CN110184241A - Vibrio alginolyticus bacteriophage resistant to high temperature and combinations thereof, kit and application - Google Patents

Vibrio alginolyticus bacteriophage resistant to high temperature and combinations thereof, kit and application Download PDF

Info

Publication number
CN110184241A
CN110184241A CN201910400869.8A CN201910400869A CN110184241A CN 110184241 A CN110184241 A CN 110184241A CN 201910400869 A CN201910400869 A CN 201910400869A CN 110184241 A CN110184241 A CN 110184241A
Authority
CN
China
Prior art keywords
vibrio alginolyticus
vibrio
bacteriophage
vap9
phage
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201910400869.8A
Other languages
Chinese (zh)
Other versions
CN110184241B (en
Inventor
乔欢
陈海
徐旭凌
黄杰
费文斌
胡怿林
何四龙
丛郁
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Phagelux Nanjing Biotechnology Co ltd
Original Assignee
Phagelux Nanjing Biotechnology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Phagelux Nanjing Biotechnology Co ltd filed Critical Phagelux Nanjing Biotechnology Co ltd
Priority to CN201910400869.8A priority Critical patent/CN110184241B/en
Publication of CN110184241A publication Critical patent/CN110184241A/en
Application granted granted Critical
Publication of CN110184241B publication Critical patent/CN110184241B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2795/00—Bacteriophages
    • C12N2795/00011—Details
    • C12N2795/00021—Viruses as such, e.g. new isolates, mutants or their genomic sequences
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2795/00—Bacteriophages
    • C12N2795/00011—Details
    • C12N2795/00032—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Virology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Zoology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Communicable Diseases (AREA)
  • Biomedical Technology (AREA)
  • Mycology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Biotechnology (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention belongs to the research and development technology fields of vibrio alginolyticus bacteriophage, and in particular to vibrio alginolyticus bacteriophage resistant to high temperature and combinations thereof and kit.The vibrio alginolyticus bacteriophage is vibrio alginolyticus bacteriophage VAP9(Vibrio alginolyticusPhage VAP9), deposit number is CCTCC NO:M 2018766.Also disclose the vibrio alginolyticus bacteriophage or vibrio alginolyticus bacteriophage composition or the application of reagent or kit in killing and/or prevention vibrios microorganism belonging to genus containing vibrio alginolyticus bacteriophage or vibrio alginolyticus bacteriophage composition.The present invention has host range wider, still has high toxicity to host strain under low concentration；And it can be preferably proliferated on non-pathogenic bacteria host；Large-scale industrial production can be achieved.The present invention can provide excellent strain resource for the application of phagotherapy.

Description

Vibrio alginolyticus bacteriophage resistant to high temperature and combinations thereof, kit and application
Technical field
The invention belongs to the research and development technology fields of vibrio alginolyticus bacteriophage, and in particular to vibrio alginolyticus bacteriophage resistant to high temperature And combinations thereof, kit and application.
Background technique
Vibrio alginolyticus is a kind of halophagia Gram-negative brevibacterium, and no gemma, pod membrane are under the jurisdiction of vibrionaceae, vibrio, It is a kind of common Marine Pathogenic Bacteria, quantity occupies first of seawater class vibrios.People can be caused wound infection, food poisoning, in The diseases such as otitis.Meanwhile it is also a kind of conditioned pathogen of the ocean waters cultivated animals such as fish, shrimp, shellfish, when environmental degradation opportunity The decline of body immunity function, vibrio alginolyticus sickly look are easily broken out.
Bacteriophage is a viroid for special cracking bacterium, and main chemical compositions are made of protein and nucleic acid, deposit extensively It is in soil, empty gas and water and organism, there is stronger specificity.Since it has powerful sterilizing ability, bacteriophage can It is used as a kind of bacterial-infection resisting preparation, many scholars at home and abroad's concern has been constantly subjected to since early 20th century.
About vibrio alginolyticus bacteriophage, Fu Hanqing et al. (" microbiology notification " 25 date of June in 2018) is isolated One plant of vibrio alginolyticus bacteriophage Ф V170, belongs to kind of an inner width by 7 plants in bacteriophage Ф V170 15 plants of vibrio alginolyticus of cleavable Spectrum；Optimal multiplicity of infection is 0.01；One step growth curve shows that incubation period is 10min, burst size 101.3；To 65 DEG C or more Temperature is sensitive.The bacteriophage has the potentiality of vibrio alginolyticus disease during treatment sea-farming.
Patent of invention CN103898015B discloses a kind of vibrio alginolyticus bacteriophage and its answering in sea cucumber disease prevention With bacteriophage has inhibiting effect to vibrio alginolyticus, generates in 12h without bacteriophage resistant strain.
Patent of invention CN107904213A discloses a kind of vibrio alginolyticus bacteriophage and the Bactericidal composite comprising the bacteriophage Object, bacteriophage can quickly inhibit the growth of vibrios in a short time, and inhibiting effect is held time length.In addition, the bacteriophage Safely, effectively, and high specificity, production cost is low, and single phagocytosis is compensated in the control process of pathogenic bacteria for bactericidal composition The deficiency of autogenic therapy.
Currently, in terms of the research of vibrio alginolyticus bacteriophage is concentrated mainly on bactericidal, for more cause of diseases The research of vibrio alginolyticus bacteriophage and its heat-resisting quantity is rarely reported.To sum up, urgent need exploitation vibrio alginolyticus resistant to high temperature is novel bites Thallus.
Summary of the invention
The purpose of the present invention is to provide a kind of vibrio alginolyticus bacteriophage resistant to high temperature and combinations thereof, kit and answer With having the characteristics that resistant to high temperature.
Concrete scheme is as follows: a kind of vibrio alginolyticus bacteriophage resistant to high temperature, and the vibrio alginolyticus bacteriophage is vibrio alginolyticus Bacteriophage VAP9 (Vibrio alginolyticus phage VAP9), deposit number are CCTCC NO:M 2018766.
Further, the vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticusphage VAP9) is strong Property bacteriophage, have one in polyhedral head and longer tail portion, major diameter L is 60~70nm, and transverse diameter W is 50~60nm, L/W ≈1；Tail portion includes contractile flesh sheath and tailfiber, and flesh sheath length is 30~40nm, and wide 5~10nm, tailfiber is with 3~4；Most Suitable pH is pH6~pH8, has patience under the conditions of 4 DEG C~75 DEG C, cultivates 8~10h under the conditions of MOI=0.001, potency is 2 × 1010pfu/mL。
Further, the vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) has Nucleotide sequence shown in SEQ ID No.1.
A kind of composition containing vibrio alginolyticus bacteriophage resistant to high temperature, the composition contain the vibrio alginolyticus phagocytosis At least one of body VAP9 (Vibrio alginolyticus phage VAP9) and other Vibriophages.
Further, the composition contains vibrio alginolyticus bacteriophage VAP7 (Vibrio alginolyticus phage VAP7), vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) and vibrio alginolyticus bacteriophage Two or more in VAP21 (Vibrio alginolyticus phage VAP21).
Further, the composition includes vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage It VAP9), further include one of each bacteriophage mutants or a variety of；The mutant at least 90% and corresponding Vibriophage Native sequences it is identical.
The reagent or kit of a kind of vibrio alginolyticus bacteriophage resistant to high temperature or vibrio alginolyticus bacteriophage composition.
A kind of vibrio alginolyticus bacteriophage resistant to high temperature or vibrio alginolyticus bacteriophage composition are bitten containing vibrio alginolyticus The reagent or kit of thallus or vibrio alginolyticus bacteriophage composition are being killed and/or are preventing the application in vibrios microorganism belonging to genus:
The vibrios microorganism belonging to genus includes vibrio alginolyticus V.alginolyticus, Vibrio anguillarum V.anguillarum, cholera Vibrios V.cholerae, non-Nonther conserved quantity non-O1V.cholerae, Fermi operator V.fischeri, vibrio fluvialis V.fluvialis, Vibrio harveyi V.harveyi, sick Vibrio damsela V.ordalii, vibrio parahaemolytious V.parahaemolyticus, Vibrio salmonicida V.salmonicida, vibrio mimicus V.mimicus, Vibrio splindidus V.splendidus, Vibrio aureus V.azureus, Vibrio gazogenes V.gazogenes, Vibrio vulnificus V.vulnificus, bamboo Pod vibrio piscium V.trachuri, damselfish vibrios V.damsela, floating vibrios V.natriegen, clam worm vibrios V.nereis, fish Enteron aisle vibrios V.ichthyoenteri, shark vibrio V.carchariae, Vibrio campbellii V.campbellii and kill prawn One of vibrios V.penaeicida or a variety of.
Vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) of the present invention is in bacteriophage The influence etc. of potency, the optimal multiplicity of infection (MOI) for cracking vibrio alginolyticus, pH stability, temperature to bacteriophage survival stability Aspect have following physiological property and the utility model has the advantages that
(1) potency (referring to table 1) with higher, wherein vibrio alginolyticus bacteriophage VAP9 (Vibrio Alginolyticus phage VAP9) infection vibrio alginolyticus best MOI be respectively 1:1000；And have high affinity and The virulent phage of cracking ability can crack rapidly vibrio alginolyticus (table 2)；The vibrio alginolyticus bacteriophage VAP9 (Vibrio Alginolyticus phage VAP9) when pH is 6-8, growth activity is impacted small；It is opposite to the thermal stability of temperature Preferably；It is suitable for 4 DEG C of low-temp storages.
(2) vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) virulence genes or not Well-founded because；There is wider host range for examination bacteriophage, to the discriminations of 180 plants of vibrio alginolyticus up to 90%.Three plants of molten algaes are mixed The bacteriophage of conjunction is to the discriminations of 180 plants of vibrio alginolyticus up to 96.7%.
(3) vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) is to non-pathogenic bacteria Cracking: only can recognize 10 plants of vibrio parahemolyticus bacterial strains in 4 plants, 5 plants floating vibrios in 1 plant and 3 plants of Vibrio aureus In 1 plant.Further, vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage of the present invention Or combinations thereof VAP9), object can not be identified exists including 7 plants of staphylococcus aureuses, 2 Hygrophilous monads and 6 plants of Escherichia coli Interior non-pathogenic bacteria.
(4) vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) and nonhost cause a disease Property bacterium interaction, can not identify 62 plants for examination nonhost pathogenic bacteria in any one plant.
(5) vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) has following advantage: It has high toxicity for stringent virulent phage and to host strain；With wider host range；In non-pathogenic bacteria place Your Majesty can preferably be proliferated；It can carry out large scale fermentation culture；Its culture solution can stable survival at room temperature, at 4 DEG C can be reserved for 12 months.The present invention does not carry out any genetic modification to for examination bacteriophage.Therefore, vibrio alginolyticus bacteriophage VAP9 of the invention (Vibrio alginolyticus phage VAP9) can provide excellent strain resource for exploitation phagotherapy, and have Good application and development prospect.The more vibrio alginolyticus of cocktail thereof cleavable of heretofore described vibrio alginolyticus bacteriophage, Cleavage rate has stronger cracking performance up to 96% or more.
(6) vibrio alginolyticus bacteriophage VAP7, VAP9 and VAP21 of the present invention or combinations thereof object can be by art technologies Personnel's record according to the present invention and common sense in the field are prepared into the medicine for being applied to disease caused by treating or preventing vibrio alginolyticus Agent or reagent.
(7) product form of vibrio alginolyticus bacteriophage VAP9 of the present invention or combinations thereof object includes but is not limited to carrier The forms such as carrying, concentration sprinkling or medicament immersion are applied to the ranges such as controlled host's body surface, life water body；As embodiment party One of case, it includes but is not limited to property of water-bearing carrier that the carrier, which carries form,；Concentration sprinkling form includes but is not limited to shrimp seedling spray It spills；It includes but is not limited to purificant etc. that medicament, which impregnates form,.
(8) vibrio alginolyticus bacteriophage VAP9 of the present invention is prepared to be applied to the fast of vibrio alginolyticus as effective component Fast detection reagent or kit.Its include but is not limited in the form of test paper, kit etc. to the vibrio alginolyticus in target sample into Row detection, or the target pathogenic bacteria in clinical sample are screened, it can effectively ensure the sensitivity detected.
(9) vibrio alginolyticus bacteriophage VAP9 of the present invention or combinations thereof object, which is produced, is applied to ring frequently as effective component The various products of border disinfection, for example including but be not limited to impregnate by liquid, sprinkling, with property of water-bearing carrier in combination use etc. in the form of pair Water distribution system, aquaculture facility or other environmental surfaces carry out disinfection decontamination, and the growth and activity of target bacteria can be effectively controlled. The liquid impregnates, sprinkling form includes but is not limited to disinfectant etc.；The property of water-bearing carrier includes but is not limited to that phosphate is slow Fliud flushing, seawater etc..
Figure of description
Fig. 1 is the vibrio alginolyticus bacteriophage VAP9 morphosis schematic diagram of Electronic Speculum identification.
Specific embodiment
Following embodiment does not limit effective range of the invention for the present invention is further explained in any manner.
In following instance, involved bacterial strain code name is the naming method number of our company.
Vibrio alginolyticus bacteriophage VAP7 (Vibrio alginolyticus phageVAP7), deposit number CCTCC NO:M 2018767, depositary institution are China typical culture collection center, and the preservation time is on November 09th, 2018, preservation Address is Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University；
Vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9), deposit number CCTCC NO:M 2018766, depositary institution are China typical culture collection center, and the preservation time is on November 09th, 2018, preservation Address is Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University；
Vibrio alginolyticus bacteriophage VAP21 (Vibrio alginolyticus phageVAP21), deposit number CCTCC NO:M 2018768, depositary institution are China typical culture collection center, and the preservation time is on November 09th, 2018, preservation Address is Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University.
In following instance,
The formula of the TSB fluid nutrient medium are as follows: tryptone 17g, soybean papain digestion object 3g, sodium chloride 5g, potassium dihydrogen phosphate 2.5g, glucose 2.5g, distilled water 1000ml, pH 7.0.
The formula of TSB (2%NaCl) culture solution are as follows: tryptone 17g, soybean papain digestion object 3g, chlorine Change sodium 20g, potassium dihydrogen phosphate 2.5g, glucose 2.5g, distilled water 1000ml, pH 7.0.
The formula of the TSA solid medium are as follows: tryptone 15g, soybean papain digestion object 5g, sodium chloride 5g, agar 15g, distilled water 1000ml, pH 7.0.
Semi-solid agar culture medium prescription are as follows: tryptone 17g, soybean papain digestion object 3g, sodium chloride 5g, phosphorus Acid dihydride potassium 2.5g, glucose 2.5g, agar 7g, distilled water 1000ml, pH 7.0.
SM formula of liquid are as follows: sodium chloride 8.5g, magnesium sulfate 2g, 1mol/L Tris-HCl 50ml, gelatin 0.25g, distilled water 1000ml。
Vibrio alginolyticus bacteriophage VAP7 (Vibrio alginolyticus phage VAP7), vibrio alginolyticus bacteriophage VAP9 (Vibrio alginolyticus phage VAP9) and vibrio alginolyticus bacteriophage VAP21 (Vibrio Alginolyticus phage VAP21) composition (1:1:1) be according to embodiment 8 method prepare.
1 vibrio alginolyticus bacteriophage VAP9 of embodiment is isolated and purified
It is taken on 20ml after acquiring Fujian marine product sample and Hainan Seawater Samples each 50ml, 5000rpm centrifugation 10min respectively 2 times of TSB fluid nutrient mediums of itself and 20ml and 2ml, are in the vibrio alginolyticus VAHN1 bacterium solution of logarithmic phase by clear liquid and degerming (108Cfu/ml) uniformly mixing, 150rpm shakes training overnight at 30 DEG C, is enriched with bacteriophage.By sample pregnant solution 5000rpm from Heart 10min, the miillpore filter degerming for taking supernatant to cross 0.22 μm obtain the filtrate containing bacteriophage.Take filtrate 50 μ l and Qi Su Main 300 μ l of vibrio alginolyticus bacterium solution is uniformly mixed, and standing 15min combines it sufficiently with the receptor of bacterium surface.By above-mentioned mixing Liquid is added 4ml and is cooled in 50 DEG C of semi-solid agar culture medium, is laid on immediately on the TSA plate solidified after mixing, to fine jade 6-8h is cultivated in 30 DEG C of inversions after rouge solidification, observes plaque growth situation.On the double-layer plate for forming plaque, use is sterile The big and bright plaque of pipette tips picking, 0.22 μm excessively of miillpore filter degerming must bite after oscillation desorption in 1ml SM liquid Thallus filtrate is inoculated in 5ml TSB fluid nutrient medium, and 0.1ml vibrio alginolyticus bacterium solution is added and mixes, at 30 DEG C 150rpm shakes training overnight, and 5000rpm centrifugation 10min is filtered after taking supernatant with bacteria filters, observes phagocytosis using double-layer agar technique Spot form.After repetitive operation 3-5 times, the consistent plaque of shapes and sizes can be obtained.It is separated from the marine product sample of Fujian To vibrio alginolyticus bacteriophage VAP9.Vibrio alginolyticus bacteriophage VAP9 generates single round phagocytosis on vibrio alginolyticus lawn Spot, center is bright, and edge has haloing, diameter 4-5mm；Vibrio alginolyticus bacteriophage VAP9 transmission electron microscope photo is referring to figure 1。
The deposit number of vibrio alginolyticus bacteriophage VAP9 is CCTCC NO:M 2018766.By nucleotide sequencing, molten algae Vibriophage VAP9 has the nucleotide sequence of SEQ ID NO:1.
The measurement of 2 vibrio alginolyticus bacteriophage VAP9 potency of embodiment
Dilution is done with SM liquid, step by step by 10 times of gradients of stoste of vibrio alginolyticus bacteriophage VAP9 (being made by embodiment 1) It is diluted to l08Times.L0 is taken respectively5、l06、l07And l08The bacteriophage culture solution l000 μ l of dilution and its host strain vibrio alginolyticus 300 μ l of VAHN1 bacterium solution is uniformly mixed, and standing 15min combines it sufficiently with the receptor of bacterium surface.Above-mentioned mixed liquor is added 4ml is cooled in 50 DEG C of semi-solid agar culture medium, is laid on immediately on the solid agar plate solidified after mixing, to agar 6-8h is cultivated in 30 DEG C of inversions after solidification.Each dilution need to do three Duplicate Samples, when counting take three of this dilution it is parallel The average of sample.Wherein, phage titer (PFU/ml)=average plaque number × extension rate
As can be drawn from Table 1, vibrio alginolyticus bacteriophage VAP9 has 10 after cultivating 12h10The potency of PFU/ml or more.
Table 1 cultivates the potency of vibrio alginolyticus bacteriophage VAP9 after 12h
Measurement of the 3 vibrio alginolyticus bacteriophage VAP9 of embodiment to vibrio alginolyticus optimal multiplicity of infection (MOI)
Picking vibrio alginolyticus single bacterium colony is inoculated into the test tube for filling 3mlTSB (2%NaCl) culture solution, 30 DEG C of shaking tables Middle 150rpm shaken cultivation 12h, obtains host bacteria suspension.Bacteria suspension is transferred to l0ml TSB (2% with 1:100 ratio NaCl) culture solution, 30 DEG C of 150rpm shaken cultivations to logarithm early period.The pure of bacteriophage VAP9 is added according to infection multiplicity ratio Culture solution (being made by embodiment 1) and host strain (MOI=bacteriophage quantity/bacterial number), TSB fluid nutrient medium, which is added, to be made respectively Pipe total volume is identical.150rpm shakes training overnight in 30 DEG C of shaking tables.5000g is centrifuged l0min and collects supernatant after culture Liquid measures phage titer.Each point makees double and answers pipe culture to be averaged, and is most to generate the MOI of highest phage titer Good infection multiplicity.Experiment is repeated 3 times.
The results are shown in Table 2, and vibrio alginolyticus bacteriophage VAP9 potency reaches highest (2.0x1010When PFU/ml), MOI For 1:1000.
The potency of vibrio alginolyticus bacteriophage VAP9 under the different infection multiplicities of table 2
The pH stability test of 4 vibrio alginolyticus bacteriophage VAP9 of embodiment
It takes sterile EP tube to be separately added into after the 900 μ l of TSB culture medium of different pH (1-14) and above-mentioned EP pipe is placed in 25 DEG C In water bath with thermostatic control, l00 μ l thallus pure culture liquid is added after equalized temperature, stands 1h at room temperature.It will after to the reaction time Sample uses double-layer agar technique to measure phage titer after doing appropriate dilution.It is repeated later respectively at 4h, 8h, for 24 hours and after 96h The above process repeats experiment 3 times.
The results are shown in Table 3, its potency is without obvious after vibrio alginolyticus bacteriophage VAP9 is to handle 96h within the scope of 6-8 in pH Variation.
Table 3 reacts vibrio alginolyticus bacteriophage VAP9pH value stability after different time
A. pH stability (the initial potency: 8x10 of vibrio alginolyticus bacteriophage VAP99PFU/ml)
The heat stabilization test of 5 vibrio alginolyticus bacteriophage VAP9 of embodiment
It respectively takes l00 μ l bacteriophage pure culture liquid loaded in sterile EP tube, is acted on respectively in 55 DEG C, 65 DEG C, 75 DEG C of water-baths 2h,24h,48h.Sample cell is taken out after action time and is immediately placed on cooling in ice bath, using bilayer after suitably diluting Flat band method measures phage titer.Experiment is repeated 3 times.
The results are shown in Table 4, and vibrio alginolyticus bacteriophage VAP9 is easier to survive at 55 DEG C in experimental group.Vibrio alginolyticus phagocytosis Body VAP9 has good high-temperature stability, and compared with the control, 65 DEG C of water-bath 2h still have 80% survival rate, water-bath 2h at 75 DEG C Still have 106PFU/ml potency.
Potency of the 4 vibrio alginolyticus bacteriophage VAP9 of table under different temperatures
The survival stability test of 6 vibrio alginolyticus bacteriophage VAP9 of embodiment
5m vibrio alginolyticus bacteriophage VAP9 pure culture liquid is taken to be sub-packed in sterile test tube, respectively at 4 DEG C, 25 DEG C, 30 DEG C It places, phage titer is periodically measured using double-layer agar technique after suitably diluting.
The results are shown in Table 5, and 50 weeks potency of vibrio alginolyticus bacteriophage VAP9 storage still have 10 under the conditions of 4 DEG C9PFU/ml； VAP9, which stores the potency order of magnitude in 12 weeks, at 25 DEG C does not reduce (109PFU/ml)；Bacteriophage VAP9 can be tieed up in 12 weeks at 30 DEG C Hold 108PFU/ml and the above potency；Bacteriophage has compared with fine melt ability host strain under each storage temperature.Bacteriophage is suitable It is stored at 4 DEG C.
Survival stability of the 5 vibrio alginolyticus bacteriophage VAP9 of table under different storage temperatures
7 vibrio alginolyticus bacteriophage VAP9 virulence gene of embodiment or the detection test of bad gene delection
65 kinds of identified virulence genes (table 6) from lysogenic phage in pathogenetic bacteria body are chosen, it is secondary by measurement Hemolytic Vibriophage VAP9 full-length genome simultaneously carries out bioinformatic analysis to it, to determine if containing above-mentioned virulence Gene.
The results show that not containing following virulence gene for examination bacteriophage.For examination bacteriophage invariably well-founded because.
The main known toxicity gene of lysogenic phage in 6 pathogenetic bacteria body of table
8 vibrio alginolyticus bacteriophage VAP9 of embodiment to vibrio alginolyticus cracking range test and vibrio alginolyticus bacteriophage VAP7, The preparation and cracking range test of VAP9 and VAP21 composition
The fragmentation pattern of bacteriophage is measured using drop method.Taking potency respectively is 1x108PFU/ml vibrio alginolyticus bacteriophage 3 plants of bacteriophages are uniformly mixed in SM liquid in equal volume, vibrio alginolyticus bacteriophage are made by the stoste of VAP7, VAP9 and VAP21 The cocktail thereof of the 1:1:1 of VAP7, VAP9 and VAP21.
The single colonie of 180 plants of vibrio alginolyticus of picking is inoculated in respectively in the test tube for filling 3ml TSB (2%NaCl), 150rpm cultivates 8h, and each strain bacterial solution is made.It takes 300 μ l bacteria suspensions to mix respectively with semisolid culturemedium and is laid on common fine jade On rouge plate, take the cocktail thereof drop of 10 μ l bacteriophage culture solutions and VAP7, VAP9 and VAP21 in the difference of plate respectively On position, it not can contact between when sample-adding each bacteriophage culture solution, in order to avoid influence test result.30 DEG C of cultures after natural air drying 6-8h observes result.Test is in triplicate.
The results are shown in Table 7, and vibrio alginolyticus bacteriophage VAP9 has wider host range.162 plants of VAP9 cleavable molten Algae vibrios, cleavage rate 90%.And the cocktail thereof liquid of VAP7, VAP9 and VAP21 (ratio 1:1:1) can recognize 174 plants Vibrio alginolyticus, cleavage rate 96.7%.Illustrate that vibrio alginolyticus bacteriophage VAP9 has wider host range, while bacteriophage chicken tail The limitation of host range when wine combination can make up bacteriophage single application, has greatly application latent in terms of Phage therapy Power.
The fragmentation pattern measurement result of table 7 vibrio alginolyticus bacteriophage VAP7, VAP9 and VAP21
Note: ++: cracking, plaque are very bright completely；+: cracking, plaque are bright completely；: it is uncracked
Breaking test of the 9 vibrio alginolyticus bacteriophage VAP9 of embodiment to non-pathogenic bacteria
33 plant non-pathogenic bacteria single colonies of the picking including vibrio parahemolyticus, Aeromonas hydrophila etc. connect respectively For kind in the test tube for filling 3ml TSB, 150rpm cultivates 8h, and each strain bacterial solution is made.Take 300 μ l bacteria suspensions solid with half respectively The mixing of body culture medium is laid on plain agar plate.5 μ l bacteriophage culture solutions are taken respectively and contain VAP7, VAP9 and VAP21 The cocktail thereof culture drop of (ratio 1:1:1) is on the different location of plate, between when sample-adding each bacteriophage culture solution not It is accessible, in order to avoid influence test result.30 DEG C of culture 6-8h after natural air drying observe result.Test is in triplicate.
The results are shown in Table 8, in our current research, vibrio alginolyticus bacteriophage VAP9 10 plants of vibrio parahemolyticus bacterial strains of cleavable In 4 plants；Its cocktail thereof can recognize 5 plants in vibrio parahemolyticus.VAP9 can recognize 1 in 5 plants of floating vibrios simultaneously 1 plant in strain and 3 plants of Vibrio aureus.All bacteriophages can not identify staphylococcus aureus, Escherichia coli and thermophilic aqueous vapor Monad.
The cross reaction of 8 vibrio alginolyticus bacteriophage of table and cocktail and 33 plants of non-pathogenic bacterias
Note: ++: cracking, plaque are very bright completely；+: cracking, plaque are bright completely；: it is uncracked.
10 vibrio alginolyticus bacteriophage VAP9 of embodiment is mutually tested with nonhost pathogenic bacteria
62 plants of nonhost pathogenic bacteria single colonies of picking are inoculated in respectively in the test tube for filling 3ml TSB, 150rpm 8h is cultivated, each strain bacterial solution is made.It takes 300 μ l bacteria suspensions to mix respectively with semisolid culturemedium and is laid on plain agar plate On.10 μ l bacteriophage culture solutions are taken respectively and contain vibrio alginolyticus bacteriophage VAP7, VAP9 and VAP21 (ratio 1:1:1) Cocktail thereof culture drop is not can contact between when sample-adding each bacteriophage culture solution on the different location of plate, in order to avoid influence Test result.37 DEG C of culture 6-8h after natural air drying observe result.Test is in triplicate.
Bacteriophage is interacted with nonhost pathogenic bacteria and is reacted, the results show that VAP9 and cocktail thereof without 62 plants of method identification is for trying nonhost pathogenic bacteria (table 9).
The cross reaction of 9 vibrio alginolyticus bacteriophage of table and cocktail thereof and 62 plants of pathogenic bacterias
Note :-: it is uncracked.
It is prepared by the fermentation of 11 vibrio alginolyticus bacteriophage VAP9 of embodiment
Picking vibrio alginolyticus VAHN1 single colonie, is inoculated into the test tube for filling 3mlTSB (2%NaCl) culture solution, 30 DEG C 150rpm shaken cultivation 12h, obtains host bacteria suspension in shaking table.Bacteria suspension is transferred to 500ml TSB (2% with 1:100 ratio NaCl) culture solution, 30 DEG C of 150rpm shaken cultivations to logarithm early period simultaneously measure its bacteria suspension concentration.Vibrio alginolyticus bacteriophage VAP9 The system of fermentation preparation is 8L, and fermentation medium is TSB culture medium.The initial pH value of fermentation medium is 7.Using flame Inocalation method inoculation, the bacteriophage of 50ml is respectively connected to its corresponding optimal multiplicity of infection ratio into fermentation medium (106) and logarithmic phase host bacterium solution (10 PFU/ml9CFU/ml).It is passed through filtrated air in fermentation process, and 3 ‰ defoaming agents are added, Fermentation preparation time is 12h.Every 2h takes the mixed liquor of 20ml bacteriophage and host strain in sterile from sample tap since fermentation In container, 5000rpm is centrifuged 10min, and the miillpore filter degerming for taking supernatant to cross 0.22 μm obtains the filtrate containing bacteriophage simultaneously Its potency is measured, method is referring to embodiment 2.To after fermentation, by whole mixed liquors of bacteriophage and host strain from sample tap It takes out in access sterile chamber, 5000rpm is centrifuged 10min, and supernatant is taken to filter through vacuum pump to sterilizing filter device It is interior, it obtains bacteriophage fermentation liquid and is saved at 4 DEG C.
As shown in Table 10, vibrio alginolyticus bacteriophage VAP9 in ferment 12h when potency up to highest, be 3.5x1010PFU/ml。 12h after fermentation phage titer by initial 104PFU/ml is increased to 1010PFU/ml or more improves 6 quantity Grade.Therefore, it is practicable for carrying out the large-scale industry preparation of bacteriophage using fermentation method.
The fermentation dynamic of 10 vibrio alginolyticus bacteriophage VAP9 of table
This specific embodiment is only explanation of the invention, is not limitation of the present invention, those skilled in the art Member can according to need the modification that not creative contribution is made to the present embodiment after reading this specification, but as long as at this Protection in the scope of the claims of invention all by invention method.
Sequence table
<110>Fei Jileke (Nanjing) Biotechnology Co., Ltd
<120>vibrio alginolyticus bacteriophage resistant to high temperature and combinations thereof, kit and application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 82456
<212> DNA
<213> Phage VAP9
<400> 1
cttgatattg cccgtaagaa cgtcgtgagc gctctacgta caacgaaatt tgaaaagttg 60
aacgaacgta aagacgaact gcatgttgtc cgtacagagt acacaggcaa gcttaacaag 120
ctcaataact caattggcat gttggaactt gaaatcaaga acctgaaagg cgccttggct 180
gaaacggaag aacaattgac taagcttcaa gagcgccaag catctgacga ttccaagttc 240
aatgaaggtc ttcgtgttcg caataaagat gtgttgaatg caaaggataa acttgcagaa 300
actcagtctg aaatccaggt gaataccaaa cttcaagacg atttgaaagc agctcgttta 360
ttgattggtg ataaagaaat caaagcggat atcgtgaagc aatacctacc gttcatgaac 420
gccaagatca acgagtacct ggaagcattg aacatctttg ttggattcga aattgatgaa 480
aactttgaag tatctatgaa cgcaccagaa cgtaagaacc agacattatt ctcattgtct 540
agtggtcagc gtgctcgtat caacctggca atcatgctgg caatgcgtga agtgggtaac 600
ttgaagtcaa gtatccagtg taacctgttg acttgtgatg aaatcttcga gtgtctgagt 660
gcacaaggtg ctgaagaagc tgtggaaatg atgaaacata aattccctga tcagaacatc 720
ttcgtggtta ctcagcgtaa cgaagaattc cgtgagcact ttgagaacaa cattcacctt 780
gaactcaaaa acggtttcac tgaaatcaaa tagttcaaaa gtttgaaagc gataaaatgg 840
tcttactggt tggtaagacc atttttattt ggagaaatat catgagtgta tcatcatatt 900
atgaacacct gttgcgaaac cctggacagc ataataatct gtcaggccaa tatgttattc 960
ttgaacatgg attcaacggt aactatatca atggtatgag tattggtcct gagtttgtcg 1020
tagagcacaa ggagcactac aacaaaccag tcttcgcaaa aattctgggt gcttggtctg 1080
atcctacaga tatggacatt catttccgtg tcgcattact cgaagacaac aaagaaaccg 1140
cacacattac tactaaacta ggtcgtatta cagagcgaca ttacttggaa caagtcaaca 1200
gtattgaaga aattcaatgt tggaccatga tttcaagaga acgttacatt gacctaatca 1260
cttacaaagc agaggacgag atcgatgccg aacacttctg agcaaactat cacccaattt 1320
gtagaaacag aattcaaaga tttttctgtt gaaaacaccc gtcgtatggt gccgtcaatt 1380
gttgatggtc taaagcgttc acaacgtaaa gccgtatggg ctatgtcaca gatgaaaggt 1440
ctggaaactg ttgagcgtgt gggtctaaaa gccgccgcga tgactgctta caaacacggt 1500
ggtaataaca tgtgtggtac agtcattaac ttggctaagg acttccccgg caccaacaat 1560
gttccattga ttcgcaagga aggccagttt ggttcgttca tcgaccatga caactcgagt 1620
gctcgttata tcgaagctga gattcatgac aactaccgtg cattcttcaa agccgaagac 1680
gatgacatct tgtcacacca gtacgaccaa ggtaaaaaga ttgaacccta ctacctatac 1740
ccaatcatac caatcgtgct cgtgaatggc gccagagcag ttggttcagg ttatggtact 1800
aaattcccag gccataaggt taacgacgtc ctgatagctg ttctggagac cttgtccatg 1860
ggcagacccc agaccaaact tgtgccaggt tacaacaatt acgttggtga tgtgatccgt 1920
aaagattctg aacaaattga aatacgtggg aagcttgaaa tcaagaatac cacaaccgtt 1980
gtgatcactg gtattatccc aaattatgat caggaaacat tcaagacaaa gcaactgatt 2040
cctaaacttg atgcgaaaga aatcgttgac tacattgacg attctaacga gactgacggc 2100
tggaacatca caattaagta tacccgtgaa gcattgagtg cccatactga agaatctctg 2160
cttgaatctt ttggtttgat taaccgtgat acgccaaact atacgctttg ggacttcgaa 2220
ggtaagatca agaaatacga ttgtcctgaa gatatcgttg tcgagtttgt tgaatggcgt 2280
ttggcccgca acgaagaacg tcgccaagcg atgttatcaa aactgcaatc agagattgat 2340
tactggacac aatatttgtg gttcctgaaa gactactttg ataatacaga taagtacaaa 2400
gcttgcaaca aacaacagat gctagagcgt ttgctgggct tgggtatcta tgacgaacat 2460
gtagacaagt tcatcaaaac ccctgcatac aaactaaatg ctgaaggcat tactaaggct 2520
gaagaagcaa tcaacaattt gaaagcagag tacaaagtgc ttcaagagaa aactccaaag 2580
tcaatttatc ttgatgactt ggagcagtta ggtcagtaca ttgaggaaca aaataatgtt 2640
gcgtaaattc ttattatggt tagcagccaa caaagttttc aaagcacatc cacttcgtga 2700
tcttatcaag caatcagagt ggacaatgtt gggtactcca ttgaacgata agtaccatgc 2760
tggcgattta ccaccaggag tcaagcctga agaatacccg gccctattgg cacgtctgcg 2820
tggcacattc aaaacaaata aggtgtatcc aatctacttg gaccggacag gtaaagttgc 2880
gattcaagca tccggtagtg acggtgagcg ttcgatccga gtgattagca tgaagaagtc 2940
ccaatacttc aacgaaatga atatcaatga gcagtggagt tactttgaac attcacctaa 3000
ccctgtttat cgtaagatgt tcaaattcat ataccgacac cttgatgtgg atatgggtat 3060
tatcaaaaac acataagtta tgatatgatg gttattacta aatactggta ataaccattt 3120
ttattactta cgataggaca acaaataatg tcaaattctt ttattaatat gcttaatgaa 3180
caacacctgg aaggtactgt tcttactgaa gctatgatta ctgaagcatt cgattcagaa 3240
ccatatgagt tccaattgga agctcctcgt gctgacaacg tgtatgctgt attcactgat 3300
gagaagggtg gtgaattccg agtacaattt tattcaggtg ttggccttgg taaaggagta 3360
cgccgtgtcc gtttgggtaa gaaaaagggc gcgtcattct ctgatgcaaa catcaagttc 3420
aataaccctc aacgcgcgat cgcaaccatg cttgcagctg cagaggaatt cttcctaact 3480
ccaatcggta agaaagcacc aggttttgct tttgaattta caaagaaagc cgctcctcgt 3540
ggtgtgcgtg ttgtacgtgc tgtcatgaag cgtaagttcc gccaaaagtt tgatgttctt 3600
gattcatcat tcagcccaga aaaaggtaaa ggtttcgtat gggtagtacg taaaggccaa 3660
gatgctgcta aagtcttctc cggtcctaag gttgccggtt taattggtgg taatgatacg 3720
gatcctatcg ttgaacctga tccagcaccg gaacctactg atgataataa actactcata 3780
aagtggaatc aaaaaactgc tgacttcatg ctcgaaaaca tccgaccaaa cttagaatct 3840
gcactatcgt tctatggtcc atgggaacag gtgggtacta ctcagaacag cgttaccatg 3900
cgtttcgaga acaatggtta tgaagccact atcggtgtca atgttgaaaa actacagtac 3960
atttttgaag gtaaattgta caagaacatc aacgaaattt ttgaacttat taaaccaatg 4020
gttcaaccta tcgttgatgt aaccgatcaa aaccatgaat ctttcgcggc tcatttgaat 4080
atgcaacagg gtattgtggc atccaaagaa gccggcggcg ttcgcatcta tttgcctggt 4140
tacaacaagg atatgttagc gacctttaca gaaggaaaag gctgggtatt acacgctgat 4200
ggtaaagctg aagaatttgt ctcaactggt gaagttaaag actttgttgt taatatgaag 4260
aacaaccttg ataagcttga tagtcttact ggtgatgaag atgaagatga agatgaagat 4320
ttcgaaggta tgcaaccaat cgatatgttt gcgggcttac tctacgaaga cttgaccgtc 4380
gatgtagatt caactggtat caactgggtg gtgacagatg acgaaggcga cttcgaagca 4440
attatccgca cggtcaatta cccgacactt aaagttgatg catacggcgt cgatcctaat 4500
catgcttacg acggcattga agaagccgca aatgctatca acgattcata tgatctgatg 4560
ttccctgagg acgacgaaga tgaagaagac gatccatcat acatgccacg tcgtatgaat 4620
gatgttaccc aatcactgca aaatgttggt tttcaggttg aaactgatga tttaaaccca 4680
atccaattga atgtttcaaa ttccagtggt cgtacggtag catcagtaat tcacaagcct 4740
acagcatttg aagttgtatc cgcatacaat gacaagcgtg aattctttga gtctgatgaa 4800
gaacacccat cacaagtaac attgggtggt attatcaagt ttgttcacaa gttgaataaa 4860
actgtcccag tcaacgagca aacagtaaac ctaggtgtca agacattacc aacgtttatg 4920
gaactgctta acgaccagaa gactattgaa gaatcatatg tcgaaatcgg taagacctat 4980
gtcggcttcg gtcgtcatgc ggatcgtggt acagtaatcg ttgatgacaa gaagaaagag 5040
attggtaaag gtttattcat catcttccat aagaagggtg aaaaacgttc ccaaaaactg 5100
gctgccaaga tttttgacca acgatataag aaggcataag tcttcaaaat aacaacttag 5160
actaatatgg gattgctttc taaatattga gagcaatccc tattttattt gtgaggaaat 5220
aatgactaaa tccttcctag aattactgag tgaaagtgat gttgaactgg taactgaagc 5280
catgatcaca gaagctttca accaagaacc atacgactac acgctagagc aaccgcgtac 5340
ggataatatc tttgcaattt tccaagagga agatggctcc gaattccgaa ttcaattcta 5400
ttcgtcggtt ggtcttggta aaggcgttcg tcgtgtccgt ataggtcgta aggccggtgg 5460
taagaactcg ttcactgacg ctggtcttaa gttcaagaat ccggcacgcg tgattgcgac 5520
catgattgca gcttctgaag aattcttcac aacacctgtt ggtaagaaag cagacggata 5580
tgcatttgag ttgaccaaga aggctgctcc gcgtggtgta cgtgttatcc gccaggtcat 5640
gaaacgcaag ttccgtcaga agttcacaac tctggactca acctttagcc ctgaagaagg 5700
caagagcttt atctgggtat tacgcaaagg taagaaagct gaagacgttt tcaatggtcc 5760
taagaacaaa ggctttatca gcgacgatgg cgaaatcact cctccaaaca aacgtgacga 5820
acagaaagaa attgctgagc tcttattccc taaaccattc aaagagatga ctggggtcta 5880
ttcaggctca tataacaatg catatcgcct ggttgttgaa tacgacggca ctttggtggc 5940
ttctaaaggt ggtacagagt ttggacgtat cgaacgcaaa gaaggtgaaa gccctcgtaa 6000
attcgtaacc cgtgcttcag ctcagcttgg tgagtggcaa actcgtatgg aaaaaggcga 6060
gtgggacgaa ggctaccgtc gttggaaaga tgatgtggtt aaacctaccg tcaaatcaac 6120
caaagaaagt gctgaagaac ttgttcaaat gcttcgagct atgggatttg aagatatcaa 6180
agacgtcagc cataaaggat ggccagatta cacattcaat gctacagcgt ttggcaagag 6240
ctacaccttt atgccaggtc gtgaagaaca ccaaggtatg tgggcattct atcctggggg 6300
ttccctctac cgcaaagatg gtgaaaccaa ctttactatt gaagaacttc gtacagaaat 6360
cgaagccatt caacgtgaag acacagaaaa caccaagact gcttttgtca atgccattaa 6420
agctcaattg attcgcacag aagttggtga tgtcaaatct gatggtcgct cgctccttgt 6480
taactgggcc gatactctca aatgggacat cagtattgaa tttgataagc ttgttaagat 6540
ggcaactgtc ggtgtaaatg gtgcaaaggt ccattttgct aatgttacaa aggtgaaaga 6600
ttttaccaat gccatcctga aagttatcga tgaagatatt cgaaaacatg aaggtgatga 6660
tacaagcgac gacgattcag aagttgttga ttcattgaca tcgatcatta agtcgatgcg 6720
tcgtactgtg attactgctg ataagcaaac agctgcaacc catagagtgg actttagtct 6780
gagcacactt gttaaaggtg ttgccaagtt atcgttcaaa actggtaaaa ttgttatcac 6840
ccgtactgat gatgagggat atgaattccc taaattgcag tttgaaagct taagtgtaat 6900
gccacgtggt gaaagcctac gtgcgatcat gaatagtggt gttgtcggtg agccagctgt 6960
ggcaatcgac ttgatttaca acagcattat tgaacatccg ttattccaca acgtgaagat 7020
gcgtggtgca ggtcgcgaaa agtctatcct ttttgaagat ccaatgtatg gagcaactca 7080
tgttattact gcggctgcta tggtccatgt gcgaccattt accagtaaca tcgcagtttc 7140
tggatacagt atgtatgtgg acttagataa gtggcaaaaa gttccaaatg aaatgggcaa 7200
aacaatcagt actaagacat ttgacgattt cgaaagtact gtacgagatg cttcgggtta 7260
caagagtgaa tccattgatt ccaatcgtaa accaacccag ttaaaacaca acgtaataca 7320
cattccacaa attaatgaga atgtgaccat caaagaaatt acgtttggcc gagcatggga 7380
aatctcatac ggtgagaagt ctaagctagt tactacgggc aaacaatttg atgaagtgat 7440
ggtagacatc cgttggcttg tggcaactca atcaaacagt gagtatcgtc agttcaatat 7500
tccagaatgg caacggttgt tgatggacaa acttacttca aacaaacatg gtttagtgtt 7560
ccgtgtacta ccacatgttg gccgtgacgg tcacgaccag gaagtcattg atgatatgaa 7620
ttacatcatc aagtttactg gtggtaagtc acgtcgtacc caagaatttg gtgtcgaaca 7680
caatgaacgt acaagcatgt actatgtcgt ttcttcatgg ggtgatcgcc aatatgctga 7740
gtttgaacat gtcgacgatt tggttgaatc catttatgcg acattactag ctggtaacgg 7800
ttggcatgat atgatcatgg caattgacga cattggcaac ggcgaataca cgttcgaacg 7860
accagagttc ttatcaaata tggatgctgc tgctattcct gactttgaag ctttgttggg 7920
tatcgaagat gacgccaaac tgaaagatat catatcattt gactcatttg acttggctct 7980
ccgtctaggt tgggaagata tgcctaaagt acgcaacaca tttaagaagg tcgttgaggt 8040
actgaaatct ggtcgtgatg ttgaagaact ccgtgaagaa atggaaaatg actacggtga 8100
agcgacatca attctaatga tggaaacgat tatggatcgt attcataaga atatcgacaa 8160
taagacgttg gaattcgttc aggatgctat tgctaacagt acattgtcta aagaagacgc 8220
tgatcaaatg ggacgtgaag ctgtagctga ttcggaccgt agtgcagtat tacacgttaa 8280
ccgcacagca ccagccgtta acctgttgtc ttcgttgtat agattatgta ataacaagac 8340
aacgtttaag cgtttcaaaa cagatgagag tggccgcgcg tactactcga atgctaccgg 8400
tgagatcgct gtaactggtc gttttgattt tgttacacta tggcacgaat ttggccactc 8460
aattgagtat agtgaccgac gcattctaaa gggttctaaa gatatcctgg agaaagaagc 8520
gacatctgct gtaaaccgtg gtagtcaagc atttgtccgt cttcgtgatg tagaacactc 8580
tggctatggt gaaaatgaaa ttgctgtcaa cggtaacttc atcaactact atactggtcg 8640
cgtgtactct aataggattg gtcaggttat ggctattcag gaaccaagta tggtccaaga 8700
agtccatgct actgagatca tctcaatggg cttacagtac ctaacagacc aagatgatgt 8760
tagcgtcggg gcatgggcca agaagaaccc acatcattac cgttttgtaa tgagtgtagt 8820
caaccaatta cataaaggac tttaatcatg attaaatttc aaatgaaact aaaagatggc 8880
acaccaattt cggttcaatt tccacaagcg aagcgacctg atgatcgtgt tgagtacctg 8940
gcatatccac tgaatcagtt ggctatcatc aaggcactca tcaagaacat tccagatgaa 9000
cgtggcatgc ttatgacaac caaaaatgca acacctgttg gtgcggtcaa cgcacttcgt 9060
aataaaggct atacagtaac aggtgacgaa gattggcaaa ttcgtccagg tgttctttaa 9120
aacttcaagg gtgacttcgg tcaccctttt tcatatctgt atttgaacat aaaatagact 9180
cagattgtta tggaagaaat actatgtcta agaaaattgc aaaatgggat catactgctc 9240
tttgccattt catgggcgaa tatgccaaaa agaaaatgaa tgcaggtctc gtatttgttg 9300
aacataaaac gatgatgacc caccaaatac ctgacgtgtt aatttaccgt cagacttgta 9360
ctattgaact tgagattaaa gtcaaccgtg ctgatttcct tgccgaccag aagaaacctc 9420
atcgcatgga tccgagcaag ggtgttggtg attaccgatt ctatgttgca ccaaagggta 9480
tgatcaaggt cgaagaacta ccagaacatt gggggttact ggaagtatct ggtaagcgtg 9540
tttataagac acatgtgcct ccatcattcc aatgtcttta tggcaatcgt gaatacctaa 9600
tgagtgccat caaacagcaa tggaaggagt tttacaaacc tgaacgttat ccaggttctc 9660
gcaaacatat acagcgttct atacgtcgcg atgttgctga tgttagtttt atctttcctg 9720
agaaagatat ctcggctgaa accagctttt tatatgctgc acatcgccag gaaattattg 9780
cttcacaaaa gggtttaatt aaacctgcta aggatatcgt gtgggttaga ccattcaatg 9840
gcgatatgat ggatgaaggg agattgttgc gatgaaccaa gggcctaata ctacagggtg 9900
tacgtgtgta ttcccatgcg gtgggacaga accaaaatgt cctagagtta aacactttac 9960
aaaacggacc agaaacaatg gcaattctcg gaagaaagac ctacgccgga gtgggcagca 10020
gaaaagctcc tgaatgggcc tgtaagaagc tttccctctt agcctatgtg ttagcatcag 10080
aaggatacat ttgtaactct gggctcgctc cacaaagcga tgaggcattc tattggggta 10140
ttaaagcgta ctgtgatgag atgaaggcgg atcttgataa attgtataag ggtttcatcc 10200
cctggaatgg gtttatgaag aagtggtatt tccacatgcc tggtgttacg ttaatccagg 10260
atccaacatt gattgacaaa gcaaccgcga tcgcaaagca tcacatgtca cagagtcatt 10320
gggatgcgtg tacggaacaa gcccaacaat tacactccag aaatagtttc cagatgtatg 10380
gtgaaaagct caataaacgt gtagggtttg tggcatgttg ggcaccacca acaggcaagg 10440
gtgatcatgt gaaaggtgga acgggccaga cggtaagaat aggcatcaaa gagaagatgc 10500
caatattcaa tatagcgact aaagatggat tacgttgggc taatgctaag atccagaagt 10560
tcctggtcag gggcggagtt ttccctaaga gtttgctcga gcacgtattt gttcagccaa 10620
caaacggcca acacgcttaa ccttttcatt acggatactt tgtttctttt tgagttcccg 10680
cttctcttta cgacgagcaa ccattttctt ttggatttct ttgtcgtgtt tgatgccggg 10740
aactaattct tcctcagtcc atttcacaaa tgcccaaccc ctttcagcgc agtaacgttc 10800
agttgcctcc catttcgcag agttgaccgc ccaagtagca cactcacgca tgtatgtttc 10860
atctttctta ttacctttgc gtggtggttg tgtctgggtt aacggcttta tctcgaccag 10920
agtaattttg agttcacccg tctcaggatc agcagtccat atcttaaggt ccatcatata 10980
acggctctga cgacctgtcc gcgggttagt atatggaatt atttcttcct cggagttcca 11040
cataataatg gcgggttcat tgtcgcagaa cttaaacgcc gctaattcat acgatgaacg 11100
gaagatgatg tcattaatat tgccgcggta cttctctgga ttaactggtt taaagtcacc 11160
cgataaccat ggaccacttg ccttatgttt agccatatcg atacctaaat aactagagat 11220
tattccaagt atttaaggtt tgatgatgct aaatttgaaa gacaaattat ttggtcgtcg 11280
tcaggaggtt aacctgacgc gtacgcaagc caaatacaat atgcttgaac ctaagactga 11340
agtaaaacag ctggtctacc catcaaacct gttaaaggct agtaacggta tgggccattt 11400
tatactgttc aacttcaact atatctctgg ttcacagttt aaatcaaaag ttaaccgtat 11460
tgagaatggc ggcaatgtgc agtcgccgtt cgctaagcct gttgtctacg ataaacagtc 11520
taactcaaca cgctcctcaa tgctgaaggt gcataaacgt tccgaggagt cagttgcgat 11580
ctacatgccg gaaaatatta gcacaactta tagcgctgac tggcagacaa ccgaacttgg 11640
cattgccgga cgtttaatca aggctactcg taactttgac aacctggatc tcaatgatgt 11700
gggtaatacg ctcacagaag aagccaagaa cctcgttact ggtgccatcc agacgttaac 11760
acctattaat gctaaagacg cggcagaact ccttacagga acaattagca atccttttgt 11820
tgaggtgtta ttcaagggag tcaataaccg agagtttaac taccagttca agtttgtacc 11880
caagaatgaa aatgaggctc gtacggtgcg tgaaattatt cgccgtttta agttccatat 11940
gatgcctgag tacaaataca aacaaaatga tagctcttac ttgttacacc catcaactgt 12000
tgacgtgacc ttcatgaaga ttaccgatca gggcgtaggt gagcgtaatg gttggttgca 12060
tcgaatgagt acttgtgcga tcagtaacgt gtccgttgac gaagcgccag aaggttacaa 12120
cgtccaccat gatgactcgc ccgttgcaac agttatggac attacattga ttgagcttga 12180
acaacttcat aagggccgat tcatagagcc aggagataca ttctaatgtt cgaacatttc 12240
ccgaaacgtt ggttcgtcga tttggaaggc gaacacttac ttatgacaaa catttgtcgt 12300
aagtttagaa taccaaagaa cttggttgat gctcaacaaa tttacttaaa atactatatt 12360
cgcgatggtg acactgcacg tagtctggcg acacgtcttt atggtgatag tgaactgttc 12420
tggttggtat acttggcaaa taatatcaag gatcctgcta acgaatggcc aaagtctgaa 12480
gaacagttga ccgattggtt agtccgtgaa tatggtcttg aaaagatgta ccagaccaag 12540
cattacatca acccttatgg tgatgttatt gatatccatg ctattaaggc ccagaagggt 12600
ttattatcga tgagtgattc tgatgtcatt agccggttca ctttggaacc tgttacatat 12660
gacaagtacc accgagatat caacgataaa cgccgtaata tgagactcat cgatcctgat 12720
tatgcagatg agttccagaa agcattggag acagtattcg atgaagacca atgatgcatt 12780
taacccatcg aacccagtta atattgagtt cttggagttt actaccaatg atggcaacaa 12840
atatgacctg aagaactttt ttattgagtt tgatatctat acgtcgattg caaatatcat 12900
ggaagcacga gtccttctta tcgatgccaa cgatattgta tccaaattcc ctatcatggg 12960
aggtgagaag attcgtttcc gattctcaac atcaggtcgt aaagataaaa caaccttgga 13020
tttccgtgtg ggttctatcg gcgaacgtat ccaaactgat aagatgaaat cactatggtt 13080
agaactggtg actgaagacc gttatatgga tgttaaacgc cgagcatctc gaggttttaa 13140
cacggtctat acagagatgg tacaaaagat cattagtgaa gccggattaa ctagaccttt 13200
gaatagtgag caaagtgttg gtattagtag tgttgctaca ccacgttggt caccattgaa 13260
tgtgtgcaag tggattgcaa ctcgtgcaat tgatggcaat aacatgccct ttgcattctt 13320
tgaagatttt aaaggttata acttcatcag tttaactacg atgattaagg gtgaagtgga 13380
agatcaattc ttcttgcaag aagctggttt caatgaggag gattacaaga agttccgtaa 13440
tattcaagat atctgggtgg aatcaaagta tgattcagtc ggcttcaact ccacagggtt 13500
aaacaaagct aaacacgttc agtttgactt taatacaaag aagattgtca aagaagaccg 13560
cagctacaag caacactttg acaagatgcc gtcaatggaa cagttccctt ataaagttga 13620
ggaaggccca gagtatgaaa cggagattat cgagtatact agaccagata acagtcactt 13680
gggacaatat aaccgaaaac ttttgaagta tttgttccaa caatatgctc taaatattac 13740
cgtatatggg gataatgaaa tcaagcttgg tggcatttac aagttcaatg tgcctgaagt 13800
tggtcctatg gtggatggtg tcagacacaa tgagaagtac accagcggca agttccttgc 13860
tgcagccatt aagcatgtca tccgaccaga cgattacaag atgaatattc gtctaatcaa 13920
ggactcatta tcagatcagt tggccgaagc ttcagaggta taatatggca aacgaattaa 13980
gccaaatcct aaaccaggtt aaagccgcac aagggacagc tgtatctgaa caaaagctca 14040
atgaaatgtc acaagccatc agtgagctta atgacaagag tgtatcccag tttaatattc 14100
ttaaccagat ggcttttact gctatttcta accttgaaaa ctcaattccg ggtaaagagg 14160
cattgattgc tgcgttctct gctggtaacc ctttggttgc cggtggtatt caacttatgc 14220
gtgatatggc agcacaacgt cgagaggcga agaagaaagc acaggcggca tacgatgagc 14280
aagtccaggt tatccaggaa gaatttaaaa aggttgtccc tgaagaaaag gcggaacctc 14340
gttcacagaa taacgaacaa caaaagttag tacctgatga acgccaacct atggagcaac 14400
caaatgtgac ggttgaaaat atcgtcccag aagctccaga acaagttgtt caagataaca 14460
ctgatattgt acagggtcta agccctttgg acaaactgga acccatccgg gagcaaatag 14520
acgacctaaa tggcattgct tatgctcaac tcaaagtttt aggcgattta tatcaagaat 14580
ggtctggtac accgagcaaa atgatcgaga tactagacga acaattgatg gaacaagaac 14640
gcctacgcaa ggataatgag cagagagcag cagacgagac gcttagggga gcagaagaac 14700
cacctaaact tccaacgcca acagatggca gcggtgtgtc ggaaaccgtt aaagatgaat 14760
ggtctccaat ccaagctgtg attatgggaa tgggtggtct ctcgtctgct gctggtgcta 14820
tgttgaaacc gttgtctaag gttcttaaaa tcttccgagt cggtccactt gcccttatct 14880
cagctgcttg ggattttggt aaagggttca taaatgctgg cgaaatcttg gataaggaaa 14940
acgttggtat ttctgatcgt atagttgctg gtgccagtga gatacttggt ggctttgccg 15000
atgttgttga ctttggtgct gggttactag gttacgatac aaactttggt cagacaatcc 15060
gtgagtctta tctcgagatt atggaaacac cattaaagta tgccaatatg gttgaggaac 15120
aaatctccaa gatttttgat ggtattgatg gttcaactaa attatcggct atacctggta 15180
tcatctggga taatattgtt ggtatgtacc acggagcaat ggctaagatc acatcttatg 15240
acctgatagg taaagccaaa gacaaactaa ctgaacttag cacatcagtc actgatagtg 15300
ttacgggatt tgttgaggat atcgagacgt caattactga ttacattaaa ggtatctcag 15360
aaacgacttc cgaattcttt acgaacttgt gggatacaag ccttgacaag atgattaata 15420
tgatcaagga tgttccgctt gttggtgagg atatgtccaa gaagcttgaa tcgatgaagg 15480
ttaaaaacca ggttgattcc gaaactttag atcctgttaa agttgaggcc gctcgtcgca 15540
agttgactga agctcgtgaa cgtaccgacc gcttccaggc aatagttgct gaacaacaaa 15600
atgaagaaac tcgcaaacag atggctcaag tgttgaacat gactactgtc aacaacaata 15660
agtcaatgac tactgtggtc aaaggtggta aacaagatgc gttcaaccca tctcgacgtg 15720
acttaggttt cgattggtaa aacaaaaagc cccagaattg ttcctggggc tttttattac 15780
atttcggata ggatattcaa tacttgagac aattctaaaa ctttagccag gttctcacct 15840
tcccagtatt gacccgcaaa ctcagcaaac acttcttctg ttacatggtt ttcgccagta 15900
atacggcttt tattgatgat ttgacaaaca tggatatttt cacctgattt gtcacacgca 15960
aaaatacgat agttgttttg gtcgtcacga ctgataccac aacatttgta aacaacatca 16020
ataccttcaa tggttaactc cgaacccagt ccaatagaat gacctgttac gtcattggtt 16080
gtgattgagt aaagtagtga gttcaccatt attttgtcta gtttttgcaa taggtatgtg 16140
gtattcatta ttttaagtct cgacggttaa tttcttggat tagccagcgt tgtgctgact 16200
catggtcaaa tacataaaag ccttgtattt gtttgatgtg atctgagatt gttggccaga 16260
acatgtcgtt cgcgtaaatc ttatcgtaca cactcacaaa ggatgtcaca cgatttaagc 16320
ccacaacgaa ccaccatgga agtttctcag caagtaaggc ctttaatagc caggtatgtt 16380
cagaacctga tgtgggacga tacaggtcca taaacgatag ccctttagct tcagcatagt 16440
caagcattga atcgttcatg aaagtacgga aacggtaggt gaagttttca ataacagagc 16500
ggaactcact agcacgttga tctgcatctt tctttagcaa gtctaataac cacatatttg 16560
gatcatggat aaagtttgcc gcaaggtgag cgatgagatg ttcttcaggc agctgcgaaa 16620
aacgtttgtg gagttttaga atctgttgac gaccacgtgc tttgtacagt gattcaacct 16680
tcagattaac ttgagaaccg tattcaaacc agtcatactt gacaccttgt ttaaagtgta 16740
gtttcatacc catgtacgtc tgatacgtag gtaaagccga ttcaaattcc atctgttccc 16800
aatgtgacag ttcaagtggt ttcttaacct gtcctttagt tcgtttgttt accatgatat 16860
ttgctccaat aagaagatga gaccatgata acacggtctc attattttga agatttaatt 16920
ggttgcttga atataagttt tttcaaaggc acccttctca atacggtaga agcctttggt 16980
gtttgataat gacgaagtcg caagcatgtc accattattc agtaccatag cttcaccttg 17040
ttcattttca aactttatat tatcgccaag ccaacgacaa gcgtatgctt cgccctttga 17100
tttgacttcc tggaattcgc ttgttaactt tggcccagta taacggtgct ggtatttacc 17160
ttttgatatc gggtaaactt caccgtcagg gttcttgata acaatatcac caggaccgac 17220
agtattctgg gtttctttct tgccgtcaat cttagtaacg ataacttcac caccttgacc 17280
caaccgtgcc tgggcgtgtc cgtatttctt gtatttttgg acacgcccca gctgttgagc 17340
aatgcggaaa tattcaagca aatccttttg ctgaactaat gccattagat tttaccacct 17400
gctgcgataa catcctgctg ttgttgaagg atttctgtgt atgtatccgc aacagactta 17460
tcaagacgag gttccgaaac gatacgcggg aagtataatg tatctggttt gtcttcatca 17520
ggattcttct ggacaccgtt acaacggatc gtgaagattt tacccatgta atattcacgg 17580
ttgttgtgca cggcttcacg gaaatcgtcg gtcataccag caacgccaga tacgattagc 17640
ccatcttcag attcaaactg gatagaacct agtgtatcag cgtgcttgcc gtttttatcg 17700
ccgtcattat agccaacaac tttgagttct acgtccatga ctagcttaac ttttaactgg 17760
tcattactgg taccgtcttt gaagaagcct ttcgggttct taacaactga accttcttca 17820
ccacgctcaa gtaactcttt aaagtgtgcg aacatttctt ctggcgtatg cacgatacgg 17880
ttttcacatt gttcaaacaa gaccggtgca ccagcctgga tttccggttc gttgttttcc 17940
aagtatgctt ctaccacttt aacaacttcg gcccaacgat gacggtactc aactttgtac 18000
tcaagtttga agaagtcttc agttggcaat gtatcccaga tgcgatactt cacaacataa 18060
ccttcaggta actctttgcc ttcttcgata agttggttga taagtccgtt gccggcttca 18120
cgttccatta gctgaccttc gttggtataa acaagaagtt caccatggaa tgtttgtgcg 18180
atgtgaccgg tccagtcaat gtctatatcg tgtacaaacg tcatacctgc cgcaatttgt 18240
gaagcaatat ttgccaggga ttgagaaaca agttcacgac cttgacgact aacccattgg 18300
aattccatcg aaggcatttt agagtcttcg ttggctgaaa tcgtaacgtg caagttatcg 18360
aacatgccat cagattttag ctgtgagaac acacccaaat cccagtccca tttttcgtaa 18420
tgcttcttgg agattgaacc acaacgcatg tatggtgagt agtaaacaag gtctggccaa 18480
atcttgttga ttgacttgat accataacca gctttgattt cacgtaatac gaaaagcttg 18540
aataacatct gtgcttcagg ttgtacagca tggtaaagct tggcaagatt tttcgcaccg 18600
acatcaccac gttgggcact ttcaatggct tcacggaacg catcgaataa tgacatacgc 18660
ttaatggatt ttagagccaa accacggacg agtgatggat caacatcagc ttggttaagt 18720
acaatacgtg agtccaggat catagtcaac aacgtagtga agaactgttc aaactcctta 18780
tcactttgga cagattttag gagttcggtt tttgccttag ctgaacctgt accctttgtg 18840
ttttgaaggg ttttaataag gtttgcggca cgaacttcat gctttggatc atagttttta 18900
aagtcaatag acatgataag tatatcctat ggttgattga gccttcatta tattctaatg 18960
aaggcatttg gaattatcgt ctttggaggg taagggtttt gaaaccgaca ccacggtcat 19020
cggttgtgta atggtcccag gtaacgcgga acccgttacg cttggcatag agtgctagcc 19080
agagtgaatc aacttcatag tgcacacggt cgtcgccatc gtcatatgga tccgtacgcc 19140
atgaaacagt tgcaatacca ccaggttcaa gggcaatggc aaataactta attgctgcca 19200
tctgttgact catggttaag tgttggaata ccgctgagca caagacgtaa tggaaggttc 19260
cggctttttc aagaagacat ttgtacaaga gttcataatc tgtatagatg tccaggacgt 19320
cgcgattatg aagaaccgac ttattttggt gtaacttgtg gaagattgaa ccaactaaat 19380
gactaggttc atagcacgtc acgcttgcta gacgttcaag catatacaaa acatccgctc 19440
ctgcaccaca accaaaatcc aggacattcg caccagttaa accccattct ttagggacaa 19500
cttgggaacg gtccaaagaa tgataaaaac gagctaaatc tacggcgttc tcgttatagt 19560
attttgtcga taacattact tattcctttt taatttggca cgcaatcctt taggttttgt 19620
gcctggtatt gtttgggttt gatgacgccc acaaggacaa gtgatagtca ctgtcattcc 19680
tgttttgcga tcacctttat attcagattt ggctgtaccg ccacatggac attgcattac 19740
tgttctcctt caaaagagac ataatcgcca taaatgtcgc cttgtttatc aacaacaaca 19800
ttctttaatg tcagtaatcc aaacaaatga gactgaatct cattagcgac ctcgtcagtc 19860
acgacacact tgatcttgtc actagtatgc atagagctga aaataaaaat gaccttgcca 19920
tcatcatcga ttggctcgtg tgtgagcaca atgactttgt atggacgaag gtcatcaggt 19980
ttcttcataa actttttgta tataaaacca ggtatcataa tgaaatctcc gttcaatact 20040
tgaatctgaa acatcatata gtggttttaa attatgaaac gttaaccagg tcagaaaaca 20100
accttatcaa tggatcgcag taataacacg tccatcactg gttctccgat ttctttgtag 20160
tttttgattt tgtcaacaac atcattcata tcgaatttaa cctggagctg acgcttatag 20220
ttgaattctt caacattaac gttttttgat gccacgagtg ttggtacgcc gtttaataag 20280
atcttaaaca ttgtaagacc tgggaaaggc gcaaccataa tttccgcatc ataagtagtt 20340
ttagctgttt ctttcttgaa aaagtttaca aaagatgcaa ggctcataat actatcctcc 20400
agaattttgc ccatcatacc gtatttagaa tcccatgtaa agaaaaaccc ctcacatgga 20460
ggggttggag ctaataattg gcagaggcct tgtttgttgt gatagttcta gagtttgatg 20520
actttcatct ctgggaacat tttttgaagc tcatcgataa gctcaacaga atcaggcttt 20580
ggttctgctg caacgttgaa gtcaacgaat acattgatat gccaagctgt catcagattt 20640
ggttcaactt gcataggagc agtcattagg atgttaccca taccattttg aacaacattg 20700
tataccaggt catctttgaa tttacgcact gtgatgtgtt ccaggtaacc gtttaccaaa 20760
gcatccgcca agtaaagcgt taggaactcg cttgacattt gacacgtagc gccatcaagt 20820
tcattcatac ccgggatagg accagtaacc atgtaaatct tgtcagcgtt tttagaaatt 20880
gcacagtcaa tgatacctag ttgtttgcct ttcaaaccct tgacagcttc tgcgaaacga 20940
ttagacttcg cttgttcgct ttcaacataa ccaccttcga taagttgaat cacttctgct 21000
agcgacttgt catttacaat aaagttgttt tgacctagta cgcttgcaac catttcgatt 21060
ttcttatcaa gtgaaatttc agttttcata atatattcct tgtaaagggt ttttagtgtt 21120
gtttcgtttg atgtgtccat tatacctaac ataaactttt gaagttttac ttaagtttcc 21180
aggcttcgtg tttaccacct ttatgccatt ccgcaatctg cattgggatc acaacttccc 21240
attggtctgg gtgcacttca gcaagctctg tacggatatg gtctgaacgg tatactttaa 21300
cagcatgttc agcatatttc agactagcaa attgtttgat gatatcccaa gttagacgca 21360
gacgagtctt ttcacgcagt gttgagtcat tttttagacg ttgtaactct gcgaataaca 21420
ttaaacggaa tcgaggaggc aaatagtgga cattcagccc gaataggtac atcacgcctt 21480
tctcaccaaa tgaaacacca tcacccagct tcatgttaaa gaaaaacact aaaggccagc 21540
catcgtacac cggtaagata ttcttccaac gtggatcgta gttatagagg aacatgcgac 21600
caatgagccc tttatccgat ggtttggtac gacgactctc cttaaggatt tgacggtgta 21660
caacttccgc tcgtaggtgg gtatcctttg tgatacgact cataaaccat ttgttcgatt 21720
cacgggtatt tcgtcgagca ttcaaccctt tagtttcgcg atatttcgaa atgtatcgct 21780
taacgatagg tggatagttt tgctcgcgtt gtaacaataa ttgttcttta gccattatgc 21840
ttaccttaaa tactttcaaa atatttagga aatgagttat gcgagagatt agaaatttta 21900
ttgctcatct gaagcaacac ggtatttcac gaaataaccg attccgtgta acaatcccac 21960
tcccaccatt agttgaccag gcacttaacc cacctggtga ccaagatgcg atatcagatg 22020
ctatcaaaaa tggcttccgt gtcgtatcaa ttctgaatgg tgatggcatt gaagcgcgac 22080
gcggtttaca gatcatgtgt tcgtcaaccg agttaccagg tgtcaacatt gatactattg 22140
atgtcaaaca gggtggtcca tcgttcaaga ttgcttcggg cacttctaag tctgatatcc 22200
agtttaactt tctgatcagt gcagactgct ttgagaagaa cacacttgac ctgtggcgca 22260
aagtcatttt tgatagtgat acccgcaaag tggcatacta tgacgattat aaggtggata 22320
ttcaaattga ggttctggat gtcaaagacc gtgttgtcta cagtatcaac ctggaagacg 22380
cttatccaat cctgttgagt ccaatcaccc tcaataagcg tgccgtagat gagatgatgc 22440
agtatcgttg tacttttacc tatgctaagg tcagcgagtt tacacagtcc caaggcatct 22500
tatctggttc ggttattgaa gaagtattgg atgatgtaca gaatggagat tggcagtcgg 22560
cattagccac ttcaagagaa cagatcctaa aagtcgcaca agggaattac actggtgttg 22620
cattggaaat gcatgggatg atttcagaga ctaccaaaca gattgttggg atgaatgcca 22680
ctgacgcaaa aggagtaatg tccaacttta ttccatctat ccaaactaac cctggaatca 22740
gttctgtgga caaaggaagt ctcactaagt tggttaaaga cttattctaa taaaaagccc 22800
cagaatcaat cctggggctt tttgatacgc ttaataacct atagcttagc tttgttctgc 22860
ttctttaagc attttttcga actcagttag ctggtcatcg ccatcatctt cttgtttggt 22920
ttctgctgca ccttgctgag caccaacatc agtctcttga cctttgtctt gaccgattgg 22980
agtatcttct tcttgctcaa caccagtaag accttccatt ttaccaggtg ctgcgccaac 23040
tgaagaatca actgccattt gagcttcaag tggtgcatct tcagggatac ccataaccat 23100
tttgaagcgt ttttcaagtt cttcataggt cttgaagttc ttgcgatcgt agaagtcttg 23160
tagcgaataa gtctgtttcc agatttcttc aatcttggct tcatcaccac cgaacagtgc 23220
gcttggttta gcccaggtta cacgagcata gtcagggctg atacaatcac ggccgttgaa 23280
cttacgcttt tcaccagtta ggttgaattc caggtccgca ccttcccaag gacagaaaac 23340
gtcttcaatc tttggatctg ttgggaactt aggagcagct gcagcatcaa taattttctt 23400
gataccgcga ccaaatttaa acaagaaaac tttaccgttg ttttcaggtt tttgtgcgtc 23460
gttaattacc aggatgttag catagtaatc cgtgtttggt agacgcttca tcaagatttt 23520
cttcagacct tcgtcgttag tttctttctg ttgcttccac agcggctgat catgcttacg 23580
tactgggcaa tcttcatcaa acgtcgccaa gcttttctcg atgtattttg aaccagtcga 23640
gccttcaaag aagtggctga taatacatgc acaaggagtc aacacagcac cttcaggaat 23700
ttcacctttt tcacggcgtt ccatatctac cattggaacc ggcaggaaac gaatagtatt 23760
aacagaaaca aattcgccag gtttactttc ttcccaagta tgtttccaaa tacgttcatc 23820
cgctttgaaa ccaccttggc cttgttgtgc cagacgctct ttaagagctt tagtgttttg 23880
acctttgttt gcttttagac gtgcaagtag tgaagacata gtagttacct tatattttgt 23940
ccatactgag gactggtttt ggtttaagtg tttgttcttg tgaactcgtt tattttacac 24000
aacaaattaa attgaagttt gaatatttag ccattcaatt ggcttggctt atcgacaagt 24060
tctacataag ccgtttcacc ttcaacaata acttcaagct tctggatgcg gttaaacggt 24120
gcaaccacag tcagatgatg agtcattgaa tcggattcct taatcatacc aggtggttgc 24180
ttttgaacat ttgctacacc cggtattttt tctttgaccg catttgaaat ccaagcacta 24240
atcattttaa aatcctctta gtttttgaac gttgtttgat taggtttaaa cggataccct 24300
cgccttccag atcattcaat aattggtcgt tgacatattg ctttgcatct tcaacgggga 24360
tatcatattc ttcgcaagcc gccaataaag actggaataa agtatcttcg ttgatattcg 24420
cttgacgctt aacatataag ctgaattcgt cagcgttgac aatagtgtta gtttgctttt 24480
tcatacatca ataaccgatt tcggccctgc ttttttgtga atgtctttca ttaggttttt 24540
gaacccttct ggcgcagccg tatggtcttt gacaccagtt acagtgccaa ctttcccaat 24600
cactaacatt acctttgatt cgccacatgt taagcaaggt tcttccgtag gcattaatcg 24660
attacggata ctcagtgttg cagaagtgac ttggttgcag ccattacatt tgtaattata 24720
cttcggcatt acaacatacc tcgcaacatg ataaggaaga atgtgaatgt ggaccacatt 24780
actgtataaa caccagcctt ctcaccaggt ttaacttgta agattaacga tttgataacc 24840
gcttctggta cgtatgtact ggtttccatt gcacgtgcct ggaataacac actgtcatgc 24900
acacgagcat ttaaccagat catgaagtgt gacatggatt tgaagaagct taaagccgca 24960
acaaagataa gtacaacttc aattgctatt tgataagcat caggattaat cactgtcgac 25020
attttctgtc tcctcagggg aataagggtt atatccacag ttgtcaatgt tatggataac 25080
acgattaacc atgaacagtt ttaacaattc tttgtactta ggatcttctt cccaaagtgt 25140
ttcaagaatt gtttggtggg atttaatccc ctgtttgatg taaaggtcgt aaatacgtac 25200
cataacaggg gatttaatgt gtccaatatc tttggccatg atgtttgtcc tattcgttta 25260
acacgtagga cattttatca cttacccatc aattgaagac ggatttcgga aaggtctgga 25320
tggcagataa ggtttgcgaa agagtcaacc aggtcatcaa ccggtttttc gtactctttg 25380
tttagaagta actgttggtg gagtttgatg ccccatagct cgttgaaaca atcaatcatt 25440
tgttctttct tggcgttacc tttaccttgg aactgcatct tgacttgagt tggcggaatg 25500
atttcataat caatattcag gttgtaaagc tcttgtttga gttggccagt acactcacca 25560
atctggaaaa cgcgattcga cttgccaccc atcgcatagc cttctaatga tgctttcgtt 25620
actttatttg attgaagtac ggcacgggcc caaagtgcga tctcacggaa gcgtggttca 25680
tcgctatcaa atttttcttg agcaaggatc acaacgttat tgaactcacc acattttgat 25740
ttggttttgc cgtagttgta ataacgtagg ttttcgtgag taagtggtag acttgaatcc 25800
cagatacaaa tagctgggcc gttcattgag tagtcgatgc cagcgtagat atcgccgtca 25860
cctaagatag gttgaatttc tgattgtgcg ttagacataa taaagccctc tagtaattat 25920
aaaattattt agagggctta aattctatgg attacacttg caagtcaggc gcttgttcag 25980
cttcttgctc tttttgttca gcaatgttgc cagccatctg gttcaccagg tccaatgcca 26040
tgttacgaag catctcatgg tgattgttcc aggcttcttc ttcaccctga tacagttggc 26100
cgtcagacgg gtcaattgca atgatgtccg ttgtgcgagc attaaattcg cgagcaatta 26160
ccgccgattc gtacgcgagc atggcagggt taccattgat attcggggcg atgtagacac 26220
caagtttttc accttcggtt tgtaccatga ccataacaga gttaaacatc atgtaaacca 26280
ttggttggtc ttcaacgctc tccagcattg caacgtcatc ccagatttgg atgagtgatg 26340
gatccacctt catgatggca acatagccgt cgcggatttc gatcgctttt ttgtaatgct 26400
cttttagacg tggatctact tttgaaatat ccattgtgat accttttggt gattgtgttt 26460
aatcgtcaaa caatgtaatg tgcttaactt gtggtttaga ttttgtcggt ttttcgactt 26520
ttggagaata agtccagtca gtagacttaa caacattctc agggtttgtg acttgacggt 26580
tacgacggta ctcgcccata tctttaatat tgatggcacc tgaacgtcgg atcgaaacac 26640
gtgttgcgtc gtcaacatca ataaacttgg ttttgataac cggaacaata acatcgtcca 26700
tgcatttgaa aattgtgctc ggtgaaataa tgagcatatc tttcatatta gagatacgtt 26760
taatagcacg actatggtca atagctttaa tgacaagtcc cttatttgga tgttcttgta 26820
aagcatcagg gttaccactg taactcaaaa tgatagaacc ttttggaaaa ccaagtgcct 26880
caatttcagc ttccatgcca acaattttaa ggttatggcg ctcggcatac gctaatacag 26940
aatccttcat ttcttgtgtc catgtgaccg gccacatagc acgaccaata tcttcggtac 27000
gtaccaagtt catgtaccca attgtgttct ctaagatgat tttcttgctc atgataatgc 27060
cctaaataat gttagttaga agtcactata ataaatgggt taaaacttat gaagaaacca 27120
ttaccaacga tgccactcgc acagcgtagt tggcattgcg cacaaaatga tactgaatat 27180
gtttaccgta catggacgat ggctgaacaa gaaatcctgg tccaggtacg tgaagaagaa 27240
acggatagaa aggacagaat cgactccatt attcaggtcg tacaaaactg tgttgagaag 27300
ccagaagact ttgatgctcg taagactcct acgttcatca tagagtcgtt gttcttgcgt 27360
atctatgcaa ttagcatggg tgaatcacat catctcaata tgcgttgtac gaacgcgatc 27420
gataaagatg gtacactcca tgaatgtggc aacaagattg aattcgacat tcctcttgtc 27480
caagcggcgg atgttaaagt ttatgacaat cataaaaaca ttatcccatt gccaggtggt 27540
tacaacctga agatgcgtta tccttgtcta tcaatggctg atgagaaacc acctaaagat 27600
gttgatgcat cctggatcat tagccgtttc ttggaatgta tctttaatga tgatggtgaa 27660
gtatgggaag cctccgacta caccagtgaa gaaatgcgtg cgtttatgcg tagcctgggc 27720
ggtgatatcc agcaacctat tatggaagac tttttcctta gtgctccaca catttattgg 27780
aaaatggcac caactaaatg tagtaagtgt ggtttcgaac acaattatga gatccggaac 27840
ttaaaccacg tttttcgcta atgttcgaaa tggacttatc ggacctatat gaattatcat 27900
atgccgcaag gatgcgtggg ataacattaa cggagttcta tcaaatggct ccatggcaag 27960
tgttaactca aattgatatt cacgactatc tcaaagaaca agagttagag cgaagaaaac 28020
aacagcagaa tgggtgatcg cgaggtcgcc catttttcat atttattaaa tgtgattaca 28080
atagggtgtt ccacccaata aatttaatat taataattaa tataaatcat taatctttaa 28140
tggtgtccca tgggacacat ggccttcggc catcattttt atatattgat cttaaataat 28200
ttaagttatt agatcaattt ataaagagaa ttacaatgct tgaagacatt aaatggtttt 28260
acggcatcat cgaaaacgtt gatgatcctc aaaaatcggg cttagtccaa gtccgaggca 28320
ttggtataca tgaactaaac gaagctatcc tgccatcgga atttttgcca tggtgtaagg 28380
ttatgacacc tgcaacaagt tccagtgtta aaggtgtagg tgttactcca acgggtatcc 28440
aaaaaggctc tatggctttg tgcattgcga tggacgaagc atacactgat aaccgcattt 28500
tgttcacatg gcaagcgggt gacatgaata ccaatgatat caccgaactt gctcgcggcc 28560
taaccaatga aatcattcgc aaaaaacttg atgagcggaa tgagtctaaa cgcactggct 28620
ttaagtgggc tgaacctcaa accaactacg gtaatgtcgt ctatccttat aaccaagtaa 28680
cagtatccag agcgggtcat gttattgagc ttgacgatac cccaaacctt gagcgtatcc 28740
atatcctaca taaggacggt tcattcattg aatatatgcc tggtggcgac gttgttgaaa 28800
aatcaatggg taaccgttat gagatttatc gacgcaacct taaccgactg atcaatggta 28860
aagaccgtaa gaatgctgct gatgtggccg tacatcatta ctctaaaacg tatcatttca 28920
attcggctga acaggcaaca ttcacatcag atactgatat gctatttgtt gcaccatcga 28980
ctattgaatt ccatacggct ttattacaga tcattggtag tgccaaaatt tcacaaacct 29040
tatatgttga cgaaattaaa gctgctcgta tttcatgtga cagcttggat gttgcatcac 29100
caatcaatgg tactgcgaag tttgctcaag ctgctgctca ggcaggccgt ttaggtggtg 29160
gagctccagc tatgggttcg tcagtgcaac cattgaatat ccgagcacag ttgaaaaact 29220
caactggtac tgagggtgat aacaacgacg ctccggcagg tagtaacaaa gcagacgaaa 29280
aagtatccca ggagcccgca tctaaacaga aatcacaaac tagtgcaaac actagtggtt 29340
caggttcaca aggctctggg gatactacag gagaccaaca acaaacaact cctggaacac 29400
aagcagaaac gattgatcag gttgtaggta aagagaacat tgatgaatgg tatcgttcgc 29460
agatacccaa cattccaaag tacaaatata accatattac gattcatacg actgcaacac 29520
cacttagtca attccatgat gtgcatacaa ttcgtcaaat gcataaagca caagggtgga 29580
gtgatattgg ttaccacgta ttgattactc gtacgggtct gatccaaatc ggtcgaccat 29640
ggaacgtgat gggtgctcat gttaaaggac ataacccgaa caactttgcc gtcgtattgg 29700
ttggtggcgt taagctttct aacggtcgtt gggttggtga taacaacttc acacaagcac 29760
agtggaagtc attacaaatt gtgattactg atgttgcccg tctgtatgat gtgccagtta 29820
ctaacatact aggtcaccga gattggagtc cggatgcaaa tagtgatggt gttcttgaga 29880
aacatgagtg gattaaagaa tgtccgtgtt ttgatgtacg ctcatgggta gcatcatggt 29940
gggatacgcg aacagtatac cgaccaacaa tgccatcagg taaaactgaa actatcgata 30000
cgaccgaacc gtctaataag ccggttgtca cggcacccga tgaaaataca gaacatagct 30060
caacaactgc gactaagttt gccgaagtat tgccggcgac agaaattagc aatgttgtaa 30120
aagaccttag tgaaagacgt gttcaagata ttgcgaccaa tattgccagg ggtattttat 30180
cttcagatgt ttcaatggtt gctgccaacg ttgctgcttt agctgctgcg gatgtgcgta 30240
atgataccgg tccaatggtg gacaaagctg ttacggcttt cgagaaatct gaagcagtac 30300
aaaagaaagt tgaagaagac ctaacgtatt tcaaaaacca atattatggt ccaagtagct 30360
ttacaccagc caaacgtcca gacggcaccg atccgcatcc tggggatttg tacttcaata 30420
caatccaaga caaagttatg gtgtatgtta ctcgtaccga tttcattggt tggacaccta 30480
ttgatggtgg tggtaaaaca tgggcaaccc aaattcagtt cgataactca ggtaccgatt 30540
taggatcaca gaacgttggt tcagcaatca aagagttaca taaactgttt aaattgtatg 30600
aacctgtcga aggtccttca caaaccggtg acggactcaa gcgccgtttc aactctccga 30660
taagccgtaa agtagatgat aaattgattt gggttttcgt tgatggacgt cgaatgaaac 30720
caacccgaga atatcaaatc gaaactgacg gcaaaatttt attgactgat gctccagcaa 30780
atggtctcaa aatcgagtca ttgtacttta acccgttgca aggtgtacta ccgttaacag 30840
gtggaacact tgtgggtcgt ttaaatggta caactttgca tatgacgggt aaatgttaca 30900
tcaaagagtt ggaagtcgaa aacgttacac tgattgagaa ctcgttacat tcgtcagacc 30960
gaacattaaa gcgtaacatc agacgtatta agaaaccatt gtctaagatt caacgtattg 31020
acggtgttag ctttaaatgg aaagagagtg acaacgacgg acatggcaca atcgctcagc 31080
aattgcaaca agttatgcca tcagcagtta aacgccataa aggtaagtta cgagtcgatt 31140
accatgcttt gatagcatta tgcattgagt caattaaagc ccttatttac cgagtcgaag 31200
cgcttgagcg gcaaggtaaa cgtattaaag agctggaaaa acgaatcgaa cagatggaga 31260
aattgatcaa tgaaaaccaa agataaccca cgtttatcat tcaagactga cgtagttaaa 31320
aatgctttgg attcttcacc tcgggtctct tttagggacc tggaggttac ccgtcttagt 31380
tggatgccta aaggtggtaa aaccctcagc atgaaagata tgttgagacg tcaaagctac 31440
agatattatt cggaccgcac aacaaaattt gattccaaca aacaaaattt gcgacaaatg 31500
gctgagtacc tatttcccac tcaaacgttg gcagccaaag ggataactgt aacctttaca 31560
aatatgacgg gcggtgtagt taaagtagac ttaacgaaat caaccgactt tgaagttgtt 31620
atggagctga cgtatttacg cggtatcgcc aagggtaaaa ttatacaagt ttactttaaa 31680
ccacaagacg agatcattga gtctactgaa aacctgtata aagtagaaat gaacggtagt 31740
gtattgaatg gattcgcggt ccacactata tatggttcta tcgacatttc ggcggtcgta 31800
agagagaagt ttgatcaacc tacaggtggt tctgataaca caaatgacga ccattgggaa 31860
gtcaaaccta cgacacctgt tataggtaaa aaatatgatt tagttactgc taccggatta 31920
caatctcacc ttggctactc cagtgaacat atccattttg ctttggctga cggtgatctt 31980
caaggcaaag tcactttggt tgcctctgcg gatacacgca tcgggaatgt ttatattcct 32040
gttaatgaac gtggtgtacg ggtaagattt gccgtttatg acctgggtca taaggatgat 32100
aaatattcaa aagcaaaact tgtggcatta tcacctatac taacaagtac gactgttgat 32160
aaaaaatatc tcagtcataa attgcgttat atgctaatcg aaaatcatga gtatcgtttt 32220
ggtttacaga tgttttcgga tcgtggtcca ttacaaccac caaacgccag gtttgtcaca 32280
gcacagttaa atacaggtag tgaccgaata cctgatgatg agtatgattt gaatatttct 32340
gaagcccaac gactcaatta cagtttagct tattatgctg aaatagaaac aaagaaaagt 32400
ggagttacta catgataggt gaattagaac gtgttgagga accaggttat cattatcgca 32460
tcgatgttgg tggtagcaaa gacgtgtttt tattgcgttt gcacaccgaa gtgtgtgagc 32520
tgtcgtttat taatgcccaa gcgttgcctt tcaactacca aaagataaca cttattttag 32580
aacaacaacg tggggcacat aaagtcacat ggcctgcaaa tgtaagttgg ccgatgaagc 32640
atatcccaat tctatccaac cagaaaggtg agattgacac gattgagtta gtcagttatg 32700
acggcggaaa cacctggttt gggtatttta ctggagcagg atttaagtac accacatctt 32760
cataggaaat accatgcatc attttaaaca tacaccaatc aataatgccc tacaactggc 32820
attattattt gataactttt tagaccgtca tgtgggtaat accgacgata acaatgacgg 32880
tctttatgtc atcgaccagg acggtgtgct taataacaac ttgcacaaag aagatcgtgg 32940
ctcctcgaat ggtgcaccag atggtacagc aacaactgaa ggccaagccc ttatgatctt 33000
gggttaccta tacatgtatg ctgcgacaga gaatcagtat tggttggata aatcgactta 33060
ttattttgag tcatatattc gcaattatta tttgcgtaaa ccaatacctg atactccaca 33120
agtattccgt tcccatatct acatcaatgg tcgtgcacca tttatcacag ccggacctaa 33180
tacttggcgt aaaggttccc gtacgggtac tcttgagtat gatatggatt tcaagaatgg 33240
tcgcggcatt gttgcatcag gttatccgga ccatggcgac gaaatcatat acataccatt 33300
tgcttttgac ggttacttaa aatggcgttc aatcgttgct gacgtattgc aacatggtag 33360
cacgtcgaaa gaaggtaaaa cgtttgatgt tgcctggtat attgattacc gtggttataa 33420
ggttgaccca gaaggtgaaa tcatttcaca agtcaacact gaaccaaaag gcacagtcca 33480
acttaaagac accacagtca acggtagata ccgcattaac tatgcttcaa acaacggtag 33540
tacaattgac cgacacgaac catttgagaa ctggcctatc tggatcccag tacgtgatgg 33600
cgaagaaggt aatgctgctg atgccgaaga atggtttacg gactgtgcag gtctcatgta 33660
ctatttcacc aaagatgaac gatacaaacg tttgtatgag tgctgtattg aaacgttgat 33720
cgaaatctct ggcgttgacc aaaatatgag attcttccgt aaagataagt cagcaggcac 33780
accgtttact gatggtatca gttatgaata ctcatactcg cctggtaatg tgagcattgt 33840
atatactcgt gataatgatg gtaacatcaa aattacaaaa gctgccgaac caaaccctaa 33900
agagtttacg gatatctcat tagagcagat tgcctcatgg ttccatgtag acaataattc 33960
gaaagttatt gttgattacg gccatactga cgcagaagcg atcacatcta tccgtattac 34020
gattatggat aagccacaaa cggaaaaggg gataatgtat gagtatgcat tcaaacaaga 34080
tacgagtacc gttaagcgtc atgagattcc aatacaacac ttcgttgaaa aggaaaactt 34140
cataactggt cgtaagtact acaaactgga tggtacatca tttgttcctt atggtgatgc 34200
aaaatttgaa aaacgtactg gtgaaaatat acgtgttgat gccgattatc gtgtagaccg 34260
ttttgggcgt gttatctttg gtaataccag ttattccggg attgtatgtg gtttctggtc 34320
gattgcatca tcaacgaatg gcacattgcc aatgggttca attacctatc gtaatgtagc 34380
ccgttcggat ccagctttag ggtctaattg tgatattgcg attgtggatg ctgatggtgt 34440
gaaatggtct atgcaactac cgtatcaacc acagtttcaa acgttctacc ttgattggaa 34500
aggtccatgg aaacgcactg atggtaaacc tggtgtcatt aaaccaggca atttgaaaca 34560
gattgagatt cagaagcatg gtagtgaaga tacgagcgca actatcgatc tgtatgcttt 34620
taatggtgtg ccaagtaagt tcgacggtaa aggtgttctc aaagagtaca acatgaccat 34680
tactggcatc aaggcgaaca caatctacac tggcgactca tttgtggaat caggtaccga 34740
tgcggctaag ttgaatgcac cttatacacc tggggtaatc ccattttcaa ccaacttctc 34800
ggttgagaca caacaggtcg actcatggcg tggtatacct taccttggat accaatatcc 34860
aatcatttgg ggtcgtgaca agaagtatga gtcgcgtatg aataatatga ccaagtttat 34920
gagagactct caaacagatt accatcgccg tcggggtctc aagggaccag taagtgctgc 34980
ttatgtttgg ccacgttggg ataatggttc gtatggtgag ttcaacacat ttgttgatgt 35040
gttctggggt agcaaacaac catgggcagg gtattattct cgttgtatgt ttgggatgtg 35100
tcgtttacga tatcatttag ttcgtactgg ccgtacacct aatgatgacg tggtccaatt 35160
taccgaggat tggattgatt atttggctga cttcatgaag aagaatgaca actggacgcc 35220
aacagtattc catagtaacc gtgaaccata tacgtatgac ggtgacttta ctggtcatat 35280
gtcagcaagt ttcctggctg gggcttgtga gtgttacctt gctggtagtc gcaatccaaa 35340
ccttgagact gtgatagaag gcatttatac acagtttgaa cgtcagctta ttaacttggg 35400
tcctggacac catatgaatg gtggttacag tgtatgggct ggaggtaatt acttctttgg 35460
ggtatggaca gaaatcctgc gtggcttcgg tgcatacatt atgtatcata aaaagaaact 35520
atacggtgat agttacacac cgccgtatga gtaaaggatg aataatgaaa gtgcaataca 35580
aagatattga tcttggtatg gctaagcacc ccgtaactgg tgatgtagcg gtgcgtactg 35640
atgtgcgtgc cattatgcag tcgattcgcg agattgtatt tactgcagct ggtgaatggg 35700
aagctgaacc tgaatttggt gtaggtatct atagcgaatt gggcgaggaa caacatcctc 35760
tgattggtga gaacttgcgt attcgaatag aacaggctgt agaacgatat gagcctcgtg 35820
ccgaaatcgc atcagtggtt attggcagag acggtgacca tggagtcaaa atccaagtca 35880
ttttctttgc tttgaactct cctgaaccat ttgaccaaga aatcgtttta gaacgtaccc 35940
gatagatatg ataaaagccc actggattga ccgtgtgggc tttttattat ctgagagtat 36000
gattatgcta aattgtccaa gaatgtcttc acatcgaagc acggacacat tttgataaac 36060
tcgtttggtg taatcttgcc gtctccgttc aggtctggac tccaatcacg atgacctttg 36120
atattaccgt ctttaattcc caaaacacct actaagtcaa tcaatacttc acgtaagcta 36180
tcgtattgct ctggtgtaaa gttatcttct gccttaccat tcgaatcgat acccccaacc 36240
aggacaatgc caagattgtt acggttatgt cccttaacat gcgcacccca tcgattaaac 36300
ggtcgaccta actcaatggt gccgtcacgt ttgatcaaaa catgataacc aatatcatta 36360
aaaccgcgtc gcaaatgcat tttgcgaatg cttgccacgt catgatcctg gtctggtggt 36420
gtagccgtag cgtggaccgt aataaatttc cattcattgc taagcaattg atgtagttta 36480
tctttatcaa gaaacattgt ttttcctcta ctgcagtttc aaatatttag ttcacgttaa 36540
tataaacacc tcaacgaaaa tactatatat tgattgttac cactcaatat taactacata 36600
ccgacaacct aaaaaggaat cacaatgcaa gtacaaaaac gcaatggtca atttgaagac 36660
ctggacattt caaaaatcac caaagtgatc aactgggctg ctgaaggcct ggaaaatgtt 36720
tcggtttcac aggttgaact taatgcgaaa atcttgtttg ctgacggcat gaagactagc 36780
gacatccaca aaacaattat taaagctgcc gctgacctga ttagccctga aactccagat 36840
tatcaatatc tggctgcgcg tcttgccatc ttccatatcc gtaagattgc gttcggtcaa 36900
tacgaagctc ctgaccttta ctctcaaatt gaacgtatgg ttaaacttgg taagtacgac 36960
aaacatctcc ttgaagacta taccaaagaa gaaattgaac aattgaacga aatcatcgac 37020
catactaaag acctgaacta tgcgtatgca gcagttaaac agcttgaagg taagtatcta 37080
gttcaaaacc gtgtaaccgg tgaaatcttc gaaagccctc agttcatttt gatgttggtt 37140
ccaatgtgtc tttttgccaa gaaaaaagat cgcatgacga aaatcaaagc gttctatgaa 37200
tcggttagca acttcaaaat ctcattgcct accccgatca tgagtggtgt gcgtactcca 37260
acgcgtcaat tcagttcttg tgtacttatc gaatctggtg atagtctgaa ttcaatcaac 37320
gcttcagctg gttcgattgt tcgttatgta tcacaacgtg caggcattgg tattaatggt 37380
ggtcgtattc gtgctatcgg ttctgaaatc cgcggtggcg aagcatcaca cactggtgtt 37440
attccgttct ggaaacactt ccaaacagca gttaaatgtt gttcacaagg tggtgtacgc 37500
ggtggcgcag caacgttatt ctacccgatg tggcattacg agtatcaatc tttggtcgta 37560
cttaaaaaca accgaggtgt cgaagaaaac cgtatccgtc atatggatta tggtgttcag 37620
attaacggct tgctttacaa acgtttaatc gatggtaaaa acatctcatt gttctcgcct 37680
tctgatgttc ctgggctgta tgaagcattc ttcagtgacc aagcagagtt tgaacgtcta 37740
tacactgaag cggaagctaa ccctaagatc cgtcgcaata ctatttcagc gattgaagct 37800
ttctctggtc taatgcagga acgtgcgtca acaggtcgta tctatgtcca aaacgttgac 37860
aactgtaaca tccagggacc attcgatgaa aaagtggctc ctgtacgcca gtctaactta 37920
tgtcttgaga tcgcgttacc aactaagcct ttggagaatg ttgatgatcc aaatggcgaa 37980
gttgcactgt gtacattgtc agcatttaac ctgggtgcaa ttgaaagtcc tgatgagttt 38040
ggtccactag ctgaaatcgt tatcgaagcg cttgatgcgt tattggatta tcaaaactac 38100
ccgttacctg cagctgagcg ttcaacaatg tatcgccgta ctcttggtgt tggtgtgatt 38160
ggttatgccc actggattgc caaaaatggt ttcaaatact ctgatggttc tgcgaacgaa 38220
gctacacacg aattattcga agcaatgcaa tatcatctga tgcgtgcatc ggttaagctg 38280
gctaaagaat atggtgcttg tccaggtttc cacgagacta aaatggctaa aggtatctta 38340
ccagtcgata cttaccgtaa aaacgttgat aagcttgttg ataacaactt gaaatgtgat 38400
tgggaaggtt tacgtcaaga cattctgaaa tacggtatgc gtaactcaac agtatcagcg 38460
ttgatgccaa gtgaaacgtc tagccagatc tcaaacgaaa caaacggcat tgagccacca 38520
cgtggatttg tatcaatcaa agcatcaaaa gatggtcaaa ttaaacaggt tgtaccggaa 38580
ctgaaaacat tacgtgataa gtacgaactt ctttggacca ttaaagacaa tacgggttat 38640
ctgactcttg tggcgattat gcagaagttt gtagaccaag caatctcggc taatactaac 38700
tacgatccga tgaactatcc tggcgataag gttccaatga agaaactgct ccaggatttg 38760
ttaacagcat atcgatacgg tgttaaaact ttgtattatc ataacacccg agaccatgca 38820
gaagacaaag accaaaatgt cgttatcgaa aaagttgatg attcttgtgg tagtggtgct 38880
tgtaagattt aattggtaat ttcaatttaa tacatggccc taaaatagta gggccattta 38940
atttaggagt aagataataa tgagtgtttt caacacaacg cacaatgacc aactcaaaga 39000
accaatgttc ctgggccaag aagttaacgt ggctcgttat gaccagcaaa aatttgaaat 39060
ttttgaaaag ttaactgaga aacaattggg tttcttttgg cgcccggaag aagtggatgt 39120
atctcaagac cgcatcgact ttgctaaatt accagaaaac gaaaaacaca tcttcatcag 39180
taatctgaaa taccagacac tattggatag tattcagggt cgtagcccta acctggcgtt 39240
gcttccgatc acttcgttac cagagctcga gaactggatt gaaacctggt cattctcaga 39300
gaccatccac tctcgttctt atacgcacat cattcgtaat gtattcacgg atccgagcat 39360
tatctttgat gacattgtgg aaaacgtaga aattcgtaag cgtgcagtgg ccgtatccaa 39420
atactacgat gatgtcatcc agttaatcgc tcgtcgtgaa gccggtcttg aagtcgacat 39480
gtatgagatg aagaaaaagc tttacttgtg tctcatggca attaacgttc ttgaagctat 39540
tcgattctat gtttcctttg cttgttcttt cgcgtttgca gagcgtgaat tgatggaagg 39600
caacgccaag atcattaaat tgattgcgcg tgacgaagca ttgcatatgt caggcaccca 39660
acacatgatc aaccttctac gtaatggtat ggatgacttc tcattcttac agattgctga 39720
agaatgtaaa gacgagtgta ttcaaatctt caaagatgcg gtgcaacaag aaaaagactg 39780
ggctgattac ctattcaaag atggcggcat gataggctta aacaaagaga ttctgtgtca 39840
gtatgttgaa tacttaacaa acatccgtat gaatgctgta ggtcttcctg ctatctacga 39900
tgtcaagacc aacccaatcc catggatcaa cgcttggctg tcttccgata acgtccaggt 39960
tgccccacaa gaggctgaaa tcagttctta catggtaggt cagattgata gcgaagttag 40020
tacggatgat ttcgcaggct ttgaacttta ataaatcgtt taatattgaa aagtgtgatc 40080
taggtcacac ttttttatta tcagatgtat tatgatgtct actcattcac aacactaaga 40140
gttattagac atgtttgcat ttgttgtcat cgtacaattg gtttttgtta ttgctggatt 40200
tgcaagttgc gaaaagcaat accgctcaga ctacggagta tttgtagttt ttgtattcct 40260
aatcgaaatt ttggtttctg tctttaatca aggacaaata tcatgagtaa acgcaaaata 40320
tcatcagcac catttactgt aacaggcgaa tatctagcta attaccttcg ccaaattatg 40380
atcgacaagg gttggcgttc cgcatacaaa tccgtaatgg aagatatcgt tggaattaac 40440
gatgcccaaa ccattgaatt cttatcgggc aaaaagacat ttgctggcga ttccaatagt 40500
ggtattgaat tcattgattg ggaagatagt gaagaaaaat ccctttactt agaagaagtt 40560
aatgaaaaat acaaaaagat ttggtgttgt gatgaactat tgctgactgt cagtgaagta 40620
gtcagagatg gtcgttatgt gtcacaactt gtcaaagggg atggtgagtg tcctaaccag 40680
ggggcttttt atcgcgaagt tgatccaaat gtgtatgcga aatattttga tgctcgtcat 40740
tatcaacgtg taacgctgga aaccgatcct gatttgcctg agcgtttaat gcttgtattt 40800
tggaaacgag ctgatgcttc tttgccgcca ccatggattg aagaatgttg taagagttgg 40860
gaagccatcc gacattatga cagttcagaa atccgtatac gtgagcttga tagttacaaa 40920
ggtttgaaat tccgagaaat ggacagacta caagctaatg gcatctatca agctgtaggt 40980
gctaatcgtg atgagcgtgt ggcactaatg gaacgacgca aagcaatact tgaccaaaat 41040
ggtgatgatt tcattgaagt tgcaggtcat aaaataccta gaaagccatt gattgcttgg 41100
ggtttacgcc gcagtgatgc caaacacctt gccgattgtg attggaaagc catctcgcct 41160
tctggtgaaa aaatgatgaa cgattgtgct gcccacactg attgggttat tggcgcaggg 41220
tttgatccag cccatttcta tcgggaccat aaagaaatca atgatgctgc atttgtctgg 41280
tgttatgatt tgaacagaaa gttatgttcc gctgacttgt tggtattgac ttcttgtcgt 41340
ggtagtgaac tacgttccaa ggttcaacaa ccacaaccta acgataagat tacgtgtgac 41400
atcgtggtca ttgaagatgc tagtacccgt tggatcaagg ttgctgaaca ggttggtaaa 41460
cgtggtggta tcataatctg tgaaaaagga ggccccatgt ctcacctgac catcaaagga 41520
caagattatg gtataaccct aatgttacac ccagaagccc gtatggtgta ccgtgagggc 41580
attgagctaa ctgtctatga aaatggcttt gctagcatta ggagcatgta atgaaacgtt 41640
ttatcttatt ggtaacagca ttactagcaa gttcttcata tgctactaca gattgtgaaa 41700
ctgtatttgg tcatgaccag ctcgtgacca tgcagaaggc ttacgattat ggtaaacctt 41760
atgggttgga atactcattg gccgccatca cctggaagga atcatcagct ggtcgtcata 41820
acgtgcggat cgaaactttc aagaatacta ccaaagtgag ttattcacca ttccatattc 41880
tattgcgcac cgcaatggtt catgaggggt gtgagaccaa acgttgtgca agtgatatcg 41940
caaataaact catgaccaac tttgactatg ccgcatctca tgcagtacta gttcttcaat 42000
tttggcagag acggcataat aacaactgga tgaaaacctg gcaggggtat aatgacggct 42060
atcaaaatag tcaggatggc cgcaattacg cccttaatat taagtataag atcgcatatc 42120
tcaaacaatg cgtaaggttg aaataatgga tcaaacttca catgcaaagg cattggcaag 42180
ttttgtcaat gttgcaaagc aagagctcga tgagttcatg catcactgtt tggaagaaca 42240
gaaaaaagaa cccgaaaatt ggcccattga tagctggaat attagcgaaa tgtgggaaca 42300
attcgaagct tggcatgact ggaagcatgg agtggaatcg tgttaaatga agacgacatt 42360
atatctgctt ttatgggcga tgctgaacca cctgcacctc cactaccctt cgacagacca 42420
ctaccagaag gttggagtcg tgaatacccg caaatggacg atcagcacat cagtaacaaa 42480
attaatcgat ttttagaaaa gcttgatcat tatctagcac atcgtgcccg aacgggtatt 42540
gaacaaatac aacatatgcc tggacatgag cgattgcttg tcattaagat ttatggggat 42600
ccaaagttgt actttgtttt ctttgacggc atcactgccg cttggcacaa ttaagcctaa 42660
atacctaaaa cgttaatttg taggaaaaag caatgcctaa tttaattgat aaattgtacc 42720
gacctgaagg tggtcgtgaa tctaacatca tcgcaaccaa taaaggttgg gttgattttt 42780
caaaagacct tgataatgac gagttgcttg tggaattcga ccaaaagggt ggtttcctta 42840
aacaccttgc cgacaacaac atcacaggtc ctggtattcc tggtttgaaa acctttggtt 42900
ataaaccgcc tggtgatatc actgctaagg gtgacgcgga tactggtggt tacgaaaatg 42960
attcggttta cgcactcact aaagccaaag cagactctac ggcagatggc gtacgttttg 43020
ctgcaactgc tggtgaccgt gtaacaaacc ttcaagttgg cttggatcct ggtgctattg 43080
cagaagttgc ggtttacgaa acatctggtt cattgccaac taaatttgtt ggtaaagccc 43140
gtgttcgtaa cacagattca aaagtcaact ggggtacagc tgttgtcgat attccattgg 43200
aaaatggcaa aacctactgt ttggttgtac attcaatcag cggtcagtta cttgaatttg 43260
ccgagccaaa tggtattgac aagtatgccg ttgtaaatgg cacattccca gcgactttgc 43320
ctggtgcaga cggcacaaac tctgattgga acgttgctgc atacgctatc gttgcggatt 43380
actcaaacgt ttaaacaatc tgaaaactaa atcaagggtg acttcggtca ccctttttca 43440
tatctggaac ctgtcttata atgattccac ctcaacaaag gaatcattat catgaagaac 43500
tatcctatta tccgaattgc tgatgtgttc aaagcattgg aagcaggtga gcacactcac 43560
ttcggtcacg gttgtaactg tcgtaaacaa atgcatcgtg attatggcgg tggtggtgtg 43620
gctgctatgg ttgccgattg gtgtccagaa gcgtttgaag ttgatcgaat gacatcagga 43680
gataaccgtt tgggcacata tacttcagct gtgaagaaag tggaatcagg taatatcact 43740
gtactcaata tgtactcaca gtacgaaagt ggccaagcgc cattattaac actcgccaaa 43800
ggcatatcca acagcctgga caaagttaaa ttgttgattg acgacaatac taaatccttg 43860
gcaatccctg cgatcggctg tggcattgct ggtggtagca tttcaatgct tgtgtcgact 43920
attcaattcc ataacattcc aaatctttat ctgtatcttc atccagacga cactattaat 43980
atcctggact tattatatgg caaagtcata gctaagtttg aagacggata tggtctgatc 44040
cgtgttatgc gtgagcctca acaaggtcgt cccttatatc gcactatggc agttgataag 44100
cgtttccact cagttgaaga cctattgtca tttgcgcgtt cattgtatgg ttcagctgta 44160
gtgaacattc caaatttaat cttggatgta gaatattcat ccctaacaac aactcaagag 44220
tattaagcac aatgccaaat ttattcatta aagctgcaga agcagtacgc atcaaaatgg 44280
gacttggtaa caatgtcgat aaacagtatc ttgatctgct tgaaaaggtg ttcaaaactg 44340
gcgaagacat tgaaggccgc aacggtatgg tacgcaaaac ctttggccat atgatggagt 44400
ttgaccatga agaaggttat ccgattccga cttcaaagaa aaccctacac ggtgcagctg 44460
ttgcggaaat gactgcgttt attcgtggtg aaagttcaaa caagaagttc atggatcttg 44520
gttgtaacgt ttggaaaggc aactgtttcg ctgattactg gatgtcaaat ccaaacaaca 44580
agaaacctgg cgatgactta ggtcgcatct atggggtaca gtggagacaa tgggatggtg 44640
gtattgatca gctcaaagag cttgttgata cgctcatgac caatcctcat gaccgccgta 44700
tgattgtcag ctcttggaac ccaacagata tggacaaaat gtcattaccc ccttgtcctg 44760
tgttattcca ggtattcgtt cgtgatggca agtatcttga ctttgcactg tatcaacgtt 44820
cgtgtgacat gttcttaggc gtgccttttg atattgttgg ttattcatat ctgcaaacgg 44880
tgatcgccaa gttaactggt tataagcctg gcaagttcaa atacttcctg gcggacacac 44940
acatttacca caaccacttt gaacaagtga aagagcagtt gggtaaccgt ctgaagacat 45000
tcccactacc aaaagccgaa ctggattggc cttcaacgga catgaacatc acattggaat 45060
cgttcagtcc tgaagatttc aaactggttg gttacaaatc acaacgtttc atcaaagcac 45120
caatgagtgc atagttcaaa tctaatatcg gggttaaaat aaccccgttc tttaaacaaa 45180
actgtctttg gagtaaaaga taatggaaac tactttcaat ctgtctcata tggttgcacg 45240
catcatcgag atgcacacaa aatttggcat tcacaatact gaagaacatg tattgtcagc 45300
cgaagaaacc aaattccgtg ttgctgcaat gtgggaagaa tttgaagaag gcaaagatgc 45360
aacttcaaaa gaagaagaat ttgacgcatt gattgacctg gcagtcttcg cacttggtac 45420
catcgatcgt atgggtattt cagatcagtt tgttccagtg ttcaacactg ttccagttac 45480
caaaaccggc catacgtttg acgatcttga aaagatgatc cacgtcgtgg catatgaaat 45540
ggaacagcaa cctacaagct tagagcaaca tgcgatcaat attgcttact tgatgcatga 45600
attgtttgct gtagttaaag caaaaggtta cggtgacatc ttcatggctg gttacaacct 45660
ggtacaagat gcaaactgtg ctaaagagct tggtgcaaac actaacaaag aaggcggtcg 45720
cggtgcattc aagattgatc tgcgtaagcc agaaggctgg aaagcacccg accacagcca 45780
attgatcgaa agcttggctt aattactcat cgtgggctcc tatgtgagcc cacatcaaac 45840
aaacttaaca aattggtaat atcctatgtc taagacaaaa ccgactggta tcatcttact 45900
tgaaggccct gacgcggcgg gcaaaacgac tcttgcggaa gcattcaaaa aagtggtacc 45960
aaatactgaa atcattcatc tgacttggtc accaaaactt gagactgtaa tgtctgatta 46020
caacattggt ctattagcac atgctgaaga attagcgaaa gaccgtttgg taatcctgga 46080
tcgccaatgt atttcgactt acgtgtatca gtcaactttc cgcccggaaa aggcgtcaca 46140
tcatccaagt ctgtatgcac aacttgcgaa atactgtcca agtatcagca tcatcttctg 46200
tctaccagag tttgacactt ggaaacagaa cttcctggac atgtgtgcat ctcgtgaaga 46260
aatgtatggt gccgaaaagc ttgaacagat gatcgaagtg taccgtcgtt acgacgtcat 46320
agctcgtgga cctcagacgt ctgaagatca gaaactaacc tggtatccag acaacgtatt 46380
gatggaccaa atcatggatg ggggtgtact gaaacatatg cattcttatg tgtatgactt 46440
cacaaaactg atggacaacg aaggtctatt gtttgcagaa ggcttcttgc ttggtcacca 46500
agcacaccaa gaaaaattca agtaaactca acatcaaaag tcaaggtctc agtattatta 46560
ctgggacctt tttattttat tggagaaata tcatggcaaa ctatcgttat ctaattggct 46620
ttggtcaacc aggctatcat tacacacttg cgggtggtcc atctgcttat atcacatcat 46680
caggcgttgg ttacaacaac aaaggtatgg acctacgtac agagcactat gccgatcact 46740
cttatggagt cgatgagtta gtgatattct tggcccagat gttgaagcgt gtatcatcac 46800
atggcaatca tatcgatctc aaccgtgtat acaatgatcg catggaatat aaattctacc 46860
gcatcccttc cggtgctatt gatctgttcg attgtgatta tcagatcaac attcatgctt 46920
atgagatcag tcaagagcag ttcattaagg atgtcaaagc cagtgatgca tggaatacac 46980
tgcgtgatgt gtttgataat cacgaccagg actaccgtga tgaaccagaa gacatcaata 47040
tccgtctcta taccaatcat attgatggga agtggattca ccatcatgac gtcctgaatg 47100
ctcctgatga accatggcaa aagacaactt tctctttacc tagtcatgtt acggttgatc 47160
aagtaaactc tcacatcaca gaaaaattgg agttgactta tggcatgtaa tccattaata 47220
caatcgattg ttgctcgtga ccttaacaat gttattggtc atggtgacaa cgagctagcg 47280
tatatcaaca aagacgacat gaagtacttc cgcactgtaa ccaaaaatgc tgtgtgtatt 47340
atgggttcaa agacgtttga aagtcttcct ggcctactaa agggtcgtga acatatcgtg 47400
ttgacatcgc aatccaaaga acaatacatg aaaacttcaa aatttgacaa agaagtattg 47460
gaattgtcag gggtcgagat cgtcaataac gttagtcaag ccttggtcgt agcccaattg 47520
atggccaaga tggaacatcg tcctattttt gtgatcggtg gcggcacagt ttaccgtcaa 47580
tttatggaac atattgatgt gtggcatgta accactaacc taacatcaaa gccattcaat 47640
aagcctgttt atttccatga atcatttgac tcaaacatta actacaaact aatcgctgag 47700
acgactcatg ttgattgcga tatcccacat tcaatcaaaa tctatcaaaa attgtaagga 47760
accactatgt tttcaggtga tatgaactta gctatcgccg accttgaaac acttgcgcat 47820
actgatgatg cgcaagttct ttcatttggt ctgtgtacag ccccagtcga taagacacta 47880
acattcaaac aattgtgtga gacgtactca atgttcctca agtttgatgt caaggaacaa 47940
gctaataagt acaaacgctc attcgatgag aacgtgatga ctgagttctg gcttaatgat 48000
agccatacga accttgaatc acgccgtgtg agtctttacc ctagtccaga ggatgtttcc 48060
attggcacta tcacagacgc catctggaag catacaagag ctctgggtat ggatatcaaa 48120
acaatggata tcggtgaccg taacgcatat gacctatcca aactgaagca tatgtttgat 48180
cagactggta gtatccaacc ctggtcatac cgtaatggtt tcgaaatcac ttctgtattg 48240
aaggcctggg gagcagaccg atatgctggt atcgatccta aagatattcc tggcatggtt 48300
taccaccatc cagtccatga tgctgtactt gattggcttc gcttccaata ccaagccatt 48360
actattggtg cattggatag gcctgaaggt tatgacctag actacatcct caataactaa 48420
taaaaaccct gatgcataac acatcagggt ttatttttat tccgaatact aaaaagcccc 48480
agaatcaatc ctggggcttt cttatatctg actaagatgt ttacttggct tcagactcat 48540
ttgactctgt gcgtcgtgta tgctcgctct gatagatacg ttcaaccaat tcttcttggt 48600
tagcagtctg gctctcgatt acttgaatca aactcattga aagcttagcc ggcaactcgc 48660
ctaatgcttg catgatcagt tgtgcttgtg taggtgttac agctagtacg tagccttcat 48720
tttggttatc catggatatc tcctgatgtt ggatgagtgt taactttaaa gggcttttct 48780
tcaacaaagc gcccaggttt gatgatgaca agctggttgt cgcttgagac tgttacaggt 48840
acacataagt catctttcaa aagccatagc ttaacaccac gtttctcggc ttcaatcaga 48900
ccacaatgcc ctgaattgcg aatatcgcgg aagacaacct tctcgatgcc tgacatccat 48960
aattctttaa cacagttagc acatggttca cgtgtaacca ctgccaaggc acctatggct 49020
gtctctgtga gacgttccag cttcttaaga gcgttaggct cagcatggac taaagatgcg 49080
cctggcgctt cacagtcatt gctatcgcca ggcattgtac cgttgcaacc ggaagcaatt 49140
ggtgtattgt atttaacaat gacacaacca actttatgtt tggtagcgtg tgataacttg 49200
gcatgagctt ccgcacatac catatgagcc caaatagtct ttggacagat gtaatccgat 49260
ttgatcatga tattctctct gtgttatttg ttgcgaacaa aagtaatgat gaaagcaacg 49320
gtcatggtga tacaaccgta taggtatgca gtggcaatat tttcagcaac ttttactaag 49380
tgtggccatg cgtcatcagc attaccaaac acgaataaaa agatgagacc aatcactgca 49440
tagataatct gggatatgat gaatccacct ccccaaatga tacctaaacc ctttaaacat 49500
ttcttcatgg tattgtcact cctgaattag gaaaatgttt acgatgaagc cccattatgc 49560
gtttggtaaa acttttgaag aatggattac ccaatctgtt aacgtatttt gcgatgatgt 49620
ctccatgaca tggcttcgac ttagtaggat catcctctcg acgacagtaa caacccaggc 49680
gcttaccaat cagtgtgggc aactgcgcca tcagcaactc gtcagccaat agatacttct 49740
cataagcagc gatcgcttgg gaacgagtca taccttcttc gaaagcataa ttaccccaca 49800
tacccccgcg accaatgtac acatcgtact tattgaagtt cttgtgcaca gctttacatt 49860
tcgccatgat atttcctcca gataataaaa aaccctgaca aatgccaggg tttcattata 49920
ttacacttca acaatttggt gctggatata gatatcgcca agttcacctt ctttacgtgc 49980
aatccagtga gcaatctcct cacgctcata aggtccagac aacttacgct caccagtctg 50040
gacaatgtgc gcttcaagca tgactgggtc tttaatcacg tccgacattt tccaaatctc 50100
ctttaatgtt ggtgatcaga acttcaaccg taccatggtc agtgtcaagc aatcgtgcta 50160
tcttgtatgt cttcttgtca atgtagtaga cattcaggtc aggattgttc ttgatccaat 50220
cttccaggat atgattacgc ttaccacgat gcgccagaac attgctcatg atgaacaacc 50280
caccagagga tgcacgattc tctgccatat caatcagctt caggtcatgt tcaaacgacc 50340
aaccatgcat atagttcgca ttagtgattg tgtatggcgg atccaggtag tcaatgacat 50400
caggaccggt acgatgcata tcaaagaatg tcatgaactc atgagagctc atatacactg 50460
atgtcaacca gcaatgggta tcgtacaacg tctgccagtg atcacttgga ttccaatcgc 50520
gattgcctgg aggattattg aaaccacctg aaccattcca acgtggtgtt gacgagaatg 50580
cacatcgcac aataccaata tccaaccatg gacaacggtc atgattcgca ttcacgaagt 50640
cacgcatcgc ccagaacgca tccttattgc ctttggttgg gccaaactgt gcttgtagct 50700
tctccaaatc cgccatcaca acatcaggcg acatatattt caggctctcg atcacacgat 50760
aacaacattt gtcgtattcg ttatagttga ccagtgcatt gacgtcttca agttcaaaca 50820
tcttcgccat attaatggac acaacacccg aaccaccaaa cacgtcattg aagataacac 50880
catccttcat tgcggaggat tgtcgcacca actccataat gtatggcaat acttctttct 50940
tatcgccatt gtaacgaagc atgttatcaa taatctccat cacatttcct tcctaattag 51000
aataaactgt aagcaaccat tatcacgcag tacggactta tgaagatcac acatcgaatc 51060
cagcgtatag acggctgcct tgtcaaggtc aacagtcgat ccggatagcg tccagtacat 51120
aaacgaacca tgggacagga cacggacaat gaactcatca tccgcttcca ggtctgaata 51180
agagacaaca accatcgcca tcagttattt cttcatctca acatggatca ggtcttgacc 51240
accaatcatg ccaacataat taaattcagc atcatcattg ccgcgtacac gacgctcatt 51300
agacgccaca cgccacacgc tatcactatg gtgagacaca tcctgatagc cggctgatag 51360
gagcacagca tattgcgcat tgatatcatc ttctttgtcg gacacagtct cagactcggc 51420
ctcataagca cgagacacaa ccgcagtacc cgtcactatg ataagcgtaa agcgacgaac 51480
gccagtaccc ttagcaatgt tatggaccaa tacttccatc atctcaccca aagcacgttt 51540
aggcaatgcc aagtgttcag cacgcaccgt aatgatatcg ccatccttga ctcttaataa 51600
atccacaccc gtagtctgtt ccaacaaacg cagaacatgc ggattgcttt cttgatcacc 51660
cagtgtatag ataggcaagt taccatgctc ataataagca ttagtgtacc cgccagtaaa 51720
gataagccca gcagtatcca tatgcttagc aatgcagtgt tcaacattac atttacgaac 51780
ggatacagta cggagttcag gaccttcttc caacgccaat acataagtta gctcctttgt 51840
ttcttcttca aatgttctca gctgatccaa gaacgcatca tagtgtttag tcatagcctt 51900
tagtcctatt gttcatatag taaaagaaaa gtggtacagc aaccattatg ccataccact 51960
ttattatgaa cttttgaaga acagactatc gagcatcatg catcttacat cgtatccacg 52020
tcttataccc gaatccctta taggttgctg tctgtgtatg cgatactatt tctgcaacat 52080
aaccagcagg cagtaatgcg gtattgatat gcgccaacca tttatgcttc tcgctcggta 52140
atgcatcacc taaacagaag atgagttcat cagggtccaa gtcatgccaa tgagatttta 52200
gatagccttt accctgatat gcatcaacgc caggaggtaa tggtaggtca gtacgagagc 52260
gtaggtattt gttaaggagg atgagtctga ttttagattt agtcttggcc attgtgcgat 52320
tcctaatcag ttaaactatt attatgaaaa tatttagcct aatccgtgga attcaaaacc 52380
aggatttgag tgtttgagag ctatatagga gtctattacc caggggaaat attgagaaat 52440
gataagaaag gtgttacaaa tgtcaggaaa ggatatgcca aatgataaga aatgataaga 52500
aatgccagga aagtaaccgc cagggagcgc cagccgacaa aatttcaaat tcgcaggcac 52560
aaccttggca ccgggaaaga gtttacattc ttcaaaagac ttttgaagtt tcgtcttaaa 52620
taaggaacca aaattgcgcc tatatagtcc gtcatcaaac aaagcctgtc acatagcaaa 52680
tcatactaca cgccaggcaa tcgttcaatc aggagctata caggagccag aacagcacgc 52740
caacaagagt aatcactatg aaacctgcaa taacatccag gatactatca taaagcttct 52800
tgtgatcgcc tgtcaactcg ccagccaggc cagcagacat gaacccgtgg aagtatccag 52860
ctatatagac gatagcctcc aggagtccta gccaataatc aacagcttcc ttagccttgg 52920
aacggaacca cgtaccgcgc ataatgccct accttaccca tacaatagat gatgaagatg 52980
atcagtgcaa tgatgaatag attctccaca caccagatca ggccttcact gaatttcttc 53040
aaaagtttct tcatgaccta tgtcctacgc caaattatcc atgagctttt gaacagttgc 53100
caatgcagat gatagacgaa tgtattcagg gcactcgcca ggcacacgag gggtatggcc 53160
atcgccattg gtttcttctt cggcaatgta ttccatgccg gccattaacg catcacgcaa 53220
ctcacataac gcagcgccag acacaggagc cgtacggact tctttattga ccagtaccat 53280
caggtcatgg atactattgc tatcgataat gcggtctacg tggtattcat caagacaatc 53340
gaatacgtca ttgaagtgtg tgccattgtc atgacgacca gatacgatca cattgttgtt 53400
ctcgatgaca atgccgtgaa cggttagtgc ctgcttgcgg atgttaacgt tattgcttag 53460
gatgcggacc agttcagccg ttagtttgca ctcgccgttg aagtaaagtt ttttgatgtt 53520
catgatgttt ctccaaatga tgttttgaga ttttgaaaag ttcaaggaaa tgatatcggg 53580
ttataaactt ttgaagtttt taactccaca gatatccatt gcttcttggg taaagttgta 53640
ataggtgatg ccatccttta cgatatccac ttccaggacg ttgccataat ccagcattgc 53700
acgaccgctg atcgtcagga acagtgtaga acccacatcc tggatgccct caaagcacac 53760
acgaccacgt ttgttcacat aagagcgtaa tggctcaccc ttaatgcaac aacgctgtat 53820
aacgctgatc atgggttgtg ttagtttaag ctttagaacc gccatatctt ggcatctcct 53880
ctggctaagt gatgatggaa tgattcacga taaggtaatg acatatccag gtggcaatgt 53940
gtatagacct ggcgagcctg acatgataga tagtcttgaa gacgttccag tgatagtcct 54000
gttggacgct cgatataaac actgcggatt cgacacttat tttcaagtac gatatcagtg 54060
atcatcacat tggttgtgcc aggcaccagg taagacttgt tctcgtccgc attatcttca 54120
aggcgatcga gttcatccca taatgccggg ttgttcttgg catcgcaaat gatgagcaca 54180
tctgaattat caggaatagg aatcattagt ccaactccaa tttatagact tgtcggaagt 54240
cctgcttatc ttcaatgaac gcatctttga agaatttcag accggaatgt tcaattgatt 54300
tgatgctagc cttgttctca cagtggacat atgccattac tgtatcaatg ccagatgcga 54360
tggcgaatgc gatatgagat tccaatgcct tcttctggat gccctgacca cgatgtgatg 54420
gatgaacgac tgtgcctgcg aacttccatg tgttccatcc tgcttccgcc tggttgcagt 54480
cttctacact ggcacgagca aagatagagt aagcaatgag cgtacgccca ctgaacacac 54540
caacaacacg ctcgcggtcc aaggcttcac ggatagacac aatatcacga ggggtgaact 54600
ggtctggagt tgtgccacgc atttgaagtt tcaacatctt gaccgcatat gaacgattca 54660
gtggtttcag tttcatccga ttagtcctcg atgaagtatt cagtgtctag gcggttacga 54720
gataagttcc aaggttggat atcttgggcc atgagattac gaaggataac caatgtatgg 54780
atcgcatcgt taatggattc aacaatgatt tcgaatggtg taaagcccac aacccagtca 54840
gtagaatttg catcacgctc ttggtaatcg tcccaattgc gatcgccaac catgaacagg 54900
tccatttcat ttctttcaat gaattctgca tcttccataa tggttaggtc caatgggata 54960
ttgtgatcag caaccatgcc acggatcatt gcagtgcatc gatcgacaga atgacaatta 55020
ccagtgaaga tgttatactg ctcacgagtc agcgtgatag gggtgttctg tgcttcttcc 55080
gcacggatag cagcttcttg ttcacggcaa gcttcaaggc cgaataggtc tacattagac 55140
atgatgattc tccatgatat ttgatgtttt gaagttcccg acaatgataa agtgccagga 55200
acttttgaag tttttataca ggttcggtaa tgaacgaact caagtcagcg aagacacgtg 55260
cgcctgtgcc atccaataca acagtggcag taggatattg gccatcggta cacacatggt 55320
ccacataacc cgtctggtct tggtatttgc cggaagtgaa taggatacgc ttacgcttgg 55380
tgatatccca taggttcgcc aagtggaatg gagtcactgt taaacgccac tcgcctggat 55440
cctcaaactc aatatcaaag tcgggtaagt ccagagtcca atagttctct ggtgattgcc 55500
aaaagttccc atcaacctta gggttttctt caaaagcgat gatgccacca cgagaaccaa 55560
tcgcaaagta gatggtgtca ttcttgatat tgcggcagct atccaagaac ttttgaagtt 55620
ttggaattga gatcatgaac ggaatgccgt aaatcttgac ctcaaccatt gggtgtttat 55680
gtactagcat tgcttgttct ccttctagtg aacgccatcc atttgatacg aattttgtgt 55740
aataggatat tgagacactt ctctaatgga tagtagtctc ggttgatttc tacgagattg 55800
ccaaggtact ttcttgggat ataacgcgga taggcttcaa caaaaccttt gaaggtcaaa 55860
tccacttcgc catcctgggt attgttggca caatcatatt tgatattaaa cccgcaagtc 55920
ttatccgcat ataacatgtt cacatcccta cgttaaaagg ttcagcacag tcatccaaag 55980
ccaatgcaat atcgccaagg aatggtaatg cttcagattg tttgcaacgc ataatccaat 56040
caaggttgtt ggcactcacc catacaatga gtggatccac actatcttgg aggtgaccac 56100
aatggttctc aaagagttct tccataaggc cagcttcatt actgacatca tgacctttga 56160
gggttaagct gcgaagaatc gcagcaattt gaagattggt aacaaagtat tccatttaga 56220
tcaccgatag gtcgcgaagc tgtttatcca caccaccaat gaacaattca gcatcgtcgt 56280
cagcaaccat gttgtatttg gcataatccc aagatttcca attatcagga atgtagtaga 56340
tgttaaccca ttcgccaaag atacgttgct gacctgagta gtttaggaag attaacgcat 56400
tgccatcgac atggagatta cggaaaacca ccaagccatt gccacggtac attaacttac 56460
cagtctcttt gtcaagagca gaaacaaaag taagtgaaga agtgatatcg acaacagtat 56520
tgccaatgtt caaaagagat ttaaccatgg tcttaacccc acttttcatc aaggttgaac 56580
acgtgaccaa taatcgcagc cacagggaag aagccaatga agaaggctag ggtaagaagt 56640
tgccaatgca tttccataat gtactccaaa gagttgttta agttgatgag gtaatagtgc 56700
caggttctat gaacaaataa cgtgatctag atcgcgcttt tataagaagt cctatatgtt 56760
tttctgaatg tcgtaaaacc ctggggtagt cggacaatcg caggactgaa tgtactatcg 56820
cgatggaagt atgggagaac gatttcgata atgccgttag cactatcaat agaaagggta 56880
tagccacgca tgtagtgcgg acggatttcc aaagccgaac ggatgcggtg atggatttct 56940
ttgagcgttt cctgggatag agtaaagcca ggcgttgtct catgacggat aactgtttgg 57000
aacatgatga ttctccatga tattaaaaag cccaggaact tatacaagcg cctgggtggt 57060
attttatcaa acatgaactt ttgaagtttt ggtataaaca ccatattgga ttaggatatg 57120
gaataccgca tctcgctggt gagtgaacgt ctggccatcg tattcatagc caatcaccac 57180
acgacctgat ttgttcaagt gtcggatgat atcatgatcg ccttgttcca ggtgcacagt 57240
gaccttatct gggccatgac gggtgatgta atacttgacc acttcatagt tcaaaggatc 57300
cgccaggagc ttatcaacaa tcctggacac gttcatacga catgcccaag tgctgatgaa 57360
ctctgcaccg tccttggtga ctctgatgaa cggattacga tgcttgagtg tcgcatagac 57420
cctggcacga accttttgtt tcatcgtttg acggcccatt tacaacttct ccaacttttg 57480
aagtttttca ttgattcggg taatctgcgg atataccttc ttgccaatga tgacactcac 57540
gacgtatagc aatacaagga acaggatttg ctcactgcca ggcaatgata atgtcgcttt 57600
cgtgaacaag acacttggcc agtagaatgg ttcatctgtc atgcccgacc agaaacacgt 57660
attcaggata attacacaca cgcccgcaat ataggttaag aacgacacaa agctttcgcc 57720
atcagaattg cgaccgccaa gttgattgat aacgatatca ttgaacttgc gagcgtgttt 57780
cttggcaata tggtctttca tgccaatctg taccatccag atttttgaga gcatgccaat 57840
gttcaggtgg attacgacgt aaaccggtgt ccagatgatg accatcatca tgatgaatgc 57900
tgctgttgat aacatttgaa gttctcctta tcgcttcttg ttaactggaa cacaagtgcc 57960
acggattgtg atcttacctt gtttgttgtc gttggcactt tccaacgctt gttcacaatg 58020
acaatgcatg atcttaacca cacggaagcg gttagcagtt gcacgatagc ggcacgggtt 58080
gataccttta actaacataa ctgtggacat gattattccc tacccttctg ataagtgtac 58140
ttgaccaaat ccatatcctt catattgcca tggaggaagt tacggagatc aacaatggat 58200
tcaaatgccc aaacgttctg aacttggtca aagaccccat agttgtcaaa caatctgact 58260
tccttcttgg catggaaata acaaatgcta tacttgcgga agtagatcac gccattgatt 58320
tctgttggga tttcaacgac tgtatgagga acacggccat tcgtcatgtt cttgtggtag 58380
cagtaatatc caagtgcacg gatatgagcc aggatttcat tgtactgctc tgtaatcgca 58440
tgattgacac gattacggtt atggctaagg gttttcatta caatgcctcc atgaacattt 58500
caagggttgg caagtgtggg aattccaatt tctgaataca tcttctccaa tctaagactt 58560
caggttttgg catatccaca ttgcaaagca gtcggttgtt catagtgcca tatgaagtga 58620
ccaacatcca accaatagca cctggaattg gttcgctatt ccaaacgaag acatcaccat 58680
tctgatcaac tgtcaaccag aatgcaatct tgccaccaga acgcgccagg ttgcgctgta 58740
cccacaattc attgtgagca ttatagaagt tccactcaat gaagttcgtg aagccacggg 58800
acaacacagg gatttctacg ccattaatag atgcattgaa tgacttaggt ttcagtttga 58860
cttctgaaaa ggctttttcc aaagcctttg caaattgaat gatagcttgt ttgctgtaca 58920
tctcagccca agtttcggca taatcagtca tcgcatagat gtaagaagta cgctcttgaa 58980
tggtgaagtc attggttgcc aaatccgcag caatcttaac agctgtagca gagccatttt 59040
cgacaatatg tgtcaggatt gcttcttctt tgtatttgaa gttctcaaca ccagttagtt 59100
tgcggatggt ctcaatagca gtttggaagt tcataacatt ttccttagtg gatgttgttt 59160
gtttcgatgg gataatgata tcaaatgacc tgaaccaata atgtgatcca ggtcacagtt 59220
tggattactt aagcaacata acttctgcca tttctttcca atctgtcatt tttgcataat 59280
ccgcaagttt caaaagtgta cggatagaca cttcacgaag ttcttgaaca ttttcccaca 59340
tccaatcaat catttcttta gttgctttat catcaacacc acgtgaagca agcagtggag 59400
tctcttgagc cacttgttgg atacgcacca tgatttcagt tggagtgtat acgccaagtg 59460
ataggtagtg tgaacgagat agcagtgcac tgaagtgtgg agacaaacga gtgccacgct 59520
caaccatgcc ttggaaatca aggttagtga tgaagacaac agtaccttgg aaatcaaata 59580
caggtggaat gtcattttct tcaagatact tacaagtaga accgtaagaa accttgcgaa 59640
cgtcgccaga atccaaagca gctttcataa tgttcaatga aacttcatct tggaagatgt 59700
tgtcagagtc atcaagaaga acgatatcat cttttgaact atttttgtac agtgtttcat 59760
acaagaatag tggagttact ttacctttga tatgttcaaa gttacggata cgaccttctt 59820
catgggcttt ggtcaattct ttttcaaatt tgtaagtttt gcccacacct ggcgcgcccg 59880
aaacgataag agcatacaaa gaatggtcga taacaccgtc agcaagcatg tcaagaacat 59940
tgaaacgttt gcggatgaac aaatcaagtt cttcaaggtc aacttctttt gtttctggcg 60000
acaaagttag aatccgcagg atgttcacta cttccaacca tttggaactg tggcaaccac 60060
aatcggtaaa cgcctgtaat cttatggcgg acataatgaa gtccagtttt aggattacgg 60120
tatacgcact gttcatagta cgacgggata gacatttact tctccaaggt ttggactgaa 60180
cgacgacgct gctttactac acgattcgca cggaccttac ttaaacgacg gttgcggcat 60240
ttcatttctt cacctcatac atcttatgcg ggcgataaag ttcaacattt gttaaaccat 60300
tgcatacaaa gttgatacca tttccaaagt tacggaattc ccaaacctga cctttttcag 60360
atttgattcg cataacagtc acatcacgca aatcgacagt gatattacaa ttgccatggt 60420
aacgactttg gaagttgatg acaccccatt ttgggctatc cattaggttg ttgatcacat 60480
gttcaaaacc acgacgcgct aacaaacgac caatgcggaa gataggacgc ttacttttga 60540
tcagattttt ctgattcatg ccttcataaa aggcaagtaa caatgcaata tttttcatga 60600
ttattcaccc aacttactgt tagcatattt gacaaaccct ttctgacgag cttgcaagga 60660
cttcagataa cgattgtttt ccaaagcttt tgtgcgaact tcttcccaca tcagtttagt 60720
tgcagtcttg atgttgtcat cttgtttgac aactttgaca tggatttcag tcaccaaacc 60780
attctttaaa caatggttga attggttagg aaccgcgaaa ttagtgtaac aatcaaattg 60840
tttaattgtt acatagttgc tgcccatatc tgcaacaagt acaacaaact ctgcatttgg 60900
gcaatgagtt atcaattcat tcataggatg ttccttatgg atgttcaaga aagagcccaa 60960
ttaagggctc atgatggatt atagagatgc aacaactttg ttaagagttt caaggtcaac 61020
acgttggata gtagtgtact tagcaccttc tttgaactca agacctagtt cgttaagaac 61080
gtcagttgct gatttacctt ttgaagggta gaatagtagt ccaaatgtac catttttgat 61140
tggagtgatt tgaagacgac cattgttaac aacaagtgag ccatatgttt gacgtactgc 61200
aggtacttta acaaccatgt cagagatgtt aagagcttca agagcttcta cagaattgaa 61260
accagagatt gctttaggct ttttgatttc aattgtttta ggctttttga tttcgataac 61320
ttcaagtttg aaacctttaa ccattgcttt cttgccgtta agaagacggt taaaaccagt 61380
agagtcgcca gtgtaacctt gctcaactgc tttgcggtag ttagccgctt tagttgctgc 61440
agtgtcgatt gcgatagtga tttggccgtt tgtgataact gttttagtgt tcataatcaa 61500
tgttccttat ggatgtttgt ttgtgttgtt tgtttcgatg aggttatagt acccaattac 61560
gagaatcaaa aacgtgacgg agatcacgtt ttataaaaga aagggaaatc tttttagatt 61620
tcccatataa gttattaatt tgcaaggcgt aaccaatcac caaattcttt caaagtcaac 61680
ggtctgtgga acgtgacata agttttggta attttcagct tggtatctat cttattgcag 61740
taagcaccct ggttgtatgt gatatctata tttgggatac cacccttgat cttacgcatc 61800
tgaacattga cggtcatatc cgcacgtatg ttgacttcat acttaatgtt ctgatatcgg 61860
aaacgaatgc ctgtatgcga tcggttactg aagtaataac attgtgtcac gttatggaca 61920
cggttaatga cgttgtgtaa ctccttggcc agaagcttac ggggaccatg accgtccgtg 61980
gcgtcaaaga agttcttgac ctgatgtgta acagccattt ccaaatcaaa cacacctgga 62040
cgcaacggtg ttgtgcgacg cttggtcata gacttcttct ggggttcagg tttagaacca 62100
aataatccaa agaaccaatt aatcattctt ttcatgataa ttacctttaa tgtcttggat 62160
gacttcattt aaaatagcca atccaacaca gtcatcttta tattggatcg ctttgttaga 62220
accccggcac acgtagtaat gtacgatcgg caacgggtta ttcttcaaaa attgtaagtg 62280
gtatggatga ccagcaatct tacgcttcat tgcagtgaag ataatgtcat ggaactgttc 62340
atgatccact ttacggttga ctggaggaag taccttcatt ttcttcgcgg cgttcttggc 62400
ttcacgacct tccatgaata gaaggtcaat gttacctgtc ttaagatact gccacaagcc 62460
ctcaatcgtg ttgtaatgca cctcatggcc attgactaca gcaacacctt cgtatcgata 62520
gaagttcgac aggatgcgtc ccacctccaa tttggattcc gaatacatgt tgatatgact 62580
gatgccttca tactgtggtg tatgcggaag caatgatggt tcccaaccga tacactcaca 62640
gatgccatct ttaatttggt gcatcatatg ttcaatgcca tcaggctcag gtcctgtagt 62700
aatcacatcg ccatcctgga cctggtcttt taacggtttg atgcgagtgt caatcattgg 62760
gaagattgac tgtcgagtaa tcttgtagta aagcatgacc tataccttat caatatctga 62820
actgaaatat gccaacgaat tctctggacc atgtttggac aacaccacaa ggattcgttc 62880
gcttggtgtc atgtaatccc aaatctttga atccaggttg atttcgtttt tctgtgcttg 62940
ggttaactgc tcccaggtct tctcaacgat accgtgagcc catgtaccct ttggataggt 63000
ttgttcatac agcttaacaa tatgaacgta ccagcccatg gttttacgct ttgaacttac 63060
agtgtcagcc atttcaacat ctccattgaa tgacgttcaa ggtttgattg catgccaacc 63120
attgtttgag gggtaatatg aataatccat cgaccattct cttttgctgg acaacaatgc 63180
cagttagatg tatggacttg caaagtgttg ttcagacgct tggcaaaaac tttcgcatgc 63240
atttcatctt cagctgtaat cgacaccgca aatgtttgca gacttttagt atcgttccag 63300
tatacaaaac cgtgtgtacg gccaacaaaa ttggcaattg actgggataa ttggactgcg 63360
acttcaccag ctaattgacg acctttgtct tcgtggacaa aagccactat gctactctga 63420
ccgtattgat ccaacatttt caaagttagg cattgttcag gtgaaggtag tgctcttgca 63480
aatgatactt gaatcatgat gtttctccaa taaaataaaa ggaccaggta ataatacctg 63540
atcccaaact tttgaagttt taaaatccac gttggacttt atgcatgcaa cacttcttaa 63600
acttcagacc gcttccacat gcacatttgt ggttacgttc ctggttctta acacgacgtg 63660
caggaattct tctttctggt acgcgaagca tacgacctag tggtttcata tccatcttta 63720
ttctcctaaa tttttcgcca tttagcaaat tcaaattcgg ctttcaaacc ctggaaggta 63780
ttgttggcaa tgtgtcgcaa caattccgga gttgatacac cattcataat atattcgttg 63840
atatctttac cgtattgttc cagtgagtgt ggtaacaagc ataccttaac accggctttg 63900
atcattttct caatacggtt aacgatttct cggttgcgtg gttgattatc tggaatatag 63960
atatcaccct tcactttcaa caaatccgca tctgctgatg ccatacagtt cggcaagaac 64020
aggctatcaa tcggaccttc aacgaccagt ttggtacggt caaagtcgat acgttccaga 64080
ccataacatt tggtggtctt ttcttctttc ttaatcgaga tatatcgcag gtcacactta 64140
ggatcgaatg aacgaccttg tgccattatc atctcgcctc gagcattgaa gaatggtatc 64200
accaaacgcg gatcctttgg acacttctca gcagacaaca tattctcagg gttcataccg 64260
atagcaaaac gctgatagtt gttggtaaag aataatatgt caaagaaacg ttttggaacc 64320
tggcgattta ccaggtagtc gtaacacata tggccagggc caagctccga aactggaata 64380
caataagtgt ccaaagcgta atcactttga acttcctgtt ttttcttgac caatgaacgg 64440
gtacgccgag tatctttgtt tgcggctttc tttggttctt cttttttctt gagtgccatg 64500
taagaaccag aattgcgttt ggtttcctgg aagctatcaa caacatatga ctggtacaag 64560
gtaggctcat aagttttaag ccataaccct aaatcccacc ctgatgcttc accgcagtta 64620
tgacatttga accgcatccg atcaatatca tgatcagcat aaaagtggcc acgacgttta 64680
ttagctttct ttttactgtc cagacatatc ggacaacgaa aacggcaaat attgtcttgt 64740
tcccatgtaa atttgtctaa catacccgac aacatgttta tgtaagactt atcaagttct 64800
atgctcataa taaaaagccc ttagaataat ctaagggcaa ttttaacttt attcaaacat 64860
ttgaggtttt acaacgttac gtccaacttc accataatcc ttatggacaa tgattgctgt 64920
cagcgtgcgg tttgaacgat aacccgcgtt attatgccat gcatcgcgac cagccaacgt 64980
attaaacact tcgacagtac atccacggta ttcctggatt tgtttatggt gtaagtgtcc 65040
tgtataccat gaacgatgtt ctgtttgacc ccaatcgctt ggacagtcat gtgccatgat 65100
ggactcaagc tcagtgcctt taacagtatg cgtatgagtc acaccgatca ggtttttacc 65160
gaaacggtgg tatgtaaata ctgaagggtt tactacgatt tcaacacgag gttcgttttc 65220
atatagacat tctaatgcca ctgccagcca ttgactacta tgttcatcat ggttaccgat 65280
tgcattgatg actttaactt tttcgtgttt ctctagtgct agatcgatac attgacgcat 65340
caggttaaca ccaatacgga taatctttgc ccaacgcgaa tcaacatcaa gggcattacc 65400
cgaatgtgct gtacggtttt gaacattgtc agaatggaag aaatcaccca ggttgacaat 65460
aagagcagtt ttggttgcgg gtgttgattg gaccagggat tccatggcag caggcaataa 65520
ctgttctgcg accttcaagt cataatccat accagtttct tcgccccatg ccatcatgcc 65580
caggtggtaa tcaccaatcg gataatggac ttcccaatct gttgtttcga ccagaggtgc 65640
aggtgttggc ataactcgtg gaacatcttc acacatcgct tgaacaactt cttgcaggat 65700
ctcgaattgt tcttcagcat cgcgttgagt cttcacccat tggagtttca gttcaccgtc 65760
tttaccgtaa agcgttgacg tacccttgac accgaagcct tcaggaactt tcttacgcat 65820
atcgtggtcc ggactccaac ctttggatgc tgccacggct ttcagtcgtt taatcgtttt 65880
ggatactgtt tggtgactga ttccaagagc acgtgcggct ttagtttggt tgccatcagc 65940
tttgataaca gcttcaagtt gttcagcctg gcgtgtagta gtacaatact ctgttagacc 66000
agacaaagcc tcttggacag gtgtgtattt cttattctta gacatctaag atttcctctt 66060
tacattaaga acctcacaat attatcttta tttacggata tgaaaaagcc tatcatttaa 66120
gataggcttg taattgtatt aaagtttgac tgcaatctct tggtccatga tgtaactggc 66180
atccaggtcc atattgccat aaactggaat ctcatcatca ggtttatcat acagacgaca 66240
acggatggtg ccagtattaa ggttgagcac aagaatctca gctgtccact ttcgcgcgcg 66300
ttccgcataa aagtcacatg tcgagtacat tggatgttca ctgagatcaa ccgttacgcg 66360
aggtaacggc aatactccag gttttggttt cttagccatt tagttagttc ctacttttgg 66420
gaatagattc atcaaatctt tcattgaaac atttgatacg ttgatgtaat ggtgtttagt 66480
gttatgctta acatcatcca tattgcgccc caggagtgca cacacaggtt cagctgccat 66540
atcctttgac atatacacca gaacagagaa aagttcgcta cggctgttat atgagacctg 66600
aacacgtcca acattagcct tacaagttac ataaggtttc ttcttcgcag gtacttttaa 66660
ttcagcacca tggatttcca aatattccac aagaccattc agtgcaggca tctcaccttt 66720
aacaattgat ggcattgtca ccttaatttt ggttggttgt ggttcaacaa cttcaatagg 66780
ttttggatta gtgatttcct caacttcaat aactttagga gttggtttag gtgtgactgt 66840
tggtttgcta ttaagagcat tgtgagagaa tttcaaagct ttctctttga tgaaaccttt 66900
tgaagaacac ttacaccaac gtaaagaacc caaacggatt ttcttatgca tttgtggtgt 66960
catgttatac tctttgtcac atgttggaca aatgtaaaca aaaggtcttg tttgtgcacg 67020
agtcttttga ccattcatta acaaagcttt atcaacatcc atattgtgat atgtcttagg 67080
ttcgcaacca aagtgacgca tcacattctt ccattctttt ccatggggtt ttgctctgcg 67140
accgtaaacc ttatgaacaa caatatgagc aaattcatga ggaatggtat ctttgatgaa 67200
ttcttcttca ttatctgcaa gaagggttct gcaaaggttg atgatgccgt tttcggtagt 67260
aaagccgtta gcgttaccag caactttacc catttctttg atattgattt ccggcatacc 67320
aaaagttttt gaaggtacga tgcggtttgc aatatcaatg tacttttgaa gaacatcttg 67380
gacttgatta acagtaagca tgacatattc cttaatggat gttgtttgtt tcgatggggt 67440
aataataaag aatcccgacc acaaataatg tgatcgggat cgcgtttttg gttattggat 67500
tattcttctt cgtagtcgga acaataatcc caggtgatgc ttgggttgat atggtccagg 67560
attatttttc ttgcgttaac ttccagtttt ggacataccc caattgactc accgttgaag 67620
aagacttcat attgtacacc aatatcttta accacatatt cagtgtcgtt atgaatattg 67680
atggcaaccc gtacatagtg tgttccaata cggtctggtg aatatccgat cgcacatagc 67740
cagttaacaa ttgatgctgc gaaaatcggg tcttttacac gatgttcagg aatgataaat 67800
tgaatatctt tggtagattc caaaatatct tgcacttctt gcggcatctt tctgaattca 67860
ctttccgaac attccacaaa taaagtattt ccgaatatat gcattttctt agccatgaga 67920
aacccccaaa ctaaacatca agaacataac aacaaagatg aagaacttat acatgatgat 67980
attggcgcca cttgcccggt caatcttcaa atcaaacatg agcattgcaa gtaacaataa 68040
actgaagtca atcggtaatc catccgggaa acttgcgata tctactatat ccaacgatgc 68100
caaaacacct attgttgaat agaaacccat acaggtgatc gtaaacaatg taatcagtcc 68160
gatgatattt gtcacacgac ctgcggcaaa gtaaatcgta cacattgcaa ggatcaacac 68220
atgtgcgatt gttaagtggt tttcaatgat gaagttaaac attacggatc tccttttgga 68280
tttcatcggc aaatttttgg atatccggcc aggtcggttc aacatcaccc caatgcttgg 68340
tacgggttgc tttgatatcc aaagatggaa tgttcacctg gacgcgtgta gtccaacagc 68400
attcttcatt gcaagggtat gttgaaacct tgaagcttgg ttcaaatcca agaacccaat 68460
tcatgaattc ttcaaagcca ttttcaactt caaattcttt ggagattaga ccaaactttt 68520
cgataccttc tttgctagga aggaacttgc ggacataatc atcacctgtt tcagacaatg 68580
aaccgatgta aatacagctg gtatccgttg atacagtcac tagacctact aaattaatct 68640
taattttcat ctttgccacc cttaattact gttggtttcc atgcttggcg tttaggttgt 68700
ttggtttctt cttgaacggc gtcaaaaagg attgcttgga gcatttcatt aacacccgtt 68760
gagatgtcaa aatgtaccca atctttttgg tcatgacttc ctgaaataga catgatgcca 68820
ccattggtaa tattgccagg gaagcgctgt ccattgaata cgcaagagta taggatatca 68880
ccggttgcag catcaattgg accatacgtg cagtcaatag taccaaagtt caaagtgatt 68940
gcctgaagcg aatgtcccga gttgagatcc tgagagtaga tacctttagg tggttgtgcc 69000
acatttttga aaggtatgtg attgaaacct gttgcaaagg tcacataaac gtatttgtaa 69060
ttcttggcaa cttcttcaat aagtgcaaga tgttgcactg agtagatatt attcatgata 69120
tttttcctta tagttagaga tttggcgttc aatccaatct tcattgagta cattacgcac 69180
caaagattct tgggctaata catcacattc caaccaggtt actaatacag cttcttcttt 69240
atcgaaaatg cattgttttt cgccaatgtc ataatattca gaaacataaa cagcaatagc 69300
atagtcttga ataccacatt ggaagtcatc aacccttttc aaatcttcat gggagaacca 69360
tgcataaatc tgatcaagac tattgaatcc gaaatacatg atgtcatagt cacaattttt 69420
gaagttttct tcatcggggt tcggatggcg ttcattgtaa cgcatatctc cccatagata 69480
ggtcgcaagc gtacttgaag acacaatctc ttttgggttg ttattggcat cgcgaccaat 69540
gactttaact ttttggtgtt gatatggacc aaggccattg agcctggatt caacacgaaa 69600
aataactttg ggttccataa catattccta tcggatgttg tttcgttagg gacataataa 69660
aaaatcccga ccacaaataa tgtgatcggg atcgcgtttt tggatttaga cttcgtgagt 69720
accttcaaca ccacgttctt tgcgttccgc agtacgcatt tcaagttctt tcaaaccaat 69780
gttgatagct ttcgcacgag tgaagtcggc tttagattca ccggcaagcg cgtcgttttt 69840
ctctttgttt gaagccagaa gacctagaga gtcgataacc aacattactt tttgaccagg 69900
ttcaaacatg tccagtagtt gggtaacctt ttgaatcatg tcttcaacag tgtaaaccgg 69960
tgtgtaaagt acgcgatcca tatcggcacc atactgttcc caaagtggtt tgcttgcacc 70020
tttctcgcta tccgcaaaga ttacgattgc ttcaggatca tgtttaagga acgcaactgc 70080
atccataacc aggaaaccag atttgtaaca gcgtgaatca ccaacaatca ttttgatacc 70140
ttcgacaaga ccaccatcaa tacgaccgga gttagcgatg ttgagtaatg gaacttccat 70200
cggacacagg atgtttttct cgccgatgcc ttcatctgaa agttgaccca ttgcatcatc 70260
catttttgca gcctgtttaa gcaacttggc catcatagga gtttttgcca taatatgttt 70320
ccttagtgtt ttctacgatt taattctttg gatacactgc cctcaaatag tgcagtacca 70380
cccagggatt caagctgttg cgatatgtca tccaaaagat ttggaagatt acccatacct 70440
aacagctcct caaaataact atgaagcccc attatagact catcttgtct tttgacaatc 70500
tgctcgagca tattaatacg ctcttgttgt cggatcgcaa gacaagtcat cacgctcgca 70560
cgctcatgag catattcaac agaatctcgt tcaccatcag tatagtaacg gatttcgctg 70620
cctggaacaa gcttgtggat atgtgagaat tcacccatgt aacgaccgag aatgaataaa 70680
tggtccaatg ccaacatgag atcaatatgt cggttatcag gatatgagat tttcatgcgc 70740
caggtattat tacgagttct ttgcatgcca tttattgttg cggggatcat gtttggggat 70800
gtatttagat atgtccacaa aaagcgtgag aacagcctgg agtgttcttc ctcatgtgac 70860
acaaattgga cattcattgt cttattcata tgtttgtacc aatacttgat aaaagaagac 70920
cagaacacca tacaggagcc ctggtcacat gtttgacaag atgcttaaat cttttcagct 70980
tgttttagtt cttcaatcca agcgataaca gattctttga tacctagttc agtttcagaa 71040
acattttgga ttttcagacg ttcacctgat gaacgacctt ccaagtacaa tgtgaagtta 71100
taacccttct tggtcggaga aacaaacaga ttagggtttt tacgagagtt tgctttgtct 71160
tcggtattga catcattaat gttaaactct agccaccgga tggcaccttc tttgttggtt 71220
gccacgaagt tgagataagt acctgcatct tcaacatctt tgatcaggtc tttatgttca 71280
ctactacggg catcaataac agcttgttct ttcttggtgt aaacacgttt cttttcattc 71340
actgtgtcat cacccaactt acccaaacca ccttggatag atacaccaag cacattttta 71400
agtgcttttt caccttcagt gagctctttt acttcttcaa cgttaacgac aactttgcct 71460
gtttcaccag gttcatcgtt atttgaagtt tcaagagttg aagtatcgac agtaacacct 71520
tttaactcat caatcttaat atcttctagg atttcggctg caccttcttc accgagttct 71580
tcgatgagta catcatcagc ttttggttta tccgaaccgt tgagaatacc agaatcagct 71640
agttcttctg aatccatgat tgtaaagcca gcaacttctt caacgttgcc ttgcaaaagt 71700
gcattaagag tgaagttacc aacctggacg ttgttatctg caagcaactg tttaactgcc 71760
tgaacgtttg attcgttgcc agtaatgtcg ataacggttt tggtgttttc atttacaaga 71820
tagttcataa tattttctca tgtcagtagt ttgtgttgat gggtgtatta aaacaagttt 71880
accaaaagaa aaacgtgatc tacatcacgt ttttgtacaa agtcattaat ttaattccat 71940
gcgggtcagc agacatcatg tagttccaga aaccaccata ttcagtcacc ggtgcgatta 72000
cctggccatg actaggacga tttaaaccaa cttccggaca tactgcaatc ttagtaccat 72060
tatccaaaca gttaaggcaa tgggtgccct tgtcatcgaa aatgatatcc aatccgcaaa 72120
ggtgtttatc accggtggaa ataaaaccat caaagatatt cgggaagtaa tgctcgatga 72180
acaaacgctt actagattcg tgtgtcggag aacaggtagt gactacaaca agcttaatgt 72240
tatgcccttc agcgttcatc ttatccactt ggctacggaa atttttcatc cattttactg 72300
cattggtaaa tggttctaca cgatcatata gatcagatag gtgccagaat tcccacgggt 72360
tgctgatgcc caagtgcgtt tgcataatgc taccgtggac gccccagtca aaatcgttcc 72420
ctgctttgag aatatcagga atacataaag gttctactag acggttctca ctgacttgtg 72480
cattacccca agtcagccaa gtccaaagag agtcagcaag agttgaatcc acatcgaatc 72540
caataattgt cggtttgttt gttaatttag ccatcggttt atccaaaaat aaaaagttca 72600
aaggcaatat taatcacctt tgaacttttg aagtttactt tttaccaaat cgtttgcgta 72660
agttggttcg tttcttccat tccatacctg tcggtttcaa gaatgtttcc agcggatcca 72720
ggaatttctt ctggaacgtc gtcttgtagt caacatactt atccatattc aaggatggtg 72780
gtaagtaatc tgggaatgcc atgtagttgt accctgttgg gtttggcact gtcaggttga 72840
ctagtttgac tttatcacca gattggattc gtgtttcgtt aacatcaagt ttgtcacaca 72900
gttggttgta caatagacct gctttcgcag cgtagtgtgc accaggttta atcttggtac 72960
ttgttggact gaccaaatat ttttccatat ctgagattga agatgctgct gcaatatcat 73020
ccacaggaag tgccatatac tcctcgcgga atttcttaac cttattcttc aattcaactt 73080
cagtgccaag caatacttct ttgtaacact cggttagacg tttacgacac cattcagggt 73140
agttggattt cttagcggca agacccatta ccttgacttt aggttttgcg taagctgtac 73200
cttccgagtt gtttaccgcc atagcgtaca tcttaggtgc tacccaaacc gcagactgag 73260
caattacttc acgttcccag aacatacgtt gttcgtaacc attaacaagg tcacacttcg 73320
cttgtgcaaa gtctttgatt ggttgatcca tatgttcttt atggaactca tccaacaagt 73380
gaacgatttc atcaacagtt ttcgtttcgt cccaacctct ctgcttaact agattgtcaa 73440
gtgtgatgta gttagagtcc gtatcaccat agaaagcata gtcgaagtta ttagttccga 73500
gtagcttgtt caggtgttcg ttgatgtaac ggatattcca acggttgatc agttgaccac 73560
caaatgtgat agcttccgca acacgcaagt caaagtaaga gctgaagtat accgaaccga 73620
ttgcaccata accagagttc atcaggatct tataacccat ctggaagacg tgttgtgcag 73680
cctcgatatc accctggaca tgtttttctt ccgcaagctt gatcatatca atgcccttca 73740
ggaacatatc ataatcaggc tcgatatcac gggcttcaaa ctcggcaccc tcacggattt 73800
ccatttcttc cttgatccat tgaactttct gttcggcagt tagaccaagt ttcttttcaa 73860
tcttacgggt tgcatataac tcacgcatta cttcagatag tacagacatc ttatcacgtc 73920
ggaagaactg catgtttggt gccatacaaa cgttatgtcg tttcaatgtt tcaaacttga 73980
agttgccaag gttaactagc tcagtaacaa tattacgagt atcgtttacc aatgacagac 74040
gtaatgcagc aagtgcttta agcttacgca tgttgccttg tttcttacaa tcttcaatat 74100
gtccatcaac ttcttcaaca agttcataga taatgtctga acgttcctgt gcgttggtaa 74160
ccagagtttc agtaccaatg ttatactgtt ggatcaagtg tggatacagt gagttcaagt 74220
cgactgagac aacccaacgg tataggccag ggacaacgtt cttaacgtaa ccgccaccaa 74280
attcaacatc accttcgtat atcttcttga tctctggtct catacccttt tcaagcagac 74340
ggttgtacat caatgctgac caaggtgccg tagtaccaag agtatctcgg tagttacatt 74400
ttgagatgta tgccagggtg aacgcaagac gcatcaggtt aagtttgttt tccaaacgac 74460
taacaagctt gatatccttg ataccatatc gcacatactt gatgtaatca tcgaagtata 74520
gagtatgcag ggatttagct tctgagtagt ccattttctt ctcgtccagc tcgtaataag 74580
caatggtatc gagtttcatg ttctcacgag taacgtatgt gaaacaatca tacaactgtt 74640
ggtagtccaa catttccgtg ccgacgaaat cataagtcgt atttttacct ttcttggact 74700
taaccgtacg gggtttgaca tgaccccaga cactgaacat cttgacccat tcttcgccaa 74760
gaatttttcg gacacgttct acaaggtacg gagcatcaaa gccctcgatg ttccaacctg 74820
tccatgcatc atattccatt tgacgccaga actggaggta gtccgccaac atttcttgtt 74880
cggtttggaa ttctttgtaa acaacatcca ggccaccgat ttcttcatcg ttctcgtcgt 74940
aacggaattt accacgcatc ttctgctctg gtaatgccca acaatacatc acaccagtat 75000
tcatatcttc atgctggatc aaagatattg gatacgctgc gttgtaatca atgaaatttg 75060
gtacacgcga ttcagggaat tctttctgga accaacgata aaagttggat acattgcttt 75120
ggtagcgttc agaaagtaac tttgttttga aagtgtgttc ttcgataact gggtgcggga 75180
acataccaga gtcgataaca ttaccatttt catcacgata ggctgtgatg acctcaatat 75240
ccacgttgct tcgacgaatt aacttatcgt caggtttcaa gcgttcgcgg aatttgtggg 75300
caatgaattg gaattcataa cggttgatac caaacattcg gtatccatac acatctttgt 75360
tttcatcaag ccactggttc gcgtcccaaa tgctatcaaa gctaacagct tttaaacgct 75420
caccaccaag acccattgca tgtggtttct tggcagatgg atcaggaata tagaactcgg 75480
gtttgaagtg atgttcgatg tgtcgtcgtt gtcctttttc gtcgcaaata cgtgcaagga 75540
tcttattacc cttgcgtgca acgtttgtat aaaaaactga tgatgtagac atgactactc 75600
ctaaggtcga tcggtctaaa tatagtaaca tgaacaaact tttgaggtta ttatgggatt 75660
cttcaaaact atacaggcaa tcttctcgga taccgggttg gttagcactg ttgggaaggg 75720
gatcgataaa gctattttca cagacgaaga aaagtcagat gcgtggatgc tattaatcca 75780
ggcttatcac ccgtttaaac ttgcccagcg ttacatcgca ctgggtttac tattcttgat 75840
gggtattggg tttattgcat cggctgtcgt gcgtattgtt ggttcagctt tgtgtgatcc 75900
tgtcatgatg cagggagcta atgataccat tatctactac aaatggtgga tcgaggattc 75960
agcttggttg gcaacaacta cgatagcaat gtttggtgaa ccgttcttgt atgctgtcgc 76020
gttcttcttt gccggtggta ccatgaatgg ggttgtagag ctcatggcca aacgacgtaa 76080
ccgtttgatg ggtatcaagc aggataaata agaaaaagcc ccaggagcga ttctggggct 76140
tttcattaca tgcctagttt ttcaactaat gacttgccgt ccgagacaaa gtcaattcga 76200
taattatcca ggaaatactt catagattgt acacggtctt tttgaacatt atcggggtct 76260
tggcagacaa aatgtgttcc gacgccaatc tcaaagttaa tccaacgttc ataaccaaca 76320
gcctggtcaa ttttttcaat caatttcttt tctgacttca ggacttgtga ctcaacgatc 76380
aatgcgattg gttggccact accacgcttt ggagtaataa ccaggacagg attctcaata 76440
ttcattttag ttactcttga ttaataaaag atttgtaatc ggttcttact ttatttagca 76500
actcgaccaa ttccgttaaa ccagcttctt caagacgttt catgcttgtc tcaacagaac 76560
gacgaatacc ttcagttgtc atgcgtttct tagcagtact actacgttgc aggataagca 76620
tactagtctg atgcaatgta cacagacctt tttctgtatc caaacctaaa acataaacca 76680
ggatattaat tgctttatcc ttgagaacat cctttgcttc ggtacgtaat cgggtatgta 76740
ggcgttccag tgcattgttt accttggtcg gttttggttg tgcttgagtt tgtttttgta 76800
aagcttttag ttccagacca ccatcaaggt gagctttcaa tttgcggaca gcctcagact 76860
ggatttgacg cactcgttcg cgggttatgg ctttaccaaa ctctttcccc atatctgcac 76920
ccgtttcttc cagggtacgg tcgccgatat taggatccaa tccataacga cgtacaagaa 76980
caaacatttc cttttctgtc aataagtcat tatcaaaggt ttcaagcacc ttaatgacct 77040
cttgtttttc caggtagtcg acatcatccg cggtgaaatc ataaagtgat tcggcttcac 77100
cgtcttcgtt gtaacgcata tcggtaccaa cactgataaa tccgctaagc gcgttgtcaa 77160
atgtcttacg actaaatttg aattctctcg gaatggcttc cttggttgcg gcaaataatg 77220
catcatcgtc catgaactca ccatgcttat cgacgtattg agcttgcaaa cgattcatag 77280
tattcatcag tttttgcaag tgaataggca aacggataat ggaattcgta ttcataactt 77340
cacgacggat acattgcaaa atccactgta ctgcataagt tgaaaaacgg taatgtttgc 77400
cattggcttc gttgatcttg aatgggtcgt acttatcagc ggcagtgagt aggccaagat 77460
tgcccatttg gatgacgtcc agaagtggca tcttgtaata caaagcccag gatcttgcat 77520
acttgacaac taaacgcagg ttatggttaa tcaatttagc gcgggcagct tcagcttctg 77580
gacccttgcc tttggtagca acagcaattt catattcctg ttcagccgtt aaaagtgggt 77640
atcggtccag ttcacgcatg taaagagtga ttgctgtgat ttcttggttt ttactcataa 77700
tatttgtgct cctaataagt ctaggaccat tatgagtggt atggaagatt tgaagatatg 77760
aaaaagcccc aggaaaaatc ccagggcttg gagaattggc gcttcctact ggacttgaac 77820
cagtgacaaa cggattaaca gtccgtcatt ctaccaactg aattaaggaa gcatatttac 77880
atattagttc atgctgtggt aaaagtaaag gaaatcacta atatgcatgt gtttagatga 77940
atcttgcatg cccatcatgc acgcgtaaag accttgatta tctttctcga gcttttcgac 78000
aagttccaaa ccatattcac tgtccaggat ggtaatatcc aggatgagtt tggtttcatc 78060
cggtgcgtca ttactaaaag aaaagtcttt aaccattgcg gcgatttggt tgatatcccc 78120
tttaccttgt tgaaatttgg tataaacagg aagaccaaaa tggctatcaa tggccgcacg 78180
caaacgtgac aggtaattat ccttttcgat tatgacgcca ttgatattag gctcatctgg 78240
tttaaaattt ggtactcgta cactcaagat agtcattgct ttactcctaa tgtgctccca 78300
ggtcttccca ggagcaccag atttaattgt tacatcattg gcgagtattg catcacatca 78360
tcatcttcgt tcgattctgg agcaatggca gcgcccgttg tgaggattgc acccgcgatt 78420
gaattggcaa attggactgc attacgcgtt acttttgccg ggtcaacaat acctgcttca 78480
atcatatcaa cgatttcacc cgttgctgcg ttgataccag tacagaaaac atttgtgtta 78540
tccgactcgg tgatatcacg ccaggctagt tcagcgttga tacctgcatt tgataagatt 78600
gtttcatatg gcgattgtaa cgcagtaaac atgatttgaa ctccaatgcc acggtctgta 78660
tggtattggg aagcagcttc agtaacacga gaaacaaact ctttatcttt tgcgaatttg 78720
agtaacgctg taccaccgcc aggaagaata ccttcttcaa cagccgcgcg ggttgcacac 78780
aatgcatcgt caactcggtc ttttttctca cgcatttcaa cttcagtggc tgcaccgata 78840
tggagaattg ctacaccacc aactagtcgc gagatacgtt tacgcataaa gtcaatatca 78900
ccgatatctt caagttgttc caggcgtgaa cgcaaagtgt caacgtgagc ttcaagtcca 78960
gtatcttcgc cttcaccatc aatgattgtg gttttgtttg gtgtaacacg cacaacagat 79020
gcacgaccaa cataaccatc aagtttatta aggtttgagt ggtgtgtttg gttttctgca 79080
tcatccagga ccttgctacc tgtcatgaca gcgatatctt ttatgtactc tgaacgttga 79140
tcgccaacgc caggagtttt aaccaacacc gcttggaacg ctttgttcat catattcgca 79200
acaaccgtat taacgaagtc cgcactgaaa tcttcagcca tgatgaaaag tggtttacca 79260
atacgttctt tgtcactgat cgcaattttt acaacgttct ggatttcacg cattgtgttg 79320
attttaccgt taaacaacag gaattgaaca tcacgaagtt cacagatgcc tgatgcttta 79380
tcggttgcaa agtatgggct ttcgtaacca cggtagaatt ccataccttc tgtgatggtc 79440
atctcatctt tcagaccacc ttcttcaacg gtgataatgc cgtctggacc aactgcttcc 79500
atagccatcg caatcatttg accgattgtt tcatcaccgt ttgctgagat agtaccaaca 79560
gagcgaatag attccaggtc gtcacattgc ttcgcatttt cttggatcgc tgtgacaatc 79620
gaatgagctg cgaaatccat accacgttta atttcaatag gacttgcacc agcaacaact 79680
gaagcataac ccgcattaac caatgcttgg gtcaatactg tagtagtggt agtaccgtcg 79740
cccgcatgat cgcctgattg tttgcttgct tctttgatca tgtttgcgcc catatttttg 79800
tatgggtcag agaatgaaat ttctttggca acagatacgc cgtctttagt aacaacaggt 79860
acttgactat gttgctgaac gaaaacaaca ttgcgaccac gcggaccaag agtcacttta 79920
acagtgtctg caaggtcgtt aacaccttca aggagtttct tttgtgcgtt atcgccaaat 79980
aagatggttt ttgctagttc tttcattttg aatactcgaa ttagtggaag atgaaatggg 80040
ccacatatag cggcccatta agattgttcg aaatgattac aggttgtaca tttcacacag 80100
acggaataac ggttgtggtg cactaggatc gcctgcaaca acacggcctt tagtaccgcg 80160
agaaaggaag tttagaattt tctgagcgcc agcccaacgg acagccatac gattttcgtt 80220
aacaaagcgc tttttatttg tgtgtttacc tttagccatg atatgtctcc taaatgacat 80280
taattgaatg ataaaattgt tcagaggttt tattatgtat cagttacata ttttgaagtt 80340
ttagttctac agctttacgt ggtgatacga agaattcatc aacagcttct acatagtgtg 80400
ttggcacact cttatttgaa ctgatcgatt ctttgagacc aaccttgtca gcccagttgt 80460
ttaatgatga accgtatatc gcaaaggccg caaaaatatt acccttgtga tcaatataaa 80520
catggtacgt tggaatgtta ctccattcat caataacgat tgacggtgca cttaacaatg 80580
catggtgccc accattttca tacaaacgag gattgtagaa acgacggaat gattcacgcg 80640
ttacactatt ttctaatagt tgcttcttgg cattgcctgt tgggataatg tttagatcaa 80700
gagaacgacg tgagcccatc tcaaattcaa cccagttttt aatccacttt ttaacagctt 80760
cttcacgagg ttcgtaaatc acaacctgac gtttagtgcc gtgaacatct cgcaataaag 80820
tattcgtgat cgcttcaatg gtaggaatgt cattaactaa ctctttacgg tctaacaatg 80880
tttgaattgt aggaatgact gtgttcatta gttgtttctt taatttagat gtactcatga 80940
taatgattct cattattaat taatattatt gatctatttg atcatacaat gtaaataaca 81000
tttgaataaa tggtgtcccg catgggacac atggccgaag gccatcattt tatataaata 81060
aatgattaat aattaattta tatttattag gtttcctgtg aaacattata aacacaatca 81120
ataaatatga acttttaatc taaatagtat tagataacta tatttggaat gatgattatg 81180
agtaataaac agaatattac tagtcgtgat tatgatggat tcttgcacga tctagcatta 81240
gcagctaaaa ctgatcctac atttaaagac tacaactttg atggttcagg tctacgcagt 81300
ctaatgcgaa tcctggcatt agcagggaac caacaagcat tttctgacaa tatgcttttc 81360
aacgaactta gtattaaaca tgctgagatg agagagaacg ttggggcatt ggcggcattt 81420
ttgtcatata ctccaatgtc atccaagggt gcaacaatga acgtagacat tactgtacgc 81480
gttccaccag gcactattag cccgccttcg agcttgacat tagaacctga ccacactttt 81540
gtcggttcaa aggatggcat tagctaccag ttttcaccaa cacaatcgaa gactgtgaac 81600
ctggccaata atgcctatac atttaccaat gtcacattga cgcaaggtcg ttgggtttat 81660
aactcattcg aaactagtgc ggccggagaa aacaccgttg aaacccatat cctaccaaac 81720
aaggatattg acattactac actaagtgtc caggttcaag cgtctaatac ctcgacggct 81780
tttgaagaat acactcgtta tacttctgct taccaattag gtccgactgc taaggtttac 81840
ttcttggagt taaacaaaga gggtcactat acaattgaat ttggtgataa caaagtagct 81900
aaacaagttg tgtctccgca agttattttg actcgagcta tggtgacgcg tggtgcaagc 81960
ggtaatgaaa ttggttctat tctaccgaca tcatcgatag gtcagttctc taccatcgaa 82020
atcaaagtta attcccgttc agccaacggc gcagatccgg agagtgtacg ttcaattagc 82080
cgcaatgcac caatcgcttt tggtaccgat ggtgtggcag tgacagctaa tgattaccct 82140
acggttttac gtggattatt cccaacagct aagattgctt catggggtgg agaggagaac 82200
gttcctgtaa tgcaagggta cacagtacta gctgtcaaac cccaaggtaa ggccacttta 82260
acagcggcag aaaaacttca gggtgagaat tatctgaatg ctcgaaatgt tggttccatc 82320
tttgccaaga ttgttgatgc tgatatctac tatattaacg ttgaatcaaa agtctattgg 82380
aatttggttc agattgggaa cagatgatcg ctagcgagaa acgtgaacgt gacaataagt 82440
tgactgccga tgttac 82456

Claims (8)

1. a kind of vibrio alginolyticus bacteriophage resistant to high temperature, which is characterized in that the vibrio alginolyticus bacteriophage is vibrio alginolyticus phagocytosis Body VAP9(Vibrio alginolyticus Phage VAP9), deposit number is CCTCC NO:M 2018766.
2. splitting vibrio alginolyticus bacteriophage resistant to high temperature according to claim 1, which is characterized in that the vibrio alginolyticus bacteriophage VAP9(Vibrio alginolyticus Phage VAP9) it is virulent phage, have one in polyhedral head and longer Tail portion, major diameter L are 60~70nm, and transverse diameter W is 50~60nm, L/W ≈ 1；Tail portion includes contractile flesh sheath and tailfiber, and flesh sheath is long Degree is 30~40nm, and wide 5~10nm, tailfiber is with 3~4；Optimal pH is pH6~pH8, is had under the conditions of 4 DEG C~75 DEG C resistance to Property, 8~10h is cultivated under the conditions of MOI=0.001, potency is 2 × 1010 pfu/ mL。
3. vibrio alginolyticus bacteriophage resistant to high temperature according to claim 1, which is characterized in that the vibrio alginolyticus bacteriophage VAP9(Vibrio alginolyticus Phage VAP9) there is nucleotide sequence shown in SEQ ID No.1.
4. a kind of composition containing any one of claims 1 to 3 vibrio alginolyticus bacteriophage resistant to high temperature, feature exist In the composition contains the vibrio alginolyticus bacteriophage VAP9(Vibrio alginolyticus Phage VAP9) and At least one of other Vibriophages.
5. a kind of composition containing vibrio alginolyticus resistant to high temperature bacteriophage described in claim 4, which is characterized in that the combination Object contains vibrio alginolyticus bacteriophage VAP9(Vibrio alginolyticus Phage VAP9), vibrio alginolyticus bacteriophage VAP7 (Vibrio alginolyticus Phage VAP7) deposit number be CCTCC NO:M 2018767；With vibrio alginolyticus bacteriophage VAP21(Vibrio alginolyticus Phage VAP21) deposit number be CCTCC NO:M 2018768 in two kinds or It is two or more；
The above bacteriophage is preserved in China typical culture collection center on November 9th, 2018, and classification naming isVibrio alginolyticus phage。
6. the composition of vibrio alginolyticus bacteriophage resistant to high temperature according to claim 4, which is characterized in that the composition packet Include vibrio alginolyticus bacteriophage VAP9(Vibrio alginolyticus Phage VAP9), it further include in each bacteriophage mutants It is one or more；Preferably, the mutant at least 90% is identical as the native sequences of corresponding Vibriophage.
7. a kind of reagent containing vibrio alginolyticus resistant to high temperature bacteriophage described in claim 5 or vibrio alginolyticus bacteriophage composition Or kit.
8. any one of claims 1 to 3 vibrio alginolyticus bacteriophage resistant to high temperature or molten algae arc as claimed in claim 4 Bacterium bacteriophage composition or the examination as claimed in claim 7 containing vibrio alginolyticus bacteriophage or vibrio alginolyticus bacteriophage composition Agent or kit are being killed and/or are preventing the application in vibrios microorganism belonging to genus:
The vibrios microorganism belonging to genus includes vibrio alginolyticusV.alginolyticus, Vibrio anguillarumV.anguillarum, comma bacillusV.cholerae, non-Nonther conserved quantitynon-O1V.cholerae, Fermi operatorV.fischeri, vibrio fluvialisV.fluvialis, Vibrio harveyiV.harveyi, sick Vibrio damselaV.ordalii, vibrio parahaemolytiousV.parahaemolyticus, Vibrio salmonicidaV.salmonicida, vibrio mimicusV.mimicus, Vibrio splindidusV.splendidus, Vibrio aureusV.azureus, Vibrio gazogenesV.gazogenes, Vibrio vulnificusV.vulnificus, bamboo Pod vibrio pisciumV.trachuri, damselfish vibriosV.damsela, floating vibriosV.natriegen, clam worm vibriosV.nereis, fish Enteron aisle vibriosV.ichthyoenteri, shark vibrioV.carchariae, Vibrio campbelliiV.campbelliiAnd kill prawn arc BacteriumV.penaeicidaOne of or it is a variety of.
CN201910400869.8A 2019-05-14 2019-05-14 High-temperature-resistant vibrio alginolyticus bacteriophage and composition, kit and application thereof Active CN110184241B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910400869.8A CN110184241B (en) 2019-05-14 2019-05-14 High-temperature-resistant vibrio alginolyticus bacteriophage and composition, kit and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910400869.8A CN110184241B (en) 2019-05-14 2019-05-14 High-temperature-resistant vibrio alginolyticus bacteriophage and composition, kit and application thereof

Publications (2)

Publication Number Publication Date
CN110184241A true CN110184241A (en) 2019-08-30
CN110184241B CN110184241B (en) 2021-01-26

Family

ID=67716330

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910400869.8A Active CN110184241B (en) 2019-05-14 2019-05-14 High-temperature-resistant vibrio alginolyticus bacteriophage and composition, kit and application thereof

Country Status (1)

Country Link
CN (1) CN110184241B (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110699330A (en) * 2019-11-11 2020-01-17 云南大学 Bacteriophage and application thereof
CN112063593A (en) * 2020-09-17 2020-12-11 扬州大学 Pathogenic vibrio phage VyZU 10474 and application thereof
CN114480305A (en) * 2022-02-14 2022-05-13 菲吉乐科(南京)生物科技有限公司 Phage powder and preparation and application thereof
CN114703149A (en) * 2022-01-28 2022-07-05 武汉格瑞农生物科技有限公司 Vibrio alginolyticus phage GVA-P21 with high fermentation rate and lasting bacteriostasis and application thereof

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU1671690A1 (en) * 1989-08-16 1991-08-23 Ростовский-На-Дону Государственный Научно-Исследовательский Противочумный Институт Strain of phase vibrionis parahaemolitici for phagodiagnosis of parahemolytic vibrios serotype 05:k15
EP1614426A1 (en) * 2003-02-20 2006-01-11 Centro Nacional De Investigaciones Cientificas (CNIC) Attenuated strains of vibrio cholerae and lyophilised vaccines containing same
CN107904214A (en) * 2017-10-13 2018-04-13 深圳先进技术研究院 Vibrio alginolyticus bacteriophage and the bactericidal composition comprising the bacteriophage
CN108070570A (en) * 2017-11-20 2018-05-25 深圳先进技术研究院 Fragility Vibriophage ValDsh-1 and the bactericidal composition comprising the bacteriophage and its application
CN108651522A (en) * 2018-04-09 2018-10-16 广州诺晶生物技术有限公司 A kind of vibrio alginolyticus phage preparation, preparation method and applications
CN110129280A (en) * 2019-05-14 2019-08-16 菲吉乐科(南京)生物科技有限公司 Wide fragmentation pattern vibrio alginolyticus bacteriophage and combinations thereof, kit and application
CN110205305A (en) * 2019-05-14 2019-09-06 菲吉乐科(南京)生物科技有限公司 Alkaline-resisting vibrio alginolyticus bacteriophage and combinations thereof, kit and application

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU1671690A1 (en) * 1989-08-16 1991-08-23 Ростовский-На-Дону Государственный Научно-Исследовательский Противочумный Институт Strain of phase vibrionis parahaemolitici for phagodiagnosis of parahemolytic vibrios serotype 05:k15
EP1614426A1 (en) * 2003-02-20 2006-01-11 Centro Nacional De Investigaciones Cientificas (CNIC) Attenuated strains of vibrio cholerae and lyophilised vaccines containing same
CN107904214A (en) * 2017-10-13 2018-04-13 深圳先进技术研究院 Vibrio alginolyticus bacteriophage and the bactericidal composition comprising the bacteriophage
CN108070570A (en) * 2017-11-20 2018-05-25 深圳先进技术研究院 Fragility Vibriophage ValDsh-1 and the bactericidal composition comprising the bacteriophage and its application
CN108651522A (en) * 2018-04-09 2018-10-16 广州诺晶生物技术有限公司 A kind of vibrio alginolyticus phage preparation, preparation method and applications
CN110129280A (en) * 2019-05-14 2019-08-16 菲吉乐科(南京)生物科技有限公司 Wide fragmentation pattern vibrio alginolyticus bacteriophage and combinations thereof, kit and application
CN110205305A (en) * 2019-05-14 2019-09-06 菲吉乐科(南京)生物科技有限公司 Alkaline-resisting vibrio alginolyticus bacteriophage and combinations thereof, kit and application

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
JUNHYEOK YU等: ""Comparative Genomic Analysis of Novel Bacteriophages Infecting Vibrio Parahaemolyticus Isolated From Western and Southern Coastal Areas of Korea"", 《ARCH VIROL》 *
MINMIN YEN等: ""A cocktail of three virulent bacteriophages prevents Vibrio cholerae infection in animal models"", 《NATURE COMMUNICATIONS》 *
YING-RONG LIN等: "Characterization of a new phage, termed ϕA318, which is specific for Vibrio alginolyticus", 《ARCHIVES OF VIROLOGY.》 *
付汉清等: "一株宽谱裂解性溶藻弧菌噬菌体ФV170的分离鉴定及生物学特性 ", 《微生物学通报》 *
周希澄等: "《遗传基础知识》", 31 August 1982, 河南科学技术出版社 *
蔡俊鹏等: "霍乱弧菌噬菌体在消除海产品中相关弧菌的潜在应用研究 ", 《现代食品科技》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110699330A (en) * 2019-11-11 2020-01-17 云南大学 Bacteriophage and application thereof
CN112063593A (en) * 2020-09-17 2020-12-11 扬州大学 Pathogenic vibrio phage VyZU 10474 and application thereof
CN112063593B (en) * 2020-09-17 2021-08-31 扬州大学 Pathogenic vibrio phage VyZU 10474 and application thereof
CN114703149A (en) * 2022-01-28 2022-07-05 武汉格瑞农生物科技有限公司 Vibrio alginolyticus phage GVA-P21 with high fermentation rate and lasting bacteriostasis and application thereof
CN114480305A (en) * 2022-02-14 2022-05-13 菲吉乐科(南京)生物科技有限公司 Phage powder and preparation and application thereof

Also Published As

Publication number Publication date
CN110184241B (en) 2021-01-26

Similar Documents

Publication Publication Date Title
CN110184241A (en) Vibrio alginolyticus bacteriophage resistant to high temperature and combinations thereof, kit and application
CN110129280B (en) Wide-cracking spectrum vibrio alginolyticus bacteriophage and composition, kit and application thereof
CN110205305B (en) Alkali-resistant vibrio alginolyticus bacteriophage and composition, kit and application thereof
CN112029733B (en) High-lytic vibrio alginolyticus phage RDP-VP-19001 and application thereof
CN109666654B (en) Novel bacteriophage, composition thereof, and preparation method and application thereof
CN112831474B (en) Wide lysis spectrum salmonella bacteriophage RDP-NSA-19050 and application thereof
CN107208068B (en) Novel Shiga toxin F18-producing Escherichia coli bacteriophage Esc-COP-1 and application thereof in inhibiting proliferation of Shiga toxin F18-producing Escherichia coli
CN112195158B (en) Vibrio parahaemolyticus bacteriophage RDP-VP-19003 and application thereof
CN107208067B (en) Novel enteroinvasive escherichia coli bacteriophage Esc-COP-4 and use thereof for inhibiting enteroinvasive escherichia coli proliferation
CN113430174B (en) High-lytic salmonella bacteriophage RDP-SA-19009 and application thereof
CN112662635B (en) High-temperature-resistant Escherichia coli bacteriophage RDP-EC-20031 and application thereof
KR101723827B1 (en) Novel Aeromonas salmonicida bacteriophage Aer-SAP-1 and its use for preventing proliferation of Aeromonas salmonicida
CN114231500B (en) Mutant staphylococcus aureus phage and application thereof
CN112876543B (en) Microcin MccY and preparation method and application thereof
KR20130028275A (en) Method for prevention and treatment of salmonella typhimurium infection
CN110452895B (en) Lysozyme from bacteriophage and gene and application thereof
CN112522214B (en) High-lytic pseudomonas aeruginosa bacteriophage RDP-PA-20001 and application thereof
KR101863593B1 (en) Novel Salmonella specific bacteriophage SP1 and antibacterial composition comprising the same
CN109136195A (en) A kind of vibrio parahaemolyticus phage and its application
CN112011520B (en) Temperate phage VneM1 for regulating coral flora and application thereof
KR20120073724A (en) Bacteriophage t10-1, my-1, and vegetable soft rot controlling composition containing the same
CN113528463A (en) Lytic and high-titer vibrio parahaemolyticus phage RDP-VP-21010 and application thereof
CN102559705A (en) EPS (Extracellular Polysaccharide) gene clusters EpsA, EpsB, EpsC and EpsD
CN114891754B (en) Fish pathogenic aeromonas phage phi A008 and application thereof
KR20130020710A (en) Bacteriophage t10-1, my-1, and vegetable soft rot controlling composition containing the same

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB02 Change of applicant information
CB02 Change of applicant information

Address after: 210000 Erbanqiao 473, Gulou District, Nanjing City, Jiangsu Province

Applicant after: Fiji Leke (Nanjing) Biotechnology Co., Ltd.

Address before: North Building 568, building 1, Jiangning Science Park longmian Road District of Nanjing City, Jiangsu province 211100 2 floor

Applicant before: Fiji Leke (Nanjing) Biotechnology Co., Ltd.

GR01 Patent grant
GR01 Patent grant