CN109402065A - One kind kind duck C hypotype fowl metapneumovirus low virulent strain and its preparation and application - Google Patents

One kind kind duck C hypotype fowl metapneumovirus low virulent strain and its preparation and application Download PDF

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CN109402065A
CN109402065A CN201810897411.3A CN201810897411A CN109402065A CN 109402065 A CN109402065 A CN 109402065A CN 201810897411 A CN201810897411 A CN 201810897411A CN 109402065 A CN109402065 A CN 109402065A
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刘佳佳
陈�峰
王占新
鲁俊鹏
覃健萍
操胜
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Wens Foodstuff Group Co Ltd
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Abstract

The invention belongs to microorganism animal virus fields, and in particular to one kind kind duck C hypotype fowl metapneumovirus low virulent strain and its preparation and application.Kind duck C hypotype fowl metapneumovirus low virulent strain of the invention, deposit number are as follows: CCTCC NO:V201823, nucleotides sequence are classified as SEQ ID NO:1, and amino acid sequence is SEQ ID NO:2 to SEQ ID NO:9.Kind duck C hypotype fowl metapneumovirus low virulent strain of the invention, by the way that S-01 plants of progress secondary culture to 50 generations of C hypotype fowl metapneumovirus are made.The present invention obtains S-01 plants of fowl metapneumovirus of time duck C hypotype for the first time and causes weak culture strain, lays the foundation for the development of Attenuate vaccine.

Description

One kind kind duck C hypotype fowl metapneumovirus low virulent strain and its preparation and application
Technical field
The invention belongs to microorganism animal virus fields, and in particular to a kind of kind duck C hypotype fowl metapneumovirus low virulent strain and It is prepared and application.
Background technique
The inclined tuberculosis of fowl is the infection of the upper respiratory tract caused by fowl metapneumovirus (avian metapneumovirus, aMPV) One of acute highly contagious disease.Fowl tuberculosis most starts to be separated in the turkey of young age, along with sneezing, tracheae There are the secretion sticked together, the clinical symptoms such as conjunctivitis in rale, nose and eye.Therefore referred to as Turkey Rhinotracheitis.In 1978 South Africa, be separated to aMPV for the first time, then also had been reported that in Europe, Asia and South America.China in 1998 have swollen it is comprehensive Report is separated to avian pneumovirus A hypotype for the first time in the chicken group of simulator sickness, 2012 again successively be separated to B, C hypotype in Ji Qunzhong. In July, 2010 is coughed what Guangdong Province duck occurred with duckling, egg-laying breed duck, and egg-laying breed duck laying rate is fallen to extremely In the duck group of feature, it is separated to one plant of C hypotype metapneumovirus, is named as S-01 plants, duck source C hypotype fowl metapneumovirus only exists France, Canada have been reported that.
Prevention most efficient method currently used for aMPV is exactly that the use of vaccine removes the poultry infected The different antibiotic of application reduces outside occurring degree and secondary infection, without more effective treatment method.Improve bio-safety Property and reinforce feeding environment management it is critically important to the generation of the disease, vaccine inoculation can greatly reduce disease incidence, can subtract Few economic loss.Fowl metapneumovirus is based on cellular immunity, currently, having been developed that many vaccine (weak poison for being directed to aMPV Vaccine and inactivated vaccine), but be only to be directed to the cell weak-toxic live vaccine of aMPV-A and aMPV-B hypotype to obtain in worldwide It is widely applied.There is the cross protections between cross protection, especially aMPV-A and aMPV-B between aMPV different subtype. But between aMPV-C plants of difference, cross-protection is less desirable between aMPV-A, aMPV-B plants and aMPV-C plants.Cause This needs to develop Attenuate vaccine for aMPV-C hypotype strain.
Duck source C hypotype fowl metapneumovirus only has been reported that in France, Canada, is directed to the aMPV-C hypotype in kind duck source Attenuate vaccine do not have been reported that also at present.
Summary of the invention
In consideration of it, it is necessary to provide a kind of duck C hypotype fowl metapneumovirus low virulent strain and its preparation regarding to the issue above and answer With the present invention obtains duck source C hypotype fowl metapneumovirus Attenuate vaccine for the first time, and weak rear strain potency is caused relatively to cause weak cell toxicant valence before It increases obvious.
The present invention is achieved by the following technical solutions:
A kind of kind duck C hypotype fowl metapneumovirus low virulent strain, deposit number are as follows: CCTCC NO:V201823；Depositary institution: China Type Tissue Collection；Preservation date: on 08 02nd, 2018；Preservation address: the Chinese Wuhan Wuhan University；Nucleosides Acid sequence is shown in SEQ ID NO:1.
A kind of kind duck C hypotype fowl metapneumovirus low virulent strain, deposit number are as follows: CCTCC NO:V201823；Depositary institution: China Type Tissue Collection；Preservation date: on 08 02nd, 2018；Preservation address: the Chinese Wuhan Wuhan University；It is described Low virulent strain reading frame includes N gene, P gene, M gene, M2 gene, SH gene, F gene, G gene, L gene, the reading The amino acid sequence of frame respectively corresponds SEQ ID NO:2 to SEQ ID NO:9.
A kind of preparation method of kind of duck C hypotype fowl metapneumovirus low virulent strain, including, by S-01 plants of fowl metapneumovirus of C hypotype Progress secondary culture algebra is 40-62 generation.
Further, the preparation method of described kind of duck C hypotype fowl metapneumovirus low virulent strain, including, by the inclined lung of C hypotype fowl Viral S-01 plants of progress secondary culture is to 50 generations.
Further, the preparation method of described kind of duck C hypotype fowl metapneumovirus low virulent strain, specifically includes:
With kind S-01 plants of fowl metapneumovirus of a duck C hypotype, African green monkey kidney cell (Vero) T25 that single layer is covered in inoculation is thin Born of the same parents' bottle, 100 times of diluted virus liquid 500ul of every bottle of inoculation；After absorption 2 hours, virus liquid is discarded, 5ml is added and contains 2% tire ox The DMEM culture solution of serum is cultivated 24~96 hours, until there is cytopathy；It multigelation 3 times, draws under poison disease vaccination A generation；So operation, continuous passage culture to 40 generations, then with after limiting dilution assay continuous purification 5 times, continuation secondary culture.
A kind of immune product of the duck comprising low virulent strain prepared by above-mentioned low virulent strain or the above method, wherein every part is exempted from (i.e. every is immunized Wei≤10 dosage) to the content of low virulent strain every time in epidemic disease product2.5TCID50；Xuan≤10 You4.5TCID50。
The invention has the advantages that:
At present there is no the reports of duck source C hypotype fowl metapneumovirus Attenuate vaccine, and the invention belongs to separate Attenuation training for the first time It supports.
The present invention cause it is weak after strain it is more stable, S-01-42P, S-01-52P, S-01-62P cause weak completely, do not occur not Stabilization.
In order to develop the vaccine of the effective prevention and control duck C hypotype fowl metapneumovirus of energy, this research is inclined by kind duck C hypotype fowl The continuous secondary culture on Vero cell of S-01 plants of Pneumovirinae, achievees the purpose that cause weak, lays base for the development of Attenuate vaccine Plinth.
S-01 plants of fowl metapneumovirus of C hypotype are passed through the continuous passage on Vero cell, pathogenic by different generations Test, strain has caused weak after 40 generations.Cause weak rear strain potency that weak cell toxicant valence before is relatively caused to increase obvious, by original 103.0TCID50/ 0.1ml becomes 105TCID50/ 0.1ml or more.The present invention causes weak rear generation (S-01-50P) immunogenicity most It is good, 102.5TCID50/ 0.1ml clinical protection can achieve 100%, cannot prevent completely toxin expelling, and 104.5TCID50/ 0.1ml faces Bed protection reaches 100%, and can prevent toxin expelling completely.Therefore by this research result selecting S-01-50P as weak poison Vaccine candidate strain (causes weak steady, in preceding 10 generation, rear 10 generation has caused weak), and 104.5TCID50/ 0.1ml both can achieve 100% Clinical protection rate and 0% toxin expelling rate.The sequencing analysis of weakening strain S-01-50P complete genome sequence shows itself and initially-separate Strain S-01-5P is compared, and there are the variation of 16 amino acid, the variation of these amino acid has very big with the variation of strain virulence Relationship.
Detailed description of the invention
Fig. 1 is morbidity duck nasal cavity.
Fig. 2 is duck nasal cavity of not falling ill.
Specific embodiment
The problem of solved in order to better illustrate the present invention, used technical solution and effect achieved, are now tied It closes specific embodiment and related data is further described.It should be noted that the content of present invention is including but not limited to following implementation Example and combinations thereof embodiment.
Particular technique or condition are not specified in the embodiment of the present invention, according to the literature in the art described technology Or it condition or is carried out according to product description.Reagents or instruments used without specified manufacturer, being can be by commercially available Etc. approach obtain conventional products.
The secondary culture of 1 duck C hypotype metapneumovirus of embodiment
Cause kind S-01 plants of fowl metapneumovirus of a duck C hypotype for duck morbidity, inoculation in nature with what laboratory was separated to African green monkey kidney cell (Vero) T25 cell bottle of single layer is covered with, it is small to adsorb 2 for 100 times of dilution 500ul virus liquids of every bottle of inoculation Shi Hou discards virus liquid, and the DMEM culture solution that 5ml contains 2% fetal calf serum is added, differs within 24~96 hours, culture to appearance Multigelation 3 times, it is next-generation to draw poison disease vaccination for cytopathy.Continuous passage culture to 40 generations, after again continuously with limited After dilution method purifies 5 times, reached for 62 generations.
Secondary culture result
F3 is inoculated with Vero cell for duck embryos yolk liquid, and F1~F3 generation cytopathy occurs without obvious lesion, F4 generation on day 4 Become, suspension cell increases, and plasomidum occurs etc..It is 10 that F4 is lower for potency3TCID50/0.1ml.With the increasing of passage number Add, virus titer becomes larger, and reaches 10 to F605.5TCID50/0.1mL.It finds in succeeding generations as the increase of passage number is thin The time that cytopathy occur in born of the same parents shortens, and cytopathy is also less by original suspension dead cell, and Syncytium formation is more, becomes Become more for suspension dead cell, plasomidum tails off.Each passage generation potency is shown in Table 1.
The measurement result of 1 S-01 plants of table different passage generation potency
Embodiment S-01 plants of passage generation pathogenicities of 2 C hypotype fowl metapneumovirus
Select by above-mentioned S-01 plants of passage generation S-01-5P, S-01-17P, S-01-28P, S-01-32P, S-01-40P, Totally 9 groups of S-01-50P, S-01-60P, Vero cell liquid control group, eye droppings collunarium infects two week old ducklings, and infective dose is 104TCID50/ ml observes clinical symptoms after immune and records the number and severity of every group of morbidity duck only, daily immune Every group of cloacal swab and throat swab are acquired within the 5th day afterwards, the swab of every 5 ducks only is placed in the EP pipe of a 15ml, into Row RT-PCR detection, every group of totally 6 samples to be tested.By lesion after observation system clinical symptoms, dissect, different generations are analyzed To the pathogenicity of duck.(RT-PCR detection method refers to Ali A, Reynolds DL:A reverse transcription- polymerase chain reaction assay for the detection of avian pneumovirus (Colorado strain).Avian Dis1999,43:600–603.)
Experimental result: F5, F17 and F32 fall ill in terms of clinic, remaining generation does not occur incidence, clinical condition Shape is mainly shown as two cough, tracheae rale symptoms, and the duck of dissect classical symptom only has mucus accumulation in its visible nasal cavity (such as Fig. 1 arrow is signified), negative control has no the appearance (Fig. 2) of these symptoms.F28 does not occur incidence, and F32 occurs Incidence illustrates that the virus weakening situation in the stage is unstable.F42 and later generation do not occur incidence, Illustrate to have caused weak.F5, F17, F32 disease incidence successively reduce, respectively 73%, 33%, 13%.Occur after each generation is immune Toxin expelling situation.Each generation toxin expelling and disease incidence see the table below 2.
2 S-01 plants of table each generation toxin expelling and incidence table
S-01 plants of fowl metapneumovirus of 3 C hypotype of embodiment cause weak generation vaccine potency experiment
Having caused in the above results, weak generation carries out immunogenicity and different potency Immunization tests, test are chosen F40, F50, F60 have been selected, specific implementation operation is the muscovy duckling of 200 1 ages in days (it is negative for crossing aMPV and detection aMPV rather) It is randomly divided into 10 groups, every group 20.40 generations, 50 generations, 60 generations each generation point, three gradients (102.5、103.5、 104.5TCID50/ 0.1ml).Vero cell liquid 0 is the 10th group of blank group.Eye droppings collunarium mode is immunized when duckling 2w.Observation daily after immune Clinical symptoms, carry out the acquisition of cloacal swab and throat swab on the 5th day, and 10 ducks of every group of random acquisition only, are put in 2ml respectively EP pipe in.The dual anti-physiological saline containing 2000U of 300ul is added in each EP pipe.The swab of acquisition carries out RT- PCR detection.Experimental group whole eye droppings collunarium mode carries out F5 for poison is attacked when exempting from rear 3w, and attacking toxic dose is 104.5TCID50.Attack poison Observation clinical symptoms daily afterwards, attack the acquisition that poison carries out cloacal swab and throat swab for 5 days, every group of random acquisition 10 ducks only, It is put in the EP pipe of 2ml respectively.The dual anti-physiological saline containing 2000U of 300ul is added in each EP pipe, acquisition is wiped Son carries out RT-PCR detection.
Experimental result: the case where not falling ill after each group of immune group is immune, in F5 generation, attacks poison after 3w is immunized, and acquires within 5 days Swab testing result show and be only immunized 104.5TCID50The other duck of group does not only have toxin expelling situation, other each generations 103.5And 102.5There is the occurrence of toxin expelling.Illustrate that 104.5 immune effects are best.Blank control group is coughed after attacking poison Etc. clinical symptoms, detect toxin expelling situation within 5 days, illustrate control group set up.Toxin expelling statistics is shown in Table 3.
3 potency test toxin expelling statistical form of table
The measurement analysis of embodiment 4, C hypotype fowl metapneumovirus weakening strain S-01-50P gene order
Above-mentioned C hypotype fowl metapneumovirus weakening strain S-01-50P carries out the measurement analysis of complete genome sequence, with initial point It is compared from strain S-01 full genome, analyzes the variation situation of its nucleotide, amino acid sites.
Experimental result: the length 14112bp (not including polyA tail) of weakening strain S-01-50P complete genome sequence, N protein Sequence (395aa), P protein sequence (295aa), M protein sequence (255aa), F protein sequence (538aa), M2 protein sequence (185aa), SH protein sequence (176aa), G-protein sequence (586aa), L protein sequence (2006aa).Pass through weakening strain S- The comparison of 01-50P and initially-separate lesion generation S-01 complete genome sequence is analyzed, it is found that its homology is relatively high, shared 16 A amino acid makes a variation.N gene does not make a variation；There are the variations of 2 amino acid sites, respectively 129Aa R- for P gene K,233Aa M-I；There are the variation 136Aa V-D of 1 amino acid for M gene；There are the variations of 6 amino acid for F gene, respectively For 60Aa C-R, 231Aa V-I, 334Aa F-L, 453Aa E-K, 485Aa E-K, 528Aa N-S；There are 3 amino for gene The variation of acid, respectively 60Aa P-L, 65Aa A-V, 85Aa T-I；There are the variations of 4 amino acid for L gene, respectively 14Aa T-P, 30Aa I-L,186Aa F-L,1756Aa I-T.The variation of these amino acid may with strain virulence attenuation of with And virus replication power becomes Qiang Youguan.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to of the invention Protection scope.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Sequence table
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tcaattcagc caaggcagat ggaaagaaaa cacatgattt gattgctctc tgcgacttcc 2580
ttgacctaga gaagggagtt ccagtgacta taccagctta cataaagtct gtgtcaataa 2640
aggagagtga atcagcaact gttgaagctg caattagtgg ggaggcagat caagctataa 2700
ctcaagctag gattgcccca tacgctggct tgatcatgat catgacaatg aacaacccta 2760
aggggatctt caaaaaactg ggtgcaggag ttcaggtaat agtagagtta ggagcatacg 2820
tccaagcaga aagcataagc agaatctgca ggaactggag tcaccagggt acaagatatg 2880
tcctgaagtc aagataaaca tagagaatac acccatctaa tcacagaagt aattccattt 2940
ctagcccact catagttaat tcctggtttg atattattta gaaaaaattg ggacaagtga 3000
aaatgtcttg gaaagtggta ctgctactgg tgttgctagc caccccaaca ggggggctag 3060
aagaaagtta cctggaagaa tcatgcagta ctgttactag aggatatctg agtgttttga 3120
ggacagggtg gtatacaaat gtgttcacac ttgaggttgg agatgtggaa aatctcacat 3180
gtaccgacgg acccagctta ataagaacag agcttgaact gacaaaaaat gcacttgagg 3240
aactcaaaac agtatcagca gatcaattgg caaaggaagc taggataatg tcaccaagaa 3300
aagcccggtt tgttctgggt gccatagcat taggtgtggc aactgccgct gcagtgacag 3360
ctggtgtagc aatagccaag acaattaggc tggaaggaga agtggctgca atcaagggtg 3420
ctctcaggaa aacaaatgag gctgtatcta cattaggaaa tggtgtgaga gtactggcaa 3480
cagctgtgaa tgatctcaag gactttataa gtaaaaaatt gacacctgca ataaacaaga 3540
acaagtgtga catttcagac cttaaaatgg cagtaagctt cggacaatac aatcgaagat 3600
tcctcaatgt ggtaagacag ttttctgaca atgcaggtat tacacctgca atatctctag 3660
atttaatgac tgatgccgag cttgtaagag ctataagtaa catgcccaca tcttcaggac 3720
agatcaatct gatgcttgag aatcgggcaa tggtcagaaa ggaaagggtt tgggatcttg 3780
attggagttt atggtagttc cgtggtctat atggtgcagc ttcctatttt cggtgtgata 3840
gacacaccat gttggaaggt gaaggctgct ccattatgtt cagggaaaga cgggagttat 3900
gcatgtcttt tgcgagagga ccaaggctgg tattgtcaaa atgctggatc cacagtttat 3960
tatccgaatg aggaagactg tgaagtgaga agtgatcatg tcctttgtga cacagcagct 4020
gggataaatg tagcaaagga gtcagaagag tgcaacagaa acatctcaac aacaaagtac 4080
ccctgcaagg taagcacagg gcgtcaccca ataagcatgg tggccttatc accactgggt 4140
gccttggtag catgttatga tggggtgagt tgttccattg gaagcaacaa ggttggaata 4200
atcagacctt tggggaaagg gtgttcatac attagcaatc aagacgctga cactgttaca 4260
attgacaaca cagtgtacca attgagcaaa gtagagggag aacaacacac aatcaaaggg 4320
aagcctgtat ctagcaattt tgacccgata gagtttccta aagaccaatt caacatagcc 4380
ctggatcagg tgtttgaaag tgttgagaag agccagagtc tgatagacca gtcaaacaag 4440
atactggata gcaccaaaaa ggggaacgca ggatttgtca tagtgatagt cctcattgtc 4500
ctgcttatgc tagctgcagt tggtgtgggt atcttctttg tggttaaaaa gaggaaaact 4560
gctcccaaat tcccaatgga aatgagtggt gtgaacaaca aaggatttat cccttaactt 4620
tagttactaa aaaattggga caagtgaaga tgtctcgcaa ggccccctgc aaatatgaag 4680
tacggggcaa gtgcaataga ggcagtgaat gcaaatttaa ccataattac tggagttggc 4740
cagacaggta cctgttacta aggtctaatt acctgctgaa tcaattactg agaaatacgg 4800
acagatcaga cggactatca ttaatctcag gagcagggcg agacgatagg acacaagatt 4860
ttgttttagg gtcaacaaat gtagtccaga actacattga taacaatgag aacataacaa 4920
aagcatcagc ttgttacagc ctgtataata tcataaaaca actgcaagag actgacgtga 4980
ggcaggccag ggacaacaaa gttgatgaca gcaagcatgt cgctctacat aatttagtgt 5040
tgtcatatat ggaaatgagc aaaacccctg catccttgat aaacaacttg aagaagcttc 5100
cgaaggaaaa actaaaaaaa ttggcaaagc tgatcattga gttgtcagca ggagtggaaa 5160
atgactccac agctaccatg caagatagtg caaactctga ttaaatgtgg agagcatggc 5220
ctgatcttcc tgaagatgag attggatgat atggtatgga ctaaaaatga attggtagac 5280
ataatttcca ctgagatagt taaagtgcat gctaatatat tcaaatgtag actagaggat 5340
atagaaatca tttatgttaa aacatttcta agttaataaa aaattgggac aagtcaacat 5400
ggagcccctg aaagtctctg gaagtggagg gatgccgatg aagacaaggc ttaatattat 5460
acttgagaag tcaatcaata aaatcttaat catcttagga ttactattaa ctgcctcaac 5520
tgtgattaca atcatactca cagtggaata tataagagtt gaaaatgaac tgcaactttg 5580
caagatggga gcagaggtgg ccaagacaac ccaagaaaca caaacactac cagtgaagac 5640
aactcctatg ctaaccagca caagatcaac taccatatcc gtcaaaacta gatcagtttc 5700
aaggactacc catcccaccg gtcccagctg ctggagagag gaggaaaagt gccagaatat 5760
cacagctaaa tggtccaatt gttttggcac atctctacct gtaagggtga attgcacagc 5820
actaagagaa ttgtgtgatg agcagctagg caatcacaca acagttcaag catcaaggaa 5880
gtgtacatgc atatatgcat taaattggga ttgtggttat gcttgagaga gaaaccacgt 5940
cagcagactt caatgagatc tacagaaaag gaactcaaat gcaaaaatca attttttagt 6000
tatttaaaaa ccatgagtat gtctgaacag tgtcaaggcc aagaaaaaca actcgagaac 6060
aggcaatcca atgattacaa tcaatcagac aaaggaaaac gggacaagtc aacatggagg 6120
tcaagataga gaatgttggc aagtcacagg agcttaaagt caaagtcaag aattttataa 6180
aaaggtctga ttgcaagaaa aaactttttg ccttgatttt agggctgatc agctttgaca 6240
tcactatgaa tataatgctg tctgtcatgt atgtggagtc aaatgaggcc ctgagctcat 6300
gcagggtcca aggaactccc gctccaagag acaacaggac aaacacagaa aacacagcaa 6360
aggaaacaac actccacaca atgaccacga caaggaacac agaagcgggg gggacaaaaa 6420
ccaccaaacc ccaggctgac gaaagagcaa caagcccaag caagaacccc accatcgggg 6480
cagacaaaca caaaacaaca agagcaacaa cagaggcaga gcaggagaaa caaagcaagc 6540
aaaccacaga gccaggcacc agcaccccga agcacatccc cgcaaggcca agcagcaaat 6600
ccccagccac aacaaaaaca acaacacagc ctacaacacc aacagtcgca aaaggaggca 6660
cagcacccaa gaacagacag acaacaacca aaaagaccga agcggacacc ccgacaacaa 6720
gcagagcaaa gcaaaccaac aaacccacag ggacagaaac aacacccccc agagcaacaa 6780
cagaaacaga caaggacaaa gaagggccaa cacagcacac aaccaaagag cagcccgaga 6840
caacggcagg agggacaaca actccacagc caagaagaac aaccagcagg ccagccccaa 6900
caacaaacac caaagagggg gcagaaacca ccgggaccag aacaaccaag agcacccaaa 6960
caagcgcaag cccaccaaga ccaacaagaa gcacacccag caaaacagca acaggaacca 7020
acaagagagc cacaacaaca aagggaccga acacagcaag cacagacaga agacaacaga 7080
ccaggacaac cccaaagcaa gaccaacaga cccaaaccaa ggcaaagaca accacaaaca 7140
aggcccacgc aaaagccgcc accacaccag agcacaacac agacacaaca gacagcatga 7200
aagaaaactc caaggaggac aaaaccacca gggaccccag cagcaaagca acaaccaagc 7260
aagaaaacac cagcaaagga acaaccgcaa caaacctcgg aaacaacacc gaggcaggcg 7320
cgagaacacc cccaacaacc accccgacaa ggcacaccac ggaaccagcc acaagcacag 7380
cggggggaca caccaaagca agaactacaa gatggaaaag cacagccgcc agacagccca 7440
caagaaacaa cacaacagca gataccaaaa cagcccagag caaacaaaca acaccggcgc 7500
aactaggcaa caacacaaca ccagagaaca caacaccacc ggacaacaaa agcaacagcc 7560
aaacaaacgt tgcaccaact gaagaaatag aaattggctc aagcctctgg cgaagaaggt 7620
atgtttatgg accctgcaga gagaatgctc tggagcaccc aatgaatccg tgtcttaagg 7680
acaacactac atggatttat ttggacaatg gaaggaattt gccagcaggt tattatgata 7740
gtaaaacaga taaaataatt tgctatggaa tctatagagg aaactcatat tgttacgggc 7800
gaatagagtg cacatgtaaa aacggcacag gactgctatc ctattgctgt aattcataca 7860
actggagtta gtccaagagc aaattagtta attaaaaaga aggaccaagt taaaaatgga 7920
tccactaaat gaaggggttg tgaatgtgta cttgcctgat tcttatttga aaggtgtaat 7980
atcttttagt gaaaccaatg cattaggatc ttgccttcta gggaaacact accttaaaaa 8040
ggataacaca tcaaaagtag ctatagagag ccctgttgta gaacacatca gattgagaaa 8100
tgctttttag actaggatca aggagaaaaa cctaagggta gtagaacccg tcaacatgca 8160
aagtgaagtc atgagaaatt cttacacctg tgaattgaat ttactcaaac aacttataac 8220
aagaagtaaa gacataagct cactcaaact tgacatgata tgtgactggt tgcaattaaa 8280
atcaacaccc gaaaatccat ctgtattaaa gtttgtggat gtaaggtgta ttcccgactg 8340
ggtaagcact tggtttagca gctggtacaa tttaaacaaa cttatattag agtttaggcg 8400
tgaagaggta gcctgcacgg ggtcaatcat atgtaaaaca atagggagca taatgtttat 8460
tatttcatcg ctcgggtgtg tcatcaaaag caacaagagc aaaagaatta gctttatgac 8520
atacaatcag gtgctgacat ggaaagatgt catgttaagc aggttcaatg caaatctatg 8580
tgtgtggatt agcaacagtc ttaacaaaaa tcaggaagga ttaggtctta gaagcaatct 8640
acaagggacc ctggttaaca aattatatga gattgttgac tctatgctca gtgtttgcag 8700
caatgagggg ttcacccttg ttaaagaatt tgaagggttc ataatgagtg agattctgaa 8760
aataacagag catgctcagt ttagtacaag gttcagaaac actttgttga atgggttagt 8820
agaccaactg gtaaagatga ggggcctaaa caggaaaagg gtttcaggca ctgtcttgga 8880
aggaaaccag tatccaatgt atgagacaac attagctaca ctaggggaag ctttaaaaac 8940
aataagactc ctggtaaaca agaacttaga caatgccgct gagctgtact acattttcag 9000
gatttttggc catccaatgg ttgaagagag agaagctatg gacgcagtca gacttaataa 9060
tgagatcaca aaaattttga aattagagag ccttacagag ttgagggggg ccttcatttt 9120
aagaatcata aaaggctttg ttgacacaaa taagagatgg cctaaaatca aaaatctcaa 9180
agtgttgagc agaaggtggg ttatgtactt taaagcaaag agttatccaa gccaattgga 9240
gctgagtagt caagatttcc ttgaattagc aggtgtgcag tttgagcagg aatttgcaat 9300
accagagaga acaaatcttg agatggtctt gaatgataag gccatctcac cacccaaaaa 9360
tttaatatgg tcagtgttcc ctaaaaatta ccttccaaca aacatcaggg aaaaattcac 9420
tgatgaaatg ttcaactcaa gtgaaaagtt gaagacaaga agggtgttgg agtactactt 9480
aaaagacaac aagtttgatc agaatgacct gaaaaaatat gttgttaggc aagagtactt 9540
gggtgataaa gaacatgtag tgtcactgac agggaaagaa cgtgagttaa gtgtgggaag 9600
gatgttcgcg atgcagccag ggaaacaaag gcaagttcag atcttagcag aaaagctact 9660
tgcagacaat atagtaccat tcttccctga gacattgaca aaatatggtg atctagaact 9720
acaaaggatt atggaaataa aatctgagtt gtcatcagta aagtcacgga gaaatgacag 9780
ttacaacaat tatattgcta gggcatcaat agtgactgat cttagtaaat tcaaccaagc 9840
ttttcggtac gagacatctt ctgtctgtgc agatgtagtt gatgaattgc atggcacaca 9900
aagtctcttc tgctggctac atttgactgt gccactgaca actatgattt gcacatatag 9960
acatgctcct ccagaaacag aaggagtgta tgacattgat aaaatcaagg aacaaagtgg 10020
gctttacagg tttcatatgg gggggattga agggtggtgc caaaagttgt ggactatgga 10080
ggctatttct ttattagatg tagtatcagt caaaaaccgt gtccaactaa catcattgtt 10140
aaatggagac aatcaatcta ttgatgttag caaacctgtt aggttgtcac aaggagttga 10200
tgaagttaaa gctgactata gcctggcagt gaaaatgtta aaagaaataa gaaatgctta 10260
caaagacata gggcacaagt taaaagaagg agagacctat atatccagag atctacaatt 10320
catgagcaag gtcattcaat cagagggtgt gatgcatccg tcaccaataa agaaaattct 10380
gagggtcgga ccatggatta atactatctt agatgacatc aagaccagtg cagagtcaat 10440
aggaagccta tgtcaagagt tagagtttag aggtgaaagc ctcttggtta gccttattct 10500
aaggaatttt tggttatacg agttatggat gcatgaatct aaaagtcatc cacttgcagg 10560
gaagcagcta tacagacagc tgagcaagac attagcaata actcaaaaat tctttggaat 10620
aacaaaagaa actgatgttg ttaatctgtg gatgaatgtt ccaatgcaat ttggtggggg 10680
tgaccctgta gtgttgtata ggtcatttta ccggaggaca ccagatttct tgactgaagc 10740
agtaagtcat atgagtgtac tactcaaggt gtatgggaaa gcaaaagagg gatcaaagaa 10800
agattttttc aaagcattat tatcagtaga caaaaataag agggccactc tgaccacatt 10860
aatgagagac cctcaagcag tgggatcaga gaggcaggca agagttacta gtgaaattaa 10920
tagagcagca gtcacaagtg tcttgagctt atcacctaac cagcttttct gtgatagtgc 10980
aatacactac agcagaaatg aagaagaagt tgggttaata gcccaaaaca ttacaccagt 11040
ctatccacac gggctgaggg tgttgtatga gtcattgcca ttccacaagg cggaaaaagt 11100
tgtaaacatg atatctggaa caaaatctat cacaaactta ctacaaagga catcagcaat 11160
caatggagaa gatattgata gagcagtctc aatgatgttg gagaatttag ggttactctc 11220
taggatactg tctgtgagcc aagatgatat aatcttgcct accaaggcaa atggtgatct 11280
gatatgttgt caagtctcca ggactttgag ggaaaggtcc tgggataaca tggaaattgt 11340
tggagtcacc tcaccgagca tagttacatg tatgaacata gtgtattcca gcagctctca 11400
actaaaagga ataacaatcg aaaagttcag cacagataaa actacaagag gtcaaagagg 11460
accaaaaagt ccttgggttg gatccagcac acaggagaaa aaattggttc ctgtttacaa 11520
tagacaaatc ctctcaaagc aacagaaaga acaactagag gcaatcggaa aactgagatg 11580
ggtttataag gggactcaag ggttaaggag gttgttggat aaaatctgca tagggagctt 11640
agggatcagt tacaagaatg ttaagccact cttgccaaga tttatgagtg ttaacttcct 11700
acacagactt tccgtcagta gtagaccaat ggagtttcca gcatctgttc ctgcttacag 11760
aacgaccaat taccatttcg acacaagtcc tgtaaaccag actctgagtg aaagatttgg 11820
aaatgaggac atcaatttgg tgtttcagaa cgctatcagc tgtgggataa gtgtgatgag 11880
tgtagtagag caacttacag gcagaagccc taaacagtta gtaatgattc cccaattaga 11940
agaaattgac ataatgcctc cgcctgtctt ttcaggaaaa tttgattata aactggtaga 12000
aaagatatca tcagaccaac acattttcag ccctgacaaa ttggacctag tgactctagg 12060
aaaaatgtta atgccttcaa caagtggagc aaagtctgat cagttttgga acaggaaaga 12120
aaacttcttc catggcaata atttggtaga gtccctatct gcagcattgg cttgtcattg 12180
gtgcgggatc ttaactgaac agtgtaatga gaacaatatt ttcagaagag aatggggtga 12240
tggttttgtt acagatcatg ccttcataga tttcaaaata tttatcggcg tatttaagac 12300
caaactattg tgtggatggg gatccagggg gggggacatt aaagatcgag acatgataga 12360
cgaatctatt gacaaattaa taagagttga caactcattc tggaggatgt ttagcaaagt 12420
gatacttgaa ccaaaagtaa ggaaaagagt aatgcttttt gatgtgaaga tcttatcact 12480
tgtaggctat gcaggattta aaaattggtt tattgaccac ctgagatcta gtgacttgtg 12540
tgaagtgcca tgggttgtca atgctgatag tgaaattgtt gaggtgtctg ctgtgaaaat 12600
atatttacag ctgctgaggg tgagttctcc cttgaggatt acagtgttga attattcaga 12660
catggcccat gctattacca ggctaataag aaggaaatca atgcatgaca atgtgccatc 12720
aatatctagg acactgtcac ctgcagagtt agctcctgta gttgaaccaa ccgtgcagat 12780
gaacttgttc cctaagatca catttgaaag attgaagaat tatgaaacta tatcagggtc 12840
tactagaggg aagttgacaa gaaattacat ggtgatgttg ccttggcagc atattaatag 12900
gttcaacttt gtattcagct ctaccggttg caaaataagt gtcaaatcat gtattgggaa 12960
gttgattcaa gacttaaatc cgacagtttt ttattttgtg ggagaaggag ccggcaattg 13020
gatggcaaga acagcttgtg agtacccaaa tacaaagttt gtttacagga gtcttaaaga 13080
tgatttagac caccatttcc ctttggaatt tcagagagtc ctagggaaca tgaatagagt 13140
tatagatggt ggagaaggtc tatccatgga cacaacagat gcaactcaga aaactcattg 13200
ggatctgaca catagaattt gtaaagatgc attactgatt actttgtgtg atgctgaatt 13260
taaagatagg gatgactttt tcaagatggt gacactatgg agaaagcatg ttttgtcatg 13320
taggatatgt acaacctatg gaacagatct ctacctgttc gcaaaatatc atgcaaaaga 13380
ggaaagcata aaacttccct attttgttag atcaattgcc acatatgtca tgcaaggcag 13440
taaattgtca ggatctgagt gctatgttct tctgacactt ggtcacccac aacaacctac 13500
catgtcacgg ggaagtacag agctccaaat taaaaatggc tgtgtgtaat gacttcagca 13560
ttcctaggaa agtagaagtg aaggctgttg aggcaaactg caaatcgctg ctttctggcc 13620
taaggacacc tataaacaga gcagaattgg atcgacaaaa gaagatgttg acactaagaa 13680
gttatcattc atcagtagca acagttggag gcagcagagt catcgaatca aaatggttga 13740
gcaagaaagc taccactata atcgagtggt tagaacatat cttgaattcc ccaaaagggg 13800
agttaaacta tgattttttt gaggcactag agaacaccta tccaaatatg gttaaattat 13860
tagataatct tggcagtgct gaactaaaaa aactgatcaa agtgacaggt tacatgctga 13920
tgagtaagaa ataataggag aaaattacct taagagatta cttataattg aggctaaaaa 13980
ctaacatatt catgctgaag gatattaaaa tgatagttgt ttaaatttag caatctgaat 14040
atgaagcaca caatcagcaa ttagttatta aaaaatagag aaactaaaat tggatgaata 14100
cggttttttt gc 14112
<210> 2
<211> 394
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 2
Met Ser Leu Gln Gly Ile Gln Leu Ser Asp Leu Ser Tyr Lys His Ala
1 5 10 15
Ile Leu Lys Glu Ser Gln Tyr Thr Ile Lys Arg Asp Val Gly Thr Thr
20 25 30
Thr Ala Val Thr Pro Ser Ser Leu Gln Arg Glu Val Ser Leu Leu Cys
35 40 45
Gly Glu Ile Leu Tyr Ala Lys His Thr Asp Tyr Ser His Ala Ala Glu
50 55 60
Val Gly Met Gln Tyr Val Ser Thr Thr Leu Gly Ala Glu Arg Thr Gln
65 70 75 80
Gln Ile Leu Lys Asn Ser Gly Ser Glu Val Gln Ala Val Leu Thr Lys
85 90 95
Thr Tyr Ser Leu Gly Lys Gly Lys Asn Ser Lys Gly Glu Asp Leu Gln
100 105 110
Met Leu Asp Ile His Gly Val Glu Lys Ser Trp Val Glu Glu Val Asp
115 120 125
Lys Glu Ala Arg Lys Thr Met Ala Ser Ala Thr Lys Asp Asn Ser Gly
130 135 140
Pro Ile Pro Gln Asn Gln Arg Pro Ser Ser Pro Asp Ala Pro Ile Ile
145 150 155 160
Leu Leu Cys Ile Gly Ala Leu Ile Phe Thr Lys Leu Ala Ser Thr Ile
165 170 175
Glu Val Gly Leu Glu Thr Ala Val Arg Arg Ala Asn Arg Val Leu Asn
180 185 190
Asp Ala Leu Lys Arg Phe Pro Arg Ile Asp Ile Pro Lys Ile Ala Arg
195 200 205
Ser Phe Tyr Asp Leu Phe Glu Gln Lys Val Tyr Tyr Arg Ser Leu Phe
210 215 220
Ile Glu Tyr Gly Lys Ala Leu Gly Ser Ser Ser Thr Gly Ser Lys Ala
225 230 235 240
Glu Ser Leu Phe Val Asn Ile Phe Met Gln Ala Tyr Gly Ala Gly Gln
245 250 255
Thr Met Leu Arg Trp Gly Val Ile Ala Arg Ser Ser Asn Asn Ile Met
260 265 270
Leu Gly His Val Ser Val Gln Ala Glu Leu Lys Gln Val Thr Glu Val
275 280 285
Tyr Asp Leu Val Arg Glu Met Gly Pro Glu Ser Gly Leu Leu His Leu
290 295 300
Arg Gln Ser Pro Lys Ala Gly Leu Leu Ser Leu Ala Asn Cys Pro Asn
305 310 315 320
Phe Ala Ser Val Val Leu Gly Asn Ala Ser Gly Leu Gly Ile Leu Gly
325 330 335
Met Tyr Arg Gly Arg Val Pro Asn Thr Glu Leu Phe Ala Ala Ala Glu
340 345 350
Ser Tyr Ala Arg Ser Leu Lys Glu Ser Asn Lys Ile Asn Phe Ser Ser
355 360 365
Leu Gly Leu Thr Glu Glu Glu Lys Glu Ala Ala Glu Asn Phe Leu Asn
370 375 380
Ile Asn Glu Glu Gly Gln Asn Asp Tyr Glu
385 390
<210> 3
<211> 294
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 3
Met Ser Phe Pro Glu Gly Lys Asp Ile Leu Leu Met Gly Asn Glu Ala
1 5 10 15
Ala Lys Ala Ala Glu Ala Phe Gln Arg Ser Leu Lys Lys Val Gly His
20 25 30
Arg Arg Thr Gln Ser Ile Val Gly Asp Lys Ile Ile Thr Val Ser Glu
35 40 45
Ile Val Glu Lys Pro Thr Ile Ser Lys Ser Thr Lys Val Thr Thr Pro
50 55 60
Pro Glu Arg Lys Asn Ala Trp Gly Glu Lys Pro Asp Thr Thr Arg Ser
65 70 75 80
Pro Thr Glu Glu Ala Lys Asn Glu Ala Thr Leu Glu Asp Ala Ser Arg
85 90 95
Leu Tyr Glu Glu Val Phe Ala Pro Thr Ser Asp Gly Lys Thr Leu Ala
100 105 110
Glu Lys Gly Lys Glu Thr Thr Glu Lys Pro Lys Lys Lys Val Lys Phe
115 120 125
Lys Asn Asp Glu Ser Gly Lys Tyr Thr Lys Leu Glu Met Glu Ala Leu
130 135 140
Glu Leu Leu Ser Asp Asn Glu Asp Asp Asp Ala Glu Ser Ser Val Leu
145 150 155 160
Thr Phe Glu Glu Lys Asp Thr Ser Ala Leu Ser Leu Glu Ala Arg Leu
165 170 175
Glu Ser Ile Asp Glu Lys Leu Ser Met Ile Leu Gly Leu Leu Arg Thr
180 185 190
Leu Asn Val Ala Thr Ala Gly Pro Thr Ala Ala Arg Asp Gly Ile Arg
195 200 205
Asp Ala Met Val Gly Leu Arg Glu Glu Leu Ile Ala Asp Ile Ile Lys
210 215 220
Glu Ala Lys Gly Lys Ala Ala Glu Ile Met Lys Glu Glu Ala Lys Gln
225 230 235 240
Lys Ser Lys Ile Gly Asn Gly Ser Val Gly Leu Thr Glu Lys Ala Lys
245 250 255
Glu Leu Asn Lys Ile Val Glu Asp Glu Ser Thr Ser Gly Glu Ser Glu
260 265 270
Glu Glu Glu Glu Glu Glu Asp Glu Glu Glu Ser Asn Pro Asp Asp Asp
275 280 285
Leu Tyr Ser Leu Thr Met
290
<210> 4
<211> 254
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 4
Met Glu Ser Tyr Leu Val Asp Thr Tyr Gln Gly Val Pro Tyr Thr Ala
1 5 10 15
Ala Val Gln Thr Asp Leu Val Glu Lys Asp Gln Leu Pro Ala Arg Leu
20 25 30
Thr Val Trp Phe Pro Leu Phe Gln Thr Asn Thr Pro Pro Thr Val Leu
35 40 45
Leu Glu Gln Leu Lys Thr Leu Thr Ile Thr Thr Leu Tyr Thr Ala Ser
50 55 60
Gln Asn Gly Pro Ile Leu Lys Val Asn Ala Ser Ala Gln Gly Ala Ala
65 70 75 80
Met Ser Ala Leu Pro Lys Ser Phe Asp Val Ser Ala Ser Val Ala Leu
85 90 95
Asp Asp Tyr Ser Lys Leu Glu Phe Asp Lys Leu Thr Val Cys Glu Leu
100 105 110
Lys Ala Val Tyr Leu Thr Thr Met Lys Pro Tyr Gly Met Val Ser Lys
115 120 125
Phe Val Asn Ser Ala Lys Ala Asp Gly Lys Lys Thr His Asp Leu Ile
130 135 140
Ala Leu Cys Asp Phe Leu Asp Leu Glu Lys Gly Val Pro Val Thr Ile
145 150 155 160
Pro Ala Tyr Ile Lys Ser Val Ser Ile Lys Glu Ser Glu Ser Ala Thr
165 170 175
Val Glu Ala Ala Ile Ser Gly Glu Ala Asp Gln Ala Ile Thr Gln Ala
180 185 190
Arg Ile Ala Pro Tyr Ala Gly Leu Ile Met Ile Met Thr Met Asn Asn
195 200 205
Pro Lys Gly Ile Phe Lys Lys Leu Gly Ala Gly Val Gln Val Ile Val
210 215 220
Glu Leu Gly Ala Tyr Val Gln Ala Glu Ser Ile Ser Arg Ile Cys Arg
225 230 235 240
Asn Trp Ser His Gln Gly Thr Arg Tyr Val Leu Lys Ser Arg
245 250
<210> 5
<211> 184
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 5
Met Ser Arg Lys Ala Pro Cys Lys Tyr Glu Val Arg Gly Lys Cys Asn
1 5 10 15
Arg Gly Ser Glu Cys Lys Phe Asn His Asn Tyr Trp Ser Trp Pro Asp
20 25 30
Arg Tyr Leu Leu Leu Arg Ser Asn Tyr Leu Leu Asn Gln Leu Leu Arg
35 40 45
Asn Thr Asp Arg Ser Asp Gly Leu Ser Leu Ile Ser Gly Ala Gly Arg
50 55 60
Asp Asp Arg Thr Gln Asp Phe Val Leu Gly Ser Thr Asn Val Val Gln
65 70 75 80
Asn Tyr Ile Asp Asn Asn Glu Asn Ile Thr Lys Ala Ser Ala Cys Tyr
85 90 95
Ser Leu Tyr Asn Ile Ile Lys Gln Leu Gln Glu Thr Asp Val Arg Gln
100 105 110
Ala Arg Asp Asn Lys Val Asp Asp Ser Lys His Val Ala Leu His Asn
115 120 125
Leu Val Leu Ser Tyr Met Glu Met Ser Lys Thr Pro Ala Ser Leu Ile
130 135 140
Asn Asn Leu Lys Lys Leu Pro Lys Glu Lys Leu Lys Lys Leu Ala Lys
145 150 155 160
Leu Ile Ile Glu Leu Ser Ala Gly Val Glu Asn Asp Ser Thr Ala Thr
165 170 175
Met Gln Asp Ser Ala Asn Ser Asp
180
<210> 6
<211> 175
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 6
Met Glu Pro Leu Lys Val Ser Gly Ser Gly Gly Met Pro Met Lys Thr
1 5 10 15
Arg Leu Asn Ile Ile Leu Glu Lys Ser Ile Asn Lys Ile Leu Ile Ile
20 25 30
Leu Gly Leu Leu Leu Thr Ala Ser Thr Val Ile Thr Ile Ile Leu Thr
35 40 45
Val Glu Tyr Ile Arg Val Glu Asn Glu Leu Gln Leu Cys Lys Met Gly
50 55 60
Ala Glu Val Ala Lys Thr Thr Gln Glu Thr Gln Thr Leu Pro Val Lys
65 70 75 80
Thr Thr Pro Met Leu Thr Ser Thr Arg Ser Thr Thr Ile Ser Val Lys
85 90 95
Thr Arg Ser Val Ser Arg Thr Thr His Pro Thr Gly Pro Ser Cys Trp
100 105 110
Arg Glu Glu Glu Lys Cys Gln Asn Ile Thr Ala Lys Trp Ser Asn Cys
115 120 125
Phe Gly Thr Ser Leu Pro Val Arg Val Asn Cys Thr Ala Leu Arg Glu
130 135 140
Leu Cys Asp Glu Gln Leu Gly Asn His Thr Thr Val Gln Ala Ser Arg
145 150 155 160
Lys Cys Thr Cys Ile Tyr Ala Leu Asn Trp Asp Cys Gly Tyr Ala
165 170 175
<210> 7
<211> 537
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 7
Met Ser Trp Lys Val Val Leu Leu Leu Val Leu Leu Ala Thr Pro Thr
1 5 10 15
Gly Gly Leu Glu Glu Ser Tyr Leu Glu Glu Ser Cys Ser Thr Val Thr
20 25 30
Arg Gly Tyr Leu Ser Val Leu Arg Thr Gly Trp Tyr Thr Asn Val Phe
35 40 45
Thr Leu Glu Val Gly Asp Val Glu Asn Leu Thr Arg Thr Asp Gly Pro
50 55 60
Ser Leu Ile Arg Thr Glu Leu Glu Leu Thr Lys Asn Ala Leu Glu Glu
65 70 75 80
Leu Lys Thr Val Ser Ala Asp Gln Leu Ala Lys Glu Ala Arg Ile Met
85 90 95
Ser Pro Arg Lys Ala Arg Phe Val Leu Gly Ala Ile Ala Leu Gly Val
100 105 110
Ala Thr Ala Ala Ala Val Thr Ala Gly Val Ala Ile Ala Lys Thr Ile
115 120 125
Arg Leu Glu Gly Glu Val Ala Ala Ile Lys Gly Ala Leu Arg Lys Thr
130 135 140
Asn Glu Ala Val Ser Thr Leu Gly Asn Gly Val Arg Val Leu Ala Thr
145 150 155 160
Ala Val Asn Asp Leu Lys Asp Phe Ile Ser Lys Lys Leu Thr Pro Ala
165 170 175
Ile Asn Lys Asn Lys Cys Asp Ile Ser Asp Leu Lys Met Ala Val Ser
180 185 190
Phe Gly Gln Tyr Asn Arg Arg Phe Leu Asn Val Val Arg Gln Phe Ser
195 200 205
Asp Asn Ala Gly Ile Thr Pro Ala Ile Ser Leu Asp Leu Met Thr Asp
210 215 220
Ala Glu Leu Val Arg Ala Ile Ser Asn Met Pro Thr Ser Ser Gly Gln
225 230 235 240
Ile Asn Leu Met Leu Glu Asn Arg Ala Met Val Arg Arg Lys Gly Phe
245 250 255
Gly Ile Leu Ile Gly Val Tyr Gly Ser Ser Val Val Tyr Met Val Gln
260 265 270
Leu Pro Ile Phe Gly Val Ile Asp Thr Pro Cys Trp Lys Val Lys Ala
275 280 285
Ala Pro Leu Cys Ser Gly Lys Asp Gly Ser Tyr Ala Cys Leu Leu Arg
290 295 300
Glu Asp Gln Gly Trp Tyr Cys Gln Asn Ala Gly Ser Thr Val Tyr Tyr
305 310 315 320
Pro Asn Glu Glu Asp Cys Glu Val Arg Ser Asp His Val Leu Cys Asp
325 330 335
Thr Ala Ala Gly Ile Asn Val Ala Lys Glu Ser Glu Glu Cys Asn Arg
340 345 350
Asn Ile Ser Thr Thr Lys Tyr Pro Cys Lys Val Ser Thr Gly Arg His
355 360 365
Pro Ile Ser Met Val Ala Leu Ser Pro Leu Gly Ala Leu Val Ala Cys
370 375 380
Tyr Asp Gly Val Ser Cys Ser Ile Gly Ser Asn Lys Val Gly Ile Ile
385 390 395 400
Arg Pro Leu Gly Lys Gly Cys Ser Tyr Ile Ser Asn Gln Asp Ala Asp
405 410 415
Thr Val Thr Ile Asp Asn Thr Val Tyr Gln Leu Ser Lys Val Glu Gly
420 425 430
Glu Gln His Thr Ile Lys Gly Lys Pro Val Ser Ser Asn Phe Asp Pro
435 440 445
Ile Glu Phe Pro Lys Asp Gln Phe Asn Ile Ala Leu Asp Gln Val Phe
450 455 460
Glu Ser Val Glu Lys Ser Gln Ser Leu Ile Asp Gln Ser Asn Lys Ile
465 470 475 480
Leu Asp Ser Thr Lys Lys Gly Asn Ala Gly Phe Val Ile Val Ile Val
485 490 495
Leu Ile Val Leu Leu Met Leu Ala Ala Val Gly Val Gly Ile Phe Phe
500 505 510
Val Val Lys Lys Arg Lys Thr Ala Pro Lys Phe Pro Met Glu Met Ser
515 520 525
Gly Val Asn Asn Lys Gly Phe Ile Pro
530 535
<210> 8
<211> 585
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 8
Met Glu Val Lys Ile Glu Asn Val Gly Lys Ser Gln Glu Leu Arg Val
1 5 10 15
Lys Val Lys Asn Phe Ile Lys Arg Ser Asp Cys Lys Lys Lys Leu Phe
20 25 30
Ala Leu Ile Leu Gly Leu Ile Ser Phe Asp Ile Thr Met Asn Ile Met
35 40 45
Leu Ser Val Met Tyr Val Glu Ser Asn Glu Ala Leu Ser Ser Cys Arg
50 55 60
Val Gln Gly Thr Pro Ala Pro Arg Asp Asn Arg Thr Asn Thr Glu Asn
65 70 75 80
Thr Ala Lys Glu Thr Thr Leu His Thr Met Thr Thr Thr Arg Asn Thr
85 90 95
Glu Ala Gly Gly Thr Lys Thr Thr Lys Pro Gln Ala Asp Glu Arg Ala
100 105 110
Thr Ser Pro Ser Lys Asn Pro Thr Ile Gly Ala Asp Lys His Lys Thr
115 120 125
Thr Arg Ala Thr Thr Glu Ala Glu Gln Glu Lys Gln Ser Lys Gln Thr
130 135 140
Thr Glu Pro Gly Thr Ser Thr Pro Lys His Ile Pro Ala Arg Pro Ser
145 150 155 160
Ser Lys Ser Pro Ala Thr Thr Lys Thr Thr Thr Gln Pro Thr Thr Pro
165 170 175
Thr Val Ala Lys Gly Gly Thr Ala Pro Lys Asn Arg Gln Thr Thr Thr
180 185 190
Lys Lys Thr Glu Ala Asp Thr Pro Thr Thr Ser Arg Ala Lys Gln Thr
195 200 205
Asn Lys Pro Thr Gly Thr Glu Thr Thr Pro Pro Arg Ala Thr Thr Glu
210 215 220
Thr Asp Lys Asp Lys Glu Gly Pro Thr Gln His Thr Thr Lys Glu Gln
225 230 235 240
Pro Glu Thr Thr Ala Gly Gly Thr Thr Thr Pro Gln Pro Arg Arg Thr
245 250 255
Thr Ser Arg Pro Ala Pro Thr Thr Asn Thr Lys Glu Gly Ala Glu Thr
260 265 270
Thr Gly Thr Arg Thr Thr Lys Ser Thr Gln Thr Ser Ala Ser Pro Pro
275 280 285
Arg Pro Thr Arg Ser Thr Pro Ser Lys Thr Ala Thr Gly Thr Asn Lys
290 295 300
Arg Ala Thr Thr Thr Lys Gly Pro Asn Thr Ala Ser Thr Asp Arg Arg
305 310 315 320
Gln Gln Thr Arg Thr Thr Pro Lys Gln Asp Gln Gln Thr Gln Thr Lys
325 330 335
Ala Lys Thr Thr Thr Asn Lys Ala His Ala Lys Ala Ala Thr Thr Pro
340 345 350
Glu His Asn Thr Asp Thr Thr Asp Ser Met Lys Glu Asn Ser Lys Glu
355 360 365
Asp Lys Thr Thr Arg Asp Pro Ser Ser Lys Ala Thr Thr Lys Gln Glu
370 375 380
Asn Thr Ser Lys Gly Thr Thr Ala Thr Asn Leu Gly Asn Asn Thr Glu
385 390 395 400
Ala Gly Ala Arg Thr Pro Pro Thr Thr Thr Pro Thr Arg His Thr Thr
405 410 415
Glu Pro Ala Thr Ser Thr Ala Gly Gly His Thr Lys Ala Arg Thr Thr
420 425 430
Arg Trp Lys Ser Thr Ala Ala Arg Gln Pro Thr Arg Asn Asn Thr Thr
435 440 445
Ala Asp Thr Lys Thr Ala Gln Ser Lys Gln Thr Thr Pro Ala Gln Leu
450 455 460
Gly Asn Asn Thr Thr Pro Glu Asn Thr Thr Pro Pro Asp Asn Lys Ser
465 470 475 480
Asn Ser Gln Thr Asn Val Ala Pro Thr Glu Glu Ile Glu Ile Gly Ser
485 490 495
Ser Leu Trp Arg Arg Arg Tyr Val Tyr Gly Pro Cys Arg Glu Asn Ala
500 505 510
Leu Glu His Pro Met Asn Pro Cys Leu Lys Asp Asn Thr Thr Trp Ile
515 520 525
Tyr Leu Asp Asn Gly Arg Asn Leu Pro Ala Gly Tyr Tyr Asp Ser Lys
530 535 540
Thr Asp Lys Ile Ile Cys Tyr Gly Ile Tyr Arg Gly Asn Ser Tyr Cys
545 550 555 560
Tyr Gly Arg Ile Glu Cys Thr Cys Lys Asn Gly Thr Gly Leu Leu Ser
565 570 575
Tyr Cys Cys Asn Ser Tyr Asn Trp Ser
580 585
<210> 9
<211> 2005
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 9
Met Asp Pro Leu Asn Glu Gly Val Val Asn Val Tyr Leu Pro Asp Ser
1 5 10 15
Tyr Leu Lys Gly Val Ile Ser Phe Ser Glu Thr Asn Ala Leu Gly Ser
20 25 30
Cys Leu Leu Gly Lys His Tyr Leu Lys Lys Asp Asn Thr Ser Lys Val
35 40 45
Ala Ile Glu Ser Pro Val Val Glu His Ile Arg Leu Arg Asn Ala Phe
50 55 60
Gln Thr Arg Ile Lys Glu Lys Asn Leu Arg Val Val Glu Pro Val Asn
65 70 75 80
Met Gln Ser Glu Val Met Arg Asn Ser Tyr Thr Cys Glu Leu Asn Leu
85 90 95
Leu Lys Gln Leu Ile Thr Arg Ser Lys Asp Ile Ser Ser Leu Lys Leu
100 105 110
Asp Met Ile Cys Asp Trp Leu Gln Leu Lys Ser Thr Pro Glu Asn Pro
115 120 125
Ser Val Leu Lys Phe Val Asp Val Arg Cys Ile Pro Asp Trp Val Ser
130 135 140
Thr Trp Phe Ser Ser Trp Tyr Asn Leu Asn Lys Leu Ile Leu Glu Phe
145 150 155 160
Arg Arg Glu Glu Val Ala Cys Thr Gly Ser Ile Ile Cys Lys Thr Ile
165 170 175
Gly Ser Ile Met Phe Ile Ile Ser Ser Leu Gly Cys Val Ile Lys Ser
180 185 190
Asn Lys Ser Lys Arg Ile Ser Phe Met Thr Tyr Asn Gln Val Leu Thr
195 200 205
Trp Lys Asp Val Met Leu Ser Arg Phe Asn Ala Asn Leu Cys Val Trp
210 215 220
Ile Ser Asn Ser Leu Asn Lys Asn Gln Glu Gly Leu Gly Leu Arg Ser
225 230 235 240
Asn Leu Gln Gly Thr Leu Val Asn Lys Leu Tyr Glu Ile Val Asp Ser
245 250 255
Met Leu Ser Val Cys Ser Asn Glu Gly Phe Thr Leu Val Lys Glu Phe
260 265 270
Glu Gly Phe Ile Met Ser Glu Ile Leu Lys Ile Thr Glu His Ala Gln
275 280 285
Phe Ser Thr Arg Phe Arg Asn Thr Leu Leu Asn Gly Leu Val Asp Gln
290 295 300
Leu Val Lys Met Arg Gly Leu Asn Arg Lys Arg Val Ser Gly Thr Val
305 310 315 320
Leu Glu Gly Asn Gln Tyr Pro Met Tyr Glu Thr Thr Leu Ala Thr Leu
325 330 335
Gly Glu Ala Leu Lys Thr Ile Arg Leu Leu Val Asn Lys Asn Leu Asp
340 345 350
Asn Ala Ala Glu Leu Tyr Tyr Ile Phe Arg Ile Phe Gly His Pro Met
355 360 365
Val Glu Glu Arg Glu Ala Met Asp Ala Val Arg Leu Asn Asn Glu Ile
370 375 380
Thr Lys Ile Leu Lys Leu Glu Ser Leu Thr Glu Leu Arg Gly Ala Phe
385 390 395 400
Ile Leu Arg Ile Ile Lys Gly Phe Val Asp Thr Asn Lys Arg Trp Pro
405 410 415
Lys Ile Lys Asn Leu Lys Val Leu Ser Arg Arg Trp Val Met Tyr Phe
420 425 430
Lys Ala Lys Ser Tyr Pro Ser Gln Leu Glu Leu Ser Ser Gln Asp Phe
435 440 445
Leu Glu Leu Ala Gly Val Gln Phe Glu Gln Glu Phe Ala Ile Pro Glu
450 455 460
Arg Thr Asn Leu Glu Met Val Leu Asn Asp Lys Ala Ile Ser Pro Pro
465 470 475 480
Lys Asn Leu Ile Trp Ser Val Phe Pro Lys Asn Tyr Leu Pro Thr Asn
485 490 495
Ile Arg Glu Lys Phe Thr Asp Glu Met Phe Asn Ser Ser Glu Lys Leu
500 505 510
Lys Thr Arg Arg Val Leu Glu Tyr Tyr Leu Lys Asp Asn Lys Phe Asp
515 520 525
Gln Asn Asp Leu Lys Lys Tyr Val Val Arg Gln Glu Tyr Leu Gly Asp
530 535 540
Lys Glu His Val Val Ser Leu Thr Gly Lys Glu Arg Glu Leu Ser Val
545 550 555 560
Gly Arg Met Phe Ala Met Gln Pro Gly Lys Gln Arg Gln Val Gln Ile
565 570 575
Leu Ala Glu Lys Leu Leu Ala Asp Asn Ile Val Pro Phe Phe Pro Glu
580 585 590
Thr Leu Thr Lys Tyr Gly Asp Leu Glu Leu Gln Arg Ile Met Glu Ile
595 600 605
Lys Ser Glu Leu Ser Ser Val Lys Ser Arg Arg Asn Asp Ser Tyr Asn
610 615 620
Asn Tyr Ile Ala Arg Ala Ser Ile Val Thr Asp Leu Ser Lys Phe Asn
625 630 635 640
Gln Ala Phe Arg Tyr Glu Thr Ser Ser Val Cys Ala Asp Val Val Asp
645 650 655
Glu Leu His Gly Thr Gln Ser Leu Phe Cys Trp Leu His Leu Thr Val
660 665 670
Pro Leu Thr Thr Met Ile Cys Thr Tyr Arg His Ala Pro Pro Glu Thr
675 680 685
Glu Gly Val Tyr Asp Ile Asp Lys Ile Lys Glu Gln Ser Gly Leu Tyr
690 695 700
Arg Phe His Met Gly Gly Ile Glu Gly Trp Cys Gln Lys Leu Trp Thr
705 710 715 720
Met Glu Ala Ile Ser Leu Leu Asp Val Val Ser Val Lys Asn Arg Val
725 730 735
Gln Leu Thr Ser Leu Leu Asn Gly Asp Asn Gln Ser Ile Asp Val Ser
740 745 750
Lys Pro Val Arg Leu Ser Gln Gly Val Asp Glu Val Lys Ala Asp Tyr
755 760 765
Ser Leu Ala Val Lys Met Leu Lys Glu Ile Arg Asn Ala Tyr Lys Asp
770 775 780
Ile Gly His Lys Leu Lys Glu Gly Glu Thr Tyr Ile Ser Arg Asp Leu
785 790 795 800
Gln Phe Met Ser Lys Val Ile Gln Ser Glu Gly Val Met His Pro Ser
805 810 815
Pro Ile Lys Lys Ile Leu Arg Val Gly Pro Trp Ile Asn Thr Ile Leu
820 825 830
Asp Asp Ile Lys Thr Ser Ala Glu Ser Ile Gly Ser Leu Cys Gln Glu
835 840 845
Leu Glu Phe Arg Gly Glu Ser Leu Leu Val Ser Leu Ile Leu Arg Asn
850 855 860
Phe Trp Leu Tyr Glu Leu Trp Met His Glu Ser Lys Ser His Pro Leu
865 870 875 880
Ala Gly Lys Gln Leu Tyr Arg Gln Leu Ser Lys Thr Leu Ala Ile Thr
885 890 895
Gln Lys Phe Phe Gly Ile Thr Lys Glu Thr Asp Val Val Asn Leu Trp
900 905 910
Met Asn Val Pro Met Gln Phe Gly Gly Gly Asp Pro Val Val Leu Tyr
915 920 925
Arg Ser Phe Tyr Arg Arg Thr Pro Asp Phe Leu Thr Glu Ala Val Ser
930 935 940
His Met Ser Val Leu Leu Lys Val Tyr Gly Lys Ala Lys Glu Gly Ser
945 950 955 960
Lys Lys Asp Phe Phe Lys Ala Leu Leu Ser Val Asp Lys Asn Lys Arg
965 970 975
Ala Thr Leu Thr Thr Leu Met Arg Asp Pro Gln Ala Val Gly Ser Glu
980 985 990
Arg Gln Ala Arg Val Thr Ser Glu Ile Asn Arg Ala Ala Val Thr Ser
995 1000 1005
Val Leu Ser Leu Ser Pro Asn Gln Leu Phe Cys Asp Ser Ala Ile His
1010 1015 1020
Tyr Ser Arg Asn Glu Glu Glu Val Gly Leu Ile Ala Gln Asn Ile Thr
1025 1030 1035 1040
Pro Val Tyr Pro His Gly Leu Arg Val Leu Tyr Glu Ser Leu Pro Phe
1045 1050 1055
His Lys Ala Glu Lys Val Val Asn Met Ile Ser Gly Thr Lys Ser Ile
1060 1065 1070
Thr Asn Leu Leu Gln Arg Thr Ser Ala Ile Asn Gly Glu Asp Ile Asp
1075 1080 1085
Arg Ala Val Ser Met Met Leu Glu Asn Leu Gly Leu Leu Ser Arg Ile
1090 1095 1100
Leu Ser Val Ser Gln Asp Asp Ile Ile Leu Pro Thr Lys Ala Asn Gly
1105 1110 1115 1120
Asp Leu Ile Cys Cys Gln Val Ser Arg Thr Leu Arg Glu Arg Ser Trp
1125 1130 1135
Asp Asn Met Glu Ile Val Gly Val Thr Ser Pro Ser Ile Val Thr Cys
1140 1145 1150
Met Asn Ile Val Tyr Ser Ser Ser Ser Gln Leu Lys Gly Ile Thr Ile
1155 1160 1165
Glu Lys Phe Ser Thr Asp Lys Thr Thr Arg Gly Gln Arg Gly Pro Lys
1170 1175 1180
Ser Pro Trp Val Gly Ser Ser Thr Gln Glu Lys Lys Leu Val Pro Val
1185 1190 1195 1200
Tyr Asn Arg Gln Ile Leu Ser Lys Gln Gln Lys Glu Gln Leu Glu Ala
1205 1210 1215
Ile Gly Lys Leu Arg Trp Val Tyr Lys Gly Thr Gln Gly Leu Arg Arg
1220 1225 1230
Leu Leu Asp Lys Ile Cys Ile Gly Ser Leu Gly Ile Ser Tyr Lys Asn
1235 1240 1245
Val Lys Pro Leu Leu Pro Arg Phe Met Ser Val Asn Phe Leu His Arg
1250 1255 1260
Leu Ser Val Ser Ser Arg Pro Met Glu Phe Pro Ala Ser Val Pro Ala
1265 1270 1275 1280
Tyr Arg Thr Thr Asn Tyr His Phe Asp Thr Ser Pro Val Asn Gln Thr
1285 1290 1295
Leu Ser Glu Arg Phe Gly Asn Glu Asp Ile Asn Leu Val Phe Gln Asn
1300 1305 1310
Ala Ile Ser Cys Gly Ile Ser Val Met Ser Val Val Glu Gln Leu Thr
1315 1320 1325
Gly Arg Ser Pro Lys Gln Leu Val Met Ile Pro Gln Leu Glu Glu Ile
1330 1335 1340
Asp Ile Met Pro Pro Pro Val Phe Ser Gly Lys Phe Asp Tyr Lys Leu
1345 1350 1355 1360
Val Glu Lys Ile Ser Ser Asp Gln His Ile Phe Ser Pro Asp Lys Leu
1365 1370 1375
Asp Leu Val Thr Leu Gly Lys Met Leu Met Pro Ser Thr Ser Gly Ala
1380 1385 1390
Lys Ser Asp Gln Phe Trp Asn Arg Lys Glu Asn Phe Phe His Gly Asn
1395 1400 1405
Asn Leu Val Glu Ser Leu Ser Ala Ala Leu Ala Cys His Trp Cys Gly
1410 1415 1420
Ile Leu Thr Glu Gln Cys Asn Glu Asn Asn Ile Phe Arg Arg Glu Trp
1425 1430 1435 1440
Gly Asp Gly Phe Val Thr Asp His Ala Phe Ile Asp Phe Lys Ile Phe
1445 1450 1455
Ile Gly Val Phe Lys Thr Lys Leu Leu Cys Gly Trp Gly Ser Arg Gly
1460 1465 1470
Gly Asp Ile Lys Asp Arg Asp Met Ile Asp Glu Ser Ile Asp Lys Leu
1475 1480 1485
Ile Arg Val Asp Asn Ser Phe Trp Arg Met Phe Ser Lys Val Ile Leu
1490 1495 1500
Glu Pro Lys Val Arg Lys Arg Val Met Leu Phe Asp Val Lys Ile Leu
1505 1510 1515 1520
Ser Leu Val Gly Tyr Ala Gly Phe Lys Asn Trp Phe Ile Asp His Leu
1525 1530 1535
Arg Ser Ser Asp Leu Cys Glu Val Pro Trp Val Val Asn Ala Asp Ser
1540 1545 1550
Glu Ile Val Glu Val Ser Ala Val Lys Ile Tyr Leu Gln Leu Leu Arg
1555 1560 1565
Val Ser Ser Pro Leu Arg Ile Thr Val Leu Asn Tyr Ser Asp Met Ala
1570 1575 1580
His Ala Ile Thr Arg Leu Ile Arg Arg Lys Ser Met His Asp Asn Val
1585 1590 1595 1600
Pro Ser Ile Ser Arg Thr Leu Ser Pro Ala Glu Leu Ala Pro Val Val
1605 1610 1615
Glu Pro Thr Val Gln Met Asn Leu Phe Pro Lys Ile Thr Phe Glu Arg
1620 1625 1630
Leu Lys Asn Tyr Glu Thr Ile Ser Gly Ser Thr Arg Gly Lys Leu Thr
1635 1640 1645
Arg Asn Tyr Met Val Met Leu Pro Trp Gln His Ile Asn Arg Phe Asn
1650 1655 1660
Phe Val Phe Ser Ser Thr Gly Cys Lys Ile Ser Val Lys Ser Cys Ile
1665 1670 1675 1680
Gly Lys Leu Ile Gln Asp Leu Asn Pro Thr Val Phe Tyr Phe Val Gly
1685 1690 1695
Glu Gly Ala Gly Asn Trp Met Ala Arg Thr Ala Cys Glu Tyr Pro Asn
1700 1705 1710
Thr Lys Phe Val Tyr Arg Ser Leu Lys Asp Asp Leu Asp His His Phe
1715 1720 1725
Pro Leu Glu Phe Gln Arg Val Leu Gly Asn Met Asn Arg Val Ile Asp
1730 1735 1740
Gly Gly Glu Gly Leu Ser Met Asp Thr Thr Asp Ala Thr Gln Lys Thr
1745 1750 1755 1760
His Trp Asp Leu Thr His Arg Ile Cys Lys Asp Ala Leu Leu Ile Thr
1765 1770 1775
Leu Cys Asp Ala Glu Phe Lys Asp Arg Asp Asp Phe Phe Lys Met Val
1780 1785 1790
Thr Leu Trp Arg Lys His Val Leu Ser Cys Arg Ile Cys Thr Thr Tyr
1795 1800 1805
Gly Thr Asp Leu Tyr Leu Phe Ala Lys Tyr His Ala Lys Glu Glu Ser
1810 1815 1820
Ile Lys Leu Pro Tyr Phe Val Arg Ser Ile Ala Thr Tyr Val Met Gln
1825 1830 1835 1840
Gly Ser Lys Leu Ser Gly Ser Glu Cys Tyr Val Leu Leu Thr Leu Gly
1845 1850 1855
His His Asn Asn Leu Pro Cys His Gly Glu Val Gln Ser Ser Lys Leu
1860 1865 1870
Lys Met Ala Val Cys Asn Asp Phe Ser Ile Pro Arg Lys Val Glu Val
1875 1880 1885
Lys Ala Val Glu Ala Asn Cys Lys Ser Leu Leu Ser Gly Leu Arg Thr
1890 1895 1900
Pro Ile Asn Arg Ala Glu Leu Asp Arg Gln Lys Lys Met Leu Thr Leu
1905 1910 1915 1920
Arg Ser Tyr His Ser Ser Val Ala Thr Val Gly Gly Ser Arg Val Ile
1925 1930 1935
Glu Ser Lys Trp Leu Ser Lys Lys Ala Thr Thr Ile Ile Glu Trp Leu
1940 1945 1950
Glu His Ile Leu Asn Ser Pro Lys Gly Glu Leu Asn Tyr Asp Phe Phe
1955 1960 1965
Glu Ala Leu Glu Asn Thr Tyr Pro Asn Met Val Lys Leu Leu Asp Asn
1970 1975 1980
Leu Gly Ser Ala Glu Leu Lys Lys Leu Ile Lys Val Thr Gly Tyr Met
1985 1990 1995 2000
Leu Met Ser Lys Lys
2005

Claims (8)

1. a kind of duck C hypotype fowl metapneumovirus low virulent strain, deposit number are as follows: CCTCC NO:V201823；Nucleotides sequence is classified as SEQ ID NO:1.
2. according to claim a kind of duck C hypotype fowl metapneumovirus low virulent strain, which is characterized in that described kind of duck C hypotype fowl The reading frame amino acid sequence of metapneumovirus low virulent strain is SEQ ID NO:2 to SEQ ID NO:9.
3. a kind of preparation method of kind duck C hypotype fowl metapneumovirus low virulent strain characterized by comprising kind duck C hypotype fowl is inclined S-01 plants of progress secondary cultures of Pneumovirinae are to 40-62 generation.
4. according to claim 3 kind of duck C hypotype fowl metapneumovirus low virulent strain obtains preparation method, which is characterized in that described The preparation method of kind duck C hypotype fowl metapneumovirus low virulent strain, including, by S-01 plants of progress secondary cultures of C hypotype fowl metapneumovirus To 50 generations.
5. according to claim 3 kind of duck C hypotype fowl metapneumovirus low virulent strain obtains preparation method, which is characterized in that described The preparation method of kind duck C hypotype fowl metapneumovirus low virulent strain, specifically includes:
With kind S-01 plants of fowl metapneumovirus of a duck C hypotype, African green monkey kidney cell (Vero) T25 cell bottle of single layer is covered in inoculation, 100 times of diluted virus liquid 500ul of every bottle of inoculation；After absorption 2 hours, virus liquid is discarded, 5ml is added containing 2% fetal calf serum DMEM culture solution is cultivated 24~96 hours, until there is cytopathy；Multigelation 3 times, it is next-generation to draw poison disease vaccination；Such as This operation, continuous passage culture to 40 generations, then with after limiting dilution assay continuous purification 5 times, continuation secondary culture.
6. kind duck C hypotype fowl metapneumovirus low virulent strain or claim 3-5 any one described in claim 1-2 any one Application of the low virulent strain prepared by the method in preparation kind duck immunoassay product.
7. a kind of include described in claim 1-2 any one kinds of duck C hypotype fowl metapneumovirus low virulent strain or claim 3-5 The immune product of kind duck of low virulent strain prepared by method described in any one, which is characterized in that institute in every part of immune product State content Wei≤10 of low virulent strain2.5TCID50。
8. the according to claim 7 kind of immune product of duck, which is characterized in that low virulent strain described in every part of immune product Content Wei≤104.5TCID50。
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111961652A (en) * 2020-07-09 2020-11-20 温氏食品集团股份有限公司 Serum-free full-suspension culture method of avian metapneumovirus and application of serum-free full-suspension culture method in vaccine
US11162148B2 (en) * 2001-01-19 2021-11-02 Erasmus University Medical Center Rotterdam Virus causing respiratory tract illness in susceptible mammals
CN115287270A (en) * 2022-10-09 2022-11-04 中国农业科学院哈尔滨兽医研究所（中国动物卫生与流行病学中心哈尔滨分中心） Subtype B avian metapneumovirus (APV) subculture attenuated strain and application thereof

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