uacaaugagg?cacguuuggc????12600
uuacgcucgc?ggucgaccca?cuacuucccc?ucuguacaac?caagguuugg?agauucacuc????12660
cgagcgcgcu?uucuucuccu?ccuucccuca?ccacaucaac?ggcaaauggg?uugcuccuac????12720
aucugugcuc?aacgucguug?cuggugcuga?ugaggaucug?aaugacucca?ccauccacuc????12780
cguccgcgcc?accgacugua?gugguaugua?cgaggucacc?gcgaccaacg?gcgaagucau????12840
cucccgacug?aacgucaugu?ucaucgacag?ugacaccacc?ucacaggcuc?ugacucacuu????12900
caucaccccg?cauucacuca?uugcuauuac?uccuuaugcc?gccgcccuca?ugguccaguu????12960
uagacucacc?aaaggcgucu?ucuccaaguc?acaccgacgu?auggucaugg?ggauugaucc????13020
agugaugauc?caccugaacu?cgcguggagu?ugcucucugg?aacguccuua?cucagcaucu????13080
gcuggaguac?gccgagcuau?acgccucugc?cagucugagg?gaucuuguca?ccgcccugcu????13140
cgacccugug?aaugucacca?ccaugcgaug?gaugaagcgc?acgcuuccca?auugugucgc????13200
agccgugugc?gagaugcgua?ccgaucccau?ccccacccug?ucccacauuc?ugaacgucga????13260
gacccccacc?acgccuguuu?ccagugccgc?cagcagccuc?aaguuguccg?aacuucaagc????13320
cgagaauacc?gcucugacau?cccagaucuc?aaccuuggag?gcccaacuug?aggcagcuac????13380
ccucucucuc?gccaccaccc?gcgaauccau?ccaacgugag?caggcggugc?auucagccga????13440
cucccuaaag?caguaccucc?augaccacgu?cugugugaac?ugucaugagg?aaccauuccu????13500
uaacgcgacc?guuggcgucg?aagccgcugg?ccagauucuc?ucugcccguc?agaaugcccg????13560
agagcgugcc?gcugauaaag?uucgucaggc?ugucagugcu?gguuucgccg?agacugucac????13620
caugcuucaa?gagcguaauc?uuucccugga?ucaacguguc?aaagccaccg?uagacgaauu????13680
gaaugccacc?gccacacaac?uccgaucugc?acuuagccgu?uugucguccg?cagagggccg????13740
uucccaugac?uugguucaga?gaaaugagcu?gcuugaaucc?caacucgucg?aagcccgcca????13800
gcucucaucc?uaucaagaca?gccaccagaa?ggagaccauc?accgcucucc?agacuacuau????13860
ucgagacagc?uuaccuuaca?ccccagucca?cuauggacaa?ggccaauuca?cuaugcccuc????13920
ccuuccaccc?guccggucca?ugaugccuga?cuucgacccg?ucagaccuuc?ucauguaguc????13980
acuuacucgu?uaucgaccuc?acgcucugcc?ucccgaccau?gauuuaagcg?cgccgauccc????14040
cacccgcccg?uauucacccc?auucccuuuu?cccaggcuaa?guccacuggc?cuucucuggu????14100
uguugccgac?ccucucuucc?gaugcuucac?ucuauucauc?guuuuaugca?cacgcgccag????14160
gaugauuacg?auugucuuca?ucccagguca?cggcuucaac?uggaacgacu?ccaccccacu????14220
uaaguacaua?gacucccgaa?uauaucgaac?ccgcaucccu?aaagauaccc?uuacccuauu????14280
cgcucccgcu?ugguucaaau?cccagcucga?acacuacguu?ggaacucacg?gcauugagga????14340
aguuuacuca?uggugccaac?gcuugaucag?uccccuuacu?aaccgauucg?uccuccuacc????14400
ccgaccuaag?ucauucgcga?aauggcucuu?guccacuccc?uccgcuaaca?uuugggauau????14460
uccucguugg?aaacucgauc?ucgcugcugc?aggcaaagcc?cccccggauc?uuuacgaugg????14520
aauacaucca?uuauuagguc?augacgccac?cgucaccaag?ucugucucgc?ucaucgcugg????14580
ucaucccauc?gucuacucac?guacacguca?ugucuuuggu?gguccucuuu?accuugccac????14640
cgauguuucc?gccuauucug?guuucaucaa?ucagucagcc?uuggacgcca?ucuucaagca????14700
cgacgcugac?cuuccguccu?cucgccgcuc?agcuguucau?auuacuaucc?ucccaaaccu????14760
gaccaacucc?cgcuccuuua?ugcucgaucu?accugacuug?acaaucgacc?cagacuaucc????14820
acuuuccgcc?uuucaugguc?auuuaacacg?uguaggacag?aacacuaccc?ggaugaugcc????14880
ccuagauggc?cuaugcuggc?gacugacacg?ugguucggcu?aaaccaacgu?ggacuccaga????14940
guucgaugag?gccuuuagac?uccuucgacu?uucccguccc?gcugcaucgg?augcucgccc????15000
uaacuuugga?accgagacgg?uguugguuca?uguagaucuc?acacuaaacg?ucgacacccg????15060
ggacaauucg?gcuccccgcg?cuccgcuuca?cguacacgua?cugaaugugc?cgcucgccua????15120
cuugacauug?auggacuuaa?aacuuucaca?augcuaucca?cuccguacag?aagaugggaa????15180
caccguaccu?ugguuccucg?ugcugguccu?gcuauccgau?ggaguccaac?uggcugguac????15240
caagagacca?guguuacuac?agacaucgau?ugccgaguua?cagcccuggu?gggaagugac????15300
cuuaaaugcu?uuccucaauc?cccaugccgu?gacggucaga?uccggaguga?uuaaagacau????15360
caugggagug?gcccucgccc?uacccaaagg?aucguauaaa?uccacguuca?ucgacgugau????15420
cacuaagaac?uugaguaacc?cggacgccau?cuucccucag?gaaacuguca?ccgacucgga????15480
cgauuuaggu?gacucgcucu?cccccaguuu?cgaaaaucag?aucauggaug?uguggcaugc????15540
ucuugggacc?gauguguugg?agcaaggugu?ccgcgccauu?cuaacucccg?gugccuacgg????15600
cgcuguguuc?cccauugaga?uauaucagga?guucuccaag?uuguaucaug?acguuaugau????15660
ccccgcacaa?cgugcucgcg?cggcuuucau?cucccaacgu?ggcaggucac?uuguuuacgu????15720
ccacacccca?uacgagauag?ucucugccaa?cgucccgaug?cagguagcuc?cuugccaaau????15780
ugcccucgau?ucaaugguca?acguucucau?ucgucacaag?cguguuggcg?gugucacagg????15840
ucaaguacug?cucgaccauu?gcuauagauu?gaugggugcg?acaacuacuc?cgcaacccgc????15900
cggacuauac?uaccggucuc?uuuuuggccc?auggcucgag?cucgcggccc?acccaacguc????15960
cacggucccc?aucuggcuag?agacagaggu?auccgcccau?acucuacgug?aagucggcug????16020
guccguugac?ggugaugagc?cucuucucau?uaacauccuc?gagggucuug?uuccgacaga????16080
cuccgucuuc?uuggucaagc?ugccccagcg?uguacccuca?cgugcauccg?ucgucguaac????16140
cgcuguuacc?gaccuuagga?ucuccuuauc?cccacccuua?cccacccgca?ugguucacuc????16200
uagcguacug?cuccccguua?caucagucgc?ucgcuucaug?gcgccuaacc?gcauucuacu????16260
cgccgggacg?ucgaucucag?uacguggucc?ugugaccugg?augacaacgu?cgucgcccgu????16320
ggcggagagc?ggaucagggc?cgucaggucc?gugaaccccc?acgagugugu?gcugauucau????16380
cguuuuaucu?ccuugcgccc?uucucaauca?ugggaaacgu?ccaaaccuca?acuaacgucu????16440
acaacguaaa?cggcgauaac?aaugcuuuca?caccaacuuc?cgagauggug?gcgucagcuu????16500
cgcccgcuau?ugaccucaaa?cccggcgugc?ucaaucccaa?uggcaagcuc?uaucaacuug????16560
aggcuggcuc?agcuccagau?ccuaccaacu?uaauuuuggu?ggucgacgca?ucagagggug????16620
acuucucgua?ccucaccaac?aacacauggg?agacacucag?caaaucggcu?uuggagacca????16680
acuccuggga?accaauguuc?agcgugacga?ugaccgguug?cgguccgcuc?aaauugggug????16740
acuacacugu?gacgaugucc?ggcuacgucg?gugccucucc?aagugacgcc?uucgauggug????16800
gugugaucga?agauggcuca?uucauuucca?gucgaagacu?gaagaacuuc?aagcugaugc????16860
ucuccaacag?gugugaagcc?aucgcaaagu?ggaacauguc?cuugaaucaa?gccaugucac????16920
uacugacucc?cgaccugcuu?gcuggaucug?guuccuguaa?guggaaaucc?gucuugcagu????16980
acaugcagaa?agugcuuccc?ucugacaacg?aggugcuuca?guacccagau?gaguucuaca????17040
ccguggcagu?cggcaaguac?cccgcgcuca?agcccggauc?cucgcccgau?acucccccac????17100
cugcugcagg?accccuuggc?gagauagcau?gugucaugaa?cgcagccagc?gccuccguug????17160
gucucaugag?uggaucaagc?gcacuccuca?ccuccgccau?ggacacccug?gcugcaaaga????17220
aucuggaucu?uguaugcgcu?gaagccccgc?ugccugucuc?caccuucacc?ccuucgcuag????17280
cgccacggga?cuacaggccu?gccuucauca?aagaugcgga?ugcccauugg?gucacgucca????17340
uaaccccgac?gacguacuuu?cgcgugacga?ccacccuguc?cgccaaaaac?uacucggucc????17400
agcucggccc?aggcgcgacu?aaaguuuugg?acaugaaucg?cauggucgac?ucagaucucc????17460
ugcucgaugu?caguggcaug?cccauugauu?ggaugucuaa?ucccgacuac?gccaccucgg????17520
ucgccgccau?cguccuguug?gagucacgcg?uuccagcuuc?agagaucucc?gccgcugagg????17580
acaucacugg?cguuuccauc?guggcaagcu?cgccgcuuuc?aaucgucaau?ucgaccguga????17640
acgugcgggg?acaacacuuc?cuggagaugc?uccacuuaag?gaccacuuuc?gaacgugaaa????17700
ccaucgccgg?aaagccauac?aucuaugguc?ucgggacucu?guugcugcug?ucccccacca????17760
ccgcgucaaa?cucacggaac?cccacucuga?uggauggucu?gcuaaccauc?acuccaaucc????17820
uccugcguga?cacgaccuac?aagggcgaaa?ucgucgagga?gaucgugccc?uccgacauuc????17880
ucggcaacca?cacuucagaa?gaaauggcug?uagcguuagc?aaacgaugcu?gucguucuga????17940
uggagaauag?cuugaaggaa?guggcugaag?ugaucggaaa?cgccgucccc?aucgcuuccg????18000
aucuugauga?cagugccacc?gcaucggucg?uaagucgucu?ggccaucacc?gagaccgccu????18060
cuacgcgcuc?acgauccaac?aauccccucg?cauucccaga?uuucggagcu?cucuggaaga????18120
aagccaagcg?cgccgccuca?cuguucgucu?cuaauccgaa?aucuguacuc?caagucggug????18180
ucccaguucu?cgcauccgcc?ggggugaucg?augcacucac?gucagccguu?ggcacaucug????18240
ucaggacggg?caacauugga?aaaggugucc?aagaugcucu?aucuauucuu?aaagcacgaa????18300
auagugugac?gaaguugagg?cagggguucu?ucucaaagau?ugaggagcuu?uggccaguac????18360
ucgaaggcua?gacguguggc?uuguccacgu?cgcgcccuag?agcccucggg?ugacgccgag????18420
gggggauauc?cauucaucgu?uuuagucaau?cauccugggg?aacacuaucu?caagcaccuu????18480
ucaguacacg?guacugcaga?ucgacagauc?uugcuguauc?aaaaccaguc?ucaccgccac????18540
cuccgaagcc?acuuccuggg?ccauuccucc?acucgcgauu?uguuguugcu?guugccucug????18600
uugcaccggc?gguuuauacc?ucguucacuc?uggacguauu?ccaagcauca?gccgaagguu????18660
ggacgugcuc?agagauaccc?ggucagccuc?agaauacaag?gugcguagca?accggaaccc????18720
aaagucacgu?guacgucgcg?uuagcaucag?ugauucuagu?gacucuagua?gucucucuga????18780
ucuggaguug?ucucggcacc?ggucucaucc?uuuggcgcau?ucauucaggc?cugagagcua????18840
uagccaacgc?ccucauuccc?ccucgcaagc?ccagucgucg?auuauccucc?cgcucguacc????18900
uguccacucc?cgaacaaguc?uagacgaugg?agucauucgc?ucucaacccu?cacgguauca????18960
gggaccccac?cagcaauuug?aggauuggcu?ucaacaagca?caucuccuac?gaccuggaga????19020
cguaucccga?gauaccaacc?ccuuccguug?acuuuauucc?ggaaugcguc?ccauccaccg????19080
aucgcuacaa?uggugauccc?gucccccuga?ucuacgaugg?ucgguugacc?ccaguuaccg????19140
guccucauca?ccuaugggaa?auugacaguc?augucgagug?gcagaccugg?ggcaaucucc????19200
guccauucuc?cccauucagc?guguggcccc?cauccacccc?uaacuggacc?aacaggaagg????19260
ccauccacgu?guucagcagu?cugucaccgu?augcguaugc?cgcugagcgc?agccaaaauc????19320
cacugucgua?ucacuuccug?aacgaucagg?gucgugacug?gggucgcuuc?ugggauuuaa????19380
uuuggcgaug?ugcucagacc?cguggcgcuc?guaucugcca?ugccagcacu?ucauucaucu????19440
ccuccaugcu?gcggcugacu?gaggaccagc?ucucaaaacu?cccagcggcu?cgggacccaa????19500
ucgaacugcu?caaugcugcu?ggcugggacg?cucuggcacu?aaacgcccua?ccacccaauc????19560
ugucccgcuc?acugaugaga?uccccuccaa?accccuccgu?cgucgucuuu?gagugucuga????19620
cggauugguu?ugacgucaug?auucguguuc?cauaugacgu?ccaacauccg?cucggucucg????19680
gucucaaccc?gugccaauuc?uggacucacc?ccuuugucgu?ucugugcuau?cugcgcugga????19740
ggcuguuggg?aggugaugac?uaggauggcg?accgcgacag?uugagaccuu?ggucucgggg????19800
auuuaguccc?cugccgcagu?cgugacuguu?auucaucguu?uuaucaucua?uggcgcgucg????19860
cacuuuugua?ggguucacuc?cgagcuuuua?cggccaaccu?ggcccuuuau?ucacugacaa????19920
cgauuaccuu?gaccucgcug?guacgaccgu?ccgcccaugg?cagaaccgca?ucacaaacau????19980
ugcgauuaac?ucuggugguu?acccgguagg?gggaggaaag?uaucccacag?ucgccucacg????20040
ccagcucauc?cucaacgcgu?ugcuuggagc?ucacguaucc?ccuuacgcag?caggagcugu????20100
uaaccaguuu?aauggcauug?cuuggagaga?cgcaccccuc?aguucauucg?ucaccauacc????20160
accagcgguc?gcccccgcgc?cuccagaucc?uccaauaugg?gucccagcug?agaacguacg????20220
ucuugacuca?aaucaguacc?cuacauacgg?gcugaaguuu?gaugcuaugu?ggccucaaaa????20280
ccaagaccuc?cacaugauga?cgaugugguc?acugacugau?cgcggcccga?uagccaugcu????20340
caccuucccc?ucccaaaaca?ucccagcaau?ggucgccacc?uccaugaagu?cccugaucgg????20400
agccuccgug?gcccaaaucg?ccauucgugc?cuaucgcuac?aguggucagc?uuccacagga????20460
aggcgugucu?auucaagccc?aagucuacca?auggcuugcg?ugcauucucu?ucggcucccu??20520
aaccggccgg?uugcaucgug?ggcgcacuug?ugaagguuuc?uucuuugcgu?acucgaaacc??20580
ugccgcaagc?caagaugaca?ugauccuucg?guggaaugau?ggaccaagag?cgagaaaacc??20640
cgucaaugac?gucaccgucu?acgucgccgc?uggcucaccu?cacuggcagc?aguccauguu??20700
gcaugucucg?uucgcuaugc?ucgcgcaguc?caccuccugu?uugcgaccga?uggcgacauu??20760
gauacgcaau?cguaaccucc?cagcgcgcuc?ucacaauaua?gccggacuga?cuggaggugg??20820
uggugcuuug?cgagaugugg?aaagauacaa?cgugccugau?cucgcuuugc?agugucaugc??20880
cgcguggcuc?gcugacggua?ucauugacgc?uggagcgcug?gcugcguacg?augcugcuac??20940
uaacgcucag?uuugucaacu?ucauggcgca?cauugucgcg?acggaagcug?ccaauccacu??21000
agcucguggu?cggauuauug?ugcaaccauu?cgcggcugga?gaugacgugg?cccccuuuga??21060
aaccgcucag?gugauugccg?gagcgcgucg?gauguuccuu?uaggugcguc?ccagggggcc??21120
aggacaaaac?cccccugcca?agaugacauu?caucguuuua?gagaucaccc?ggaguagaca??21180
uggcaacccc?aucagcuaaa?cucaucaccc?aagagaacac?cgagcguguu?acacgcuugc??21240
ucgagaacua?uccgacauua?agucugaccc?uccgacagga?caccuccaac?cguggagcga??21300
uugaauccaa?cuacagugcc?ucuggucccg?cugguucccu?gagacuccug?aaccccaucg??21360
cugugaacaa?gaacccacau?cagccuggcu?acucucgccc?ugaccccgag?gcuacacgcc??21420
cucccccccu?gcgcacccug?aucaacgcug?gccuugaugc?ugcucugcaa?cauggcucuc??21480
agugcccagu?ggaccaagcu?cugcguguau?ucaucgagaa?ggccugcccu?cauuggcacg??21540
cugagguacu?ucccaaugac?uggacucgug?ucugcccccu?gacucuggcg?ucucgcauca??21600
gucuugcugc?agccggguuc?gaccccaaca?cucuugaccu?ccaggcgcca?ucauccacau??21660
caauggucau?cgccuacacc?uccaaggugc?uuggccuaug?ugccgaucug?gccaacucau??21720
cccucggaua?cuuccagaca?uccgcugcug?augcuauccg?caaccccgac?ucacucgccg??21780
ucgucaucac?ucaguauggu?uacgagacaa?aagguuucca?gcgacaggau?gcccccaccu??21840
gcaucucccc?uaacgaccug?gacccuaagu?acgacaucgc?cuggcugucu?gccaucguga??21900
uccugaucgc?cuaccagcug?gagcucgacc?ugacugcagc?aucacugucc?accacugacg??21960
ugaauuccau?guccacucac?aguaccgcug?ucgagacucu?cauguucaag?augaaguggc??22020
uagcccccuu?cuccuccagg?aucaugcauc?ucugcgccgc?uaaugcggcu?aacccauucc??22080
gcagcuucag?cgagauguuc?uugauguggc?agaaacccac?gaaguaugug?cuucccaaca??22140
ucaccaugaa?acugucaggc?agguuccugg?agguuaucgc?ugauggcucc?gaguuguuca??22200
gugucuccag?cucgcgggug?ggagguaacu?aggcacggcc?auaggccuac?ugccucauuc??22260
ucuuuauuca?ucguuuuaua?aagcacaccg?guaaacacca?uggagaccaa?accaauucuu??22320
ccaacuauug?cgacccaguu?cugugacuca?cucguccgcg?gucauuuauc?cggagcccau??22380
auugauggac?aaugguccac?ccaccuggau?gaccccgaac?ugcuaacuga?cggcgguuac??22440
aucaucugug?ccugcugcuu?caaagugcug?cugaauuggg?augggccaac?ggcauacauc??22500
acccacgagu?gucacgauuc?ccauggagcu?cgucgcaccg?gacgccaccu?cgcaggaaac??22560
cugcugcgca?ugcaagacac?aauccaacgc?acuguucaag?acggguuccu?cgcgcugcgc??22620
agccgagacu?cacuggccaa?agccgcaucc?ggugccggua?augagauggc?cgcugaggcc??22680
uucgaagaca?ucaaggaaac?caucaagaag?gcaagaaccg?guaaagucgc?caagguaaug??22740
agucuagacc?gcaucugcgg?ugcagucaac?ugugaucgcg?cucucacuuu?cuaugggaag??22800
gaucuggcag?accauccgcu?ugucacuggu?gcuaccgcac?ucaagaccga?gcuggaggag??22860
gugacuggug?agaagcuugc?gcgagugagg?cagaccgugg?ucgguggucc?aguugacguc??22920
aucguucaga?ccgaugcgau?gcccaucccc?auucuauacc?gaccggccau?ggaaaugauc??22980
gagccagucg?ucuccaaagg?ugcacgugcc?aucugggcuc?acgccauucu?gaaccagagc??23040
ugcacgaaac?ugagcaccgc?cugcaagcag?cgcgccuaug?gucgagucgg?gguuaaucug??23100
gccaaucucu?cucucuccag?cgccaacaag?agcagcgcca?cacguuguuu?caaggggugc?23160
acuccagagc?uguucgcgcc?ugugggugag?gauuaggucu?cccgguggcu?cgggauugcc?23220
caccccauug?ccguggcguu?uggugugcuc?cauucaucgu?uuuagaagac?aucggagcag?23280
gaauggcuca?ggacuugauc?aacauuuccc?aauuggccaa?caaacuuucc?cacaucaccg?23340
uuaacaaugc?agaacgacuc?gucaccggcg?ccgaacucac?cucccucgaa?acaagacucg?23400
agcaguccau?caaggccgca?aaguccagug?ucgaucguca?ccucacgcgu?cacgcugcag?23460
acccauuugc?ucaugagcac?acuucuggag?gugccccccc?ugccacugcc?uucaucccua?23520
cugucccacg?ccagagcuau?cggcuuucag?cuaugccagu?cgugcuccac?ucuugcgggg?23580
gccacagcuc?uuccgugucu?gugccgacua?agaucauugu?cgaggcaacu?caaugcacca?23640
uuacucucca?uacccccgcc?uucgacuucg?cccacguccc?ugccaccguc?cccggaggaa?23700
agcucuuuau?aaagguggac?aaagguuccu?ggcaugacac?cgccuucgga?gcuuucaacc?23760
ccgcugccua?ugaccagcug?cucacuguag?ccagacgucu?caaguccacu?gagguuaacg?23820
ucaaguggua?uggugguuca?ggcccuuccc?gaugcgaccu?gcgcucgaca?ggcagacgug?23880
cucaaaucca?cgcuacggac?aaccucauca?uguucgagcu?gccaggugug?gauuccgccg?23940
gugcauaccc?cgaccugacc?uguuggccaa?ucaugggcgu?auuugaguag?agucuugagu?24000
ccgccuggcg?uggccgcgua?gacccgaugu?cugccauuca?uc????????????????????24042
<210>2
<211>198
<212>PRT
＜213〉turbot reovirus
<400>2
Met?Gly?Asn?Thr?Ile?Ser?Ser?Thr?Phe?Gln?Tyr?Thr?Val?Leu?Gln?Ile
1???????????????5???????????????????10??????????????????15
Asp?Arg?Ser?Cys?Cys?Ile?Lys?Thr?Ser?Leu?Thr?Ala?Thr?Ser?Glu?Ala
20??????????????????25??????????????????30
Thr?Ser?Trp?Ala?Ile?Pro?Pro?Leu?Ala?Ile?Cys?Cys?Cys?Cys?Cys?Leu
35??????????????????40??????????????????45
Cys?Cys?Thr?Gly?Gly?Leu?Tyr?Leu?Val?His?Ser?Gly?Arg?Ile?Pro?Ser
50??????????????????55??????????????????60
Ile?Ser?Arg?Arg?Leu?Asp?Val?Leu?Arg?Asp?Thr?Arg?Ser?Ala?Ser?Glu
65??????????????????70??????????????????75??????????????????80
Tyr?Lys?Val?Arg?Ser?Asn?Arg?Asn?Pro?Lys?Ser?Arg?Val?Arg?Arg?Val
85??????????????????90??????????????????95
Ser?Ile?Ser?Asp?Ser?Ser?Asp?Ser?Ser?Ser?Leu?Ser?Asp?Leu?Glu?Leu
100?????????????????105?????????????????110
Ser?Arg?His?Arg?Ser?His?Pro?Leu?Ala?His?Ser?Phe?Arg?Pro?Glu?Ser
115?????????????????120?????????????????125
Tyr?Ser?Gln?Arg?Pro?His?Ser?Pro?Ser?Gln?Ala?Gln?Ser?Ser?Ile?Ile
130?????????????????135?????????????????140
Leu?Pro?Leu?Val?Pro?Val?His?Ser?Arg?Thr?Ser?Leu?Asp?Asp?Gly?Val
145?????????????????150?????????????????155?????????????????160
Ile?Arg?Ser?Gln?Pro?Ser?Arg?Tyr?Gln?Gly?Pro?His?Gln?Gln?Phe?Glu
165?????????????????170?????????????????175
Asp?Trp?Leu?Gln?Gln?Ala?His?Leu?Leu?Arg?Pro?Gly?Asp?Val?Ser?Arg
180?????????????????185?????????????????190
Asp?Thr?Asn?Pro?Phe?Arg
195
Claims (4)
1. one kind is separated the full gene of turbot reovirus, and its sequence is the nucleotide sequence shown in the SEQ ID NO.1.
2. isolating protein, its sequence is the aminoacid sequence shown in the SEQ ID NO.2.
3. the described a kind of application of the full gene of turbot reovirus in the viral detection of drugs of preparation that separate of claim 1.
4. the application of the described a kind of isolating protein of claim 2 in preparation inducing cell fusion rotein medicine.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104569391A (en) * | 2015-01-30 | 2015-04-29 | 肇庆大华农生物药品有限公司 | ELISA (enzyme linked immunosorbent assay) detection kit for grass carp bleeding disease virus antibody and preparation method of kit |
| CN110283883A (en) * | 2019-05-08 | 2019-09-27 | 湖南农业大学 | A kind of primer and method for unknown RNA mycoviruses genomic clone |
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2010
- 2010-04-27 CN CN2010101623459A patent/CN101864435B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 《BMC Genomics》 20110620 Ke F et al. Turbot reovirus (SMReV) genome encoding a FAST protein with a non-AUG start site 1-13 1-4 第12卷, 2 * |
| 《环境与健康学术研究讨论会论文摘要集》 20071001 柯飞 等 养殖大菱鲆中一种球形病毒的分离及超微观察 全文 1-4 , 2 * |
| 《高技术通讯》 20030930 史成银 等 大菱鲆病毒性疾病研究进展 99-105 1-4 , 第9期 2 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104569391A (en) * | 2015-01-30 | 2015-04-29 | 肇庆大华农生物药品有限公司 | ELISA (enzyme linked immunosorbent assay) detection kit for grass carp bleeding disease virus antibody and preparation method of kit |
| CN104569391B (en) * | 2015-01-30 | 2016-05-25 | 肇庆大华农生物药品有限公司 | A kind of GCHV antibody ELISA detection kit and preparation method thereof |
| CN110283883A (en) * | 2019-05-08 | 2019-09-27 | 湖南农业大学 | A kind of primer and method for unknown RNA mycoviruses genomic clone |
| CN110283883B (en) * | 2019-05-08 | 2023-03-14 | 湖南农业大学 | Primer and method for unknown RNA fungal virus genome cloning |
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