CN108179136A - Coliphage Esc-COP-9 and its enteropathogenic E. Coli Proliferation Ability purposes - Google Patents

Abstract

The present invention relates to a kind of 9 (numbers of being commissioned of Styloviridae phage E sc COP detached from nature：KCTC 13131BP), it is characterized in that, the genome for carrying with specific killing Escherichia coli ability and being marked with sequence number 1, and the method that enteropathogenic E. Coli infects is prevented and treated using the bacteriophage as the composition that active ingredient is included the invention further relates to utilization.

Description

Coliphage Esc-COP-9 and its enteropathogenic E. Coli Proliferation Ability Purposes

Technical field

The present invention relates to the phagocytosis that a kind of separation from natural environment can kill Escherichia coli after ehec infection The method that body and utilization prevented and disposed enteropathogenic E. Coli infection using the bacteriophage as the composition that active ingredient includes.Tool Body is said, is exactly related to a kind of characterized by carrying with specific killing Escherichia coli ability, the genome marked with sequence number 1 Slave nature in the Styloviridae phage E sc-COP-9 (numbers of being commissioned that detach：KCTC 13131BP) and utilize with described Bacteriophage prevents the method disposed after the infection and infection of enteropathogenic E. Coli for the composition that active ingredient is included.

Background technology

Escherichia coli belong to intestinal bacterium section, as gram-Negative bacillus, catalase (Catalase) positive, oxygen Change enzyme (Oxidase) feminine gender, facultative anaerobic bacteria, it is most of all to reduce lactose.Escherichia coli are divided into from serological angle Cell body (O), flagellum (H), pod membrane (K) antigen, and it is pathogenic related to Escherichia coli known as these antigens.It is pathogenic big Enterobacteria refer in Escherichia coli it is a small number of obtain viral Escherichia coli, it is usually big according to the type etc. of pathogeneticing characteristic, toxin Cause is divided into enterorrhagia type Escherichia coli, the malicious type Escherichia coli of intestines production, intestines invasion type Escherichia coli, intestines pathological form Escherichia coli, intestines Gather type Escherichia coli this 5 types.The domestic animal that enteropathogenic E. Coli can infect a variety of ages is allowed to fall ill, and cardinal symptom is Diarrhea, case fatality rate is also very high caused by extremely serious dehydration.Diarrhea caused by this enteropathogenic E. Coli Disease is the principal disease spread in the almost all of raiser in China, is thus quite big to the loss that farming industry is brought 's.

Under normal conditions, in order to which the method that the infectious disease to enteropathogenic E. Coli is prevented and treated and taken has Utilize the method for vaccine and antibiotic.However using the treatment of antibiotic because causing under its lasts during the increase of drug-fast bacteria Drop.Simultaneously as strengthen the regulation that antibiotic is used animal, therefore a kind of in addition to antibiotic there is an urgent need to develop Other effective ways.

Recently as the counte-rplan of bacteriosis, the utilization of bacteriophage (Bacteriophage) receives very big pass Note.Especially since people tend to close environmental manner so that everybody to the degree of concern of bacteriophage than ever all It is high.Microscopic organisms of the bacteriophage as bacterial infection, generally also letter are called antibiotic (Phage).Bacteriophage is infecting (Infection) after bacterium, after the cellular proliferative of bacterium, proliferation, when progeny phage dissociates outside bacterium, just pass through It destroys the mode of the cell wall of host, that is, bacterium and kills bacterium.The bacterium infection mode of bacteriophage has very high specificity, Therefore the bacteriophage type that specific bacteria can be infected only is limited to a part.That is, specific bacteriophage can only infect specific category Interior bacterium.Therefore, specific bacteriophage can only kill specific bacteria, without being had an impact to other bacteriums.This bacteriophage Bacterium specificity only can to target bacteria generate antibacterial effect, without in environment or in animal body normal flora produce It is raw to influence.Under normal conditions, the existing antibiotic for being widely used in bacterium disposition can while have an impact various bacteria.Therefore, The problems such as causing environmental pollution or interfering the normal bacteria clump of animal.In contrast, since bacteriophage is only to specific bacteria Effectively, the problems such as therefore normal flora in interfering bodies will not be generated using bacteriophage.As a result, using bacteriophage with using antibiotic It compares, it is not only safer, and it is also relatively very low to bring in the possibility of side effect.

Bacteriophage be Britain in 1915 bacteriological Twort to staphylococcus (Micrococcus) bacterium colony by certain What the reason of being transparent this phenomenon after substance dissolving found when being studied.In addition, French bacteriological d' in 1917 Herelle has found certain object that there is dissolving shigella dysenteriae (Shigella dysenteriae) in dysentery patient stool filtrate Matter, and the research by being carried out to this, are independently found that bacteriophage, and can swallow bacterium this connotation from it and be ordered It is entitled " bacteriophage ".Hereafter, it is found that dysentery bacillus, salmonella typhi, cholera bacteria etc. are a variety of for pathogenic bacteria successively Bacteriophage.

Since bacteriophage has the special ability for killing bacterium, after bacteriophage is found, research staff expects energy A kind of effective scheme of reply bacterium infection is enough developed, and has carried out a large amount of correlative studys.But Fleming has found mould It is widely available with antibiotic after element, the research of bacteriophage only in a part of East European countries and the former Soviet Union is carried out, is maintained A slim chance of survival.But, after 2000, due to the increase of antibiotic resistant bacteria, it was found that the limitation of existing antibiotic.With It and is continued to increase as the research and development possibility of existing antibiotic substitute matter, bacteriophage is of concern again as antiseptic. Particularly, recently with the regulation to antibiotic usage is strengthened from government aspect in global range, people are to the pass of bacteriophage Note degree further increases, and uses it for the example in terms of industry and is also gradually increasing.

As previously mentioned, bacteriophage is very high to the specificity of bacterium.Just because of there are this species specificity, so even if right Belong to the bacterium of same (Species), bacteriophage often also only can play antibacterial effect to part of it strain (Strain).Separately Outside, with the difference of object bacterial strain, the antimicrbial power intensity of the bacteriophage thus played in itself also can be different.Therefore, to obtaining Obtain the method controling effectively for particular kind of bacterium, it is necessary to ensure that there are a variety of useful bacteriophages.In order to open Effective bacteriophage for enteropathogenic E. Coli is sent out using method, is also required to ensure to be directed to Escherichia coli and provide to resist certainly The diversified bacteriophage of bacterium effect.Further, in a variety of useful bacteriophages ensured, it is also desirable to from antimicrbial power intensity Or bacteriophage of the level screening with comparative advantages of antibacterial range is used.

Invention content

Problems to be solved by the invention

Therefore, the present inventor is dedicated to researching and developing a kind of combination that can be used for preventing or disposing enteropathogenic E. Coli infection Object is utilized the bacteriophage be capable of and detached in the slave natural environment of selectively killing Escherichia coli, and is researched and developed using the composition Go out to prevent or dispose the method for enteropathogenic E. Coli infection, by unremitting effort, isolate symbol from natural environment finally Close the bacteriophage of above-mentioned condition.Meanwhile in order to distinguish the bacteriophage of the separation and other bacteriophages and its specific property And after ensuring the gene order of its genome (Genome), have developed the combination using the bacteriophage as active ingredient Object, and confirm that the composition can be effective for preventing and disposing enteropathogenic E. Coli infection, so as to complete the present invention.

Therefore, the object of the present invention is to provide a kind of Styloviridaes detached from natural environment (Siphoviridae) phage E sc-COP-9 (numbers of being commissioned：KCTC 13131BP), which is characterized in that it carries with sequence number 1 The genome of label has the ability of specific killing Escherichia coli.

A kind of enteropathogenic E. Coli it is a further object of the invention to provide composition and using the composition Prevention method is infected, and the composition is made with the phage E sc-COP-9 that Escherichia coli can be killed after ehec infection It is included for active ingredient, and the composition can be used for preventing the infection of enteropathogenic E. Coli.

A further object of the present invention is, provides a kind of composition and utilizes the enteropathogenic E. Coli of the composition Infect method of disposal, the composition using after ehec infection can by Escherichia coli kill phage E sc-COP-9 as Active ingredient is included, and the composition can be used for the infection of disposition enteropathogenic E. Coli.

A further object of the present invention is, provide it is a kind of using the composition to prevent and dispose pathogenic large intestine Disinfectant for the purpose of bacillus infection.

A further object of the present invention is, provide it is a kind of using the composition to prevent and dispose pathogenic large intestine Drinking water additive for the purpose of bacillus infection.

A further object of the present invention is, provides a kind of pathogenic big by preventing and disposing using the composition Feed addictive of the infection of enterobacteria for the purpose of improving feeding efficiency.

The method that subject solves

The present invention provides a kind of Styloviridae phage E sc-COP-9 (numbers of being commissioned detached from natural environment： KCTC 13131BP), which is characterized in that carry the ability with specific killing Escherichia coli, the gene marked with sequence number 1 Group.Meanwhile it prevents and locates by the use of the composition included using the bacteriophage as active ingredient the present invention also provides a kind of The method for putting enteropathogenic E. Coli infection.

Phage E sc-COP-9 is deposited with South Korea's bioscience after being detached by the present inventor, on October 17th, 2016 With Biological Technology research institute (number of being commissioned：KCTC 13131BP).

In addition, the present invention provides a kind of disinfectant, drinking water additive and feed addictive, to can be used for preventing and The phage E sc-COP-9 for disposing the infection of enteropathogenic E. Coli is included as active ingredient.

Phage E sc-COP-9 included in the composition of the present invention can effectively kill Escherichia coli, thus to by The prevention (preventing from infecting) or treatment (disposition infection) for the disease that enteropathogenic E. Coli induces show effect.Therefore, originally The composition of invention can be used for the prevention and treatment of the disease that are induced by enteropathogenic E. Coli.

The meaning of terms such as " preventing " or " prevention " for being used in this specification：(I) prevents enteropathogenic E. Coli from infecting； (II) inhibits the infection of enteropathogenic E. Coli to develop into disease.

The meaning of terms such as " disposition " or " treatment " that is used in this specification：(I) by enteropathogenic E. Coli to being induced Disease inhibited；(II) makes all behaviors mitigated by the state of an illness that enteropathogenic E. Coli induces an illness.

" separation ", " separation " or " separation " in this specification refer to, kinds of experiments is utilized from nature Method isolates bacteriophage；And guarantee to distinguish and the thing of the feature of the specific bacteriophage with other bacteriophages.Meanwhile also Including using biotechnology by bacteriophage multiplication for use in industrial field.

It is object usually used when producing preparation to allow the carrier used in the pharmacy included in the composition of the present invention Matter, including but not limited to：It is lactose, glucose, sucrose, D-sorbite, mannitol, starch, acacia glue, calcium phosphate, alginates, bright Glue, calcium silicates, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate Ester, propylparaben, talcum, magnesium stearate and mineral oil etc..The composition of the present invention may be used also other than mentioned component Including：Lubricant, wetting agent, sweetener, fumet, emulsifier, suspending agent, preservative agent etc..

In the composition of the present invention phage E sc-COP-9 is included as active ingredient.Wherein phage E sc-COP-9 packets Containing 1 × 10¹Pfu/ml~1 × 10³⁰Pfu/ml or 1 × 10¹Pfu/g~1 × 10³⁰pfu/g.Preferably, comprising 1 × 10⁴pfu/ Ml~1 × 10¹⁵Pfu/ml or 1 × 10⁴Pfu/g~1 × 10¹⁵pfu/g。

The composition of the present invention is easily implemented according to the technical staff with the technical field of the invention Conventional wisdom Method can be manufactured, Huo Zheke using pharmaceutically allowing the carrier used and/or excipient that preparation is made by unit capacity form It is manufactured with being packed into the container of large capacity.Its dosage form is either oil or the solution in aqueous medium, suspension or emulsion Form or extract, powder agent, granule, tablet or capsule form can also include dispersant or stabilizer.

The composition of the present invention can be achieved to add for disinfectant, drinking water additive and feed according to the difference of occupation mode Agent, but it is not limited only to this.

In order to improve this efficiency using in purpose, addition other strains can also be directed into the composition of the present invention The bacteriophage of antibacterial activity is provided.Alternatively, it is also possible to add the bacteriophage for other types that there is antibacterial activity to Escherichia coli. Even having the bacteriophage of antibacterial activity to Escherichia coli, in the intensity of antimicrbial power or the level of antibacterial range, each other it Between all have differences.Therefore, it is carried out appropriately combined to play its effect to the maximum extent.

Invention effect

Utilize the sense of the enteropathogenic E. Coli of composition included using phage E sc-COP-9 of the present invention as active ingredient Dye prevents and method of disposal, with it is existing based on the mode of the chemical substances such as antibiotic compared with, advantage is it to Escherichia coli Specificity is very high.It means that it will not have an impact other useful normal floras, while can also be used to pathogenic The infection of Escherichia coli is prevented or is disposed, and the side effect being generated by it but it is very small.Under normal conditions, if using antibiosis The chemical substances such as element will also result in damage to common normal flora, eventually results in the immunity of animal and reduces, so as to show Go out with a variety of side effects used.In addition, due to the present invention be separation nature in already existing bacteriophage as composition Active ingredient, therefore it also have the advantages that very in person so, environmental protection.In addition, even if antibacterial work can be played in bacteriophage The strain of property is identical, for the performance of antibacterial effect, in the intensity or antibacterial range of antimicrbial power【It is being subordinated to Escherichia coli Various bacteria bacterial strain (Strain) side, bacteriophage to individual bacteria bacterial strain play antibacterial activity range.Normal conditions Under, bacteriophage is merely able to play antibacterial activity to a part of bacterium bacterial strain (Strain) for belonging to same strain (Species). That is, even if belonging to same strain, indivedual different bacterium bacterial strains can have differences the sensibility of bacteriophage】Level exist it is poor It is different.Therefore, compared with having other bacteriophages of antimicrbial power to Escherichia coli, the present invention can provide the antibacterial with otherness Effect.It means that there is very big difference in effect caused by can making in practical industry application.

Description of the drawings

Fig. 1 is the electron micrograph of phage E sc-COP-9.

Fig. 2 is to show that phage E sc-COP-9 kills the experimental result of Escherichia coli ability, and transparent part is subjects Bacterium is by bacteriolyze spot finally formed after bacteriolyze.

Specific embodiment

In the following, the present invention will more specifically be illustrated according to embodiment.But cited embodiment is this The example of invention, the scope of the present invention are not only defined in these embodiments.

Embodiment 1:Separation can kill the bacteriophage of Escherichia coli

When separation can kill the bacteriophage of Escherichia coli, the sample ensured from natural environment is utilized.In addition, for biting The Escherichia coli of thalline separation are separated in advance by the present inventor, and are pathogenic by identification (identification) Escherichia coli.

In the following, the separation process of bacteriophage is described in detail.In the TSB of the ratio inoculation Escherichia coli according to 1/1000 (Tryptic Soy Broth) culture medium (casein digest, 17g/L；Soybean digest, 3g/L；Glucose, 2.5g/L； NaCl, 5g/L；Dipotassium hydrogen phosphate, 2.5g/L) in add the sample (sample) of collection together, then shake under conditions of 37 DEG C Swing culture 3-4 hours.It after culture, is centrifuged 20 minutes with 8,000rpm, recycles supernatant.By Escherichia coli according to 1/ 1000 ratio is inoculated into the supernatant of recycling, then shaken cultivation 3-4 hours again under conditions of 37 DEG C.In bacteriophage In the case of having included in the sample, the quantity (Titer) to ensure bacteriophage can fully increase, and the above process is weighed in total It implements again 5 times.The above process is repeated after implementing 5 times, then culture solution is centrifuged 20 minutes with 8,000rpm.Centrifugation point From rear, the supernatant of recycling is filtered with 0.45 μm of filter.By using the conventional of thus obtained filtrate Spot test (Spot assay) investigation is with the presence or absence of the bacteriophage that can kill Escherichia coli.

The spot test is implemented in accordance with the following methods.Escherichia coli are inoculated into TSB cultures according to 1/1000 ratio After in base, one night of shaken cultivation under conditions of 37 DEG C.E. coli broth 3ml (the OD that will be thereby preparing for₆₀₀1.5) to apply Smear (Spreading) to TSA (Tryptic Soy Agar) plating medium (casein digest, 15g/L；Soybean digest, 5g/L；NaCl, 5g/L；Agar, 15g/L) on.By the plating medium after smearing on clean bench (Clean bench) It places about 30 minutes or so, dries baste.After drying, the 10 μ l points of filtrate that front prepares are dripped to and are coated with large intestine On the plating medium of bacillus, and placed 30 minutes or so and made it dry.It, will be by the operation of above-mentioned drop after drying Plating medium quiescent culture one day under the conditions of 37 DEG C.Then, it investigates and whether generates transparent ring in the position that drop has filtrate (Clear zone).Generation has the filtrate of transparent ring, it is possible to be judged as that it contains the bacteriophage that can kill Escherichia coli.It is logical Cross this investigation, it can be ensured that the filtrate comprising bacteriophage, and the bacteriophage has the killing ability for Escherichia coli.

The separation for implementing pure bacteriophage using the filtrate of bacteriophage contained and can kill Escherichia coli is confirmed.Pure phagocytosis The separation of body employs conventional bacteriolyze spot analysis (Plaque assay) technology.Specifically, by shape in bacteriolyze spot analytic process Into a bacteriolyze spot with Jing Guo sterilization treatment tip (tip) recycling, be then added in E. coli broth, It is cultivated together under conditions of 37 DEG C 4-5 hours.It after culture, is centrifuged 20 minutes with 8,000rpm, obtains supernatant.It is obtaining Supernatant in E. coli broth is added according to 1/50 volume after, then cultivated 4-5 hours under conditions of 37 DEG C.For Increase the quantity of bacteriophage, the above process at least implemented 5 times or more, finally centrifuged 20 minutes with 8,000rpm, Obtain supernatant.Bacteriolyze spot is again carried out using the supernatant of acquisition to analyze.Under normal conditions, due to the separation of pure bacteriophage not It can only be completed by above-mentioned 1 process, therefore entire abovementioned steps are repeated again using the bacteriolyze spot of the formation.This process It at least repeats to implement 5 times or more, it is ensured that the solution containing pure bacteriophage.By convention, the separation of pure bacteriophage carries out It repeats, until the size of bacteriolyze spot and all similar shape of formation.Confirm finally by electron-microscopic analysis Whether bacteriophage has purely been detached.During electron-microscopic analysis, aforementioned process is also repeated, until confirmed purely Until separation.Electron-microscopic analysis is implemented using conventional method, this is briefly described below.Pure bacteriophage will be contained Solution is stained on copper mesh (Copper grid), then carries out negative staining with 2% uranyl acetate (Uranyl acetate) After (Negative staining) and drying, pass through its form of transmission electron microscope observation.The pure bacteriophage electronic display of separation If micro mirror photo as shown in Figure 1, judged by morphological feature, can be confirmed the novel Phagus of acquisition in long-tail disease Malicious section (Siphoviridae) bacteriophage.

It has passed through the process of following purification containing the pure phage solution confirmed in this way.It is overall according to solution is accounted for The volume of product 1/50 adds E. coli broth into the solution containing pure bacteriophage, is then further cultured for 4-5 hours.Culture Afterwards, it is centrifuged 20 minutes in 8,000rpm, so as to obtain supernatant.In order to obtain the solution containing sufficient amount bacteriophage, This process is repeated to implement 5 times in total.0.45 μm of filter will be utilized finally by the supernatant for centrifuging acquisition After being filtered, conventional polyethylene glycol (Polyethylene Glycol are implemented；PEG) precipitation process.Specifically, to PEG and NaCl is added in 100ml filtrates and reaches 10%PEG 8000/0.5M NaCl, is then stood under conditions of 4 DEG C It 2-3 hours, is then centrifuged 30 minutes in 8,000rpm, so as to obtain phages object.By thus obtained bacteriophage Sediment buffer solution (Buffer；10mM Tris-HCl, 10mM MgSO₄, 0.1% gelatin, pH 8.0) and 5ml makes its suspension, It is referred to as phage suspension liquid or phage solution.

By the above process, the pure bacteriophage by purification is obtained.The bacteriophage is named as phage E sc-COP- 9.Hereafter, South Korea's bioscience and Biological Technology research institute (number of being commissioned are placed on October 17th, 2016：KCTC 13131BP)。

Embodiment 2:The genome separation and sequence analysis of phage E sc-COP-9

The genome of phage E sc-COP-9 is detached as follows.The separation of genome be utilized with The phage suspension liquid that the identical method of embodiment 1 obtains.First, in order to remove the Escherichia coli that may contain in suspension DNA and RNA is added to DNase I and the RNase A of 200U into 10ml phage suspension liquid, then in 37 DEG C of item respectively It placed under part 30 minutes.After placing 30 minutes, in order to eliminate the activity of DNase I and RNase A, 0.5M ethylenediamine tetraacetics are added Acetic acid (Ethylenediaminetetraacetic acid；EDTA) after 500 μ l, then 10 minutes are stood.Then, it is existed again 10 minutes are stood under conditions of 65 DEG C.Then, in order to disintegrate the outer wall of bacteriophage, after adding Proteinase K (20 ㎎/ml) 100 μ l, It is made to have been reacted under conditions of 37 DEG C 20 minutes.Then, 10% lauryl sodium sulfate of addition (Sodium dodecyl sulfate；SDS) 500 μ l then allow it to be reacted under conditions of 65 DEG C again 1 hour.After reaction 1 hour, to the reaction solution Middle addition 10ml is according to 25：24：1 composition ratio is by phenol (Phenol)：Chloroform (Chloroform)：Isoamyl alcohol (Isoamylalcohol) composition mixed liquor and stir.Then, it is centrifuged 15 minutes with 13,000rpm Make its layering, upper strata is then taken from the layer of separation, add the isopropanol (Isopropyl of 1.5 volume ratios thereto Alcohol it after), is centrifuged 10 minutes with 13,000rpm, makes gene group precipitation.After recycling precipitate, add into sediment Add 70% ethyl alcohol (Ethanol), then centrifuged 10 minutes with 13,000rpm again, sediment is washed.It will wash The recycling precipitate washed after implementing vacuum drying to it, then is dissolved in the water of 100 μ l.The above process is repeated, is obtained Obtained the genome of a large amount of phage E sc-COP-9.

In Seoul National University's agricultural and Life Science Instruments center (National Instrumentation Center for Environmental Management) carried out using thus obtained genome using illumina Mi-Seq instruments under Generation base sequence analysis (Next generation sequencing analysis), obtains phage E sc-COP-9's Genomic sequence information.Finally, the size that the phage E sc-COP-9 genomes of analysis have is 51,102bp, full gene Sequence sequence number 1 marks.

Based on the phage E sc-COP-9 genome sequences of acquisition, online BLAST (http are utilized:// Www.ncbi.nlm.nih.gov/BLAST/) to its similitude between currently known phage genome sequence (Similarity) it is investigated.The survey showed that by BLAST, the genome sequence of phage E sc-COP-9 and shiga The sequence (GenBank Accession .KC710998.1) of bacterium bacteriophage pSf-1 has the high similitude (homogeneity of comparison：93%).But It is, with phage E sc-COP-9 with cricoid genome on the contrary, Shigella bacteriophage pSf-1 is with glm gene group, two Person has very big difference in terms of the form of genome.Opening deciphering frame on phage E sc-COP-9 genomes The number of (Open Reading Frame, ORF) is 89, and in contrast, Shigella bacteriophage pSf-1 is 94, it has There is the opening deciphering frame number different from phage E sc-COP-9.

According to the fact that can be concluded that, phage E sc-COP-9 be with the existing bacteriophage reported not Same novel bacteriophage.At the same time, it is generally the case that if the type of bacteriophage is different, the antimicrbial power that can be provided Intensity and antibacterial range are also different, by the fact that may determine that, phage E sc-COP-9 is capable of providing have been reported with existing The different antibacterial effect of other bacteriophages.

Embodiment 3:Phage E sc-COP-9 kills the Capability surveys of enteropathogenic E. Coli

The ability that enteropathogenic E. Coli is killed to separation phage E sc-COP-9 is investigated.Kill Capability surveys It is according to implementing by way of whether generating transparent ring the spot test that is proposed in embodiment 1 investigation.For killing ability The enteropathogenic E. Coli of investigation is to detach and be accredited as enteropathogenic E. Coli by the present inventor, has used 10 plants in total.Phagocytosis Body Esc-COP-9 be directed to as experimental subjects 10 plants of enteropathogenic E. Colis in total 9 plants have killing ability, have generation The experimental result of table is as shown in Figure 2.In addition, phage E sc-COP-9 has also been investigated to bordetella branchiseptica (Bordetella bronchiseptica), enterococcus faecalis (Enterococcus faecalis), enterococcus faecium (Enterococcus faecium), streptococcus mitis (Streptococcus mitis), streptococcus uberis (Streptococcus uberis) and the killing ability of Pseudomonas aeruginosa (Pseudomonas aeruginosa), the results showed that, Phage E sc-COP-9 does not have killing ability to these strains.

From the above results, it can be seen that, phage E sc-COP-9 has outstanding killing ability to enteropathogenic E. Coli, to more Several enteropathogenic E. Colis can play antibacterial effect.It means that phage E sc-COP-9 can be used as to prevent and locate It puts the active ingredient of the composition for the purpose of the infection of enteropathogenic E. Coli and uses.

Embodiment 4:The experimental example of the infection of phage E sc-COP-9 prevention enteropathogenic E. Colis

The 1 × 10 of 100 μ l are added in into a hose of the TSB culture mediums equipped with 9ml⁸The bacteriophage of pfu/ml standards Esc-COP-9 solution is only added to the TSB culture mediums of equivalent into the hose of another TSB culture medium equipped with 9ml again.So Afterwards, the culture solution of enteropathogenic E. Coli is added in into each hose, it is about 0.5 or so to make its absorbance at 600nm.Add It after adding enteropathogenic E. Coli, moves the hose to and shaken cultivation is carried out in 37 DEG C of incubator, and observe pathogenic large intestine bar The growth conditions of bacterium.As shown in table 1, in the hose added with phage E sc-COP-9 solution, pathogenic large intestine bar is observed The growth of bacterium is suppressed.On the contrary, in the hose for being not added with phage solution, the growth quilt of enteropathogenic E. Coli is not observed Inhibit the phenomenon that.

【Table 1】

The growth inhibition of enteropathogenic E. Coli

By this result it has been confirmed that the phage E sc-COP-9 of the present invention can not only hinder pathogenic large intestine bar The growth of bacterium, and with the ability killed.The conclusion therefore deduced that is, phage E sc-COP-9 can be used as with Prevent enteropathogenic E. Coli infect for the purpose of composition active ingredient and use.

Embodiment 5:Utilize the example of phage E sc-COP-9 treatment enteropathogenic E. Coli infectious diseases

The therapeutic effect for pig of phage E sc-COP-9 is investigated, which induces by pathogenic large intestine Disease caused by bacillus.The weanling pig 4 of 25 days after birth is divided into one group, is always divided into two groups, then separately raises it It supports in experiment raising pig house (1.1m × 1.0m), experiment has carried out 14 days.Ambient enviroment is controlled under lagging facility System, the temperature and humidity for making pig house keep constant, all pig house ground are cleaned daily.Since experiment day the 7th My god, all pig is made to have taken orally enteropathogenic E. Coli solution using oral cavity infusion tube all pigs.Oral pathogenic large intestine bar Bacterium solution prepares as follows.After enteropathogenic E. Coli is cultivated 18 hours using TSB culture mediums under conditions of 37 DEG C, Only recycling thalline.Then, it is adjusted to 10 with physiological saline (pH 7.2)⁹CFU/ml.From oral enteropathogenic E. Coli Next day starts experimental group to be made (to inject phage solution daily according to the mode identical with injection enteropathogenic E. Coli solution Group) pig take orally 2 times 10⁹The phage E sc-COP-9 of PFU, and the pig of control group (group of unimplanted phage solution) is not cooked Any processing.The feed and drinking water provided for control group and experimental group is all identical, after injection enteropathogenic E. Coli Start to have investigated diarrhea situation occurred by object of all experimental animals daily.The investigation of diarrhea situation occurred is according to measure abdomen Rush down what the mode of index was implemented, Scours index measures (normal according to conventional excreta consistency (FC) score is measured：0, soft stool：1, It has loose bowels：2, severe diarrhea：3) mode is implemented, and the results are shown in Table 2.

【Table 2】

Scours index measurement result

By this result it has been confirmed that the phage E sc-COP-9 of the present invention is for treating because of enteropathogenic E. Coli The infectious disease of induction is also highly effective.

Embodiment 6:The manufacture of feed addictive and feed

Utilize phage E sc-COP-9 solution manufacture feed addictive, it is ensured that in per 1g feed addictives containing 1 × 10⁹The phage E sc-COP-9 of pfu.The manufacturing method of feed addictive is as follows, and maltodextrin is added into phage solution It is freeze-dried after (50%, w/v).Finally, it is ground into fine-powdered.Drying process in the case of the manufacturing process in question May be used be dried under reduced pressure, heat drying, air drying.In order to carry out contrast experiment, the feed addictive without bacteriophage is pressed It is manufactured according in the following manner, i.e., using the buffer solution (buffer used when manufacturing phage solution；10mM Tris-HCl, 10mM Mg SO₄, 0.1% gelatin, pH 8.0) instead of phage solution.

By the 2 kinds of feed addictives manufactured by the above method respectively according to weight ratio and 1,000 times of foster pig's feed Mixing has been finally made 2 kinds of feeds.

Embodiment 7:The manufacture of drinking water additive and disinfectant

Drinking water additive or disinfectant are that occupation mode is variant, dosage form be all it is identical, therefore, both It adopts and manufactures in a like fashion.Utilize phage E sc-COP-9 solution manufacture drinking water additive (or disinfectant), it is ensured that Contain 1 × 10 per 1ml drinking water additives (or disinfectant)⁹The phage E sc-COP-9 of pfu.Drinking water additive (or disinfection Agent) manufacturing method it is as follows, manufactured according to the phage E sc-COP-9 solution and well-mixed mode is added, it is ensured that system It makes and contains 1 × 10 in the every 1ml buffer solutions used during phage solution⁹The phage E sc-COP-9 of pfu.In order to be compared Experiment, the drinking water additive (or disinfectant) without bacteriophage manufacture in the following way, i.e., directly using manufacture bacteriophage The buffer solution used during solution is in itself.

By the 2 kinds of drinking water additives (or disinfectant) manufactured by the above method according to 1,000 times of water of volume ratio It is used as final drinking water or disinfectant after dilution.

Embodiment 8:Confirm the feeding effect of pig raising

Whether have to the feeding effect raised pigs using the feed, drinking water and disinfectant manufactured in embodiment 6 and embodiment 7 Improvement is investigated.Particularly, this investigation is the angle implementation from case fatality rate.To every 10 points of 30 piglets in total It is one group, is always divided into 3 groups and (uses the group-A of forage feed；With the group-B of drinking water supply；Group-the C disinfected), experiment is real It has applied 4 weeks.Every group is further divided into the group formed by 5, each group is further divided into (small using the group of phage E sc-COP-9 Group -1.) and group without using bacteriophage (group is 2.).Piglet as this subjects is the wean son of 20 days after birth Pig, by the feeding piglet of each Testing Team in the independent pig house of difference separated at regular intervals, each group situation as shown in table 3 It distinguishes and indicates.

【Table 3】

Small group differentiation and expression in swine rearing experiment

If with forage feed, just fed with differentiation of the feed manufactured in embodiment 6 in table 3 according to conventional feed Mode is fed.If fed with drinking water, just with differentiation of the drinking water manufactured in embodiment 7 in table 3 according to normal Rule drinking water feeding patterns are fed.If disinfecting, one week 3 times are sterilized in turn with existing disinfection way.Sprinkling The date of disinfectant of the present invention just carries out disinfection without common disinfectant, and result of the test is as shown in table 4.

【Table 4】

By result above it has been confirmed that fed with the feed and drinking water that manufacture according to the present invention and according to the present invention into Row is disinfected can effectively reduce case fatality rate during pig is raised, and the conclusion therefore deduced that is to use the present invention Composition can be effectively improved pig raising effect.

In the above description, the specific part of the present invention is elaborated.Skill with industry Conventional wisdom For art personnel all, it will be appreciated that detailed description above is only the preferred embodiment of the present invention, the scope of the present invention is not only defined in this. Therefore, the actual scope of the present invention should be determined according to claim and its equivalent.

【It is commissioned number】

Entrust to the care of organization names：KCTC

It is commissioned number：KCTC 13131BP

It is commissioned the date：20161017

<110>Yin Telun biotechnologys Co., Ltd.

<120>Coliphage Esc-COP-9 and its enteropathogenic E. Coli Proliferation Ability purposes

<130> DFP16KR2908

<160> 1

<170> PatentIn version 3.2

<210> 1

<211> 51102

<212> DNA

<213> Bacteriophage Esc-COP-9

<400> 1

atattgaaac agcttttgac cttgcgaagc agtctgataa aatggcggcg cagcgtaaat 60

ctgttaaggg cgattctttc gagtctggca agagtgaaaa aaccgactcc cttgatccga 120

ctgcgcgttt gaaaaaactt aaataaggag aaataaacat ggctacaatt ggagcaagtt 180

attttgctgt aatgtctcgt gcgttacctg gtcaggtttc tgacacttca gcttataata 240

ttgatggcgc ttgtgttctt gagaatgcgg aaggaaacca gaacaaaaac atttttgttg 300

gtgttgcagt ccagcatggc ggcgttgatg ctatgggcaa taagctaatt aagccgatgg 360

cggcaagcgg caaggcttac ggtgttgcaa ttcgatccca cttccagacg acttcggctg 420

acggtcgaat gatttatgag tctggcggcg gaatcaacgt aatgactgag gggcgtgttt 480

ggatgttggc taaagacgct tccgcaccag cgttcggcgc tacaatcaaa ctgtctgatg 540

atggtcaggt tgataccgcg tctggtacta tcgaaaccaa ctggatcgcg actggtgatt 600

tcaccaagtt tcaagacttg aagctggtag aggttcaggt acaaaaagtt ccggcagttt 660

cagcagttta ataaatatgg cgctcactga gcgccttttt tatatttggg gttatcatga 720

gtgattattc agtcattgtt ggcgctgagt cgccagggtg catggtcgat tcttcaataa 780

tggtgattaa cggatctttc gtgtgcggag atgaatttat tgagccaggt aagattgtgc 840

gactatcaca tataacaaat ggatacaaaa tagcaagtgt taatggggat ctcgttattg 900

gtgtggcagt aaggccgccg gactcgtgca tatatgagga gggtgatccc gttagtgtag 960

ttagtagagg tagtgtgtgg tgcttaactt ctgagagcga agcgccagaa tatggagata 1020

aagtatttgt aacgcaagat ggagatgcgg catacagtga aggcgacttg ctgattggtt 1080

ggatctttac tggcgaacat gtaaaggttg atcatgagtc atatattgtc ggggtaagca 1140

ttagcgcata acacaaaatt atgttgcacg tttaacaaaa aatgctatca ttggggcgtt 1200

acatactaac agcctttata tatggaggaa gaaaatggaa aaagaaaaat ttgatcagct 1260

agatgcggat attgtttcta gctatctggc tactcgcggc gttaagggtg atgcttctga 1320

tatgggtatc tggacggcgc aagagcttca caagatccgc tctaatgctt acgagaaaga 1380

atatccggca ggttcagcgc ttcgtgtatt ccctgtaact aatgagcttt ctgatactga 1440

taagactttt gaatatcaga tgtttgacaa ggttggttac gcaaaaatta tcgctgatta 1500

caccgacgac ctaccgaccg ttgacgcgct gatgtcaagc gaatttggta aagtgttccg 1560

cctgggtaac gcgttcttga tctccattga cgaaatcaag gctggtcagc gtactggcaa 1620

gagcctttcc actcgcaagg caaacgcagc gcaaaacgct catgatcagg aagttaaccg 1680

cctggtgttt aagggttcaa agccgcataa gattctttcg gtgttcgaac atcctaacct 1740

gacaaagatc gtgtcaactg gctggttaag tgctgatgaa aacactaagt tcccgcaggc 1800

agcatctgat gaactggaaa aggctattga aaccattcag accatcacca atggtcagca 1860

ccgagcaacc aacatcctga ttccgccatc tatgcgcaag gttctttccg ttcgaatgga 1920

aaacaccact gaaagttatc tggaatactt ccagaaacaa aacagcggaa tcaccattga 1980

ctcaatcgca gagcttgagg atattgatgg cacaggaacg aaaggttgcc tggtatatga 2040

gaaagacccg atgaacatgt cgatcgaaat tccagaagca tttaacatgc ttcctgcgca 2100

accaaaagat ctgcatttca aagttccgtg tacttcaaaa tgtactggcc tgacgattta 2160

ccgtccgttt acgctagtgc ttattactgg cttgaaaaaa ccaagttaat cgtataacat 2220

tggggaatcc ttaactggat tccctttttt ttattggagg ttatgaaatg gctaaagaaa 2280

aaactgtaac gctaaaggtt tccggcgtgt gccttatcat tgttgatggt gagcattatc 2340

atccaggcaa agagtttgac gtagaagaat caaaattgaa aacaagcgcg tttgaatatc 2400

taattgcaaa aggcgatctt gaggttaaag acaactccgc attgaacgaa gaaatcaaac 2460

agagcgcagc aagcaaacgt aaaaaagacc cgcgagaggg caagagcaaa gccgaacttg 2520

aagacggcgg catctattaa taaagagggc gcttatagcg ccctttgtca tatctggagg 2580

aaacaaaatg gttgatgaat ttttcactga cgcagaaatc acgcagcaaa tcgtgaaatt 2640

agctccaccg atgaaacaga ttgatccaga tttaatggtg gcatggattg atctagcaaa 2700

ggaatttgtt tgcaagaagc ggtttaaaga atcatatcct aaagcggttg ccctttatac 2760

tttgcacctt atgacgcttg atggagctat gaagcaagaa ggggaaggcg tagagagtta 2820

ctcacggcgc gtggcgtcat tttccctgac tggtgaattt agccagacat ttgatcgcgt 2880

gtccggcgat tcaagcggaa aagagattag acaaactccg tggggcaaaa tgtatgagac 2940

gttaaataga aaaaaaggcg gtggatttgg ccttgtgact ggatttcgga ggcgctgcaa 3000

atgaactata acgagatcgc aagaatggca accgaaggaa ttaacttctt tagtgatagc 3060

aacggtgagt ttaagtgcat aacgcagcgt ggaagcgttg aaataatcgg aggcgaagaa 3120

gtacaaaagc cagagattag tgtaatgatt aaaggtttaa ttcgctcacc aaaagtacgc 3180

gaggttgacg gcgagacaat acgagtaaac gataagctgg gagttttcaa caataatgtt 3240

gaaattaaga acgggtatca tattgatgtt gatggtgaat tatatgttgt agttgaagca 3300

agaccaatca gacaaaccaa tgtcacagtt gcttatcggc ctattttgag aaggatttcc 3360

gtgcatggct aaaaactaca cgataaggga atttcatggc aacattgatg cgtggataaa 3420

cgccgttgat aatggactaa aagattgcgt ggaattgttc gccgaaaaag tacacacaga 3480

tttggttaag cgttctcctg tagatactgg tcgttaccgt gcaaactggc aggtaacagc 3540

aaacaatccg ccattgtatg cgcttaacca gtacgataag cacggcgaca aaacgatcgc 3600

ggaaggcaag cgcgccatat acgcaatatt acgaggtggc ggagcagtaa gagccattta 3660

tttttccaat atgcttattt atgctaacgc gcttgagtac ggtcattcaa aacaagcgcc 3720

agcgggtgtg cttgggattg tagctgttaa gttgagatct tatatggctg aagcaataaa 3780

agagtcgagg gctaaaaatg cactatgaat taatgctatc ggcgcgtaag gcgctggcga 3840

ctgaatacga aagcagattc atgatcgctt atgaaaacgt agagtttacg ccacctggtg 3900

acggttcgcc gtggttgaag tttgactatg ccgaagttga cacagaatac ttgtcattgg 3960

atcgcaaatg cgtttcttat atcggaatga ttcaggttgg tattgtgttt ccaccaggtt 4020

acggaactga caggccgcgc gttctcgcta aagagattgc gcaattcttt tacgatggta 4080

aaatgctgga gcatggttat atatatgagg gtgcaagagt tcacaagccg ctcaagagtg 4140

aaagcggttg gattctccca ataaggtttt atgttcgaat tgaaacaaag gagtaaaaat 4200

tatgcattta ccaaatggat cgcaaatttt cgttgagagc aatcgcggaa gcgaaatcca 4260

agcaacagca gtgtcaaacg ccaaagatcc tgttttcact gttgcatctg gcggcactgc 4320

gtacaagaaa ggtgattatg tgatcatcac tgcgtcttct tggggtaagc tgattgatcg 4380

tgttatgcgt gtaaaggcta acggtgaaga aacaagcgtg actcttgaag gcgtagatac 4440

tactgatcag aacgttttcc cgactggcgg cactgcatct tttgccaaaa ttgatgcatg 4500

gactgagatc ccttgcgttc aagatttgtc gcaggacggc ggcgaacagc aatactacac 4560

ttatcagtgt ttggcagacg atcaggaaca gcagttacct acctataaga gcgccgtttc 4620

tcttacctac acttttgcgc acgaatacga taacgcgatc tatccgctat tgcgttcagc 4680

agatgaatca ggcgaagtaa ccgctcttcg catgtatgta ccaaaagcaa aagagatgcg 4740

ttgctgggct ggtgtgcttt cgtttaacga aatcccgcag accacagtta acgaaatgga 4800

aactgtttcc ctttctgtat cactgaaagg tcgattcacc tttctaccct ctcaagtaga 4860

gtaataacaa ggggcattgc gccccttttt ttgttactgt acaatcatga tacacgattc 4920

ataatcaatt cttttaacaa aaagtgctat caaggagaaa gaaatgtcaa aaatgaaatt 4980

gacgattggg ccgcttcctg actttaaatt accagtgaaa tttgcaatgc ctaacggaga 5040

agatcaaaca attatcttca cagttcgcca ccgcaaaaca agtgagattc acgagcgcta 5100

cacgtcagat actccaatga gcgatgttga aatgatcacc ttccttgctt ccggttggaa 5160

tcttgatgat gaatttaacg aagaaaatat taaacagctt cttgattact acccagcgac 5220

agcaatcgga ttaactagcg catacatgaa agcgcttgcg gggcagcgag taaaaaacta 5280

aaaagggcgg tttacctgtt ttatcagaaa ccgccgacag atgcagagct tgaggccgtt 5340

ggccttacaa gggcagacta tgaaggagaa gatccgccag aggttatatt tgatgaaagc 5400

atgatgcaat catgggatat attttgcgca atgcaaacgc aatggagatg ttcaggcggt 5460

ggcgcttacg gatttgatta taacgtcttg cctatgcttt ttgagattta caaggttgag 5520

gatcgcgaga tggcgctaaa cgacttgcga atcatggagc aaaaagcact tgaaatgatg 5580

cattcaaaat aagcgcctac gggcgctttt ttattacctg gaggattaat aatgtcagaa 5640

caatacgcag gcttgacgct tggggttgat gtttctcaac tcaacaatgc tgtaaagtct 5700

ttgcagcaat tcaaaaaggc aaacgacgat gcaaagggaa gcgttgagag ttttgttgat 5760

tcagaggttg ttgcaagaca aagagccaaa caactggctg aggaattggc aaagcagaag 5820

caagaattta aagcaattca gtcggcaata gatccgacag caagcaaaat ggataagttg 5880

cgccaggctg cgacacagct tgatgcgctt tggaaaaaag gaattgtgcc ggatgataca 5940

ttctttgagt taggatcaat tcttgagacg cagcaaaaca aactgatcgc aactaaaaag 6000

gcgctaactg aagaaggccg cgcggcgctt gaggaagcaa aaaacaaggc aagggcagag 6060

gctgaggcaa gaaagtttat tgcggcattg caggcgcaag ctgacgcagc ggaaaaaaca 6120

aaatcagagc ttgtcgaaat gagagccgca caacttggag ttagtgcaga ggctgcgcca 6180

tttattgcaa aaatgaagga gcaggaaaaa caagcctcaa aacttggcgt ttctatgggg 6240

cagtacaagc aagcaatggc gcaattgcca atgcagatca ctgacgttgt tacttctctt 6300

gcttcaggaa tgccagtatg gatgatcgca atccagcagg gcgggcagat caaggactca 6360

ttcggcggtg ttgctaatac ttttaaagcg ctaatgacat ttgtcacgcc gctaagtgtt 6420

ggaatgacgg cgctgactgg tgctcttggg tatgcagcat ataacgctta caaagccaac 6480

gcacaactga aggagatcac aaaaaccgtt caggatgcaa caggtctttc cggtgagttt 6540

gcagagcgga tcgcaactgg aatccaggcg ctatctgaca agacaggaga gagcgcagac 6600

aatatcgcaa aggcttacat aagcactaag gatggagcaa gcgaggctat acaaaaactc 6660

gtcgatgttg gatttaccta tgacgaagca aaggcaaaag taaacgaata caaaaacgca 6720

tcaagttttg taagtcttaa caatgaaatt gctgatcaca aaaacaaagt tcttgagctt 6780

ggcgattcat ggtatgcggt cttaaaagca aaacgtgatt atgcttcacc gtccggcggg 6840

ctgttgggga aagagcttgg ttatgtgaat ccaatgctgc aatttgcgaa gaacacctat 6900

gaggatatag gaaagatagt aaaagatgcg aataaggata tggcggaacg agcagcacgc 6960

atcgacagag aaaatctttc attaaacaga gttcgagcgg cgcaagaggc tttaaacaaa 7020

gccatagagg atcagaagaa cgtagcaagg acagcagacg aagagttgaa aaagcgcgcg 7080

gctgagaacg tggagttcag gcgaaaagag cttgaagagg cgaagaaggc gcagcagcaa 7140

aaggagaaag ttggtggagt tgtaaaagca ccaacagagc agcttgataa ggagctatat 7200

gttctcaagg cgcaacttga aacgctgaaa gagcaccgaa ctgtaaatga tgttatttca 7260

cgccagaggc aatctttgtg gagcattgaa aagcagatcc agatccttga ggaagcgcaa 7320

agtaaacgca agctgacaaa agcggaacag gcgttactaa acgagcaaaa agcagttatt 7380

gctatggcaa aagaaaaggc agagataggc gatcagattg ttttgcaaca gagaaagaac 7440

aagcaatacc aggaagggct taaattcatt cagcaaacat cagatgctat tgacgctatg 7500

aacttgcgac agtctggagc tacagatctg caaatccagc gagagcttga gttgaagaag 7560

ttgcgaactg attatgttgc tggcggcggt agtatcgacg atgaaattta tcagcagatg 7620

gaagcaaagc tgaaggagta ttacgccact gaggatcagc ttagaaataa ctggattgca 7680

ggagcaaaaa acgcatggga ggtttacgga caagatgcaa tggatatgta cggcaacgtg 7740

caagatattg caagcgaggc gctcaacggt ctaaccaacc aaatggctac attccttgca 7800

acaggaaagg caaactttaa gagcttcgca acgtcaatca tccaaatgat tatccagatg 7860

atcacgaaga tggttatctt taatgccata tctggtgcga ttggaggtga cacttggacg 7920

attggaagcc ttcttaaaaa tgttggattt gcaactggtg gatacactgg cgacggcggg 7980

aagtatgagc cagcaggtgt tgtccataaa ggcgagtttg tcatgacgaa ggaggcaacg 8040

aagaggatcg gagttggaaa tctttacaaa atgatgcgcg gttatgcaaa tggcggcgtt 8100

gttggcggca cttcatacac tggcggcgga gtttcatctg gagcaacaaa tcttaatatt 8160

ggtgggatta gcgttgatat taacaacgga aacgatccaa aagggttgga gactggcgta 8220

aaaatgattt tcactgatat gattaagcgt tcctgtacgc agggtgggga agtttacgaa 8280

tttgttatgt ctaagcgggg gtgatagtga aacttgagca attcaaatgg tgcacgcaaa 8340

cgcaaggagg cggcggcact atgaccacat caaataacga cagggaaatc tcatttggta 8400

atggttacac gcaggttgcg tcaggagggt ttaacactgt acgcagggag ttttccattg 8460

tctatgtggg taatgattac cgagatgttg ttgacttcct gaatggtcac aggttgaagc 8520

cgtttttatg gtttatgcct gacggtcaac ctggtctatt cagggtaaaa tctggtagcg 8580

ttgggttaac tccaatatca gcaaccgttc aggaagtgaa agcaacattt actgagcaat 8640

ttacatcaat gcaataattc aagccgcctt tgtgcggctt ttttattgat gatacaatgt 8700

acgaaaggag gtgcaattat ggctaatgaa acaacaggtc gcgcggatct tgaaaattgt 8760

ctgcaaagcc tttaccctgg cgagattatc acgctaattg agattgacgg tacaaagttt 8820

ggcgcaaaca tttaccgcat acacaatgag aatatatcat acactgcgga agaattatta 8880

caggcgcgag aaactggagt tcttcctccg aaagagatta cattccgtgg cgaggtttac 8940

ggcgcgcgcc cgtttggaat atctggaatc aacttcacaa gcaacggaaa ggctgataag 9000

ccacaattga tactatcaaa ccttgatagc caggtgagcg cgatgattcg caactttaac 9060

ggcatgatgc aagctaaggt tacaatatgg atcacgccag cggaattaat ggggaaagat 9120

ggcagcatta aagatggagc ctcaagaaag ctggtttact acattgagcg accaagccat 9180

tacaatagaa tgatggcgaa atttgacctg acatcgcctt atgatatgga tgggataatg 9240

attcctccgc gaataactca aagcgtttgt tattgggcgc agcgtggatg gtatcgaagc 9300

ggaaaaggtt gcggatacaa tggatcgcga atgtttgaca aagacaacaa tcctgtaacc 9360

gatccatctc aagacttttg cgccggaact gtaacagcgt gcaaacttcg ttttggtgca 9420

gataagcagc ttgattttgg cggcgcacca gtagcaagcc tgttaaggag aaatcagtga 9480

tgattagtgc aaagataaaa cttgaaataa tgcagcacgt aaaagatgaa tacccgcgcg 9540

aggcgtgcgg ggttatcact caaaagtcgc gggtacagaa atatcaccgc ataacaaacg 9600

tgcatgatga tcctgaaaat cattttgaga tgggcgcaac tgaatacgtt gaggcgtgtg 9660

aaagcggtga gcttattgct gttgtgcata gccacacagg agacggggca agtacaattc 9720

caagcgccca tgacacatgc atgtgtgatg aaatgggtgt ttcatgggtt attgtgtcat 9780

ggcctgaagg tgatatgaga atcattgagc ctgaatctcg tcctctgatt ggtcgcccgt 9840

ggtcgcttgg ggcgtatgat tgctgggggc ttattatggc atggcacaag caacatggcg 9900

ttatcctgaa tgatttcagg aaaccatatg aatggtggaa gcctgagcac ggcgaaaatc 9960

tttaccagga aaattatctg aaagagggtt ttgttgaaac aggagagccg ccaaaacctg 10020

gtgatatggt tatctttcag ctttccgcgc cagtgtggaa tcatgcaggc atttatcttg 10080

gcaacaatca gttgcttcat catgcctttg gtaagttgtc gagggttgat ttgtattctg 10140

gatggtatca ggaacatgca aaaatggttt gtagacataa ggatctgaaa tatgactttg 10200

aaggtaatta aattatctgg atctttaggc cgaagatttg gagtatttca taagctggct 10260

gttgattcat acccagaagc aatacgcgca ctatcttcac aggtggaagg attcaaggac 10320

tacatgcaaa gcgaagtagg atcgcgaatg cgttacgctg tatttgttga cggtaagaat 10380

gtaggacagc acgatgaaaa agcgtggcaa tgtgcaaagg aagtgaggat tattccaatt 10440

ccaacaggtt caaagtctgg cgggttgttt caggttgttc ttggcgcggt tattatggcg 10500

actgcatttt tcactggcgg ctcttcactt gcgttaatgg gtgcttttgc atcgtctgct 10560

tttatgatgg gtggagcaat ggtacttggc ggagtaatgc aaatgatctc accgcagcaa 10620

ggaggatcaa gattgtcatc ccaatcagca gaaaacaagc cgtcttatgc tttcggtggc 10680

gctgttaaca caacggcagc aggataccca ataccattgc catatggtca aaggaccgtc 10740

ggtggagcta tctggtcggc tgggagttat gcagaagata aggcttaata taaaagagtc 10800

gcgcgttgcg cggctttttt tttgcccgta taattcaaca aatcaaatag cacaaaaggt 10860

gaaaaagcat ggctgaaaat atgataacgg gcagtaaggg tggatcatca aaaccttatg 10920

tgccaaaaga gatggaagat aacctgatct caatcaacaa aatcaaaatc cttcttgccg 10980

tttccgatgg tgagtgcgat ccagatttta cacttcgcga tctgtatctt gatgatgttc 11040

cggtaattgc agacgacgga actgttaact accagggtgt gaaagctgaa tttcgacctg 11100

gaacgcagac gcaagattac atccaagggt ttactgacac atcaagcgaa gtgacgctgg 11160

cgcgtgacat tacgacgaca aatccttatg taatttccgt aaccaacaaa acattatcgg 11220

ctatcagaat caaaatgcta atgccaacag gcattaagca agaggataac ggcgatcttg 11280

tcggcgttaa ggttacttat gctgttgata tggctgttga cggagactct tacaaagaag 11340

tattgctaga caccatcgaa ggtaaaacgc gttccggtta cgacagaagc cgaaggattg 11400

accttccggc atttaatgat cgcgtattgc ttagggttag aagggttacg gcagacagcg 11460

catcatctcg cgttactgat ctgattaagc tacaaagtta cgctgaggtt attgatgcaa 11520

aattccgtta tcctctgact ggtcttgtat acgttgaatt tgacagtgag ttgttcccta 11580

accagatccc taacatttca atcaagaaga aatggaagct gattaacgtt ccgagtaact 11640

acgatcctgt tttgagagag tatcatggtt catgggatgg tactttcaag aaagcttggt 11700

cgaacaatcc ggcgtgggtt ctttatgaca ttatcacaaa ccagcgatac ggattagatc 11760

agcgagaact tggtgtgcag gttgacaaat ggagtcttta cgaagcggcg caatactgcg 11820

atcagaaagt gccggatgga aaaggcggta cagagccgcg ttatctatgc gacgttgtga 11880

ttcaaagcca gattgaggct tatcagctta ttcgtgatat ttgctcaatc ttccgaggca 11940

tgagcttttg gaatggcgag agcttgtcaa tcgtcattga taagccgcgc gatccgtcgt 12000

acatcttcac caatgacaac gttgttgatg gtgattttca gtacacaaca gcaagcgaaa 12060

agagcatgta cacgcagtgc aacgtgacgt ttgacgacga acaaaacatg tatcaacagg 12120

acgtagaggg cgtattcgac acagaggccg cattgcgctt tggatacaat ccgacaagca 12180

taacagcgat cggatgtaca cgcaggagtg aggctaatcg gcgcggtcga tggatactaa 12240

aaaccaactt gcgcagcact acggtaaact ttgctactgg actggaaggc atgatcccat 12300

caataggtga tgtgattgct attgctgaca actttcacag cagcaacctg aagttaaacc 12360

tgtcagggcg cgtgatggaa gtttccgggt tgcaggtatt cgttccgttt aagattgatg 12420

ctagaccagg tgatttcatt atcatcaaca agccggacgg taagccagtt aagcgcacaa 12480

tctcaaaggt aagcggtgac ggaaaaacca ttgagctaaa cattgggttt gggtttgaag 12540

ttaaacctga cacggttttt gcaatcgacc gcactgatat tgcattgcag caatacgttg 12600

taacgagtat cggcaaaggt gatgatgatg atgaatttac atactccatc acggctgttg 12660

aatatgaccc gaacaaatac gacgagattg attacggagt aaacattgac gacagaccaa 12720

cctcaattgt ccagcctgac acaatggcag caccggaaaa tgtgcaaata tcctcatact 12780

cgcgaattgt ccagggtgca agcgttgaaa caatggttgt gtcgtgggat aaagtacctt 12840

acgcatcgct gtatgaaatg caatggcgaa aaggtgatgg caactggctg aatacaccac 12900

agacagcaaa caaagaaatt gaggttgaag gtatttattc aggaaactac caggtaaggg 12960

ttagatctgt ttctgcttcc ggttcaacgt cgccatggtc cagaattgtg acagcttcac 13020

tgactggtaa ggtaggagag ccaggcgcgc cagttaactt aactgcatca gataatgagg 13080

tgtttggcat tcgtgttaag tgggggatgc cagaaggtag cggagacacg gcatacattg 13140

agcttcatca gtcgccagat ggaacggcgg aaaactcaag tctgttaacg ctggtcccat 13200

atccacaata tgaatactgg cacgggacgc ttccggctgg tcatgttgtc tggtatagaa 13260

tccgcagcgt tgacagaatc ggcaacgttt ccggatggac tgattttgtt agaggcatgg 13320

cttcagatga tgtggaggct gttttaggcg atattcttga taagattttt gataccgaag 13380

caggaaagga tctgaaagag aatgccattg atagcgcaaa caaaatcaag gatcaggcac 13440

aaagcatcat tcaaaacgca ttggcgaatg atgcagacgt taggattatg aggaaggaaa 13500

acggaaaacg caaagccgag tttagacaat caatccaatt gatcgcaagt gaaactgagg 13560

cgcgcgttac cgcaatgacg caacttaagg ctgaatttga cgaggaaata actagcgaag 13620

taacgagact cgatcaggca attgcaacag aatcggaaac gcgagcaaca gccattgagg 13680

aattgaaatc acagattggt gatgatattc aggggcaatt aacgcgggtt gaggaagcga 13740

ttgcaaacga aacggaggcg cgcgtttcgg ctgatacagc attaacagcg aagtttggag 13800

atgttgaatc agcgcttaca gaaaaacttg attcatgggc tggcgttaat ggagttggcg 13860

cacagtacgc aatgaaactt ggattgacat acaacgggca gaagtacagc gccggaatgg 13920

tcatgcagtt atcaaacagc gctcaagggt tgatttcaca aatccttttt gatgctggaa 13980

gatttgctat catgacgagt tctaccggag ggtcgtatac attgcctttt gtggtcgaaa 14040

ataaccaagt tttcattaac agcctgttag tgaaagacgg atcaatcaca aacgccatga 14100

tcggtaactt catccaatca aacaattacg tttggaatca aacaggatgg aggcttgata 14160

aaaacggcac gtttgaaaat tatggttcta ctcctggaga gggagccatg aaaatgacaa 14220

acgaaacgat cagcgttagg gatgcaaacg ggcgcttgcg tgttcagatt ggtaggctta 14280

ctggtacgtg gtaaaatcaa agcggggcat ttgcctcgca tttatggagg tttaataatg 14340

gctgaatatg gtgtttcaac gtgggatgct aacgggaagt ataacaacta cggaatcaag 14400

ccagtttccg ttgttggtgt cattagcctt gccgctggtc agacaagcgg atcgtggagc 14460

tttagcattc caagtggttt taaggttggg tatgttgttt cacttgatga aggtgccagg 14520

ggagttggaa gggaaatagt agcatctgga aacacaataa gaattaatcc tacatcgtca 14580

gttggtgcaa atagatattc ttcttcaaag tgcgagttag ttgtttttct tgagagggct 14640

taaaatggct gaatatggag caatgttatc tttatcaaat gggaatccat ttatcacacc 14700

aaaatcaacg ccattttgct tatatggcaa gtacacttat tcatcttcag gtacttctgc 14760

ctatcatagc gcaagcgcaa atataccagt aaaccaatct tatccatgca tggcatttat 14820

aaaaacaaca aatacacaac agccaactgc attaatagct tacaggaatg gtggtaatat 14880

ttatgtaaac ggaggaaatc catatggtca atcatttaca atgactgttt atatatttgc 14940

catattccct caaactctac caaaatatgg tatggctata tgggatgcaa gcggcaagtt 15000

agttttaact aatgaaagca gggtgctaac agatcttgtt acaattggta ctcctggttc 15060

ttcaggtgga acgaatatag atcaaacgtt atctggttct tatgccgttt gcccttcaag 15120

gcttggggcg gttataggta tgggcgcttc tgatatatat acatcatgca ggtataatgg 15180

ttcaagcaca aggattggtg cagcaaggac aacgccagga acaggttcta taacaaataa 15240

cggaaattca ataattgcca tcaagactga tatttacgac caataaaaag gggctttcgc 15300

ccctttattt ttatttacag actgcaatca gcgtttttaa agttgtcaat gcttacccac 15360

tgaaaattaa acgggtagcc agcattaaca agcgtgcgat ttccatctcg ttttactcca 15420

aaaatagcaa ctgaatactg catgccacca ttttcataaa tagccgtgca ttcacgttta 15480

ggcatatttg cgcaaccagt catggcaacg gctgccatca aaatggcgat taactttttc 15540

atcttggtat ctccttcgtt gttgttggtt gcaactatac acgatcttgc ggttggcgtt 15600

tagcaaaaag tgctattctt gttctttttg ttatctggtg ttctcaaatc tgcggagtct 15660

taaatttagc tcaaaaagcg aacaacaata gagcgcaata catacagata aatatataca 15720

tacatataaa tcaataagtt agtagtatat atattcatgt ttgttcttta ttgttatcta 15780

tgttttgtgt gattttgtgt gtctggtgat ttttctgtga ttatgcatgt gtcattggtt 15840

atgtgtgtct atatatctat atccggcgca aattgaggaa caagagaaca aaaagacaac 15900

aaagatattt atcaatcact tacatgtaag ttttgttatc caagatagct agaacgttgc 15960

tagaacagaa aaaacaaaat aactattgac taaacgcatc aaatagctac aatgcagaca 16020

taacaaccaa tgagggtaaa tcatgaataa gtttatggcg tacaccagcg aagaaatgag 16080

taatgagctt tatcacgatc cagaagcgtg gactgctgat tatgttagtg gatcaagcct 16140

tgcggagatt tacagcacat gcccggcggc gtggaagttc aagccgcgtg acgataagag 16200

taaggcgcta gtttttggta cacagtcgca taccaacttt gaaagcaagg agcttttcga 16260

aaaaacctat cgccgtgcgc cagcgccaga agactttaag gatctgataa ccagccaaac 16320

ggcgcttgct gcaaagctga aatcatttgg cctgaaaggt acaaccggaa agacttaccc 16380

cgaacttata aaaatgatgg ttgactgcgg agaagacctt aacgtcatgt ggttaataga 16440

catgatcgca gaaagccagg ccagggccga tggggttcag ttaatcgagg ctaaagatta 16500

cgattcgtgc gtggcaatgc gccaggttct ggaatcaata ccggaacata acgcatgtat 16560

gaatagcaag acggcacaac gtgaattatc gttgttcggt gaaattaacg gcgtaaaagt 16620

caaagtgcga tgcgatcacg ttgacgtaac aaagaacgtc accgcaacgc tgattgatgg 16680

ttacgatgaa aaaggtcagc cgatttgccg agatattatt tatccagagg ccattgtgat 16740

cacagactac aaaactacaa tgagcgcgaa tccggctgag tttatgcgcc tggcttacaa 16800

tcacggctat tatttgaaaa tggcgttgca atgtgattta ttcagaaaag cgtatccaga 16860

agaaaagcga cctatagttg tgcgactgtt agcgcaagag aagaaagagc cttatttgcc 16920

gttggctttc cgcatgaata gcgagcaatt gaaaatcggg cgcattcagt atatgagcgt 16980

aattaaccag tttgccatgt gccagcaaca tgacgtttgg ccttcgtatt caaacggcga 17040

gccggaggtt tgccttgata ctcctgattg ggtgcgacgc cagtttaagc aatatcttat 17100

ttaaacagca caaatagcta aacaaatgaa aaatgcggtg ttataattca acgcataagt 17160

taacaacaag cgtgaggaaa taaacatgga aaacaaacaa gtgtcagagg tggcagttca 17220

tatcaacaat tttgcaactg gcatggctat gttgcttcgt gactttgtag cgccacttga 17280

tccaacggca ggcgaagaag aaatggagta catcagaaaa gtaattgatg cggtcgataa 17340

cgtcgtactt gtcgcaacaa tgagcgaaga caacgaggaa gcaatcaagg ctgtaaaaga 17400

aagttctgat cgcatgatgg aaaatctgat taaattacac accgaagaag aaacaaagca 17460

ttaacagcaa taaggcggcg aaagccgcca tgaattacgg gaatcaatca tgaaactatc 17520

agagcaattc gacaaggttt taccagcact gcataaggcg cgcagtttgt ttgtaaaggt 17580

taagaaggat aagcaaaata cacacttaaa aaaccgttac gctacacttg atgccgttct 17640

tgatgcaatc actccagcat tgaacgataa cgagctaatg ttgatgcagg atatgattga 17700

aagcgaacaa cctaacagaa tcaaggttga gacaacagtt cttcatgttt ctggtcagtg 17760

ggtgaaattt tacgcagagc taccaatcgt taaaaacgat cctcaaggtg tcggatctgc 17820

attcacatac gcgcgtcgtt atgcagccgc cgcagcattt ggcttaagcc aggcagacga 17880

tgacgcacag attgccgtta aatctgcaaa cgactggaag cgtgacattg agaaatgcga 17940

aagcgtaaac gaattgcaag aagtgcttaa atcagcatgg aaggcaagcg atccggcaag 18000

caagcaggta attaaagagc attacgagaa gcgcaaggca gagcttgaga tcggcaaagc 18060

acgaggattt aatccagcgc aaccaaagca aaatcttgca tcgccagaag ttgacacaaa 18120

aaacgacgag caagtaaaat cgcaaagtat cactgatttt gaatgattta caggggcgat 18180

tgcgcccctt ttttacagga gaataaaaca tgcatattat tactggtgag atccgaaaag 18240

agccgcgagt aaagcaaatg cctaacggca gcacacttta tgtggttgaa ctttcagagc 18300

gatacaaaga taaagatgga aactggcaat atacaaacta cagtttcttt tttaatgcta 18360

aaacagaagg tcttaagggt tggtatgatg aggcgttcca ggttggcaag gttatttctg 18420

tatcttgcga cacgctgcgc attgaaacgc gtgaatataa cggaaagatg tattcaagtt 18480

taatgccagg aggatttgca aatcttatct tcagtcagcg cggggaaagc cagcaacaat 18540

atcagcagcg aacgcaaggg ggatggggcc aaccacaaca acagaatcaa ccgcaacaac 18600

cgcaacagcc aagacaaagc aatcaacctc ctatggactt tgatgacgac attcctttct 18660

gatcaacaag ggggcattac gccccttttt cactttcagc catcccaaga acggcaataa 18720

ccttaacagc aattcgcttt gcaagttcgg tttgatgagc gttaagtttt ccagtggtca 18780

ttattgacaa cattccggtc atacttccaa gcccgttaaa atctgacatt gccgatttca 18840

tgattgatat tggtgagtga ccttcatcag ctattgtatg cgctctctct gacaggtctt 18900

taactacctg atctattcct tcacttttcg tcattttctt ttccttcagg agttgattgc 18960

tgttctggca ttacaggcca cgggctaatt gattcccttt ctgctactgg cttttcaggt 19020

tcaattccta aaaactttcc tacaaactca ccaacaaaat ttgagctatc gcttacataa 19080

acatgcatcc tgtctagatc tgcaacagta acgctatttg gtgcgatcat gatcctctgt 19140

tggtgaaaac cgatatacat cacattagcc ggacgctcaa gagtccccat atttggcgca 19200

attacgccca ttatccaaca ataatgaatg ttgccatcaa gcggctcaca tcctgcgtta 19260

agccagttta ctttctggtg tgggaatgcc attgatgaat cagtagcatc acgccgtaac 19320

gaccatttaa cgccgccaat ctcaatgatt tgtttcggcc tcatcccgtt gaattgagtt 19380

acgccagcgc cattttgatt aatgttttgc atatcggttc accttgttac attgatttca 19440

ttttacgcaa tgcgcagata caagcggcgg caagatcggt taattctttc tctatgccat 19500

gatgcgagcc gtctgccttt tcggtcatta gctctttgta ttccatttct acaatcgaca 19560

ttaagccgcc tggcttgtca atgtgcgcgt cccaggtctt aggatggtgc ttagttcttt 19620

caataacact tcccataact gaatcatgtt tgctatctcg ctcataatca tcatcatgat 19680

catcactccc gattgaaact cgcggcaaat gaatgcgcgg tattttatag ccagcataac 19740

ggatcttcat tatctcgccc tcttgatgaa aagcgccccg aaaggcgctt gatttattat 19800

cagccgcaac agccagtagg cggagtctgc ggagtaggta acttgaagtt caccaattgc 19860

tctacctggt tgattttgca attcagtgcg gcggtctggt ttgcctgaga tagagcaaaa 19920

cgcgcttcct gcaactgagc tttcatatcg caaatcaaaa cagcctgaga atcagcaaac 19980

tgttgacgaa tcagatctcg cgttgcgtta ccttgtcgct caatgttcag gttagtctcg 20040

cagcaacatc tctccgctgc aatctgtgcc tgataggaac gctcaagagc attaacatca 20100

gcagcattaa tcgcagcaac cgtctggtta gtgccgctaa caatcgcggt attcagtccg 20160

gcgaaacctt gcaccgatgc aagcaggttt tgcgtattct ggctggtaat gccgttaaag 20220

gttgatgcgg cggagcgctc aacggcaagg tttgtggagt tttggccttg cagagtttgc 20280

agccccaggt tgtttacgcc tgtctggatg ttattaatac catcaagaac agcgttagca 20340

ccgacagcaa cagcagcgcc atcaccacca tagccaccac gaccattaaa gccgttacca 20400

aaccatgacc caatcaggcc accaacagca ccgccaagac cagccgcgcc agcttcgcca 20460

ccaaaaccac caccagtagg gagtaaagtc atatctgaca tagtaaattc ctctttgacg 20520

ttaaaaaaat taaaaaaagt ttgcaacatt tatgttacga caaagagtat gcgcccaatt 20580

accaggaata gatataaaat atttgtaagc gatatgtaag caactggcat aaagacgaaa 20640

aaagcgcagc atgagccgcg cttgagttaa aaatattgtt atttattgag cagtgcagca 20700

acagccgcct ttaattcgtc tatctccttt tgctggtttt cgatcattcc tcccatgtaa 20760

accatagcgc aagccatatc agccatgatc acgttgttat caagagcaag cgtatcatct 20820

gctttgtcaa ccacaatttg acgatctaca tattcttcac cttcatcatt aacgcctata 20880

acatctgcaa aaatgtcccc gccaggtatt agctttacat attcaggatc tatttttagc 20940

agatcctggg ctatcatccc gaatcgctgg atgttgcttc cattgtattt aaacattgtc 21000

ggaagccact gcataacgcg atcatagctt tcttttccat ctgtataaat tatgtcgtgc 21060

tttaaatctc tgtcagagtt agcggccttt tgaaaaatga aatttccact tgatgaccat 21120

gtggttatat cgtttgaggc tggctgaaac tggaagttac agaaaaaact accatcacca 21180

cgaagtctta acgagcaaaa aggccaagcg ttattgcctt gtgggataag accccatgtt 21240

gcgcgaattg gataaccacc tgttgattga gtaccccatg atagacctgg cgcccatcca 21300

ccatcattgt taggaataat ctcgccccaa acaagagcat taccccaata gttaggatca 21360

tcccagccca tttgctgggt tttgccagtg tattgagcag aatgcgttct gccgtcaggg 21420

taaaacttgt gcatatagct ttgcacgcca tgtttataca gttcaatccc aaacgactca 21480

atttgtgcag caccgcctct gatcaaacca gcgatagcgt aatcgttata ccagttgtaa 21540

cgacttacac cagtccatga gttcattggt ggattcatga ttgaatatag ttgtgacgga 21600

ttgtgttcat tcccgtctac ataaatacca gccccaggag ctaaggcaat acttttcgct 21660

cctgaaaaag caccattttc attaaactga taatactgtg ataacccatt atgaaatacg 21720

cctattgtat gctttcctac gcctacttga gtctcaaaat aagactgaga agacccaata 21780

tcttcgcctt gtgtactctt taattttgaa tggtatatac ctgcgtttgc agtagctgca 21840

tttccagttt cattttcaac attaaggctt aggaattttg gtgcatgttt ttcgccaagt 21900

cctatattgt ttctgatgcc atttgcgtca atagctccag taccaccaga agaaatagct 21960

aaaggcacag tttcaaatgc accagatcca gaaatatctc tctgaaaaac aagaagacca 22020

tcatttctga taacaagcct atattctcta tttctagagt acaagatagt atgtacgtaa 22080

tcctgaaaaa atgaatcaat actaagcgtc tggcgagcag attcaatgtc agaaagatct 22140

ttaagattct gatccttatg taatatattt tcaggattta cctgttgcgc ccactgtcga 22200

gcttcatcac gcgcagtttc agcgccagcc tttgcattat ttgcatcaac ggcatgattt 22260

ccagcagcat cacgcgccaa tacagcctga tcgcgagcgc cgaaagcatc ttgtttaatg 22320

gcttccgttt cgattatagc tgaatcttta atctgttgcg tttgcgcctt gattgcgtca 22380

gtgtctgatt taatctggtt tgtttcagca acagcagaat cctttatctg ttgagtgttg 22440

tttttcagtt gttcagtagc agcgcgatca ttggccgttt tctctgcatc acttttaacc 22500

tgattagcca gatcttgcag ctgctttagg tcaaagtttt tgaaaaattc aatagcttcc 22560

tcgatctgag tttctttgct ttgatagtaa cgcagcgttt ccgcaacatc ttgtgctagg 22620

ccgtcaacag tgattgagtc atgcagcaaa ataatgtagt tgcttcgccc aatttcagcg 22680

ccgccagttg tgatggcgcg aatttgtgta tcgctgataa cttcactgat cacggccgct 22740

tgaaatggag catcaatgaa aaagattgtt gcaccagggc gaatgagcgt tagttgctct 22800

cgccattttg tgtcgtatcc agtgatataa ccttgcgcgt ccattgacgc ttgtcctgtt 22860

ctgtaaattg ccatgcgttt tctcccgtgt tgcatgttaa tgtcaatgca gataatagca 22920

tttgctaagg aaaaaaaaaa ccccgcaatg cggggtaaaa gtagcagtga catttagaga 22980

agagattcag gtttgtaaac cttcctctct atcccttgcc tgtaattatc atgaatcggc 23040

atcataattt caagtcctct ttcctcggca gcaatgatcg cgtcacggtc tgtactctta 23100

cagacaactc gcattcttct gttgccttta tacctgtaag ctacaatttc aatgttagtt 23160

gcatcaaagc aggcccaaac ctcctgacca gttgcaaggt gaatatgttg cgcatcaatc 23220

cagtcaacgc aaagatagat cgtcttatct gtgcttcctg ttctaacaac cgacccccgc 23280

gtgtagtcct ttgcgagaaa agacttatta ccttcttcat caatgaaaag cacattgcac 23340

atttcatcat caatttcatc tgaatgtacc aggtaacatg ggagcgcgtg aatgtatcga 23400

ccatctttaa cgccgcaatc ataatgacca tcttcaggcg ggtatattcc ttcatatgtg 23460

cttaacggcg tgcgatcaaa ttttagtgtt tttgacataa cgcgctcgca gcattcatgg 23520

cttgcctgtt tcccaaacga ataaccagaa tcacgagata ctcgcttatt tgccttgatt 23580

ctatagtcct gcggaacttt tccaagaaaa cggcccaaaa tattgatcac ttcgctgtaa 23640

ggttctccgg ttagcttcat gatccaacca attccggtat cattaccgca cacgttgcag 23700

atcgcaccgc catcgccttt tttatcgcca agtttatccg tccatctgaa gcgatccttt 23760

ccgccgcagt gggggcaaga ttgatgcttg ccgttaaaaa cttcatttgg aagaccgcaa 23820

atactttgta gcgcctcacg ccacaatcct tgcatatatg ggagaacatc ttccttttga 23880

taaaccataa agttttcgtt gttcatatgc acctcaaaca aaaagcctga cagatgataa 23940

catcatgcca ggcattgttt ttagcaatta gtgctttcag gtttatacag gaatccgcca 24000

ggttcataat ctgacattct taccactcgc aacatttcac gcttatcaca tctctttgtg 24060

agcggttttc cattggaatc aaatcgcaga tctggacggc agaatgaagc gcgatatcct 24120

tgacagccat actttttgta atcgttatgc accttttgag cgccagctgc tgaaatcata 24180

ccgcgaactc gccactgatt tacagtttgc tggctaacgt taagacgttt tgccattgct 24240

gacacgctgc cgtaatactc aataagaata tcaagccgtg cttttagtcc ggcgcgcact 24300

tcatctttta gaacataata acctgtcggt cgcttgcgct tcttcttgtc cttgcctcgt 24360

cttgttccgt tattgccatt aattgtgcgt ttgtcaatct taccagttga tactgcgatt 24420

cgctgttctt tcattattca ttctccaata gcattttttg ttaaacgatg atttaatgat 24480

tgtgtattat acacgccaaa tggcattact caaaggtttt agaatgattc caaacattga 24540

aaagcaaata tcagcacttg gtgaagcggt aattaaatcc attcaggagc gattcactgt 24600

tggagagatt gtgccttacc cttaccagtg cgttgcatac gctgagatcg caaagcgtat 24660

gaaaaattat gagcatccat tctttgttaa agcgtccgta tcggctggca aaacattgat 24720

gtttgcaatg gttgctcacc agtgccggaa aatgggcttg aaaatgatgg tccttgctcg 24780

acaggctgag attgtggatc aggattctga agagataaca aatcttggtg tgccgaactc 24840

catctattgt gcaggactga agacaaaaag cgcatacttt ccgatcgttg ttggttctga 24900

gggaactgta gttaatggac tgtttaaagc gcttggcgac tatgttccgc acgttatcgg 24960

gatcgacgag tgtcaccagg tggattggga agaccttgcc gacgcaatag agaaagacga 25020

gtcattttta caaatgacaa ctaagaaagg cgaaaaagta ccaaatccag attatgacat 25080

aacaaaggga agcaggaaca gaaatacaga gttcctgatt ggtgaggatg gcttgccaat 25140

ggagggaaca ggccgcacac aatacactgt cattatcatg gaaatgatgc gccgctgccg 25200

gaaaacatat ggtcatgaac tgcgcatatt tggcatgact ggatcggagt ttcgcggcgt 25260

agtcccgatc ctggtagaag acaaaaaaca gaaaggattc tggcgtgaac aggttactaa 25320

tattgacacc aactacctga tcaaattcgg ttctgttgtg ccaacaaatt ttggcgacgt 25380

tggcgatcta gggtatgacc tttcagagtt tgaggcatcc agtgaggatg gcgttgcaga 25440

ctttgatgct aaaacgcttc gcaggatgga acaaaaaatt catgaagaag cgaccatgac 25500

aaagcgcatc atggcgaaag ttcacgagat ctgtaaaaac cgcaacggcg tacttgtgac 25560

atgtgcagga gaaaggcatt gcaaagaggc agcggcagca ttgccacctg gaacgactta 25620

caggatcatt actggtaaga ctggcgacaa tcagcgcaag gagtggttga aggaagcata 25680

cgaagggaag gtaaaataca tattccaggt tcaggcgcta actactggcg ttaacgtgcc 25740

gttttgggat acgtctgtta ttctgcgtaa gattggatcg ctaactttgc tgattcagct 25800

tttaggacgc ggaatgcgac ttcttaagaa atggcataag gagcaaggct tcaagaaaga 25860

tgatcacctg gttatggatt tttcaggaac gatggacgag ttaggtgagc tttattttga 25920

tccgatactg gaacaggcgc aacatcagaa gcgattcagg aacggtaaag atccgaagcc 25980

ttgtcctgtt tgcggaactc ttaatagcta ttatgctcgt cgttgtatgc acgtagacga 26040

aaacggcaac agatgcgagt ggttctttaa attcaggaca tgtgatgatc agattgaccc 26100

gcgcactaaa aagattattc agcgcggttg tggaacgaag aatgatattg cggccagggt 26160

atgcagacat tgcgacatgt cattgattga ccctaacgag aagttaagcg gaaagcacta 26220

cacaaaaaac gactggttcc aggtgcaatc tttctcggtt gacatgacga aaaaccagaa 26280

agggataata tttaattatg agctttccga tggcatagac actttcaggg cgagagagat 26340

atttttccct gaatctgaga gccaaatttg ccgcgcaaaa tggcgaagcg ttgctcttaa 26400

gcacatacca gatcgccgtg tcgctggcat ggtggcgagc tatcgtaatg cacgtaaaat 26460

catgcaatat gtgaatcaca tcatgccacc atcgcgtgta actcaccgca agactagcaa 26520

gggtgaagat aatttgtaca agaaggactt taattatggc aatgactgat aaaggcgatt 26580

atctggagtt ttacgaaaga gatccgacgg atacgctaaa ggaggagtca caccagatcg 26640

gggcgtttca atggttaact tatgctcatc ctgaatttct tgcctggcat actaaaaatg 26700

agggcgacaa ggggatcgcc accgcaatga tggatcagca agccggacta gttaagggcg 26760

taagtgattt catcatcctg attggattaa aaggccgcta tccttttgca gcaattgaga 26820

tgaagcgcgt caataagtca ggcaagggga aggcttcacc agtcagcaag gagcaaaaag 26880

ccttcttgcg ccgtgtgcgt gagcttggcg gattcgctgc cgtaacatac ggatataagc 26940

aattcatgat cgcggtagaa tacatgatga aatagcactt tttgttaaaa caggcgcgaa 27000

ggattgcgcc ataataacac cagttcaaca agcaacaggt aattaagaaa tgagtaaaga 27060

aactgaagtg acatttgaac aaatagaacg cgaaacattc attggcaatg ctcttgctac 27120

tggaggccat taccaagccg ttaaaccaaa tcaatattat aaagtcactg gcaaccgcta 27180

taacggcagc aatacgccgg acattgtgcg cgatttgtgg tctacgccgt atgagcttgt 27240

agcatggatg gaaagcgaat acggtgatta tgacatcgac gcggcggcaa gcaaagaaaa 27300

tgctgtatgt gaaaaattct acagcaagga aacaaattgc ttaaagcgtt ggtggggtag 27360

taacaagcat atatggctga atccgccgta cagcaatata acgcctttcg ttaagaaagc 27420

gattgagcaa atggagcaca ataaccagat cgacattctg ttaccttgcg acacatcgac 27480

aggatggttt tatgaggcac aacagaaagc agcggaaatc atctggatca ctggcgaagt 27540

ttaccaggag gacggaacag aatattcccg cactggacgc ctggcgttca cttcagcgct 27600

tacaggtaaa ccagttcagg gaaacaacaa aggcagcgtt attttcatca tgcgcgaact 27660

taaagaaggt gagcagcaga aaacgcgata cgttaaaatc agcgacattt gcccgtcagt 27720

ggcagagcgt cgcgcacgca aacggagcta aaaccatgca gaaagaaaga agagttttac 27780

ttaatgaaga tggcgaattt cttttgtatc gcgctatggt atgcgatgca cttgataaaa 27840

accctaacgt gaaaaagatg attgattgcg atccgtggga attttcaagc tcattagata 27900

tgagctttga ggagacaaaa aaactcccac ttgaaaaatg gcacgaacaa atcagaaatg 27960

acatgcaaga atttgtcgat tcgtggaatt ttcataacgg aatgatccac caatagcact 28020

ttttgttaaa acgcccggcc ttgtgtcggg cataattatt tcatcaaaac gaacaagagg 28080

aaaacaaaat ggttgtttac gatccacgcg cttttaaaat tgctcaagaa gtttcacgcg 28140

aatctgttgc tggtggttcg gttaacggct atcagtttga ttggtcggca gcaatgaccc 28200

tgcttaaggt tgcatatggt cacgcgccaa ttgaaacagc ggaggaatat tataagcatg 28260

aaggttgaac aaggtaggca ggccgtatgg gatcacgcaa aggagtgcgg gatctctgaa 28320

gacatagcca ggatagcaaa atattttgat attgctgaca taagcattat cagcggcgac 28380

aaaatgacat ttctcaatga acgaccgcgc aagatgcatc gcgtaccagc aataccaaca 28440

aagattgatt tcaaagaggc tatggcgaaa attcgcgagc cgcgcaaata ctacaaatga 28500

ggattattat catgtggcgt ttgttacttt tgcccttacc tgttatgatg gcgatctcta 28560

ttgtatacgt tgtcataatg agataaggag aatttttgat gaagcaaatc aaaatcacag 28620

atgaacaatt tatcaatgag cgcaagcagg gaaagacata taagcagatc gctaatgagt 28680

atggtatgaa cgttcgcagc attgagcgac gcgcggcacg attagcaaag caaggtaaag 28740

ttacaaccat cggatcacct ggttttggtg ttactggaga atcaaagcta attgataaag 28800

atggcaatgt ggttatgaca tggattaaga caagtaagga ccgcgaacag ttagaagcat 28860

taatgcaggc cgctatggat gcatttagcg aagaagttcc tcgacttgat ccacagccag 28920

aatcacaaaa ggattatagc gagacgttat cactgtatcc gatctttgac atgcacttgg 28980

gagcaatggc gcataagcat gaatgcggcg agaattggga tacagcaaca gcagagcgcg 29040

taatgaataa ttttattgat tattccatcc agtgcgcgcc ggatagtgaa aaagctgttt 29100

tgctgattgg cggtgatatg cttcacagcg acggactgga agcggtgaca cctgcaagcg 29160

gtcacgtatt agatcaagat agtcgatacg caaaacttgt ttatgttgcc atccggtcag 29220

tgcgtcgagc aatcacaaag ctgttatcaa aacacaaaaa cgtcgagatt cagattattg 29280

aaggaaacca cgaccaaagc ggcatgatct ggttgcgcgc agcaatggcg gcagcatatg 29340

agaatgagcc acgagtgcat gttgatgtat ctccgcgagt agttcatcac acacaatatg 29400

gcaaaacatt cctggcatac catcacgggc atactatccg caaacctgaa acattgctca 29460

tgatgtgcgc ggcagactgg cgcgaagact tcggcaattc aaaaatgatg tatgctcacg 29520

tagggcattg gcatcatcaa acagtaaccg aaacaagcct gggcattgtt gaagtgcata 29580

gcactatggc ggcaaaagat gcatatgctg cgcgcggtgg atggcgttct cgtcgccgtg 29640

cggctgttat tgtttacgat aaagaatacg gagaagtagg gcgatttatg cattatccag 29700

aaatggctgg ttgaactttt aatcattaaa atataaatat aaatcatcaa tttaattggg 29760

cgtggcattt tgcttcgccc tttttttatt cctttttgtg cgttcaatgg ttaggataca 29820

atcaattaat tgttgacttt tatctacagg agatctaatc atgaaagatt ttttaaacgc 29880

tgcaacttcc ggcactggcg gcgcgtcaat cactggcgca gtaactggtc aaacaactat 29940

cgcaatagcc agcttagttt tgatggctgc atttggcatg tggggcgctt atcttcgttg 30000

gcgtgatagt aaggcgctac gtgatgcgct tgaatgcggt gatattaaga aagctattga 30060

gatcagaggg aaataatgag tataaaacaa agagtgaccg ctgcggcttt tggcgttgct 30120

cttgccctta cttctccgct actggaggaa atagaaggag taaaacataa gccgtacaaa 30180

gatattgctg ggatatggac ggtttgcgcc ggaataactg gacctgacgt aatacaaggc 30240

aaaacgtaca cacaaagaga gtgcgatgca ctactggcaa agcatatcaa gatcgcaaag 30300

gatgaagttg ataagcgtgt aaaggttgac attcctgata caatgcgcgc agcaatgtat 30360

tcattcacat acaatgctgg aactggagct tttagaaatt caaccatgct taagctgatt 30420

aatagcggtc gttacatgga agcgtgcaat cagttatggc gatggacaaa atacaccaat 30480

ccaaaaacag gaaagaaaga aacgtctaaa ggactgcgca accgccgcgc ggtggaattt 30540

aaatattgca ttaaggatct gtaatcatga gaaagttatc agcaatcgcg atcgctgtta 30600

ttttgtctgg atgttcaagc gttacgccgc tgaccggatt aatcggtagt aagccggaaa 30660

tcacagcaca ggcaggagcg gagaacgtaa agcagaacgt tggcgttacc gcaaagcagg 30720

acaccagcac aaaacaggaa actacaataa aggaatctgc ggttgataag gtggacactt 30780

ccagcaagaa ggatttcact acgtcaacca ttcaggcaaa caccataaag gctgataaaa 30840

tccaggttgt gcagggtaat aacggcagat ggtacgaccc cataataata tgcgtggtag 30900

tgttccttgt gctgatttgt ctttactgga gggaaaagaa aaaggaggct taacgcctcc 30960

tttgttttat agtgaatcca tcaaatcttg aaagctgttt ccgctataat cattctcgac 31020

aagaatgtaa gcacctttct tctttatatc ccataacatt tcattgtatt cgtaacgatt 31080

tatttcaaac gtgctgccat cttctcttgt cgcagttcct ttgtcgttga aaacatcaaa 31140

agttacattg atgatttttg tctctttctt tgttactttg atagccattt ttaaaatcct 31200

catattcgtt tcgatgaaac aaatataccc gattcgagat cgggagtttt aacaaaaagt 31260

gctattcaat acgctttaca gtaaccaaca aaacgccgtc ctcatcacac aaattatgct 31320

ctgatatgtt aaacgctcgc atgtcagaat gaattagcgt taacattgcg tgcatgaaat 31380

cgtcatgtga gattgaactt acggcgcaga acttcaatat tttgtctatt agcatttgac 31440

gtaaacaatt cttcatcgct catttctccg atccttaaaa catcccttac ccattcgttt 31500

tgcatacctt ccaggctgta aagttcaaac gagaaggggc gatttgctga tcctttctta 31560

caccagaact ttccaccaac tccatcaaga taaccagtgg tcatcattct tgcgcaaaac 31620

tcttttgaaa cggcggttgc aaacattctc ggcgtaaatc ctgcgaccct tgcgagccgc 31680

tcacattctc tatgctgaaa tataaatttt gctatatgtt gacgtgtaaa ccattcgaat 31740

gactcacacc agcgatacag atccagaaga aacataatta ccccagcaat gttggattca 31800

cgaaaacaac gccattaatc acacaaatat aatttctttc ctcaagcata ggaagcagtt 31860

gcgtttccat gcgcttcatt acgccagcct gaccaacaaa cggcttaact tttcgtgcag 31920

cctcatatat ggcgcgcaca ttcaagatcc ccttgttagc cttaccgtgt cgagtgataa 31980

tctcgattag tttacccatt tcagcatcat caccagcata gccagaagcg ttggcggcgc 32040

taatgtatgt tttgctcaac tcgctaaaca tcaacagtgc ttcctgaacg gtttcaacct 32100

caatctcttt cgacttttga gggtatccgc caggattaaa ccaattgcgg atagtgtgca 32160

aaaccgaagc aatcctgatc acctgcttat ccattttgcc aagagcaccg cgcagcattg 32220

tatgcgaata ctttccgcca tcagcaaggt gcggctccat ctcctgacga gccatgttaa 32280

gaactcgcat tgctgatttg ctgatcttaa gctggatgtt tgattcactc atgatgttat 32340

ggatcagctt gaaatattgg cttttcagtt ccccatcaac aggctcataa gtagaatcac 32400

cgttatcatc aacaaacttt ctacgaccta aaaatgattc ctcacgcaca agcaaataac 32460

gctcacttac accgataccg cgcgatcctg catccatgat ggcattgatt gtttcatcct 32520

gggcaattac cgccatacag ccaagagctt taaaactcat attgttgtca gcgtttgcac 32580

gagctataga aacatgaccg tgatcccatg cttttaatac aagttcgctg ttggttttac 32640

gttcgctatt ggcatatgtc agacctaaaa ggctgttaat acttgttgcc tcgtctgaaa 32700

tcacagcaaa gttaccctgg cgattgttga ttcttgcaag tccttccggc gttgtgtcgg 32760

acacagggaa aacgatgtca cacatctttt ccagtttttc ttccagatct tctttctcct 32820

ggtacagaga ttccatatca gcgccggagc gctcgctttt catctctttt gcaagacctg 32880

ccaactttgc ctgaatcttt tttctttcct tctttcgctg atcattaatt ctttccactt 32940

cagcaaccat aggagcaagt gccaggctgt taacggcaga tttaccagtt gatggaggct 33000

ggcttgtgac aacgtaaaga gcggttggtt gatccgttcc gtgatactca accgtgaagc 33060

gtccaagcat tgcagcagac acgcagccga taaaatgcat atatgccgat gattcaggga 33120

actgaactga tcgagcaatg ttgcgcgcca acttaccaac aacatcaaca tcattgccaa 33180

gagaaattac agggtagcga tcgtttccgt tattaacatc attaacctca ccccaaaagc 33240

tggacgattg acggtatccg tttgcctgaa tggacacgcg caccggagac aatccttgtg 33300

catcagctaa ttcgataatt tgttttggtg ttagtttatt ggtttcaaac gaaaaattca 33360

ttgatgataa ctcctttttt gatgcggcaa taataccgca ccaatgctac ccgcatttaa 33420

caaaaaatgc tatttgatcc gctctatgct aactaagatt ttttctggtt cccttgcgct 33480

atagcaatgc gcggttacgg aatcagcaaa atcaaagtaa agtgatttgt ggtaagaaag 33540

accaatcagc catccaacca tcgttttctt tattggcctg actatatcgc caggcttgaa 33600

ggtttttgtt gccgaatgag tcaccttata gcgatattca gccttgatca cttatacata 33660

gcctcaaaca aatgacagcc accagcggaa aatccaactt cttcgcggta tagcgtccag 33720

cggtcgccag attcatcaaa aacataacca gcaacagcgc caagcgcgcg gccttgctca 33780

atctggtagc gcttcccttc tttgaaatgc tttttacacg ggatcttgtg atctacaaag 33840

gtgcatttaa tcgtttttgt cttgatatga cgatagtcag actcaacagc gttttgccac 33900

ttgctaccct ttgtaatgct ctttacctga aattgttcca actcgatcac atcatcgcta 33960

ccagcaattt tttcgtatac agctttataa cgacagttca tcaaaacgtt ggttttattg 34020

cagataactt tcatcgttta tttctccttt cgtttcggtg aagtgattat gccaggcttt 34080

tacacctggc ttttaacaaa aagtgctatt ttgcgtttgc ttcaaagacg gcgcgagcaa 34140

aacctcgcgg agtgagggag cggatcatct ttgtgcgaga tgattttccg ccagttttgg 34200

cccaaccagg attatccttg tcatcctctt taggaaggat tgttgcgcgc ggaggaataa 34260

caaatccgtt tccagtccaa aggcatgttt tcttaaagta ccgatcccgc gccggaataa 34320

tatcagggaa gtcgggatgc ttatcatctt ccggcagata gccagcataa tcgcacggat 34380

ggaacgtata atcaggcttg cgccataacg ttgacagctt gcctacagga ttctctatca 34440

tgtacggaac accgaaataa tcagcgatat gcgccgcaat tttgcaagtg tcggcggctt 34500

catattggaa gttaggattt tccatgcgct ttttagccca atgacgagat ccgctatttg 34560

ctaattgtgt gcaaggcgga aatgccataa tgaaatcagg cttaccccaa ttttcaagtt 34620

ttgcggtata tttgaatagc tcatcaatcc acacattaac gtaaaaaata ttgtcgtgga 34680

ttatgcgcac tgattggtaa tcgccatgat ctgcatcatc gtaattaaag caaatgcatt 34740

tatatccagc atcagcccaa tctttaacgg ccttaccaga tccatcaaac aaagagaata 34800

ttaaaccttt atacatttca tcacctcaaa acggaaagca tctaggacag ttaggatcaa 34860

aattgcaacc acaatcatta acaatcatag ttggatctgc aaatgcagcg ccgcattcat 34920

ggtccatatc catttcaccc aacgcatcat caagcgtcct gattttatat gcgacttcaa 34980

gcgcggcctc tttattcagt ccggcatctt ctgctttttg cagtcgctca aaaaacccgt 35040

cttgactcat ttaataaact ccttaataac agcaataata attacaagcc cagcagtaaa 35100

aacaagcaat ccgacaaggc caatcataac accaaataaa atatcaagca tcttaatcat 35160

ttattcacca ttgataaaac gggaggcggc aacctcccat tgattaatca tacagccttc 35220

cagctttcaa taaacatgct gttttcttcc agcagtttaa tcaattcttc gcgtgtatat 35280

ttgcggattt tactacctgg atgtgagcca acaacaaaca cacctttacc actcgtgatg 35340

gttaactggt tgattccgca agatttatcc atcttaatgg cgacgcggtg atgtgcatca 35400

acagtgtgca tgatgttagt gactttagct ttcattatgc gatcctcatt gcacttagtg 35460

tagggcatta tatttacttc aaattctcga tcttgttctt tccaattgag ttgtgaacgc 35520

gatgttcttg cgtctcttta tctccatgaa catcaatata ataaccaggt atggcagtga 35580

aaaaccaatt tccacgccag aaaaaataaa ggccgtgatt gcttcctttc gcgcatttgt 35640

ttgcattttt aggtattgga agtccgtgta ctctccgcaa ttctacattc ttgttaaaat 35700

ttggcattac acaacctcca taaaagtatt gaatccacgc agtttgaaag cattgccatg 35760

tttaacaacg ttccatgtca ggccggagtg atacgcaacc atatcctcaa gatcaatggt 35820

gatgacagtg cctttcttaa agattccttt ctcgtgagtg ttgtcacgct gtacaacttt 35880

aacatcaact tgattttcca tttttaaatc ctcgttcgtt gtcgatgcgc ttaatatacc 35940

aatcagcagc gcatcggttt taacaaaaag tgctattcaa caacagatcc agttaaagag 36000

ataagctgcc ttgtgatctt gtgtctgcga tactggcgcg caacaaccgg aagattttca 36060

ccgctttgat gaccgaagca gatcggcaac tcaccctcaa gagagcggaa gccgccaaat 36120

tcaagatccc gcactggctt tgagcgccac gcttcaataa atccgcgttc gtcacgcgtg 36180

atatagcggt atatgttagg gaggggcaat tgtatcccct caaaatccac agtatcggta 36240

tagctgtaaa acttagaact catagtcaac aaccccaaat tcgttaacaa tttgcgcctt 36300

atagaagccg ccatttttcg ccaggttgta gcaggccgct acagtttcaa attcgcgcgc 36360

ttccggctgg ctgttttcgt gttcccaggt aatcagagta accattttca aatcctcgca 36420

ttcgttgtcg atgtgtgtaa tatatcggat tgcgaggatt gagttttaac aaaaagtgct 36480

atttagtcca ggagttcgat ttcgcgctcg ccagttaacg gatctttaat ccagaatcca 36540

agcgcctcac cgtaaatacc gccatagaat ccaagcgccg acagatcctc acctttaaca 36600

aataccagat ccccgctacg gcttacgtga gatacgttaa cgataaccgg aaaggtgcga 36660

tccttaaatg caggccgcag agcgtaacac ccgttattga gcaaacgtgc tttgcgctta 36720

actggaatag gtgcattgat tcgcgccatc tttatttctc cttcattgtt gatcgtgttt 36780

aacggccatc agaacatcac aggactcgtt aatgaattta gccatgcgtt tatcatcacc 36840

aacagggatt agcgcgttaa actcgcgttt gctactgaat accgtcactt tatgaaacga 36900

tgcgaacggc atcgacggga actcgactgt ttgcatcaag atccgcaatc ccatcttttt 36960

ggcccttacc tgggcctcgg caagttcaaa atccacttcg ctaatcattt cttgatctcc 37020

tcagcaataa acttcaggac tttgcgcaaa aacacttcat tattgatggt cgctggcaat 37080

tcctctttca cttcttgcgg cagatagata ttgccgtgcc aggtcaaaca atcgtagttg 37140

atacgataag agcaaagcca ctcgcaatcg ttatctgctt cccaaatgca gagttgcaga 37200

acatcgccaa aattctcaac agtcaggtcc agaccatgac gcttggcgaa gttaaaagtg 37260

gttttgctga tgttcatttg ctttctcctg atgggtatct cgtttcgatg tgtgtaatgt 37320

acccgatcag gatgattcag ttttaacaaa aagtgctatt tctcaactgg cgattattcc 37380

agctattcca ttaatcagga ataacaagaa taactcattc gcgatcacct ggattgtgag 37440

actttgagtt tgatcaattt taatcaagtg attaactttt tgtgtggagg ggatagagaa 37500

acgcggtcct cttgcatcct ttccggcaat cctggcgcgc aacatgttca ttatgtttcc 37560

agccgttcac gcaaataagc ctccgcagtt cttacacgtt tcatgatgtc atgcacacaa 37620

aagaaacaaa gtaaaatcta tataaataca cattaaatca ataagttagt tatatatgta 37680

tttatatttg tttcttattg ttcttgttgt gtgatggtta ttggtgtatg ttgcgatttt 37740

tgttttggtg gatattgatt gtttattgtt cgatttggtt ggtgatcatg attgctcatg 37800

gttgcgtatg attcttgatg tatttatgta tatatccgcg aaaattcgtg aacagccaca 37860

cagccaaaaa acatggcgag gattcttttg aaatcatgtg cttaagtgtg aattttgttt 37920

ccgcgcaatc tccacacgta tagcaaacaa ttagaacgct tgacatagac acggcgcggt 37980

ggcaacattg caatcaccaa caaacaaggc aaataggagt tacactcatg actatgacaa 38040

gtaaagaatt ttgcgatatg ttgcgagtta atgcagcctc actagctaac ggggatgtta 38100

gcgaactgca aatcaaggag cttgacttac aatgtaatat tgcgccggaa gacttgagga 38160

aaattaacgc ctcatacatc agaacgatta tgaatcgaac gccggaggtg aaagctgttg 38220

gcgcggtgaa ggttgagcgc gttcgaggtg atgcggatat tcctgattgc tatcgcgtaa 38280

ccattaacca gaatccaaag cgccgcttgc tgacagatga agatctgcca aaacttgaaa 38340

gtaagtggcg agcaaagttt attgctcaac tactgaaaac acagccgcga atcagtgatc 38400

tggaaggtga gcgcctggaa ggtgcagcaa tcgcacttga gcgaatgaat gaaatgctca 38460

acgagatggt aaaggtgggg gatgaataat gaataaggca ttaattggag ttttgattgc 38520

aattgccacc ctgaccggat gcgaaaaggc atctgaatca acagttgtcg taggagtgga 38580

gtcagatttc aaagtcggtc gcctgtttac tgttgatgga tgcactgcgt accgattcta 38640

tgacaacggt cgcgcgatct attacaccaa ttgtaatggc gcaactgata gcacataccg 38700

cagcggcaaa aacgtaatgc accaacaagt cacaacagat attaaatagc actttttgtt 38760

aaaactcaaa aacgataacc aggcataata gcctcatcga aacgagattg aggaaaacaa 38820

aatggcacgt cgcatcacca aagaccttaa agtcctgaat aaagaaaacg tagttaaaat 38880

cctggttatc tggggctaca acgaagaaac agcaaagcag aaggtagagg caggttatga 38940

cctggctgta aaagcaatgc caaacgacga cgccaaaggc attgcaaact atgtagcttt 39000

cttttaatta gcaggtgaat aaatgaaaac gaaattagtc cataaatcag aaatcaagat 39060

cggcgacact gtgattcaca acagcgagct tagaacggtt ggcaaagagt caatacgcca 39120

cgatgaattt atggggatct tgttgttcgg tgactcttat cggcttggct ataaaatggt 39180

tgagcttgtg gaagaggtaa aattctaaat agcacttttt gttaaaactc aaaaacgata 39240

accaggcata atacatcaca tagggcggca cggcgtcgcc cagttaaacc aatcaggaga 39300

aagtaccatg ttaaaattgg ctgacatcaa attcccgatc accttcgaat ctcgcggaat 39360

tggccattac acgttcacag ataagaacac ttgtcataag ttttggtcgc atagcgacaa 39420

gccagagtta tcaactatgg aaattgagca cttcattgat gcgcataata gcctttatgt 39480

gaaaacgaag gactatggct acgcaatgcc aggatacact tgtttgcact acaaaccata 39540

taaaggccaa gtgttaccag tgcgctcaac tgagcgatca gagattcata cggtcgagat 39600

gaaaacctgc gactacaaaa agctgtggtg cttaagtgtg gacaatatca agtatccgat 39660

cacgttctac acggttggta attcaacgat tcgcttcaca tcgcgcgaca atggcgttta 39720

tagcgatggt cagaaatcgg agttgaaagt tgatttcttt gttaatggtc acaacaaatt 39780

caatccgcac aatgaaaagg ataaggtgta ctacgtggaa gatattacaa aacacttgtt 39840

tggagaaaca ccattcggcg gcggaaagac gtcaattcaa caaatcattg gcgaggagat 39900

ccacaaggca gccgttaaaa tgaaagcgcc tggaggctgg cgtattgaca agagcggctt 39960

tgccaatcaa gaaatcgtaa cgcagggcga aatcgaagac atgccgctcg gcgcttcaat 40020

catcggcatc agagacaata acggcgactg gaatgatgtt tcattccctg tcaagcggat 40080

cgagttgcac gaagacatga tttgcatcat ggcgcatcat ggtagctata aggacaaacc 40140

attcgccgca gagcttaaca tcaaacgcgg aacgattgtt aagtggaagt ttaaagtgta 40200

aatagcacga attgttaaaa ctcaaatgtg gggccaagta taatggcctc actttcaacg 40260

aaggggataa aacgatgtac attagcaaga agatgaaatg cgtttccgtt aattatggtt 40320

gtgctggaat gttcaagcct ggcgaaatct atacggcgca aaagttgaaa tcatcaaccg 40380

gatcttttta cgtcagcaac agtaaagggc atcgcatgtt cctgaacggc ggcgaaggca 40440

ctaaggttat ggcgcacgct atggtgattg ctgaatttga ggaagtgaaa gatggaaaca 40500

aataagcgcg ttaaacacaa gtttattgat aatcatggcg atgcacttga ggtatacaag 40560

ttcggcgatc gcgtgtttat tgatggcacg tttcaaggtg gaagcatgcc ggattttagg 40620

tcaatactca ccattgacca ggtgcgcaag ctggcggata agttaaatga tctggcagat 40680

gaaatcgaat acaagcaaat tcattttaat tgataggttt gataatgaaa aaatacgaat 40740

ttgaactttg gggcagcaaa taccattttt ccacgagcaa gccgattgtt atcgttgacc 40800

tggacggaac gttatcagac ggatcgcatc gcttgcacct gcttccgaaa gaagatttgc 40860

accttacaga aagctggtct gagttcaata aggcggctgt tggcgattcg ccgattaata 40920

gcacggttgc agtggttaac ggtttgtgga tgtcaggatt tgccatcgtt atcctgaccg 40980

gacgcagcga cgaagtgatg gcggatacct gcaagtggct tagtgaaagc ggcgtcaagt 41040

acgacgcgtt aatcatgcgc cgcaaggaag acaaccgcaa agacacgatc atcaaagaag 41100

aagtattgcg cgctattggc ctggacaaca ttgtgtgcgc ttttgatgat tcacctaacg 41160

tggttaagca cttccgcagc ctgggaatca caacctatca ggtcacggaa tacgataagc 41220

cacataatca tatccaatcg catggcgtgg aggaattgca gaaatgatcc tgaaagtagc 41280

atatgctaac cctcgcggct gcaatatata cggctttaag gttggcgcga tggtcaacgc 41340

caggataaac gaaaaaggcg tattagttgt caagtctcct aaaactggca tggatatgcg 41400

catcaaggag aacggcggaa gattgtttat caggtcaatg gctggtcaaa cgctcgcaac 41460

gttcacaaaa tagcactttt tgcaaaaaca gattgtcggg cttgcggtat agtaagccca 41520

tcgacaacaa acgaggatta aatcatggtt aataaagttg agtttttcaa agcggtagac 41580

aagattgacg gcgcgcaaag tttctttgct atccattcca ttgacggcga agacgttgca 41640

tacaacgtga ttagtcgaga cggtatatca aaacttagtg acctggtggg ttatattgtg 41700

gcgaacggga aatgtgcatg gcccgatttg ccacgagagc cgcgccgccg agacatgcaa 41760

aatccagtcc tgctttacac gctggtaatt gaaggagtca acaaatgatt tggttacttg 41820

tatttttggt ggttttcttt tatatttcag gcttattcat cttccgtgcg ctggtgaaaa 41880

catgcgattg cacggataaa gatcagcctc ttgttctgat gttctggttt gtgtggtttt 41940

gggtggcgct ttaccagatt gtacgggatg aaagcggctt taagtggtga ttcttactat 42000

agcgcattca cgcagtgcgc tatgttgaga taacaacatg agatcatgaa gatgcgaaac 42060

ttcgaaaaaa atcgtatcca aaaagaagcg atacagctac cacgagcacc aagagatgaa 42120

tcaacgctat cgcaataagc gccagagcaa aggtaaacat tccggtaaac atggagaaaa 42180

tcaatatgaa acgagacatt atcatcctta acggaccgcc aggagttggc aaagatacgc 42240

ttgcggcata cttaacgggt catcgttacg cagcggttaa agcgtcgttt aagcaaccga 42300

tgtttgatat tgcattctcc atgcttggcg tttaccgata tgacgagttc attgatctgt 42360

ataacgaccg cgagcagaaa gaaaagcctc aagcaatctt gcaaggtaag tcgccgcgac 42420

aattcatgat ctggattagc gaagaagtca tgaagccagc ttttggcgag caatatttcg 42480

gaaatcgcat ggttgagcaa gtacacgaga tgtacagaga tttggctgtt gtaattagtg 42540

atggcggatt ccctgaagaa atcaagccgc tcgttaaggc aggccatgag gttcacattt 42600

gccgactgca tcgcgaagga ttcacgtttg atggagattc gcgaaactat attgacctga 42660

gcggatacca tcacagagtc aaacattacg acttcacaat gactgatggc gagccagaaa 42720

agactgttga cgagatcatc aaaactgtcc agtggaaaca cattaagatc aaatagcacg 42780

aattgctaaa agcggggcgc aatgcctcgc tataatgcaa acaccaatca acgagaggaa 42840

atcatcatgg tagagttaaa cgacattcgc gttggaacaa aatttcgcgt aacctgggca 42900

gataaatatt gcggagttag caagggccag gttgttactg tggatagtat ttatcgcgga 42960

tgcagcaaag actttcgccg cccacgcatc aagaatggct atatcattac tcgacgtttt 43020

ggctttgagt gttattgcgt agtggcgact caaggagttt tgattgagct tcagcgaatt 43080

agcgatcatc gcggctgtca tgtcaaaaca acgaagatac catctcagcg atcgcgatac 43140

gatgcgcgac ggatgcgccg actcgctcga aatgcgatca agttcaagaa gcatggtggt 43200

aacttttacc gaatgtacaa agggattgcg cgaaatgctg gaaaataaca acaacgcaac 43260

caacccaaag ccgccatttg attttcatga aatcatgatt gtgttattct tgattctctt 43320

agttgtttat tcagtgaggg ccgcaatatg gtttttaatg cactaaaacg aatctttaaa 43380

cctgaatacc ggatcgttgc ttgcacagaa tgcaagattt attatctgca acgccgccgc 43440

gttattagtg gggattggga atatctgacg gatgatgctt tcggcttttg ggctatggag 43500

ttcaatagct acagcgaagc gcgtgagtac gctaaaaagt tgcacaagtt cagaccttac 43560

aagctgcgtt tgctggttgt caaaaaatgc gatgtgtggg atcacgtatg aaaagagtaa 43620

ttataacttt tattgctctt atcgctggtt ctcacttggc ttatgggtat agcggcattg 43680

tcgagtcaat cggcggaatg attatcttcc tggctggcgc atgttacggc tacggcgatg 43740

gatttaaaaa tggcaagcgt tacatcctga ctggcaagaa ataatgttaa ccgcgccgct 43800

tcggtggcgc taaaaggagg aaacaaaatg gctaaaagca atcgtaaaat gttgattaac 43860

gcattcgaaa aggcagcgct tgagcgtggt tggaatgact catttcacac agcaaactgt 43920

atccgccgtt acggattcaa gaattgccgc tcctgggcgc gtcaaatggc ttcatggtat 43980

gatcttgatg cgcattacct tgatatggat tgcgccctgg ttgaaatgat tgagcaggca 44040

gcgttagagg atcgaccatt aacgcaatcc gattttgacg attttgttcg cgatgaaatc 44100

tattacatga gctaatagca ctttttgtta aaacttccgc gagggcattt gttaaagtgt 44160

cctcattgaa gcgaaacaga ggaaaacaaa atgaaaattc gaatcaccag ggttgacatt 44220

gaccgcaatg acggaagcat tacccttgag caatgcggat tcaaagttgg tgacattgta 44280

gaggttgacg gattctttcg cgatggttct tactgcgtgt tagcaatccg caacacggaa 44340

gaaattcgaa tcggtgataa catcggcgta aaccatgacg aatgcgaggt agtagaggaa 44400

tgatcacgat taacctttca gaagagcaag caaagaaact cctgcatgct gtaggttctc 44460

gtgcactttg cggatcaact gatgaaatga tgattgatca tgaggttgca agggagcttt 44520

ttactcagct tgagcaaaaa ttctcaccac gtccgactga cacggcagaa actgcaaaat 44580

ggcttgctga gaatggatac acggctttaa aagttcacaa ctggaagcga aaaacgaagc 44640

gttggagaat gctggatttt aattaaaatt gaatgcctca tacagcgaac aggacgcatt 44700

ttgacgaggc attttttagg taaggtgatt caatggataa ctacataaaa ccgtcgcagt 44760

ggtgcgcaca gaagcaagag gaagcattag agcgcggcga tactgaaaca gcaatgcact 44820

atttcgaaat gtacaacttg tggaagggta gaggtctgta atgtttggaa tgaacgaggc 44880

gcaattcaac gctgcaaaac gccaggctaa gaagtgcggc gaggcaatga aggcggatgt 44940

tgaaaagcgc gggaagtatg ttgatgaagt catgaagggc attattgccg aacattatca 45000

gccagttgcg ccaatgctga caattaccca attcatctgg ttgtgtggtt atcttcgtgg 45060

tagatgggga aacgcatttg accgcgaatg attgacaacg atttcctgat tggttagcat 45120

attcagcgtt tacataagag cggcattagt gccgaacggg tggcgcggct gccacgcaac 45180

aacggcgata tgcaggtttt taccgtaaga cgtatcgcct ttttaatagg tgaatatatg 45240

gaaaagtaca aattaacttt catgtttgac aatatggtga tgaatccctt aaatggttac 45300

gcttgcaata gcttttctgt taccgtgaat gcggatgaaa ttgaggatga tggcaaattt 45360

atcctaatag gtgattacat ttatcctatt cacaatatca attgcatcaa acgcgagatc 45420

gtaaaagatg aagtacaatg taagattgac gatcaggcga atggggcgta attgctcgtc 45480

atgtagacag agttttgaat gtgaggttga ggcgcgcgga gagcttgagg cagcagcaag 45540

agcaaaagag ttgtccggcg ctaatcaaga cactcaccag ttttcaatag atcttgtaag 45600

gaaaatatca tgagttctgt aattattggt ttggttgctt taatcattgg tgttctaatc 45660

ggctttgtcg ctgtatcttt cgtggtggct gttggtgttc gctataagtc aaaaaaaggt 45720

gaatttgctt tagcgttttg ggatgaagag cagaaagttt ggcaggttcg cggtcactat 45780

ttatcaatcg gtggcaggat tcacagcact atgaaaacaa atccagaaaa agtaaaatac 45840

aagtattgac caactacccg ccaaatggcg ggttttttat tgcctgcaat ttgtctataa 45900

tgaggctttg ttataaagga gggcgcatta tggctaaaaa cgttgagcca gcagtaaagg 45960

cttgcaattt taaaaaacta tataataagc aatatggcga tatagcaaaa atgacaaagg 46020

cgcacaagta cacgccggaa caggtgtttg atctggctgt tcgttacttt acgtgggctg 46080

aggaaaacca tatacaggct gcggaaacgg catcctttca gggtgacgta tacgaaagca 46140

agattcacaa gccgcgcgtg ttcaccttaa acggattccg attattcgct ggcctttctg 46200

cctccgtgct tgaaaaatgg cgacgcgagc caggatttag cgatgttatg gactttatcg 46260

acggcgttgt ttacgagcaa aaattccagc ttgctgcaaa caatattgtt aacgccggat 46320

tcattggtaa agagatcggc attgaaaagc cagccaccgt taccattgag aataacacaa 46380

gcgccagcgt tgacgctgta acggcggagg aagtcaaaga ggcggtaatt gacattctgg 46440

agaagatata atgctgattt gggaagactt aacagcggcg caaaaacgcg ccattaagga 46500

aatgagcgaa tattcattcg aaaagatgat ccgcatttgg tttcaacttc tccaggggca 46560

aaggttttta ggaaactggc atttcagcta cctatgcagc gaagttgaaa aaatcatcaa 46620

aggcgagtca cagaatgtaa ttttcaacat aacgccaggc agtggtaaaa cagagatatt 46680

ctcgatccac atgtcacctt atgcgtcatt gaaaagtaag aaggtgagaa atcttaattt 46740

gtcgtttagt gatggacttg tccagcagaa tagcaaccgc attaaggaga ttattggttc 46800

tccagaatgg caggagctat ggcctggaaa gttggcaaag gcgagcgcaa aggatttgat 46860

tgtaacagat ggcggcaagg ttaaattaca agtcaactcg cgttctattg gtggtcaggt 46920

tactggtctg cgtggcggct acatggacga tggttttact ggtatgctgg ttcttgacga 46980

tccagacaag ccggatgaca tgctttctaa ggtgaagcgc gaagccggac acatgcgcct 47040

aaagaacacg gtgcgatcgc gtcgaatgaa ggacaccacg ccaattgtga tggttcaaca 47100

acggctacac gtcaatgatt caacgtggtt catgacaaac ggcggcatgg gcggcatcca 47160

gtttaaggtg gttagtattc cggcgctcgt tactaaggaa tatcgcgaaa cgcttcctga 47220

ttggttaaag cctgagtttg atcgcgatgt tttgtccagt aatccagtga tgattgatgg 47280

tgttgctcat tattcgttct ggcctgctaa ggaaagcgcg gaggaattac ttgcacttcg 47340

aaatgctgat ccgtacacgt tcgccagtca gtatcagcaa cagcctgttg cgcttggtgg 47400

taacgtgttt aaaacagagt ggttccagta ttacggcagc agcgagaaat gcacgctacc 47460

aaaaccggat cgctttgaat atacatttat cacggttgat accgcgcaga aaacaggtga 47520

gcttaacgac tactccgtga tctgttattg ggggatgtat aaagatcgtg tctacttcat 47580

tgatggcgtt cgcggaaaat gggaagcacc ggaccttgag acgaattttg ttgcattcgt 47640

taatcagtgc tggaagcgca ataaagaatg cggaacgttg cgaaggattc atgttgaaga 47700

taaatcatct ggcactggct tgattcagag tgcagcgaaa aaaatcatga tcaagatcaa 47760

tcccgttcag cgcgataaag acaaggttac gcgagcaatg gacgcagcgc cagtaatgcg 47820

cgccggacgc gtagcacttc cagaatccca tcctatgctg gctgagatac tagctgaagt 47880

tgcggcgttt acattcgatg atagccatcc tcatgatgat attgttgata acatcattga 47940

tgcggttaac attgaaatga acatggcaga tgatccggtt ggaagaatga aaaaattggc 48000

aggtttgcgg aacaaatagc acgaatcgac aaaccaatat ataatcaaag gctggaaaat 48060

ccagcctttt ttattggagg aaagataatg agtaaatcac gaatcgttaa ggctgatggc 48120

tataacgaga tctttaaagg cgagaattgc aaatcgcgag tagagcaacc attctacatg 48180

caatcaatgc catacaagac acttgctgac ttttacgaaa aagacggcct ggcaaaaagg 48240

gttattgacg tagtaccaga ggaaatggtt tcccctggct tcacggttga cggcgtggca 48300

gacgaggcag cattccgttc tttgtgggat gagaagcgat tgaatgcaaa aattattgat 48360

gcgctttgct ggtcgcgttt gtttggtgga tctgcaatca ttgctcttgt tcaggatggg 48420

cgagcgctca aatcacctgt aaagcctggt gccatgttgg aggatgtgcg agtttacgat 48480

cgctatcaga ttcgcgttga agcgcgcgaa actaatccac gcaaagttcg ctatggtgag 48540

ccagtgcttt acactgtaac gccaggcgga gacttgccgg aataccaggt acactacacg 48600

cgagtttgca ttatcgacgg tgagcggata cctaacgcaa agcgacgcag caatgacggc 48660

tggggtgagt cagttcttaa caagcggctt gttgaggcta ttgttgatta taactattgt 48720

gaaacattag cgacgcaatt attgcgaaga aaacaacagg cagtatggaa ggccaaaggt 48780

cttgctgact tgtgcgatga tgaagaaggt gttagcgcgg cacgcttgag acttgcgcag 48840

gttgacgatg agggcggcgt aggcaaggcc attggtattg atgctgagga tgaggaatat 48900

gatgttctta actctgacat tagcggcgtt gattcttttc tggaaaagaa aatggatcgc 48960

atagtttccc tttcaggtat tcatgaaatc atcctgaaaa ataaaaacgt tggcggcgtt 49020

agcgcaagcc aaaacacggc gcttgaaaca ttctataaac ttattgagcg aaaacgcgtt 49080

gaggattaca agcctatcct tgaatttcta ttaccgttca tcattagtga gcaggagtgg 49140

agcattgagt tttcaccttt gagcgtgcct agcgataaag atcaagctga gattctgaat 49200

aagaatatcg actcaatcag caaggctatt gatggtcagt tccttgatgt tgaagaagcg 49260

cgcgacacgt tgcgagcaat cgccccaagc gttaaactaa aggacactaa taaaatcaag 49320

ctgccggagc caactgagcc ggagccaggc acacagggga atgagtaatg aaagtgaaag 49380

gcgttgttaa acaatggcgc tttcctgaag caagcgagcg gcaatttagc cgctcaattc 49440

aggaggcaat cagagatctt gtagttctca tgcgcaagcg aacaaaagcg atgaagtttg 49500

acgcaacaga taatgagatc aacagcgcag aagatgaaat taacagcatg gcaactgacc 49560

ttatagctgg cattgtttcc actcttcctg caatcgcgtt gaccatttat aaattcaatg 49620

cgaagcagtt tatcaatgtt gccaagtcaa ctggcggaaa ggataatacg gcggtaattg 49680

ttttgatcgc ggttggcgct aacgctaacg aggattggta tcagacgctc tacggtcaat 49740

ggcacgggct tacagaatca tcgctaagga agttattcac aaacattgtt tctgattggt 49800

caacaaacat tcgcaatgca aactttcgcg gaagcaatga taaacaggtt aatgatcttg 49860

cagaaaagag atttgctgtt tatagctctt ggggtaaaac gagatcagaa aacattatcg 49920

gagcgtggaa tagtcgcctg atgcgccagc gcctttatga tgctaaggta actcattatt 49980

tctggcatgg aatgttagat gacagagagc gcttgcagca tgtgttatgg gaaggtaaaa 50040

gaatagcgct tgatgcaatt catgatttcc ctggtgagcc gtggggttgc cgttgctggg 50100

cgattccaga ttggaatagt aaaggagaat aattaatgaa ggcaaaacaa agatttgata 50160

gcgtccaggt taaggcgcat tttgatgata atggcttttt agtggatcgc ccaattgtgg 50220

cgcgaattgg tttgcaggag taccgcacac cgtatggaat cagacgagag ttcagaccag 50280

cgtccgaggt gttcaaggct gattcccttg ccacattcgc aggtaagccg atcactatcg 50340

gtcacgtcac ggtaacgcca gacaatgcgg atcaggttgt tgttggttca tgcgctggtg 50400

ctggtgttcc aaatggtgtt ggcgttgaag ttccgttaag tatttactca aagcgagcaa 50460

ttgagagcgc aaaaaagaaa gacacggcgg aaatttcagt tggttacaca tcaattgata 50520

ttgataaacc aggttggggc aataataaaa ctggcgatta cttgtttgag gaagacatta 50580

aggaagactg gaagcctgat tcccctgatt gggttaaatt tgacgccatt cagacgaata 50640

tccgtgttaa tcacattgca cttgtctttc gtggtcgtgc tggcattgcc aaattaaatc 50700

ttgatagtga acaagatttt ccgtatagtg acgaggttct aaatgacaaa ggagatgagg 50760

aaatgacagt aaaaattaaa cttgatggcg cggtagaatt tgatgttcca aaggccgttg 50820

ctgatcatat tgaggcgtta aaggcagatg caaaagctgc aaccgaaaaa gctgatggcc 50880

ttgaggccga gcgcgacgcc ctgaaaacta aagtcgatgg cattccggcg cagattgcag 50940

aagctgttaa gaaggcaaag actgacgcag aagaacacgc caagctagtt gcagatgcaa 51000

ctgaaattgg catcaagtgt gacggcttgg acgctaaagc gatcaaagtt gcttacgtca 51060

aagaagtcac tggcgctgat atttcagaaa aagccgatgc at 51102

Claims (5)

1. a kind of Styloviridae phage E sc-COP-9 (numbers of being commissioned detached from natural environment：KCTC 13131BP), It is characterized in that：

Carry the ability with specific killing Escherichia coli, and the genome marked with sequence number 1.

2. a kind of, for preventing, enteropathogenic E. Coli from infecting and the composition for the treatment of enteropathogenic E. Coli infection, feature exist In：

The composition is with phage E sc-COP-9 described in claim 1 (numbers of being commissioned：KCTC 13131BP) it is active ingredient It is included.

3. according to claim 2 be used to prevent enteropathogenic E. Coli from infecting and treating enteropathogenic E. Coli infection Composition, it is characterised in that：

The composition is used with manufacturing the purposes of feed addictive, drinking water additive or disinfectant.

4. a kind of prevent that the side infected as caused by enteropathogenic E. Coli is infected or treated as caused by enteropathogenic E. Coli Method, it is characterised in that：

Including：It will be with the phage E sc-COP-9 (numbers of being commissioned described in claim 2 or claim 3：KCTC 13131BP) The step of composition being included for active ingredient is supplied to animal in addition to a person.

5. according to claim 4 prevent from infecting or being treated by enteropathogenic E. Coli as caused by enteropathogenic E. Coli The method of caused infection, it is characterised in that：

The composition is supplied to animal in addition to a person with feed addictive, drinking water additive or disinfectant applications.