KR101971572B1 - Novel Salmonella Enteritidis specific bacteriophage and antibacterial composition comprising the same - Google Patents

Abstract

The present invention relates to a novel bacteriophage KFS-SE having a killing effect specific to Salmonella enteritidis, a pharmaceutical composition for preventing or treating infectious diseases caused by Salmonella enteritidis including the bacteriophage, an antibiotic composition, , A feed, a disinfectant or detergent, and a method for preventing or treating an infectious disease caused by Salmonella enteritidis comprising administering the bacteriophage to an individual. The method for the prevention or treatment of infectious diseases caused by Salmonella enteritidis using the bacteriophage KFS-SE of the present invention is more effective than the conventional antibiotics such as Salmonella enteritidis alone and has excellent lytic activity and physicochemical environment It has the advantage of being strong. In addition, the bacteriophage KFS-SE of the present invention has an advantage of being able to solve the problem of antibiotic-resistant bacteria caused by misuse of antibiotics, the problem of remaining antibiotics in food, and the problem of a wide host range. Therefore, the bacteriophage KFS-SE of the present invention can be used in various fields in the field of prevention or treatment of infectious diseases caused by Salmonella enteritidis, composition for antibiosis, composition for feed addition, feed, disinfectant or detergent, It has the effect of contributing to the improvement of competitiveness and the public health.

Description

[0001] The present invention relates to novel Salmonella enteritidis-specific bacteriophages and antibacterial compositions comprising them.

The present invention relates to a novel bacteriophage KFS-SE having a killing effect specific to Salmonella enteritidis, a pharmaceutical composition for preventing or treating infectious diseases caused by salmonella including the bacteriophage, an antibiotic composition, , A disinfectant or detergent, and a method for preventing or treating an infectious disease caused by Salmonella enteritidis comprising administering the bacteriophage to an individual.

Salmonella spp. Is a generic term for fungi with a certain similarity, which in 1885 was recognized by DE Salmon and co-investigator Theobald Smith in commemoration of the discovery of porcine cholera bacillus, (1924), which is the name given to it. Salmonella is a common bacterium commonly found in water, soil, insects, plant surfaces, kitchen surfaces, animal feces, uncooked meat, poultry, and seafood, and is widely distributed in poultry and pigs.

Salmonella is a gram-negative, tuberous anaerobic bacterium belonging to the intestinal bacteria and is a bacterium that does not form apoa. Salmonella is a pathogenic microorganism that causes diseases such as food poisoning, gastroenteritis, sepsis, enteritis, salmonellosis or typhoid by infecting various livestock including pigs, cattle, chickens and others as well as humans. In more severe cases, pneumonia, arthritis, Lt; / RTI > Salmonella spp. Was serotypes more than 2,500 species reported to date, species that host specificity of the animal (eg, Salmonella Thai Phi (Salmonella Typhi in person), Salmonella two Dublin in cattle (Salmonella Dublin), pigs the number of salmonella cholera device (Salmonella choleraesuis), Salmonella flow column (Salmonella Pullorum) and Salmonella Galina volume (Salmonella gallinarum)) as a species no host specificity of chicken (Salmonella typhimurium (Salmonella Typhimurim), Salmonella Entebbe utility disk ( Salmonella Enteritidis), among which the serotypes causing human food poisoning through livestock are the latter.

These Salmonella strains infiltrate into cells and multiply and infect when infected with animals. Therefore, it is difficult for antibiotics, medicines, and other probiotics to penetrate and act, and Salmonella isolates are also often resistant to antibiotics and medicines It is difficult to prevent. In particular, Salmonella can cause serious economic losses to farms by causing high or low rates of infection in domestic or poultry because they cause vertical or horizontal infection through contaminated individuals.

Currently, antibiotics are mainly used to prevent and treat infections of Salmonella enteritidis. However, in many fields, problems related to antibiotic resistance have been caused, and the cause of the infection has been identified as abuse of antibiotics and the use of antibiotics has become insufficient. As a result, antibiotics used as growth promoters in industrial animal feeds began to be banned. In the EU, in 2006, Korea began regulating the use of antibiotics in the second half of 2011. Consumer demand for safe food without antibiotics is also increasing, and the use of antibiotics will gradually decline. Therefore, research and demand for control methods based on environmentally friendly materials that can replace antibiotics are expected to increase. One of these studies is to use bacteriophage, a virus that exists only in natural environments and kills specific bacteria.

Bacteriophage refers to a bacterium-specific virus that targets and infects only specific bacteria and inhibits and inhibits the growth of infected bacteria. Of the Staphylococcus bacteriophage origin by British bacteriologist Twort 1915 years cocci (Micrococcus) colonies began a study from the melting phenomenon transparently by something. In 1917, the French bacteriologist d'Herelle discovered that there was an effect of dissolving Shigella disentriae in the filtrate of heterozygous patients, and by studying this, they discovered bacteriophages independently and eaten bacteria Called bacteriophage in the sense of. Bacteriophages have the ability to kill bacteria by infecting bacteria, multiplying inside bacterial cells, and destroying the cell walls of the host bacteria when the progeny bacteriophages come out of the bacteria after proliferation. The bacteriophage infection method of bacteriophage is highly specific, and it is advantageous that a specific bacteriophage kills only a specific bacterium and does not affect other bacteria.

The research using bacteriophage has been continuously studied only in Eastern Europe and the former Soviet Union due to the emergence of antibiotics, but recently many problems of existing antibiotics such as the appearance of super bacteria with multidrug-resistant due to abuse of antibiotics have been highlighted, Friendly materials that can be used in the future. In particular, development time is short compared with antibiotics, development costs are low, and superiority in terms of high selectivity, persistence and side effects is recognized.

Therefore, it is absolutely necessary to study a novel bacteriophage for preventing or treating infection caused by Salmonella enteritidis, which causes enormous economic damage to the poultry and animal husbandry industry. Therefore, bacteriophage having a lytic activity specific to Salmonella enteritidis, It is necessary to develop the technology for the application.

Accordingly, the present inventors isolated bacteriophage having a lytic activity specific to Salmonella enteritidis and analyzed the morphological and genetic characteristics of the isolated bacteriophage to confirm that the bacteriophage can selectively kill Salmonella enteritidis, Thereby completing the invention.

It is an object of the present invention to provide a bacteriophage KFS-SE (KCTC13147BP) having a killing ability specific to Salmonella enteritidis.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating infectious diseases caused by Salmonella enteritidis including the bacteriophage KFS-SE.

It is still another object of the present invention to provide an antibiotic composition, a composition for feed addition, a feed, a disinfectant, or a detergent comprising the bacteriophage KFS-SE.

It is still another object of the present invention to provide a method for preventing or treating an infectious disease caused by Salmonella enteritidis, which comprises the step of administering the bacteriophage KFS-SE to an individual other than a human.

In order to achieve the above object, the present invention provides a bacteriophage KFS-SE (KCTC13147BP) having a killing ability specific to Salmonella enteritidis.

The present invention also provides a pharmaceutical composition for preventing or treating infectious diseases caused by Salmonella enteritidis, which comprises the bacteriophage KFS-SE.

The present invention also provides an antibiotic composition comprising the bacteriophage KFS-SE.

In addition, the present invention provides a composition for feed addition comprising the bacteriophage KFS-SE.

The present invention also provides a disinfectant comprising the bacteriophage KFS-SE.

The present invention also provides a detergent comprising the bacteriophage KFS-SE.

In addition, the present invention provides a method for preventing or treating an infectious disease caused by Salmonella enteritidis, comprising the step of administering the bacteriophage KFS-SE to an individual other than a human.

The method for preventing or treating infectious diseases caused by Salmonella enteritidis using the bacteriophage KFS-SE of the present invention is highly specific to Salmonella enteritidis as compared with a chemical substance such as existing antibiotics, and is infected with Salmonella enteritidis It has an advantage that it is excellent in lytic activity and strong against physicochemical stimulation by proliferation. In addition, the bacteriophage KFS-SE of the present invention has an advantage of being able to solve the problem of antibiotic-resistant bacteria caused by misuse of antibiotics, the problem of remaining antibiotics in food, and the problem of a wide host range. Therefore, the bacteriophage KFS-SE of the present invention can be used variously in the field of preventing or treating infectious diseases caused by Salmonella enteritidis, antibiotic composition, composition for feed addition, feed, disinfectant or detergent, There is an effect that can contribute to health improvement.

Brief Description of the Drawings Fig. 1 is a graph showing the result of checking the activity of the bacteriophage KFS-SE for Salmonella enteritidis.
2 is a diagram showing a genetic map of bacteriophage KFS-SE.
FIG. 3 is a graph showing the morphological characteristics of bacteriophage KFS-SE through an electron microscope.
4 is a diagram showing the results of SDS-PAGE of the protein pattern of bacteriophage KFS-SE.
5 shows the results of confirming the pH stability of the bacteriophage KFS-SE.
6 is a graph showing the results of confirming the thermal stability of the bacteriophage KFS-SE.

Hereinafter, the present invention will be described in detail.

The present invention provides a bacteriophage KFS-SE (KCTC13147BP) having a killing activity specific to Salmonella enteritidis.

In the present invention, the term " bacteriophage " is a bacterium-specific virus that infects a specific bacterium and inhibits and inhibits the growth of the bacterium. The genetic material of single or double-stranded DNA or RNA is a capsid protein The virus is surrounded by the virus).

The bacteriophage KFS-SE of the present invention has excellent lytic activity against Salmonella enteritidis. The bacteriophage KFS-SE was found to belong to Siphoviridae having a head (71.429 nm) and a relatively long tail (211.429 nm). As a result of protein pattern analysis, 120 kDa, 64 kDa, 48 kDa, 38 kDa 35 kDa protein and showed similarity to the previously reported bacteriophage base sequence as a result of the base sequence analysis but it was confirmed that KFS-SE was a newly isolated bacteriophage due to the absence of 100% identical bacteriophages in all the fragments . In addition, the bacteriophage has excellent stability against heat and pH.

The salmonella enteritidis-specific lytic activity, acid resistance, basic basicity and heat resistance of the bacteriophage KFS-SE of the present invention can be used for the prevention and treatment of infectious diseases caused by Salmonella enteritidis, and for the effective use of the bacteriophage KFS-SE It is possible to apply various temperature and pH ranges when applied to various products including as a component.

The bactericidal activity of the peel KFS-SE of the present invention is maintained in the range of pH 2.0 to pH 11.0, preferably pH 3.0 to pH 11.0, but is not limited thereto.

The bacteriophage KFS-SE of the present invention maintains the lytic activity within a range of 2 ° C to 60 ° C, preferably 4 ° C to 60 ° C, but is not limited thereto.

The present invention also provides a pharmaceutical composition for preventing or treating infectious diseases caused by Salmonella enteritidis, which comprises the bacteriophage KFS-SE.

Since the bacteriophage KFS-SE of the present invention has a killing activity specific to Salmonella enteritidis, it can be used in a pharmaceutical composition for preventing or treating infectious diseases caused by Salmonella enteritidis.

In the present invention, infectious diseases caused by Salmonella enteritidis are concepts collectively referred to as infectious diseases that occur epidemic or acute, and may be at least one selected from the group consisting of salmonellosis, food poisoning and enteritis. The salmonellosis includes symptoms of infection by Salmonella and infection by Salmonella.

Infections caused by Salmonella can cause diseases in humans or animals by contaminated food, water, utensils and various infectious agents. Salmonellosis has gastrointestinal symptoms such as fever, headache, abdominal pain, diarrhea, nausea and vomiting after a latency period of 12 to 36 hours. Dehydration occurs, and infants and elderly people can become severe, and acute symptoms disappear in 1-2 days. Anorexia and diarrhea last for 7 days. Sometimes it starts with acute enteritis, and it causes sepsis, abscess, gastritis, arthritis, cholecystitis, pericarditis, pneumonia, syphilis and pyelonephritis.

The pharmaceutical composition of the present invention contains 10 ³ to 10 ¹⁰ PFU / mL of bacteriophage, preferably 10 ⁶ to 10 ⁹ PFU / mL of bacteriophage. The term " PFU (plaque forming unit) " used in the present invention is a unit in which a bacteriophage digitizes a frying pan appearing after the death of a bacterium.

The term " prophylactic " of the present invention means any action that inhibits the disease or delay the onset of the disease by administration of the composition.

The term " treatment " of the present invention means all the acts of improving the symptom of the disease or inhibiting or alleviating and beneficially altering the disease upon administration of the composition.

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier.

As used herein, the term " pharmaceutically acceptable carrier " means a carrier or diluent that does not irritate the organism and does not interfere with the biological activity and properties of the administered compound. Examples of the pharmaceutical carrier which is acceptable for the composition to be formulated into a liquid solution include sterilized and sterile water, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

The pharmaceutical composition of the present invention can be used as a method of applying or spraying on a diseased site, or may be administered through nasal spray, oral administration or parenteral administration. In the case of parenteral administration, intravenous administration, intraperitoneal administration Administration, intramuscular, subcutaneous or local administration.

The appropriate application, spray and dose of the pharmaceutical composition of the present invention may vary depending on factors such as formulation method, administration method, age, body weight, sex, disease symptom level, food, administration time, administration route, Response sensitivities and the like, and usually a skilled physician or veterinarian can easily determine and prescribe dosages effective for the desired treatment.

Examples of formulations for oral administration comprising the pharmaceutical composition of the present invention as an active ingredient include tablets, troches, lozenges, aqueous or oily suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or elixirs It can be formulated into a zero. A binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin, an excipient such as dicalcium phosphate, a disintegrating agent such as corn starch or sweet potato starch, Calcium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. In the case of a capsule formulation, in addition to the above-mentioned materials, a liquid carrier such as a fatty oil may be further contained.

The parenteral dosage form containing the pharmaceutical composition of the present invention as an active ingredient may be formulated for spraying, such as an injection form such as an intravenous injection or an intramuscular injection, or an aerosol formulation capable of being inhaled through a respiratory apparatus . For formulation into injectable formulations, the compositions of the present invention may be formulated as solutions or suspensions in water with stabilizers or buffers in water, and formulated for unitary administration of ampoules or vials. When formulated for spraying, such as an aerosol formulation, a propellant or the like may be formulated with the additive such that the water-dispersed concentrate or wet powder is dispersed.

The present invention also provides an antibiotic composition comprising the bacteriophage KFS-SE.

In the present invention, the term " antibiotic composition " means a preparation which can be given to an animal in the form of a drug to kill the bacterium, and collectively refers to an antiseptic, a bactericide, an antibiotic and an antibacterial agent.

Since the bacteriophage KFS-SE of the present invention has a higher specificity for infectious diseases caused by Salmonella enteritidis than conventional antibiotics, it does not kill useful bacteria, can kill only specific pathogens, does not induce drug resistance or resistance, It can be used as a new antibiotic with a longer life cycling than conventional antibiotics.

In addition, the present invention provides a composition for feed addition comprising the bacteriophage KFS-SE.

Antibiotics for feed additive used in animal husbandry and fisheries are used for the prevention of diseases. The administration of antibiotics for prevention is problematic because it increases the possibility of producing resistant bacteria and antibiotics remaining in the livestock can be delivered to humans. If antibiotics are absorbed into the body through meat or fish, they may cause antibiotic resistance and spread the disease. In addition, since there are many kinds of antibiotics fed in feeds, there is a problem that the probability of occurrence of multidrug-resistant bacteria is increased. Therefore, as a new natural food additive antibiotic material which is more environmentally friendly and solves the problems caused by the use of existing antibiotics, -SE can be used.

In addition, the present invention can provide a feed comprising the feed composition, and the feed of the present invention can be prepared by separately preparing the bacteriophage as a feed additive, mixing the feed into the feed, adding the feed directly to the feed, Can be prepared by direct addition. The bacteriophage in the feed of the present invention may be in a liquid or dry state and may be used according to the intended use. Drying methods include, but are not limited to, air drying, natural drying, spray drying and freeze drying. The bacteriophage of the present invention may be mixed in powder form in a composition ratio of 0.05 to 10% by weight, preferably 0.1 to 2% by weight of the feed weight, but is not limited thereto. In addition to the bacteriophages of the present invention, the feed may further include conventional additives that can enhance the preservability of the feed.

In the composition for feed addition of the present invention, microorganisms other than non-pathogenic microorganisms may be further added. Examples of microorganisms that can be added include lactobacilli such as Bacillus subtilis capable of producing proteolytic enzymes, lipolytic enzymes and glycosyltransferases, lactobacilli having physiological activity and organic resolution at anaerobic conditions such as bovine, ( Lactobacillus sp.), Filamentous fungi such as Aspergillus oryzae , which increase the body weight of livestock and increase the milk yield and increase digestion rate of feed, and Saccharomyces cerevisiae and yeast such as S. cerevisiae .

Examples of the feedstuffs containing the bacteriophage KFS-SE of the present invention include cereals, rootstocks, food processing Busan Logistics, algae, fiber products, pharmaceutical Busan Logistics, fats, oils, Include, but are not limited to, proteins, inorganic materials, fats, mineral oils, fats, single cell proteins, zooplankton, and residual foods.

The composition for feed addition of the present invention may contain a binder, an emulsifying agent, a preservative, etc. added to prevent deterioration of quality. An amino acid, a vitamin, an enzyme, a probiotic, a flavoring agent, A nitrogen compound, a silicate, a buffer, a coloring agent, an extracting agent, and an oligosaccharide. In addition, a feed mixture may be further included.

The present invention also provides a disinfectant comprising the bacteriophage KFS-SE.

The disinfectant containing the bacteriophage KFS-SE of the present invention having a specific killing ability against Salmonella enteritidis can be usefully used as a disinfectant for hospital and public health to prevent hospital infection, and can be used for general disinfectant, food, Disinfectants, poultry farms, buildings such as farmhouses, shafts, water bottles, litter, stagnation, transportation vehicles, dishes, etc.

The present invention also provides a detergent comprising the bacteriophage KFS-SE.

Since the bacteriophage KFS-SE of the present invention has a specific killing effect on Salmonella enteritidis, it can be used also for washing the surface of skin or parts of the body of an animal which is exposed to or exposed to Salmonella enteritidis .

The present invention also provides a method for preventing or treating an infectious disease caused by Salmonella enteritidis, comprising the step of administering the bacteriophage KFS-SE to a subject.

The method of the present invention includes the step of administering bacteriophage KFS-SE to an infectious disease caused by Salmonella enteritidis, such as at least one species selected from the group consisting of food poisoning, enteritis and salmonellosis. Infectious diseases caused by Salmonella enteritidis, which can be treated in the present invention, are not limited to the general symptoms of food poisoning such as vomiting, nausea, abdominal pain, hemorrhage, chills and sepsis, acute enteritis, abscesses, gastritis, arthritis , Cholecystitis, pericarditis, pneumonia, pyoderma, pyelonephritis, and the like. In freshwater fish, it also includes bacterial sepsis, such as swelling and protrusion of the eye, protrusion of the anus, skin and muscle ulcers, blood vessel hemorrhage, and ulcerative dermatitis.

In the present invention, the term " individual " may include both humans and animals other than humans, and preferably refers to livestock such as pigs, cattle or poultry, but is not limited thereto.

The term " livestock " is a concept that refers to useful animals that are refined and improved by humans to co-live with humans, including birds, fishes, insects, and mammals. The livestock are pigs, , Horse, duck, and dog, but is not limited thereto.

The term " poultry " is a concept that refers collectively to animals belonging to the domestic livestock, and the poultry may be selected from the group consisting of chicken, duck, pheasant, quail, ostrich, goose and turkey, but is not limited thereto .

In the method of the present invention, the bacteriophage KFS-SE or the composition can be administered to an animal in the form of a pharmaceutical preparation, or mixed with feed or drinking water of a livestock and fed to the animal.

In the method of the present invention, the administration route of the bacteriophage KFS-SE or composition can be administered through various routes of oral or parenteral administration as long as it can reach the target tissues.

It is obvious to those skilled in the art that the appropriate total daily dose of the bacteriophage KFS-SE administered to the method of the present invention can be determined by treatment within the scope of sound medical judgment. The specific therapeutically effective amount for a particular individual will depend upon the type and extent of the response to be achieved, the age, weight, general health, sex and diet of the animal, the time of administration, the route of administration and the fraction of the route of administration, It is preferable to apply differently depending on various factors including drugs used or co-used and similar factors well known in the medical field.

Terms not otherwise defined herein have meanings as commonly used in the art to which the present invention belongs.

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

Example  One: New  Isolation and culture of bacteriophage

1.1 Salmonella Entertidis ( Salmonella Enteritidis ) And screening of single bacteriophages

Sputum analysis was performed to screen and isolate bacteriophages specific to Salmonella enteritidis. 25 ml of washing water obtained from the chicken embryo processing process of a chicken processing factory in Sangju, Gyeongbuk province was prepared as a sample solution, and a soft agar overlay method was performed using the same. The soft agar overlay method refers to a method of observing that a bacteriophage lyses by using a host cell growing in a top-agar (pasted on a solid medium using 0.4% agar).

Concretely, 25 ml of the washing water sample and 1% shaking culture of Salmonella enteritidis ATCC 13076 (OD ₆₄₀ = 0.135) were mixed in 225 ml of TSB medium and cultured at 37 ° C for 18 hours. Then, Centrifuged for 10 minutes, and the supernatant was filtered with a 0.25 [mu] m filter. Then, 4 mL of TA soft agar (0.4% (w / v) agar, nutrient medium, NaCl, MgSO ₄ , CaCl ₂ ) and Salmonella enteritidis ATCC 13076 shake culture solution (OD ₆₀₀ = 0.465) 200 mu] L was added to the mixture, and 10 mu L of the sample solution was added dropwise thereto, followed by incubation at 37 DEG C for 18 hours to confirm formation of lytic group. The result of confirming the formation of the fryer was shown in Fig.

As shown in Fig. 1, a bacteriophage forming a fungus was identified, and it was confirmed that the bacteriophage was a bacteriophage having a specific fungicidal activity against Salmonella enteritidis.

Further, since it is known that one kind of bacteriophage exists in one bacterium, it was attempted to separate a single bacteriophage from the formed bacterium. Specifically, the lysing medium was placed in 900 μL of SM buffer solution (50 mM Tris-HCl, 100 mM NaCl, 10 mM MgSO ₄ ) and left at room temperature for 2 hours to obtain a bacteriophage solution.

1.2 Bulk culture and purification of bacteriophages

In order to mass-culture the Salmonella enteritidis-specific bacteriophage identified in Example 1.1 above, Salmonella enteritidis ATCC 13076 was used to culture a large amount, and the bacteriophage was purified therefrom.

Specifically, salmonella enteritidis ATCC 13076 was shake-cultured and dispensed into 3 mL of TA culture solution (nutrient broth, NaCl, MgSO ₄ , CaCl ₂ ), and cultured for 2 hours. The bacteriophage obtained in Example 1.1 was divided and cultured for 2 hours, followed by centrifugation and filtering to isolate only the bacteriophage. In the same manner, the amount of TA culture was increased (3, 8, 20, 200, 800 ml order). Then, polyethylene glycol (PEG) and 1M NaCl were added thereto so that the final concentration became 10% (w / v), and the mixture was allowed to stand at 4 ° C for 16 hours and then centrifuged at 8,000 rpm for 20 minutes at 4 ° C. ≪ / RTI >

The purified phages were suspended in SM buffer (50 mM Tris-HCl, pH 7.4) at a CsCl density gradient ultracentrifuge (density gradient: 1.20, 1.30, 1.40, 1.45, 1.50, 1.70 g / mL; 22,000 rpm; 100 mM NaCl, 10 mM MgSO ₄ ) and stored at 4 ° C until use.

The newly isolated bacteriophage was named "Bacteriophage KFS-SE" and deposited on November 7, 2016 in the Korean Collection for Type Cultures (Korea), and deposited with the deposit number KCTC13147BP received.

Example  2: Bacteriophage KFS Gene analysis of SE

In order to analyze the entire gene of the bacteriophage KFS-SE purified in Example 1, the bacteriophage was extracted using Phage DNA Isolation Kit (NORGEN, Canada). The extracted gDNAs were analyzed using the Illumina MiSeq platform and the open reading frames (ORFs) were analyzed using GeneMarkS, Glimmer and FgenesV software. Genetic analysis results are shown in Table 1, and a genetic map of bacteriophage KFS-SE is shown in Fig. The entire gene sequence of the newly isolated bacteriophage KFS-SE is shown in SEQ ID NO: 1.

Query Name Query Length Subject Name Subject Length Alignment Length Query Start Query End Subject Start Subject End Hsp Score Hsp Expect Hsp Identities Percent Match gene_1 | GeneMark.hmm | 95_aa | + | 614 | 901 95 gi | 509139264 | ref | YP_008058158.1 | helix-turn-helixdomain-containingprotein [EnterobacteriaphageChi] 95 95 One 95 One 95 490 7.61E-63 94 0.989473684 gene_2 | GeneMark.hmm | 861_aa | - | 942 | 3527 861 gi | 509139265 | ref | YP_008058159.1 | putativeprimase [EnterobacteriaphageChi] 861 861 One 861 One 861 4592 0 854 0.991869919 gene_3 | GeneMark.hmm | 81_aa | - | 3524 | 3769 81 gi | 509139266 | ref | YP_008058160.1 | hypotheticalproteinchi_043 [EnterobacteriaphageChi] 93 81 One 81 13 93 426 2.15E-53 81 One gene_4 | GeneMark.hmm | 138_aa | + | 4048 | 4464 138 gi | 526176475 | ref | YP_008240301.1 | hypotheticalproteiniEPS5_070 [SalmonellaphageiEPS5] 138 138 One 138 One 138 670 5.32E-89 137 0.992753623 gene_5 | GeneMark.hmm | 109_aa | + | 4526 | 4855 109 gi | 526176474 | ref | YP_008240300.1 | hypotheticalproteiniEPS5_069 [SalmonellaphageiEPS5] 141 109 One 109 33 141 564 3.10E-73 108 0.990825688 gene_6 | GeneMark.hmm | 435_aa | + | 4878 | 6185 435 gi | 451937753 | gb | AGF88050.1 | hypothetical protein SP124_00330 [Salmonella phage FSL SP-124] 445 435 One 435 11 445 2313 0 435 One gene_7 | GeneMark.hmm | 198_aa | + | 6240 | 6836 198 gi | 414090197 | ref | YP_006990314.1 | hypotheticalproteinSPN19_067 [SalmonellaphageSPN19] 198 198 One 198 One 198 1039 3.26E-143 198 One gene_8 | GeneMark.hmm | 679_aa | + | 6903 | 8942 679 gi | 414090196 | ref | YP_006990313.1 | putativeDNApolymerase [SalmonellaphageSPN19] 679 679 One 679 One 679 3660 0 678 0.998527246 gene_9 | GeneMark.hmm | 95_aa | + | 8944 | 9231 95 gi | 414090195 | ref | YP_006990312.1 | VRR-NUCdomain-containingprotein [SalmonellaphageSPN19] 95 95 One 95 One 95 507 2.05E-65 95 One gene_10 | GeneMark.hmm | 509_aa | + | 9224 | 10753 509 gi | 414090194 | ref | YP_006990311.1 | SNF2domain-containingprotein [SalmonellaphageSPN19] 509 509 One 509 One 509 2739 0 508 0.998035363 gene_11 | GeneMark.hmm | 189_aa | + | 10740 | 11309 189 gi | 414090193 | ref | YP_006990310.1 | putativeterminasesmallsubunit [SalmonellaphageSPN19] 189 189 One 189 One 189 1008 6.49E-139 189 One gene_12 | GeneMark.hmm | 693_aa | + | 11293 | 13374 693 gi | 414090192 | ref | YP_006990309.1 | putativeterminaselargesubunit [SalmonellaphageSPN19] 691 691 3 693 One 691 3732 0 691 One gene_13 | GeneMark.hmm | 84_aa | + | 13385 | 13639 84 gi | 414090191 | ref | YP_006990308.1 | putativehead-to-tailjoiningproteinW [SalmonellaphageSPN19] 84 84 One 84 One 84 450 4.51E-57 84 One gene_14 | GeneMark.hmm | 559_aa | + | 13636 | 15315 559 gi | 269975281 | gb | ACZ55505.1 | phage portal protein [Staphylococcus phage SA1] 559 559 One 559 One 559 2982 0 557 0.996422182 gene_15 | GeneMark.hmm | 423_aa | + | 15357 | 16628 423 gi | 526176464 | ref | YP_008240290.1 | putativeproheadproteaseClpP [SalmonellaphageiEPS5] 428 423 One 423 6 428 2246 0 422 0.997635934 gene_16 | GeneMark.hmm | 134_aa | + | 16658 | 17062 134 gi | 526176463 | ref | YP_008240289.1 | putativedecoratorproteinD [SalmonellaphageiEPS5] 139 134 One 134 6 139 696 6.29E-93 134 One gene_17 | GeneMark.hmm | 354_aa | + | 17075 | 18139 354 gi | 525972490 | ref | YP_008239955.1 | capsidproteinE [SalmonellaphageFSLSP-088] 354 354 One 354 One 354 1887 0 354 One gene_18 | GeneMark.hmm | 97_aa | + | 18201 | 18494 97 gi | 414090186 | ref | YP_006990303.1 | hypotheticalproteinSPN19_056 [SalmonellaphageSPN19] 97 97 One 97 One 97 480 2.92E-61 97 One gene_19 | GeneMark.hmm | 121_aa | + | 18497 | 18862 121 gi | 414090185 | ref | YP_006990302.1 | hypotheticalproteinSPN19_055 [SalmonellaphageSPN19] 121 121 One 121 One 121 618 1.45E-81 120 0.991735537 gene_20 | GeneMark.hmm | 208_aa | + | 18862 | 19488 208 gi | 414090184 | ref | YP_006990301.1 | hypotheticalproteinSPN19_054 [SalmonellaphageSPN19] 208 208 One 208 One 208 1084 7.49E-150 208 One gene_21 | GeneMark.hmm | 167_aa | + | 19485 | 19988 167 gi | 414090183 | ref | YP_006990300.1 | hypotheticalproteinSPN19_053 [SalmonellaphageSPN19] 167 167 One 167 One 167 871 9.81E-119 167 One gene_22 | GeneMark.hmm | 379_aa | + | 20002 | 21141 379 gt; < RTI ID = 0.0 > 379 379 One 379 One 379 1982 0 379 One gene_23 | GeneMark.hmm | 153_aa | + | 21238 | 21699 153 gi | 414090181 | ref | YP_006990298.1 | hypotheticalproteinSPN19_051 [SalmonellaphageSPN19] 153 153 One 153 One 153 788 1.57E-106 153 One gene_24 | GeneMark.hmm | 65_aa | + | 21744 | 21941 65 gi | 414090180 | ref | YP_006990297.1 | hypotheticalproteinSPN19_050 [SalmonellaphageSPN19] 65 65 One 65 One 65 338 1.10E-40 65 One gene_25 | GeneMark.hmm | 1431_aa | + | 21934 | 26229 1431 gi | 414090179 | ref | YP_006990296.1 | putativetapemeasureprotein [SalmonellaphageSPN19] 1431 1431 One 1431 One 1431 7430 0 1426 0.99650594 gene_26 | GeneMark.hmm | 562_aa | + | 26235 | 27923 562 gi | 414090178 | ref | YP_006990295.1 | hypotheticalproteinSPN19_048 [SalmonellaphageSPN19] 562 562 One 562 One 562 2988 0 561 0.998220641 gene_27 | GeneMark.hmm | 272_aa | + | 27933 | 28751 272 gi | 414090177 | ref | YP_006990294.1 | putativetailassemblyprotein2 [SalmonellaphageSPN19] 272 272 One 272 One 272 1437 0 272 One gene_28 | GeneMark.hmm | 76_aa | + | 28763 | 28993 76 gi | 414090176 | ref | YP_006990293.1 | putativetailassemblyprotein1 [SalmonellaphageSPN19] 76 76 One 76 One 76 382 5.82E-47 76 One gene_29 | GeneMark.hmm | 79_aa | + | 28993 | 29232 79 gi | 414090175 | ref | YP_006990292.1 | hypotheticalproteinSPN19_045 [SalmonellaphageSPN19] 79 79 One 79 One 79 421 7.86E-53 79 One gene_30 | GeneMark.hmm | 1296_aa | + | 29222 | 33112 1296 gi | 526176449 | ref | YP_008240275.1 | hypotheticalproteiniEPS5_044 [SalmonellaphageiEPS5] 1296 1296 One 1296 One 1296 6908 0 1289 0.994598765 gene_31 | GeneMark.hmm | 246_aa | + | 33112 | 33852 246 gi | 526176448 | ref | YP_008240274.1 | hypotheticalproteiniEPS5_043 [SalmonellaphageiEPS5] 246 246 One 246 One 246 1290 6.92E-180 243 0.987804878 gene_32 | GeneMark.hmm | 335_aa | + | 33862 | 34869 335 gi | 509139295 | ref | YP_008058189.1 | hypotheticalproteinchi_072 [EnterobacteriaphageChi] 335 335 One 335 One 335 1764 0 334 0.997014925 gene_33 | GeneMark.hmm | 320_aa | + | 34880 | 35842 320 gi | 509139296 | ref | YP_008058190.1 | hypotheticalproteinchi_073 [EnterobacteriaphageChi] 320 320 One 320 One 320 1681 0 319 0.996875 gene_34 | GeneMark.hmm | 339_aa | + | 35856 | 36875 339 gi | 509139297 | ref | YP_008058191.1 | hypotheticalproteinchi_074 [EnterobacteriaphageChi] 339 339 One 339 One 339 1795 0 339 One gene_35 | GeneMark.hmm | 393_aa | + | 36938 | 38119 393 gi | 712913904 | ref | YP_009101130.1 | hypotheticalproteinCHI_35 [SalmonellaphageChi] 409 393 One 393 17 409 2072 0 389 0.989821883 gene_36 | GeneMark.hmm | 709_aa | + | 38129 | 40258 709 gi | 525972472 | ref | YP_008239937.1 | hypotheticalproteinSP088_00180 [SalmonellaphageFSLSP-088] 709 709 One 709 One 709 3550 0 671 0.946403385 gene_37 | GeneMark.hmm | 112_aa | + | 40322 | 40660 112 gi | 525972471 | ref | YP_008239936.1 | lysisproteinB [SalmonellaphageFSLSP-088] 112 112 One 112 One 112 567 4.78E-74 112 One gene_38 | GeneMark.hmm | 237_aa | + | 40664 | 41377 237 gi | 451939535 | gb | AGF89680.1 | lysis protein A [Salmonella phage FSL SP-099] 237 237 One 237 One 237 1243 5.01E-173 237 One gene_39 | GeneMark.hmm | 93_aa | - | 41383 | 41664 93 gi | 526176440 | ref | YP_008240266.1 | hypotheticalproteiniEPS5_035 [SalmonellaphageiEPS5] 56 56 38 93 One 56 287 1.77E-32 53 0.946428571 gene_40 | GeneMark.hmm | 153_aa | - | 41759 | 42220 153 gi | 509139229 | ref | YP_008058123.1 | hypotheticalproteinchi_006 [EnterobacteriaphageChi] 153 153 One 153 One 153 798 5.75E-108 148 0.967320261 gene_41 | GeneMark.hmm | 299_aa | - | 42217 | 43116 299 gi | 509139230 | ref | YP_008058124.1 | hypotheticalproteinchi_007 [EnterobacteriaphageChi] 299 299 One 299 One 299 1615 0 296 0.989966555 gene_42 | GeneMark.hmm | 237_aa | - | 43106 | 43819 237 gi | 451939530 | gb | AGF89675.1 | hypothetical protein SP099_00155 [Salmonella phage FSL SP-099] 237 237 One 237 One 237 1251 2.79E-174 233 0.983122363 gene_43 | GeneMark.hmm | 150_aa | - | 43821 | 44273 150 gi | 414090160 | ref | YP_006990277.1 | hypotheticalproteinSPN19_030 [SalmonellaphageSPN19] 150 150 One 150 One 150 785 3.46E-106 150 One gene_44 | GeneMark.hmm | 103_aa | - | 44270 | 44581 103 gi | 451939528 | gb | AGF89673.1 | hypothetical protein SP099_00145 [Salmonella phage FSL SP-099] 109 103 One 103 7 109 539 5.84E-70 102 0.990291262 gene_45 | GeneMark.hmm | 93_aa | - | 44578 | 44859 93 gi | 414090158 | ref | YP_006990275.1 | hypotheticalproteinSPN19_028 [SalmonellaphageSPN19] 93 93 One 93 One 93 485 3.73E-62 93 One gene_46 | GeneMark.hmm | 170_aa | - | 44940 | 45452 170 gi | 451939526 | gb | AGF89671.1 | hypothetical protein SP099_00135 [Salmonella phage FSL SP-099] 170 170 One 170 One 170 880 5.17E-120 168 0.988235294 gene_47 | GeneMark.hmm | 71_aa | - | 45442 | 45657 71 gi | 451939524 | gb | AGF89669.1 | hypothetical protein SP099_00125 [Salmonella phage FSL SP-099] 73 71 One 71 3 73 368 5.49E-45 71 One gene_48 | GeneMark.hmm | 64_aa | - | 45660 | 45854 64 gi | 414090154 | ref | YP_006990271.1 | hypotheticalproteinSPN19_024 [SalmonellaphageSPN19] 64 64 One 64 One 64 332 8.54E-40 64 One gene_49 | GeneMark.hmm | 272_aa | - | 45865 | 46683 272 gi | 525972458 | ref | YP_008239923.1 | hypotheticalproteinSP088_00110 [SalmonellaphageFSLSP-088] 272 272 One 272 One 272 1382 0 257 0.944852941 gene_50 | GeneMark.hmm | 228_aa | - | 46763 | 47449 228 gi | 414090152 | ref | YP_006990269.1 | putativeN-6-adenine-methyltransferase [SalmonellaphageSPN19] 228 228 One 228 One 228 1217 1.95E-169 228 One gene_51 | GeneMark.hmm | 366_aa | - | 47449 | 48549 366 gi | 509139241 | ref | YP_008058135.1 | hypotheticalproteinchi_018 [EnterobacteriaphageChi] 366 366 One 366 One 366 1875 0 349 0.953551913 gene_52 | GeneMark.hmm | 201_aa | - | 48546 | 49151 201 gi | 509139242 | ref | YP_008058136.1 | hypotheticalproteinchi_019 [EnterobacteriaphageChi] 224 224 One 201 One 224 988 4.83E-135 188 0.839285714 gene_53 | GeneMark.hmm | 248_aa | - | 49148 | 49894 248 gt; < RTI ID = 0.0 > 248 248 One 248 One 248 1312 0 245 0.987903226 gene_54 | GeneMark.hmm | 91_aa | - | 50084 | 50359 91 gi | 526176423 | ref | YP_008240249.1 | hypotheticalproteiniEPS5_018 [SalmonellaphageiEPS5] 91 91 One 91 One 91 492 2.41E-63 91 One gene_55 | GeneMark.hmm | 77_aa | - | 50364 | 50597 77 gi | 526176422 | ref | YP_008240248.1 | hypotheticalproteiniEPS5_017 [SalmonellaphageiEPS5] 77 77 One 77 One 77 396 3.87E-49 77 One gene_56 | GeneMark.hmm | 358_aa | - | 50600 | 51676 358 gi | 269975298 | gb | ACZ55522.1 | recombination-associated protein [Staphylococcus phage SA1] 358 358 One 358 One 358 1890 0 355 0.991620112 gene_57 | GeneMark.hmm | 113_aa | - | 51657 | 51998 113 gt; < RTI ID = 0.0 > 113 113 One 113 One 113 574 3.38E-75 107 0.946902655 gene_58 | GeneMark.hmm | 111_aa | - | 51985 | 52320 111 gi | 414090144 | ref | YP_006990261.1 | hypotheticalproteinSPN19_014 [SalmonellaphageSPN19] 111 111 One 111 One 111 596 1.69E-78 111 One gene_59 | GeneMark.hmm | 146_aa | - | 52307 | 52747 146 gi | 509139249 | ref | YP_008058143.1 | hypotheticalproteinchi_026 [EnterobacteriaphageChi] 146 146 One 146 One 146 750 6.88E-101 144 0.98630137 gene_60 | GeneMark.hmm | 123_aa | - | 52819 | 53190 123 gi | 414090142 | ref | YP_006990259.1 | hypotheticalproteinSPN19_012 [SalmonellaphageSPN19] 123 123 One 123 One 123 644 1.91E-85 122 0.991869919 gene_62 | GeneMark.hmm | 86_aa | + | 54160 | 54420 86 gi | 526176416 | ref | YP_008240242.1 | hypotheticalproteiniEPS5_011 [SalmonellaphageiEPS5] 85 85 2 86 One 85 417 4.56E-52 78 0.917647059 gene_63 | GeneMark.hmm | 179_aa | + | 54439 | 54978 179 gi | 414090140 | ref | YP_006990257.1 | hypotheticalproteinSPN19_010 [SalmonellaphageSPN19] 179 179 One 179 One 179 937 2.53E-128 176 0.983240223 gene_64 | GeneMark.hmm | 160_aa | + | 55196 | 55678 160 gi | 509139254 | ref | YP_008058149.1 | hypotheticalproteinchi_031 [EnterobacteriaphageChi] 160 160 One 160 One 160 825 6.39E-112 154 0.9625 gene_65 | GeneMark.hmm | 67_aa | + | 55690 | 55893 67 gi | 414090137 | ref | YP_006990254.1 | hypotheticalproteinSPN19_007 [SalmonellaphageSPN19] 67 67 One 67 One 67 352 1.17E-42 67 One gene_66 | GeneMark.hmm | 182_aa | + | 55904 | 56452 182 gi | 451937694 | gb | AGF87991.1 | hypothetical protein SP124_00035 [Salmonella phage FSL SP-124] 182 182 One 182 One 182 956 4.21E-131 181 0.994505495 gene_67 | GeneMark.hmm | 73_aa | + | 56449 | 56670 73 gi | 414090135 | ref | YP_006990252.1 | hypotheticalproteinSPN19_005 [SalmonellaphageSPN19] 73 73 One 73 One 73 373 1.05E-45 73 One gene_68 | GeneMark.hmm | 389_aa | + | 56796 | 57965 389 gi | 509139259 | ref | YP_008058153.1 | hypotheticalproteinchi_036 [EnterobacteriaphageChi] 384 389 One 389 One 384 1985 0 373 0.958868895 gene_70 | GeneMark.hmm | 90_aa | + | 58195 | 58467 90 gi | 451939503 | gb | AGF89648.1 | hypothetical protein SP099_00020 [Salmonella phage FSL SP-099] 90 90 One 90 One 90 478 3.63E-61 88 0.977777778 gene_71 | GeneMark.hmm | 68_aa | + | 58470 | 58676 68 gi | 414090133 | ref | YP_006990250.1 | hypotheticalproteinSPN19_003 [SalmonellaphageSPN19] 68 68 One 68 One 68 345 1.20E-41 67 0.985294118 gene_72 | GeneMark.hmm | 83_aa | + | 58669 | 58920 83 gi | 451937689 | gb | AGF87986.1 | hypothetical protein SP124_00010 [Salmonella phage FSL SP-124] 83 83 One 83 One 83 412 2.29E-51 80 0.963855422 gene_73 | GeneMark.hmm | 83_aa | + | 58913 | 59164 83 gi | 414090131 | ref | YP_006990248.1 | hypotheticalproteinSPN19_001 [SalmonellaphageSPN19] 83 83 One 83 One 83 401 1.08E-49 82 0.987951807

As a result of gene analysis, KFS-SE had double-stranded DNA and its total dielectric size was 59,715 bp, and its GC content was found to be 57.14%. As a result of analysis of ORFs, bacteriophage KFS-SE had 73 ORFs as shown in Table 1, 21 of which ORFs (28.8%) had distinctive functions and the remaining functions were unknown.

Example  3: Bacteriophage KFS Morphological characteristics of SE

To observe the morphological characteristics of bacteriophage KFS-SE, transmission electron microscopy (TEM) observation was performed. Bacteriophage KFS-SE was loaded onto a copper grid, followed by negative staining with 4% phosphotungstic acid and drying. Electron microscopy was performed in Kyungpook National University joint lab habit, and the shape of bacteriophage KFS-SE was observed with a voltage of 100 kV using H-7100 (Hitachi), and the result is shown in FIG.

As shown in Fig. 3, the head (71.429 nm) and the relatively long tail (211.429 nm) were observed in the bacteriophage KFS-SE, so it is presumed to belong to Siphoviridae.

Example  4: Bacteriophage KFS Protein pattern analysis of SE

To analyze the protein pattern of bacteriophage KFS-SE, 40 μL of the purified bacteriophage KFS-SE solution at a concentration of 1.93 × 10 ¹¹ PFU / mL and 10 μL of the 5 × SDS sample solution were mixed, boiled for 5 minutes, Respectively. The results of SDS-PAGE and protein electrophoresis are shown in FIG.

As shown in FIG. 4, it was confirmed that the bacteriophage KFS-SE is composed of main proteins of 120 kDa, 64 kDa, 48 kDa, 38 kDa and 35 kDa.

Example  5: Bacteriophage KFS Investigation of the specificity of -SE for pathogenic strains

Specificity studies were conducted to confirm the lytic activity of other pathogenic strains other than Salmonella enteritidis of bacteriophage KFS-SE.

Specifically, the strains of the following Table 2 were each cultured to obtain a culture solution, and then it was confirmed whether or not a lysosome was formed through a soft agar overlay method using each culture and purified bacteriophage KFS-SE. The results are shown in Table 2.

Name of strain Whether the sputum is formed Salmonella Enteritidis ATCC 13076 ○ Salmonella Typhimurium ATCC 19586 X Bacillus cereus ATCC 13061 X Bacillus subtilis KACC 12680 X Bacillus cereus JCM 2152 X Vibrio parahaemolyticus ATCC 43996 X Listeria monocytogenes ATCC 19116 X Pseudomonas aeruginosa ATCC 10145 X Shigella sonnei ATCC 9290 X Shigella boyish NCCP 11190 X Staphylococcus aureu s ATCC 25923 X Escherichia coli O157: H7 X Escherichia coli K-12 X Escherichia coli BW 25113 X Yersinia enterocolitica ATCC 23715 X Klebsilla pneumoniae ATCC 13883 X Aeromonas . hydrohphila ATCC 7966 X Aeromonas . hydrohphila ATCC A3 X Aeromonas . hydrohphila ATCC A5 X Aeromonas . hydrohphila ATCC A6 X Aeromonas . hydrohphila ATCC A7 X Aeromonas . hydrohphila ATCC A8 X Aeromonas . hydrohphila ATCC A9 X Aeromonas . hydrohphila ATCC A10 X Aeromonas . hydrohphila ATCC A11 X Aeromonas . hydrohphila JUNAH X

As shown in Table 2, it was confirmed that bacteriophage KFS-SE has no lytic activity for pathogenic strains of other genus such as Bacillus cereus and Aeromonas hydropila.

Therefore, it was confirmed that the bacteriophage KFS-SE of the present invention is a bacteriophage exhibiting lytic activity specifically on Salmonella enteritidis.

Example  6: Bacteriophage KFS PH stability investigation of SE

To confirm the pH stability of the bacteriophage KFS-SE, 900 μL of TSB medium in various pH ranges (pH 3.0, 5.0, 7.0, 9.0, 11.0) and 100 μL of bacteriophage KFS-SE at a concentration of 1.93 × 10 ⁶ PFU / mL After that, it was allowed to stand at room temperature for 1 hour. The activity of the remaining bacteriophage KFS-SE was measured using a soft agar overlay method using KFS-SE for 1 hour. The measurement results are shown in Fig.

As shown in FIG. 5, it was confirmed that there was no significant difference in the activity of the bacteriophage KFS-SE in the range of pH 3.0 to pH 11.0.

Therefore, the activity of the bacteriophage KFS-SE of the present invention was maintained in a wide range of pH 3.0-11.0, and it can be seen that the bacteriophage KFS-SE has excellent pH stability in the range of pH 5.0-11.0.

Example  7: Bacteriophage KFS -SE thermal stability investigation

In order to confirm the thermal stability of the bacteriophage KFS-SE, the activity of KFS-SE was measured at various temperature ranges (4 ° C, 22 ° C, 37 ° C, 50 ° C and 60 ° C).

Specifically, 1 mL of the bacteriophage KFS-SE solution at a concentration of 1.93 × 10 ⁹ PFU / mL and the TSB medium solution were mixed and incubated for 1 hour at 4 ° C., 22 ° C., 37 ° C., 50 ° C., . The activity of the remaining bacteriophage KFS-SE was measured using a soft agar overlay method using KFS-SE for 1 hour. The results are shown in Fig.

As shown in FIG. 6, it was confirmed that the bacteriophage KFS-SE maintained stability at 4-60 ° C.

Therefore, it was confirmed that the bacteriophage KFS-SE of the present invention is a bacteriophage having a relatively high heat resistance, so that it can be stably formulated against the heat generated during the bacteriophage formulation when it is used as a feed additive.

From the above experiment, it was found that the method of preventing or treating infectious diseases caused by Salmonella enteritidis using the bacteriophage KFS-SE of the present invention is highly specific to Salmonella enteritidis as compared with a chemical substance such as existing antibiotics, And it has the advantage of being strong against physicochemical environment. In addition, the bacteriophage KFS-SE of the present invention has an advantage of being able to solve the problem of antibiotic-resistant bacteria caused by misuse of antibiotics, the problem of remaining antibiotics in food, and the problem of a wide host range. Therefore, the bacteriophage KFS-SE of the present invention has the effect of contributing to the competitiveness of the aquaculture industry and the livestock industry and the improvement of the public health.

Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

Hereinafter, formulation examples of pharmaceutical compositions for the prevention or treatment of infectious diseases including bacteriophage KFS-SE of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically explained.

Formulation example  1: Manufacture of medicines

1.1 Manufacture of Injection

Bacteriophage KFS-SE 10 ⁶ PFU / mL

Sterile sterilized water for injection

pH adjuster

(2 mL) per 1 ampoule in accordance with the usual injection preparation method.

1.2 Liquid  Produce

Bacteriophage KFS-SE 10 ⁶ PFU / mL

20 g of sugar

20 g per isomer

Lemon incense quantity

Purified water was added and the total volume was adjusted to 1,000 mL. The above components are mixed according to a usual method for producing a liquid agent, and then filled in a brown bottle and sterilized to prepare a liquid agent.

1.3 Manufacture of tablets

Bacteriophage KFS-SE 10 ⁶ PFU / mL

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

BRC KCTC13147BP 20161107

<110> Kyungpook National University Industry-Academic Cooperation Foundation <120> Novel Salmonella Enteritidis specific bacteriophage and          antibacterial composition comprising the same <130> KNU1-291-1 <150> KR 10-2016-0153400 <151> 2016-11-17 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 59715 <212> DNA <213> Artificial Sequence <220> <223> Bacteriophage KFS-SE (KCTC13147BP) <400> 1 ggtgcgcaga gccgctttct cggaaaatgg gccggatctg ccatagcccc gctttgcgaa 60 atcaataact tagccgaaaa accgaatttc tcccgtaggg agtctccaaa tcgcggtggt 120 gacgggggtg cgggcggaac ggcgctttag ccccgcaatc ggttgattgt attactgttt 180 tacccggtgg gcgggtgctg accaagtcca cggcagcagt atcaggttac aagttacaga 240 cgggttacaa gtttttcgct ttgtaacctc tgactatcta actgttttta ctacgttttt 300 cttctataaa gttacaagtt acaaataaag tatagtaata gtccagaaat agagaaaacg 360 aggtaaaaat gtgaatttta tatagggggg aattttaccc gggcgctaaa ccacttcggt 420 ttgtaacctg aatacccgga tcgctgagag ccgcgccgcc actggcctga acatggttac 480 aaacggcgaa aatggtatgt aaccttttgc ccgttttggc cgttttcttg cgtttcaata 540 acaaatttgt tatacgtctt gacctgaatc ctgctataat ttatttgtta taaacagcaa 600 aaagaggtca aaaatgcagc acgatgaatg ccgtgaaacg ttgatggatt tcaaacgaca 660 attacagaca catttgaagg cgattttacg tcgtcagggg gtcacacaga gcgcgcttgc 720 gacgatgatc ggcgtacacc agtcaagatt aagtttcatg cttaacagtg acgcagacag 780 agtttcgatc gataaactca ttgaagcgat agtgaaactg gatggcgtgt ttatggtgaa 840 aatcggagat caaacagaca taaatcgagt gcggtacaag gagtgcgaag acaacgtttg 900 acagacaaaa ataaggccgg atttcccggc ctttttatcg actaaacttc gtcttcatcg 960 tcaaacgggt ctggcttttt gccgaccagt ttttcgactt ctgcgtcatg ccgttttaac 1020 gtagtctgca cccatgacga gttaagtttc cccgcgtcat cgaatgcccc tttgacccgc 1080 ttccggacat aaacgcggcc agattcgccg ccaaatttga ccacgccgcc cggcttgaag 1140 cccaattcca taagcatgtt tttcaggcgg ttcgtcttcg gcatttccac gtctgtcatt 1200 tcgaggaatg cgtttttcag ggacggcacg aagatgacgt cgcggtgtac gcccggcgtc 1260 gtcgtcac gaagcatttg tgcgattgcg ccttccacgc catcgtcaga cgcgtcggcc 1320 atgaccgcga atgcttcggt gtcctgcggt gcgtgtcctt tggcatcgaa atcctttccg 1380 atctcccaat cgaggaacca ctttttgata gcaggtgcgt ggttttcgat tgcccggtac 1440 acgtttttga agaagcgcgc gcgttcagtc ttcgacccgt aaacccgctc catgtccaga 1500 tgattttctg cctgcgttga caccaccaga taacgacgct ggttttcatc gagtggcaat 1560 gcgttgaagt ggttggtgaa catcattttg ctgctggtgt tcactacttc cttcggcttg 1620 cggccctttg gctcaatctg gaagcggtcg tttgtgatga attccttctg gcgttccacc 1680 gcgctgtacc gatcgccttt gtcgtagatc tcttccacga ttttcaggat gtggccctcg 1740 gcccaatcgc tgaaacggcc attcatcacg gtgtttgaca catagcccac gttcgacgcg 1800 ccgagcatcc gcgacatgat gatgccgagc gtcgttttac cagaaccatg cgcaccacgg 1860 atcagaagcg agtagttgat gcgtttcgtt gggtgctgaa taacccacgc catccagtcc 1920 agaacatagc ggcgttcctt ctcgttcggg aaaagcacga tgaagaagtc tttgatgatc 1980 tccagcgcct ttttgtcgta cggagaaagg tcagtcgcgg cctccggaat ggattcaggg 2040 tcaaaagagt tcagccacgt acgcccacgg tcgtcaataa agaattccgg cccgatcacc 2100 ccctctttcg gcttccacac agattccggc atgtcaccgt gcatcgccgg gtagtacatc 2160 gcgtcccaaa cgataggaac cgggcggact accgacgcat agtggacagg cttgaaggct 2220 tcttcctcaa acacgcattt tttcgcaaaa gcgacgtcga aagcgccttt tgtgatgtaa 2280 tcgcccgtgg ttttttcgat aaaactgtca tttgccttcg agaaaacgaa gtttttcagc 2340 cactgcggag tctcttttga cccgtaatcg aagcccaaaa gctccctttt gtcaccagca 2400 gtgaggcgga agttgttgat cctgcggtat gcttcggccg caatctggac gatggccttg 2460 cgggtaagcc cgaaaatctg cgcttttttg aatgattcgg cccacgcatt ccagtcattc 2520 gcctcggtac actcggcgaa agcctcgcgg tactcgatgg cgcgttcttc ctgctcggct 2580 ttctcgatgt cctgcacttc tttgatgatg gtgccgagcg tgatcagtcg agtcgagttg 2640 tgcgcgaagc ccttatccca cttgtattcg aaatcacctt cgtcgaagtt gtgtgcctgc 2700 atggcccatt cgcgggcaat ggccttcgcc tcgtcctgat cgcggcagga aatttgcagc 2760 gcggccagta cggtgatgta gttctcgtaa tcttccgggt tcgggtattt caggacgatg 2820 tcgcgcagtt catcgtacgt gccgtcccac tttttgacgt cagacagcgc ggcccaatcg 2880 tcatcgtctg ccgaaccgat agactcataa ccgttcattg gccgtttgac cattgtccag 2940 cccaagtcct tcgcgtatcg gtcgaattcg gcggcgatct cgcgagcggt atcgaggctg 3000 atctcttcga ggtcaaggtc ggcggcgttg ttcagcggat tctcgccatc aatccagtaa 3060 tagtcgcgct tggtgtccgg gtggataccg tacgcaacga actgctggcc gtcggcgagg 3120 atttcaactg catggcgctg gccgaagtcg tcttcccaca cgccggattt cactttggcg 3180 aacgggtttt cggttcggta caggtagagt ttcttcggct cccggccgat acgacatggc 3240 gctgcaccta cgttctcggc gatccagtct gccatgtgcg ccacggcgtc ttcatcgtaa 3300 acgtcgatgt caaccgccgg ggtgtatttg gtcagaatcc cgatgccgtt attcgtatag 3360 cgtttcgccc aacgctggat ctgctcctta tcattgattg ttcgttccca atcttcaccg 3420 atagggcgtt tgcccgcacc cttcttatcg gcgtccgggg cgtagatagg tacgatggtg 3480 tacccattcc cccaaagcgt ttcgccgtac tcttgccagt agttcattca aggatttccc 3540 ctgtttcgct gtccgttttg atccatgaag ggttagtcag ccaatgcgga cggatgccgc 3600 agtgagggac agctttgcaa actttttcgg ccacgctgct cgttacgttg ttctgcacgg 3660 cccataacaa tgtcgggtaa ctgatcttcg ctgccttcgc cagttcggcc attctggcgt 3720 tcgggtctac ctccaaagcc gcttgcatca tgagaaattt gacccgcatc cggtcgtcgt 3780 ttaaagccca atctggaaat cgcatagtga tgccctcgcg taatgtgcgc gtgttataga 3840 tttgcggata ttatatccgg gcttgacatc tgtcaagttg gccagttatt cgggtaccaa 3900 aggcgcttta tgtccggaaa acaaagggct tgcggtaccc gaataaaaaa tttctaaaaa 3960 agtgtttgac agtgattcgg cgtttgggcc atacttcatc ccgtcaacac gacacgacct 4020 tttaaacact taacggagta gaaaattatg tctggtatct tcgaacaact gctggctgaa 4080 caacaaaaaa ccaacaccct gctggaaggc gttctggcag cactgaaagg cggcgcggtg 4140 aacacttccg ctgacgctgg tagcgaaact actgaaaaaa ccaccaccac caaaggcggt 4200 gctaagggcg gcgcgaaagg cggtgcgggt aaaaccgaaa ctaaagcggt caagcctaaa 4260 cacaccaaag acgaagtagt cgcggctgtc gttgcggtta aagacgcgtt cggcgcgccg 4320 gaagccaaga aaatcaccgc acacttcggc ctcgccaaag tcgcggaagc gaaggaagaa 4380 cacttcgacg aaatcgtcga aatgtgtgaa gccaaactgg ctgaaaaaga tgaaggcggt 4440 aacggcgaag aagacgacgt ttaatctttc gccccgcatc ggctcccttc ggggggccgt 4500 ttttcacacc aacggagagg caattgtgac cagcattact ttcaccaacg tagcaacatc 4560 cggcggcgca gttctcgcgc gcacttgggc ggagtttcgt tttcaaatcg gtgatcgcga 4620 tgtgctgttc agagttactt cacctgtttc gaaacacgaa gcgccgctgc tcgtaaccca 4680 tgctgcatcg ggctatcgag tgtgcgacgt ctggtgggac gacgtggcga agactcgctt 4740 taacgcaggc cgcaataatt acagcggggc aggcgctagg gcattggtag cgacgattga 4800 gcgagtcggc gaacaccgat tcctcaatgc gataaaggcg tatccgaatg agtagtaagt 4860 cactgttagt ccgcatcatg cgggaattcg gcggtggcgg ccactcaatc ttcgcgcctt 4920 ctgcgtcagc gcgttggatc gcatgttctg gaagcctgat agccgggctt ttcgagcatg 4980 acgaaaccat ctacgaggcg gcggaaggca cggtggccca cggaatcgcc gaacaatggc 5040 tgcgaacgga tgtccggcca acccatttaa tcggtacggt cgagacgatc acggaaggcg 5100 gtgtcagcca tgacatcccc attactcggt caatgctcga ctcggttcag gagtacgttg 5160 actggtgccg atttgaagag ggtgaaatgt tcacggaaat ccgggtctgg tttaccgacc 5220 tgatgccgcg agcgaacccg gatgaacctg acgaagaccc gatcgagttt gtcgctcagg 5280 gcggaacggc ggataatatc atcatccgcg accgaacctt gatcgtgacc gacctgaaat 5340 acgggactgg cgttcaggtt ttcgcggaag ggaacccgca ggcgctactt tacgcgtacg 5400 gtgcgtaccg cgctttctcg gatgaatacg agtttgaccg tattgtcatc cggatcgcgc 5460 agccgaggct cgaccatttc gatacgtggg aaatcacgat tgacgaattg ttagagttcg 5520 cggagtatgc ccgggagcga attgcagcgg catggagcct taccgcgccg cgccgcgcat 5580 ccctgaaagg gtgtcgcttt tgccgggcag cacataactg cgcagcgata gcgtacatga 5640 tggaatgcgc agtgggcggg gatgtggaat tccttgaagc tgaattcggg gaggaagaaa 5700 tgtcagtatt gcgtgatgcg ctggcgcagg agtataaatt ccggcgggcg cagttcggaa 5760 acctgacaac cgagcagatg gcgaagatcc tgccataccg caaggtggtc gaaaactggt 5820 tctcgcgcct tgacttcgaa ctggaacgcc gcgcaatgaa cggcgagaag gtaccgggtc 5880 aaaagctggt ggagtcgcgg acaaaccgta aattcgccaa tgagaaagat gcgatcgcgc 5940 tgttccggtt cctcgatatt gacgaggaca agtacatcga acgcaaactg cgtaccccgg 6000 cgcagatgga agaagtgctt agggatgaat tgggcgtgtc ccgggctggc gcgccgaaca 6060 tcatcgcggg gatcgtctac aagccggaag gcaagccgac actggcaccg ctgaccgata 6120 agcggccacc actggacggc aagtattccg gggcgtggga cgacgaagat gatgacgaag 6180 tgtaaacccg taaaaccgta gaaagtaaac ccgtaaaaca gtaaaccgag gatattaaga 6240 tggccgagaa attagtccct gcgaaaaaag tcaagaatgg cgtgctgtac aaatctggtc 6300 acatcaagat ttctaacgta cgtgcttctt acccgcacct cgataagccg tacggcggtg 6360 aagacggcgg cgagccgaaa tactcgctga cgcttttgat gccgaaagac agccacggcg 6420 agatcgagaa aatcatccgt gagcagatcg aagtcaccaa gaagaaccac aagacaggcg 6480 cgctgaaagt cgccccgtcc atgttgttca tcaaagacgg tgataccgat tttccggata 6540 aaccggaatg cgaagggatg tgggttatct ccgcacgcga aagcaaacgc ccggatgtgt 6600 ataacatcga gcgcgaagag ttgactacct cttccgaaat tctggaagag atctacggtg 6660 gttgttgggt gtcggttgtc atccgtccgt ggtcgcagga aaacaaattc ggcaaacgcg 6720 ttaacgccaa cctgatttcc gttctgaaac gcaaagacga cgagccgttc ggtgaaggtc 6780 gtgtggacac gtccgacgcg tgggatgaag acgaagattg ggaagacgaa gtgtaatgta 6840 cgccctcggg cggtaataca aggcccgccg cgtgcgggct tttttcatac aggagcgtta 6900 acgtgaccga ttgcattaat ctggactatg aatcccgaag ccgggtgaac cttaaaacga 6960 acggattcga ccgatattcg aacgacgaat cgactaaaat tctgatgggc gcatggtcta 7020 tcaatggcgg agcggtgcag cacatcgacc ttcaccgagg gaagaagttg cccggcgaac 7080 tgcgtgaagc gctggaagac ccggccgtcg agaagtgggc ctttaacgcg cagttcgaac 7140 gcgtaatgac ccgccgggtg ctcggcatca agacgccgta taaatcgtgg cgctgtacga 7200 tggcgctggc gtacatgctc ggctttgcgg gcgaccttct ggccgtcggc acgcagatcg 7260 gcttgccgga agagaaactg aaagacccgg aaggcaagcg cctgatcgac ctgttcagca 7320 agccgcagcg cgtcacgaag aaaaacccgt ttgagtggcg caatgaagcg accgacccgg 7380 aagactggtg gcacttctgc aaatataacg tgcgggacgt tgaaaccgaa ctggcgatta 7440 aagcccgcct gatgcggtac ccggttctgg atgccgaatg ggacatgtac gcgctggatc 7500 agttcatcaa cgatacaggc gtgatgattg accgcgaatt cgcgtactcg gcgctggcgc 7560 tcgcagaggc gcgcaaaccg cagattatcg aagaaatggc cgacatcact ggcctgaaca 7620 accccaattc cccggcgcag ttaacaccgt ggctcaaaga gcgcggctat ccgttcagcg 7680 accttcgcag tgacaccgtc aacaaggtca ttcgcgaagc cgatgaaaac ggcgttgacc 7740 cggatgcgat ccgcgtgctg cggatgcgcc tgaatagcgc gaaaaactcc ctgtcaaaat 7800 atcagaccat gattgacgcg gctggcgaag atggacgatt ccgcttctcg ctgcaattcg 7860 ccggggcaag ccgcacgaac cgttgggctg gccgccgtat tcagactcaa aacctgccga 7920 gaacgccgaa gctgatcgag gacgtgaccg acctgtccat cgttaacaag atgatccacc 7980 agcgcgacct tgatgcgttg acgctgtatg tgggcgagcc gatggacgcg ctggtgggct 8040 gcattcgctc cgccttcatc ccgacgccgg gccacaagtt catcgttgcc gacctgtcgt 8100 ccatcgaatc cgtggtcatc gggtggctga caaactgcaa atggttcatg gacacgctgg 8160 cggccaagca cgacctttac cgttcattcg cggctcactg gctgcacctg ccgtacgaag 8220 aaacgaagcc acaccgcccg aaagccaaac ctgctaccct cggcgcgggc tatcgcctcg 8280 gcggcggcca catggacgag aatggcaaga agaccggact gtggggctac gccgaaaaca 8340 tgggcgtaca catgacgcag aaagaggcag aagagtcggt acaggcgttc cgcgaccttt 8400 gcccggaaat tgtgcgcgca tggtatgacc tcgaaaacgc ggtattcaag gttatccgta 8460 cccgtcaggc cgtcgtatgg ggctgtctga tcatcgaata cacaaaacct ttcctgacca 8520 tccgtctgcc gtctggccgc aaaatgtatt acttccgtcc gcgtctggtc gagcgccaga 8580 tgaccgttca gaaagggccg aagaagggtg agaagtacac aaagaccaac ttccagtacg 8640 aaggcaaggt ggaaggcagc ggcaccaagt ggggcaagat ttacagccac ggcggtaagc 8700 tggtggaaaa catcgttcag gcgctggcgc gcgacgtgct cgccgaaggt ctgaaaaagg 8760 cgcaccgcat gggcttccgg atcgtcatgc acatccacga tgaaatcgta accgaagtgc 8820 cggaagacag cccgctgacc gtggacgacc ttatttcctg catggcggcc gaactaccgt 8880 gggccgaagg gctaccgctt ggcgctgccg ggtgggaagg ctatttctac cgtaaggatt 8940 aacatggccg agccgataat cagagaatcg aaggtcgaaa agcgctgctg cgaatacgcg 9000 caggggcgcg ggtggtgggt gtcaaaattc acctcccccg gcaaaaaagc cgtcccggat 9060 cgcgtgctga tccgtggcgg cattgtgctt tttatcgaat tcaaacgccc cggcgaagaa 9120 ccaacggcgc agcagtatca ccgccacaag gaaatgcgca aggccggggc taacgtgaca 9180 tgggtggaca attttgcaga ctttaaaaac tacattatcg cctttgagta gagcgctcgc 9240 gtactgcctg cggacggtta aactgaaccg cagcgacatg cacgactatc aggacgaagg 9300 cgtccagttc atcaaagacc atccgtactg cggcatgttc gtagacctcg gcttgggtaa 9360 gactgtcatg accgggacgg ccatgctcga cctgatcgcg gacggcgaaa tcaacaaagt 9420 cctcgtcgtc gcgccgaagc gcgtggcccg taccgggtgg ccgacggaat tcgacgagtg 9480 gggccacctg tgtttctaca agatgtcggt catcgccggg aacgccaaag agcgcgcagc 9540 ggccgcgagt aaggactgcc atttttacac gatcagcgtg gataacattg cgtggctgtg 9600 tgagcacttc aagacgaaat ggccgttcga cgccgtggtg ctggacgagt cgagcatgtt 9660 caaatcccac acctcgcagc gcttcaaact gctgcgccgg gtgcggaagt acatcaaacg 9720 gatcgtcgag ttgacggcca cgcctgcggc cgaagggtac atgggtatct tctcgcaaat 9780 ttacctgctg gacgaaggcg agcgctttgg ctccaccatc ggcggttatc aggaaaacta 9840 cttcacgcag aaccgctaca acttcaaatg gaaacttcgc cccggtgcgg aagacgagat 9900 tattcggaag atttctgata tatgtttagt catgaaagcg gaagactacc ttgacatgca 9960 cgaaccgaat ttcgttcctg tcccggtcga gttagacggg gagacggcgg agcggtaccg 10020 catgatggag gaagagagcc tcgtcgagat tatgccggac gatttcgatg aacatctgga 10080 tgacccaatc gtgatcgaag cggagcaggc cgcgtctttg cagtcgaagc tattgcagat 10140 gtgctcaggc ttcatctacg acacgaaaat cgttggcata acgcaagacg acaaggtcat 10200 taagcagaag gacacgtaca ggctgcacga cctgaaattc gatgcactgg aagaactgct 10260 ggaaaccacg ctggccgata aaaacgtgct gatcgcctat cacttcaagc cgaccctcga 10320 acgactgaaa gagcggttta aagatctggt ggtcatggat gatgacggta agtgcatcaa 10380 gaagtggaac gccggaaaga ttcgactgct ggccgcgcac cctcagtcag cgggacacgg 10440 tttaaacctc cagcatggcg gccacgtcat cgtctacatc gacaatccgt ggtcactcga 10500 acgcttcctg caattcaatg gccgattgca ccgtcagggg cagaagtacc cggttacgat 10560 ctaccagttc aaagccatgc tgcgaacacc gaacgggctt ctggccgaga cggcggacga 10620 cgtagtcata caggcgctgg tcgaaaagga agacgtgcag gacgcgttct tcgaactgct 10680 cgaacgtatc aaagggcgca tcaaacgccg caagaaatcg aaaaataaag gggtatggga 10740 tgacgaagag taatgcgcca ctggcaaccc ggaatcgccg ctctaatgcg ccggacgccg 10800 ataccgaagc gatgatcttt cagggctgca acatcacgca actggcgaag ctgtttcgca 10860 tggagcgccg ggacatcacg ccgaaaatca tggatgtccc gcctgtcggc gagcgtggcg 10920 ggtacccgat ctacgcggtt cacgaagtgg ctccgtacct cgttaagccg ctgtacgatg 10980 tcgagacgta cctgcgccgc atgaatttca aggatctgcc gaaggagttg tcgaaagagt 11040 tttgggccgg gcagcgtgca aagcaggact tcgacatcaa agcgggaaac ctgtgggaaa 11100 ccgagaaggt gatcgagcac ttcggcgagg cggtgaaggt gctgcgcatg tcgatgttgc 11160 tcattcccga cacgctggcg cgtcaggccg gactcagcga gccgcaacgt caggtcatca 11220 cgtcatccat tgacgccatg ctgaacgacc tgtcgagcgc gctgatcgat aagttcaacg 11280 gcgacgaggt ggatgacgat gaagtttaag agcatcgggc agattatcag ttcggtggcc 11340 gagcagttac ggccgccaat gcgcatgacg gtggccgacg cggccgccaa gtatcgctac 11400 gtcaaccaac ccggcgcata cgtcggggat tggcttaact cgacgacgcc gtacatggtc 11460 gagccgatga acatgatgaa cagccgggcg tacgacaaga tggcctttgt cggcccggcg 11520 cagtccggta aaaccgatgc gctgatcctg aatagcattg tctattcggt gaaagttgac 11580 ccgatggatc tgatggtcta ctgcccgacg tcaacggccg cgcgcgactt ctcgatgcga 11640 cgtgtagacc gtcttcaccg ccatagcccg aaggtgggcg aaatgctgat gaagaaccgc 11700 gacgccgaca acaaattcga caaacactac gtcacaggca tcatcctgac gttgagttat 11760 ccgtcagtaa ccgaactggc cgggcgtccc gttggtcgca tcatcatcac cgactacgac 11820 cgtattgacg acgacatcgg cggcgacggt aacgccttcg acctcgcatc aaaacgtacg 11880 acgaccttcg ggtcgtttgc catgtgtgca gccgagtcat cgccatcgcg cccgattaaa 11940 gacccaagct ggattaagcg cacgccccac gaagccccgc cgtgtgacgg gattgtcggc 12000 ctgtacaacc gtggcgaccg ccgccgctgg aaatggccgt gtccgcattg cgaccagtat 12060 ttcgaaggta ccttccagct tttgaagtgg aacacgaaag acgccgatgg caaccctcta 12120 accaacctcg aaaagtcgga gacagtgcgc atggcctgtc cacactgcgg ctgcgaaata 12180 gacccggttg ataagtatga aatgaacctg tgggggatgt gggtgccgga aggctgtcat 12240 gtcaacgagt tgggccaact ggccggggta ccgatgcgct cgtcgttcgc gtccttctgg 12300 cttcgcggaa cggcggccgc attcatcaca tggcagaaac tggttctgaa ctacctcgat 12360 gcgatggatg actacgagcg tacgatgtcc gaagagtcct tgaagaaatt ctggaacaac 12420 gacatgggcg aaccgtacgt gccgaaggcg atcgagacgg tacgcgttcc ggaagcgctg 12480 aaagcccgcg ccgagaagtg gccggaacag acggtgccga agaatgtgcg gttcttggcg 12540 gccacggtgg acgtgcagaa gcatagcttt gaagtgtcgg tgcatggtgt ggcccccggc 12600 tacccgttcg acacctatct gatcgaccgc ttcaacatca cgaaatcgcg ccgactcgat 12660 gcgatggcg acccgaaaat cctgcacccg ggcgcgtacc tcgaagattg ggatctcatt 12720 gaaaccgaag tgatgaacaa gacgtatccg ctgtccgata attcgggccg ttcgctgccg 12780 atcaagatga ccgcgtgcga ctccggtggt gaagccgggg taacgggcaa cgcgtacgaa 12840 ttcttccgga aactgcggaa ggaagggaaa accgggcgct tctgtctggt gaagggcgac 12900 ccgaaagcga acaacccgcg cacccgtgta gcgctaccgg actccaacca gaaagacaag 12960 aaagccatcg cacgcggcga cgttccggtg ctgatgatta actcgaacgt gatgaaagac 13020 agcctgaacg gccgactgga cgtcgtggag ccgggcaaag gcatgtacca catcccggac 13080 tggatgggcg acgcggtatt cgccgaactg tgtacggaga cgcgtgacga gaaaggctgg 13140 cacaacccat cccacgcccg aaacgaaacg tgggacttgg catattattt attgggaatt 13200 tgcgtcagtg gccgcgtgct cgggattgag cagatagatt gggataaccc gccatcatgg 13260 gcggccgagt gggacgataa cagcatggtt attttaccgc agagcgatga acctattgat 13320 gcaccgaaag ataccgaata cagtttcgag aaactcgcaa tggcgttagc ataggagaac 13380 caccatgaca ccagaagaat gccgcgcgca atatcgcctc atgctcaaag aagcgatgga 13440 tgcgtaccac caattaaacc tcggcggctc ggtgcgcgtc gtggtggatc agaacagtga 13500 acgcgtggaa tataccgcag cgaaccgaca aagcctatgg gcctacatcg ttcgactgca 13560 aaacgcgatt aattcggaca acccttgcgc ggccttcatg ggcctaccaa gttcaccagc 13620 aggattctta ttcccatgac cgagaaaaag agaagcacta cacaacgggc gaagaaagcg 13680 gcaaagaccg ccgacgtcgc tacgttggac gccacgccgc agaatccttc ggcactcggt 13740 ggaggcttag agggggctga acggaacacg cgtgaaatgt tccgctggac gcctgccatc 13800 atttcaccag accagcagat cgcccaagac ggtacgctgg cgctttcgcg tgcgcaggac 13860 atcgtacaga acgacggcta cgcatttggc gcggtggcga ttcaccgtga cagcgtcgtc 13920 ggctcgcagt acaaactcaa ctcgaaaccg aattcgctgg tgctcggcgc gccggaaggt 13980 tgggcggatg aatttcagga ggtagtcgag gcgcgcttca acatggtggc cgagtctcct 14040 gaaaactggt tcgatgcccg ccggatgaac acgctgaccg ggcttgtccg tctggccgtt 14100 ggcggcttca tcatgaccgg ggaagttctg gcgtcatgcg agtggatgaa gccgaaggga 14160 acccgtatgc agcgccgtcc gttcggcacc gcgatccaga tgatttcgcc gtaccgtctg 14220 tccaacccgg acaacatcat ggacgacaag tatctgcgct ccggcgtcaa actggatgaa 14280 atgggcgctc cgatcggcta ctggctgcgt aaagcgttcc cgggcgaccc gactgacctc 14340 gaacaatggc gatgggaata ccagcccgct cgcttcgatt ggggccgacg tcggatgatc 14400 cacatcatcg aagcgctgct gccgggccag actcgcggta tcagtgaaat ggtcgcagcg 14460 ttgaagcaga tgaagatgac ccgaaacttt caggaagtca cgctgcaaaa cgccatcgtc 14520 aacgcgacct acgcggccgc catcgaatcc gaattgccgt ctgacgtggt cttcaatcag 14580 atggggatgg gacagacgcc tttcggcgac atcctgaaaa cctacatggg gagtctggcc 14640 gagtacatcg ccggatcgaa aaacatcgca atcgacgggg cgaaaatccc gcacctgttc 14700 cccggtacga aactgaaaat gcagcccgca ggaacgccgg gcggagtcgg aaccgattat 14760 gaagaatcgt tgctcaggaa catcgccgca tcactcggcc tttcgtacga gcagttcagc 14820 cgcgactata cgaagacgaa ctactcttct gcccgcgctt cgatggctga aacgtggaag 14880 tacatggaaa gccgtaaaaa gctggtagcc gaccgtttcg cgtcgatgat ttatacgttg 14940 tggttggaag aagaggttaa cgctggtaat gttccactgc cgccgggctt cacttggcgc 15000 gacttctacg acccgatgaa acgtgacgcg ctttgcaacg cagaatggat cggcgcgagt 15060 cgcggccaga tcgacgagaa gaaagaaacc gaagccgcta tccttcgcat caagaacggc 15120 ctgtcaacgt acgaagcaga aatcgcccgc ctcggtgggg acttccgcga agtattcaaa 15180 cagcgcgccc gcgaagaagg catcatcaaa gatttgggcc ttgatttctc cggcaagatg 15240 gtcgaaggga cggaggcaag cggttcaact ggcagcactg gctcggacaa caacgaagag 15300 gatacgaagg aatgagcaac cacgtaagag cgagcgtctc aatggcgctc agtcgaatga 15360 acggccagcc gatcgccatt cgcgagaatg acccgacgct gctgaccaac atgcagcaga 15420 tgatgtcctg cgatgacgaa gattttcagg agcgcgctga acagcagatg cgcgaaaacc 15480 tgtgtgcagc gtacggcgtt ggccgaccgt ctgcggacaa gcctttcgcc ttctcgaacg 15540 gcctcgccat catcccgatt cacggcaccc tgattaaccg ctatggcggg tattactacg 15600 gctacgtgac gggctacaat tttatccgca gtcagcgtaa tgcagcgctg gccgacccgg 15660 atgtcgaggc tatcatctac gacgttaact cgaacggcgg ggaagcagca ggctgtttcg 15720 agctgtcaca ggaaatgttt gatacacgcg gcgagaaacc ttcgctggcg gttgtcgatt 15780 caaactgcta ttctgcggca tatgcgttgg cgagcgcagc ggataaaatc gttgtaacgc 15840 cgtccggcgg tgccggaagc atcggtgtta tcgccctcca catcgacatg tcgaagatgt 15900 tggcggacat cggtatcacc gtgaatatca tcaagtccgg ggagcacaaa gccgacggca 15960 acccgtacga accaatgagc gatgaagtac gagcagactg gcaagcgagc gtagattcaa 16020 tgcgtaccga cttcgtcaat cttgtcgctc aaaacaggga tttagatccg aaagtggtgc 16080 gcgacaccga agcgatgtgc tataacgctg gtgaagcgat gtcaatcggt ctaattgacg 16140 cggtatcaac gccttcgaag gctgttaccg aattcctcaa cgggccgtcc gataacagcc 16200 cggatcactc aggagcgaac gcaatgagct ttacgcaaga agaaatggac gcagcccgcg 16260 cagaggcggc gaccaacgaa cgaacccgca ttgccgggat catgggctgt gatgcagcgg 16320 ctaaccgtac caaactggca aaccacatcg ccttcaacac gaatatgtcc gttgaagacg 16380 cgactgcaat gctgaccaat tccgccgaag aaccgaaagc cgctgttacg actaccgtcg 16440 ttgacgctgc tgcgccgggt aaaaccggaa ccgaagcgaa gggtgacagc ccgttcgatt 16500 cagtgatggc aagcgcggaa catcctaacg ctggtgccga tgcaggcaac gagcaggaag 16560 gcgcgaaaga aacggctggc ctgatggcgg caatgaccgc tgtcgctggc aacaacatgg 16620 ctaagtaata ggggatcttc ggatgaatct tttgactatg atggccgcct ccagcctgcc 16680 taactatctg gctggaaacg gcgacctcgg ctcttgggag ccaacccaaa tcttcgcagg 16740 tgaagcggat attgtgaccg aaggcggtgc cgctggcgct gacatcgaga tctatcaggt 16800 tatcgcgaag aacgccgctg gcgctatggt gccacatgac ccgactgcga ccactggcac 16860 gtctccggac gaagtaccag caccgcagtc tgtggctatc ggtatcgcag cgcagcccgc 16920 gaagtccggc cagaacgtgc cgtactacat cggtggtgta ttcaaccacg ctgccctcgg 16980 ttggcacgcg tcgcttgata cgctggcaaa acgtcaggcg gtcttcgacc gtactaacat 17040 ccacatcggc aatctgtact aaggagcagc gcaaatggct ggactttaca ctacttacca 17100 actgctcgaa gtacagcgca agctgaaaac gctgcctgcg ttcttcctgc aatggttccc 17160 gcgccagatt aactttcagg aagacatgat tgccttcgat aaagttattc aggacgtaac 17220 ccgcgtcgct ccgttcgttg cacccaacgt acagggccgc gtgatcaaag aaagcggcta 17280 caacacgaag acttttaaac ctgcctacgt gaagccgaaa catgtcatcg accctaacat 17340 gatcatcccg cgtcagccgg gtgaagcgtt gggtactggc accctgtcta tcgcgcagcg 17400 ccgcgaccgt gtcatcgcat atctgctgat gaaacaccgc gccatgcacg aaaatacgtg 17460 ggaatggatg gcggcgcagg ctgcgcaata cggttacgtt gacgttcagg gtcaggacta 17520 cccgctggtg cgtgtagact tcggccgtga tgcagcgctg accatgacaa ccgactggac 17580 tgccagcggc gtcaccctga tggacatgat tgctgacctg cgtgacggtc aacgtctggt 17640 gtccgataag tccatgtccg gcaccgtgat ccgcgactac gtcttcggcg gcgacgcttg 17700 ggatcagttc gtcaaggtcg gcggtaaaga actgtggggc aaagacggcc tgatggactc 17760 aaccatccgt gggtctgaaa ccaacgttac ccgcctgtgg gacgacgtgg aaggcgttca 17820 gtacatgggc gaactggtag gggctaacgg tgctggccgt atgcgtattt gggtgaacac 17880 tcagaaatac cgcgaccaga acgaccaaga acagttcctg atgaagcaga aggcggtaat 17940 gggtatttcc tcggcgatcg agggcgtacg ttgcttcggc gcgattctgg acaaaggtgc 18000 gggctatcag gcgcttgatt acttcccgaa aatgtgggat caggaagacc cgagtgtgga 18060 atacctgatg tctcagggcg caccactgat ggttccggcc gacccgaacg cgtcgttcct 18120 gctgaccgtg atgtcctaat aacccgtaat tcaaacctac gatccgcctg taccgggcgg 18180 gtcgctttga ggaaagagcc atgccaaaac gtaaagttct ccagaccgtc atcgtttacc 18240 gcgacggcgc gcgtatcaaa ccgtccatcg gcgagatctt cgatttcaaa caagccgaac 18300 tggacagcat caacaaaatc aacccggatg ccatcggccg cccggtgatg gaagttgacg 18360 ttgaaaatca ggctgcacag gaagccgcag cgcagcaggc gaaagctgac gcgtccaaaa 18420 aagacgagaa ggctgacggt gctaagaaaa acgccaaaga cgataaaaag tctggcgcgg 18480 aagatgaagt ctaatcatgg cttctaactt cgccgtaatt aaagcgaagg ctcgcaggga 18540 cgtacatgcg tccctgtctg tcgctgctcg ctatgagaat tatgcgcagg atgtcatcgt 18600 agacgacatt agcgtccgct ggcacaacaa gatcgccatc atgggcgact tggaaaacgg 18660 tggttacgct aacgttatcg agggcatcga gcgcatcatt tttatgcgtg acgagttaaa 18720 tgcaaaaggg atcatgctgt ccgaaggcga tagcatcatc atgacggccg aggggtacga 18780 gaacgcccgt ttggtactca aaacgcagga acctatcgtc gggccagtgg aaatcgtctg 18840 gcaggtggcg agggccgact gatgcctgtt aacgtgttag cgatcggctc cgacgagttg 18900 aaagggtatc tggaacaggt accggaaatc gctaacaatt cgatccgcat ggcgattaat 18960 agcgtggctg ctggcaaggg catgaccctt atcaagaagt cgatgaccga cgagattgcc 19020 ttcccaaccg ggtatctcaa cgcagaccgc ctcaagctga ctaagcgcgc caccacatcc 19080 aacctcgaag ctgtcattgt cggccgtaag cgcgccacca gcctcgcccg tttcgttaca 19140 ggcggcgcaa tggtgacgaa tagcaaacgg cccggcggtg tacaggtgcg ggtgaagaag 19200 ggcaagacga cgtacctgaa aaatgcgttt cttgtgcggc taaacaaagg cgcgagcctc 19260 accgaagacc agtataacgt cggtctggcc gtgcgacttt ctgccgggga atccctgtcc 19320 aataaacgct cgcagcacaa atcatggctc gtaccgggcc gggtggcatt gctgtacggg 19380 ccgtccgtgg atcaggtctt tgcggaagtg tccggcaccg tcgcgccgaa gattggcgac 19440 atggtggccg ctgaattcca tcgtaacttt gcgaggctgt ctaaatgagt caaaggcttg 19500 acattcttaa agcattgaca gcgcaccttg aacagatcac cattgagaac ggatacgcct 19560 acgacctgaa aggtaaagtg tatcgcgggc gcgaccgatt cggcgcggac ttcacgtcca 19620 aactgccaat cgtgtcgatc ctcgaagcaa aggccacgga ctacggagcc tttgccaacg 19680 aggaacagac cgtacgaatg gacgattggg tgctgctggt gcaggggtgg gtcaaagatg 19740 acccgagaaa cccgaccgac ccggcgtacg aactgctggc cgaggtggaa aagcgactgg 19800 cgatgttggt ggcgaaggac gagaacggac aaccgatgta tccggccctc taccgcttgg 19860 gtgggaagat tgctaaactc acactcgcgc agcccgttgt caggccaccc gaagacggct 19920 tgtccgatac agcgtttttc ttcctacctg ttagggtagg actcaaagtg gacattagga 19980 atccttaaca ggagaaacag aatgaacgat aactaccaaa acaactacgt ggtaggtcgc 20040 ggaacggtct actttgaccg attccaagac ggcactaacc gtaagacggg tgaaatgtat 20100 ttcggtaaca cgccggaatt caccatcaac accgattccg aaacgctgga tcactactct 20160 tccgatcacg gtatgcgcgt gatggatgcg tccgtcctgc tggaagcgtc tcagggcggt 20220 acctttacct gtgacaacat caacgcggat aacctcgcgc tgtggttcct cggtgaagta 20280 tcgaacacca ctcagacgca gcagaccgac gcgaaagaag tgttcaaccc gatcatgcgc 20340 ggccgttact atcaactcgg caccactgac gataacccga cgggtgttcg cagcgtgact 20400 aacttccaga tggtcaaggc tgacgcgtcc atcgcgattt ccgttggtac aggcgacatc 20460 acttctatcg tgggtgccac tgtcgttaac ccggcgggta actacgaaat tgatctggaa 20520 gcgggccgta tctacatcga gccggattca gccgacctcg cgggcaacgt gcagatcgcg 20580 gttcagtacg acgttgatgc gcagaaacgt acgctggtca ttggcaagtc caacatggtg 20640 tacggcgcgc tgcgcatgat ctccgataac ccggtgggtc tgaacaagaa ctactacttc 20700 ccgaaagtgt ccatcgcgcc ggatggcgat tatgcactga aaggcgacga ctggcaggtc 20760 atgtccttca ccttcaaggc catgcagctt aacaacatca cgcagcgcgt ctacatcgac 20820 atcgtcgaag cggccgcagc ggttgacccg accacgcaac gcaccatcga gatttcgcct 20880 gcatcgacct cggccgcatc cggtggcgca ggtgtcgttt gtacggtgac tgtccgtgac 20940 ggcactggca cagcagttca gggcgatgca gtgacgttta ccaccgtttc cggtgcgacc 21000 gttacgccta acagcgcgac gaccgggtct accgggacgg cgaccaccac cgtcaaccgt 21060 gccgccgctg gtaccgcgac tgtcaccgcc acgctggcga atggcaaagc ggccacgacg 21120 ggtaccatta ccttctcgta atacatgcca aactgataaa gcgccttcgg gcgcttttct 21180 tttatctgca agtatgtgct acattaacga cggattagcg caaaggagat ccgtcaaatg 21240 tcattagcag attacgcacc agaaatcgaa gaaatcacca tcccggccag acacggtaaa 21300 aaggctggct cgtttaacgt ccgtgggcta tccttccacg acatttccaa aatcatccgc 21360 gtccactatc acgaccttga tggcctgttc gacctgtacc agaagactgc cggggaagac 21420 ctgacggcca tcgctacagg gcgcttcgcc gtgtcgctgg tatcggacgc gccggggctg 21480 gtggcacatg tcatcgcgct ggccgccgac gaagaagccg aactggaaaa ggtacagacg 21540 ctgccgcttc tggtgcagtt cgacgccttg aaaaagattg ctggtctgac gttctcggac 21600 gttgaagaag taaaaaagat gttcgctcag gtaatggagc agatcggcaa gatgaagggc 21660 gaagaggcgg ggtcgaaacc cggtcaaagc gcgaaatagt tctcgacttc cacaataacc 21720 ttcgtgccac ggtgtccttc ctcatgtccg agggacaccc ggccgcccgg cattacccgt 21780 taggatattt gtggtcagaa acgaaaatcg cgaagcgccg ggtaaacgcg ggttatgtca 21840 cccaaagttt actgatgcag tcctgtatcg gcgctgtact caacggcaag aaaggcggga 21900 aagagtataa aaaactaatc aaggaattat ctgatggcta acagtaaaga cgtcgagcta 21960 cgaatccggg cgagggacta cagccagaag cccctaaagg ctgtgaccag tgccatcgag 22020 cagatggcga aggcgcagga cgaacagcgt aaagcagccg agcgcggtga agtctcgacg 22080 cgtgatctgg aagccgcata caaaaagctg gaaagtgccg ggaaccaact gcttaaactc 22140 aactcgctga tcgaggtgtt caaacggcag aatcaggcca tgacggaggc gacggccaag 22200 accgaaggac tccgccagaa acaggccgat ctccagaaag cctacgactc caccgagaag 22260 gttacgcaga aacaggaacg ggcgctggcc cgggtgacgc gtcaggtcga agccgcgacc 22320 cgcgcggaag cgactcaggc cgaacgtgtc aaccgggcta cccgcgacct cgaacgatac 22380 gggatcgaga cgtcgaaagt tggggcggcc caagcgggta tcgtcaccag cgtagcgcag 22440 gttaaccgtg tcctcgaacg tcaggacgaa atcatttcaa cgtctgccgc agcagccgca 22500 caagcgaagg tcattcgtgg attgcagcag caggcagatc aggcaatggc gacggcgcgc 22560 gggtaccaaa ccctcggccg cgtggtgcag caggcaaccg ggcaactcgg gccgcttggc 22620 acgcagatcc agactatcgt cagcccggca gaagcggcgc gccgcaccct gtccggcctt 22680 gagaatcagg tacacggcgt tactaccgaa ctggcccgta acagtaagga agtggaaaac 22740 gtcgcacaga aagtccgaat gctcaacgag gcgaataaaa ccgtctcggc gctggcgcag 22800 caaatcgacc tgtaccgcca gcaggtgtca acccttcgga acgcccgcac tgaataccaa 22860 caagcgcgac aagacgtcat caagttagca cagcagatgc gcaccgcgac aactgatacg 22920 gt; atgcgaaaca cggccacggc cgcccggtct acccaagccg cgttacgagc ggcgcaggtg 23040 gacacgcgta acctgtccga tgcggaagcc cggctgatct ccaccagcca acaatcggcc 23100 gcagcgctta acacgctgtc caccgcgacg aaccggaaca gccaagcggc gcgcgacggc 23160 tcgaaggcgt ggtcactgtt ccgagacgag ggccgtacca cgctgtcctt cctgcaacgt 23220 attcgcggtg aagtgctggc gctgacgacc acttacatcg gttttcaggg ggctatcaat 23280 ctggcgggag gcgcgatcga cgcgtacaag aaccgtcagc aggcgatggt caaaattgcg 23340 aacgtggtgg gtaacagcca agcggccatc aacaaagagt gggaatacat ggtcggtctg 23400 gcgaatacgc tcgggatcga catcaccacc ctgtcacagt cgtacacgaa attcgccgta 23460 tccgcgaagg cggtcggcct gtcattgcag gactcaaagt tcatcttcga aagcgtagcg 23520 aaagcgggcc gtgtattcca cctgtcacaa gatgacatgg aaggcgtatt ccgcgcgctg 23580 gaacagatgt tgtcgaaggg tcaggtgtat gcggaagaac tgcgcggcca attgggtgaa 23640 cgtctcccgg cggccttcgc gctgttcgca aaaggcatgg acatgaccac cgccgaactg 23700 atgaaggcga tggaaaacgg ggaagtcact ggcgaggcgg ttatcaactt cgcccgcgag 23760 caggccaaag cgattgacgc tcaactcgcc acggcgcaga aaggtgtcga tgcgatggaa 23820 gcgcgcgccc gcaacgccat gaacgcgttt caactggcat tggccgacgc aggctttatc 23880 gaagcgtacg tgcagatgct taacaagatc accgacttcc tgaacagcga ggatggtcgg 23940 gcggccgccg ttaaactcgg cgaggcgttc ggcacgttgg ccgacgctgt gacgtggtgc 24000 atcgagaacg tggacacgct gatcaccgcg ttaagcgttc tggccgggct gaaagtggtg 24060 cagttcatcg gcggcatgat ttcgggcctt aaaaacctgc tgccgctgtt cagcacgctc 24120 ggtaaaatcg gcgatggcat tatcacgatg ctggaaggtg tcgcggccag aatgattgcg 24180 gcacaagggg ccgtagggct tctcggcatc gcgttgaaag ggctgacccg gttaatcccg 24240 attgtcgggt gggcgctgct ggcgtacgac atcggtgcca tcatgtacga ccagtcgcag 24300 accttccgcg aggcggtcaa cgcgatcatc cgggacttca agaacctcgg gaaccaactg 24360 gcagccgtcg tcgagtccat cccggcgctg ctgtacgatc tggcggtatc ggtgctgcgc 24420 cctattacca cgatgttcgc cgacaccaca caggcgatta ttaagtggat cgcggatgtc 24480 ctgaaactga tcccgggcgt tggtcaaggg ctgtccgagt gggccatgtc cattggcgac 24540 gacctgacga aagagcagcg tggcttcctc gaatcgactg gccgcatttg ggacaacgtc 24600 aataaacagt gggtgaaact gaacgacgaa atggtcgcga agaacgccga tgcgactgac 24660 aagattcgcg ggcaggtcaa ccaactggcc gccgacatgg ccgctatcac gaagggcgaa 24720 ggcttccagt tcacccaaga cccgggaaca ggcgtcacga agcgcagccg cgagatcgcg 24780 ggcctgacga aagagttagg caagatggaa gaggcggcca aaaaagcgga cgtcgcggcg 24840 cgtaaagccg agcagcggaa gaacctgtcc ggccgcctcg ccatcattga cgaggaattt 24900 gcgccgcagt acgcccgcgc taagtcgatt ggcggctcgg agggtgacgc ccttaccaaa 24960 cgtcttgacg ccgttgtggc cgcgcgcaag aaggcagaaa cgaccctctt caactcgcag 25020 caacgtacga cgggtggtat caagaaacag gaaaacgcat tacaggcgct gatcaacaag 25080 tacaacgagt tgaatgccgc cgtcggcgtg aaggaagtta agattgaccc gaatgccacg 25140 tttgacgacc gtctggccgc taaactggcc gccgtcaata cgcagtacga tcagctgatc 25200 gccaagtcga agaaactcgg ggctggcggg cagaatctgg ccgggcagtt cgaagacctg 25260 cgcaaacgca atctggaata cgcgaccacg caggccaaac tcgaagaact gaaacgcatt 25320 gaggatcagt taaactcgca gcaggagacg aagaaaaacc tgctggacga aatcaatgcc 25380 aaacggcagg ccggggttat ctccgaaaat gaagccgtcg cgcagacggt agccttgtat 25440 cagaacatga atacgggcat cgcgtcgtcg gccgagcagc ttgacgcctt cgcgcagaag 25500 attaaagaca cgatgtcgcc ggaagaattc agccgcatca tggcgcagat tgcatccgtc 25560 aaagccgggc tggtggacgt cacaggcacc ttcacgacga tggacacgac tgtcgttcag 25620 ggcgtgctgg acggaatgag cacggcgctg tcatccatcg ttgacgaaat ggcgttggtc 25680 gtggccgggt cgcaaagcat tggcgatgcg ttctcaaacc tcggcgtggc cgtggcacgg 25740 ttcttcgcgg acttcttgca gaagatcgca atggcaatct tgcagcagat ggcgctcaac 25800 gcactggcct ctatgggggg tggcatcggt tcggcggccg ttgcattagg cggcaccgtg 25860 gcgaaacaca atggcggcac cgtcggcagt aagacaactg gcggcacgca gatgaagggc 25920 gggatcagcc cggccatgtt cgcgaatgcc ccgcgcttcc acgacggcgg tctgccgggc 25980 ttgcgttctg acgaggttcc gaccattctg caaaaaggcg aacaggtgct gtcgaaggat 26040 gacccgaaca acgtgcttaa ccaatcacgc ggaggcggcc agacggcgca gtcgccacaa 26100 ggtcttcgct tcgtgctggt ggatgatagg tcgaaagtgc cggaagccat gaacacgccg 26160 gaaggagaaa aggcggtgat gcagattctt caacgcaacg taccgacgtt gaaaaactta 26220 gtgggataat aacaatggct ttgagcggac tgcgccccgc gcgcgaacag tttaaccaag 26280 caatctacga cgcccttaac ggggcgtacc agctttacgt tcaggacgtc ttcggtcagg 26340 ctgcggccaa ccagacgtac tacatggcga agtggatgaa gttgaaggcc gggtcttata 26400 cggcggtcat gtatgtcgat gactccggca ccctgtctat cgatcatgcc gtcgtcgcga 26460 ctgcggccat cggcaccaat ccgaacagcg gggagtttac gatctccgcc gacggcgtgt 26520 accgctttga ctgcatctac tccaacgtcc cggccgacac cccggcgtac atggcattcc 26580 agctaatcct tgacggccag accgtcgagg ttagccgggc caacgacttt atcgcggaca 26640 tcgtgccgat cccggataag gcattggggc cgaagccgcc atactccgac gacgtccgcc 26700 tgacgtaccc ggtgttcctg ccgctgccga actggaaaga cggcgtcacc gagcgcatcg 26760 agtggcaaac tgacgtcatg atttccgagt ccggcgcaga acagcgacgc ccgatccgcc 26820 tgcaccctcg ccgctcattc gaagcgacgt tcctgcgttg ggaagaaaag cgtacgctgc 26880 tcgatacgac catcgcgggc gtcggccagt ctccgttact cctgcctttg tggcacgaca 26940 tgacagccac ggaaaacgat gcgccagccg ggtcggtgga catcttcggc cagttccggg 27000 tgaaagactt caacgtgggt gacgtggtca tgttcaaccg tggcacgacg tgggactatg 27060 agacgaacat catcgccggg ctggacgttg acgccgggca tatgacgctg acgtttggcc 27120 tccagtcgga cacgccgcga ggcacgcgcc tgtatccggt tcgcgttgcg cagatccggg 27180 aggcgatgaa cggccagcag atgaccgata gcgtgtcaca gacccaagtc cggttcttct 27240 gcaccgagaa ctatgacctg acgccatcgt ggtcagactt ccctgtctac acccgtaccg 27300 ggctgcacat cttcgtcctg ccggaagatt ggggatcgtc caatgaaatc acgtctgacc 27360 gcctgaccta caatttcgat aaccagtccg gcccggtggt cattgttgac ccgggcggcc 27420 agaactacgg caccgtgaag aagtcgtaca ccattaacgg ccgcacagcg gatcggcagt 27480 tccggcagat cctgttcgcg ctgcgtggcc gcaccaagac tttccacctg ccgctcgata 27540 cgaatgactt cattctgtcc cgcgacatca acccggccga cggcgcgctc gtagtccggc 27600 gctgcggtta tactcagtac atcggcggga cgcaggaaac gaaacgtgac atcatggtcg 27660 aactgtacga cggcaccaga atcccgacca ccatcatttc gagtcgtatc gttggggatg 27720 aagagtggct attcctgtca cagtccatcc cggcgacgtc cagaaacgac gtccggcgca 27780 tcggatatat ccccgtagcg cgattagatg ttgacggaat cgaaataaaa cggttgactg 27840 actcggcggg cgtaagtcaa gtttctctga ccttcaagtt tttcgacgat cggaggattg 27900 caacgccctt accattatca taagagggca ttatgtctta caacattatc gagacgtcca 27960 atgacaacgg cagacccgtc tttatgtacg agtttcgcct gctggataaa tactggcgtt 28020 atacctccgc agacgctaaa gtaagtgcgc tggggagcat ttgggaaccg atgggagtgt 28080 ccgacgacgg gattaaacaa acgggcgagg ctaagacgga tgcgctaaat ctgacgctgc 28140 caaactcgaa tcccgtcgtc ggcctcttca tcgggactcc ccccggctcc cccgtaaccc 28200 tgactatccg acggatgcac cttgacgata acgacccggt ggtctgttat gtcggcaccg 28260 tcgatagcat caatcagggt gaaaacccta ccgtagcaac tgtcacctgt tcaaccctgt 28320 cagccacaat ggacagaaat ggcctgcgcc tttcgtggag ccgtggttgt ccgcacgcgc 28380 tttacgacgg ccaatgccgc gtcaacaaag aagccttccg cgtggatgcc accatcctca 28440 ccgtcggcgc gggtaccgtt acagccgctg cctacgccac ccggccagat gggtacttcg 28500 cgggcggctt cattgagtgg atcgacccgg tgtacggcgt ggagcggcgc ggcatcgaaa 28560 cccataccgg gaacaccatc accatctttg gtaccgtaga cggcttggcc gggggctaca 28620 tcctcaagac ttacccggga tgcccgcgta ccagtacggc ctgtgatacc atcttcaaca 28680 atctggcgaa tttcggcggt atcccgtcgc tgccagaccg ttccccgttc gacggcaacc 28740 caatctttta aggagaatcg ctatgtggtg ggcgttagca atgctggtgg cctcggtgct 28800 gatcaacgcg gccctcgcgc cgaagcctgc atcggcaaaa ccagcaacaa ttcaagattt 28860 cgacatcccg caggtgaagg aaggcacgcc gcagtcggtg gtattcggcg aggtgtggac 28920 agccgactgg caggtattgg gcttcggtaa cttccgaacc aaagcagtta aagcgaaaca 28980 ggcgaaaaaa taatgagcag acacgacgag ccgcaggatc agccgcgcat ctttatgcgc 29040 catgcccgga aactcgggta ttgcaaaaac gggtcggagc gcttggccga gcgcttcggt 29100 atcaccttcg aacaattcct acgggaaggc tacccggtag cggaagccat gaagtccgca 29160 aacccgctgc tgcgtaaagc cgcagccgtg gcccaacaag aatgggacga ggcgcacgca 29220 catgggaagt aaagccaaga agattacggt tgggtacaaa tactacatgg gcctgtttat 29280 gggccttttc cgtggcccgg tgaatgagat tgtcgagatc cgcgtgggcg accgcaccgc 29340 gtggacaggc tcaatcactg gcaacacaac catccagatc aaccgcgaag acttgttcgg 29400 cggcacgaag gcggaagggg gcattgatgg cccgctcgaa gtttatatgg gcgcgccgac 29460 gcagaccgtc agccagaagt taaagaacat gctcggcggc cgccagccgg aattccgtgg 29520 tgtcgttacc gcgtatttcg acggcctgat ctgcgccatg aacccatacc cgaagcagtg 29580 gaaattcaag gcccgccgct ctacggccgg gtggactggc ggcgtctggt atccggaaaa 29640 atgtctggtc aagatgcagg ggtacgacgg gcagggcaac cagcacgaaa tccatgccat 29700 gaaccctgcg cacatcctgt atgagtgcca gtcgaacttc gaatggggcc gtggcctgtc 29760 acgctcgctt atcgacgata cgacgttccg ccttgtggcc gacaccctgt ttaacgagaa 29820 cttcggcctt tgcatccgat ggaaccgtca ggacacgctc gaatcgttca tgcagcttat 29880 cctcgaccac attggcggtg cgatgtacgt cagcaaagtg acgggcaaac tgtccctgcg 29940 gttgatccgg aaggactacg acttcgacac gctgccgatc ttcgacacgg attccgggct 30000 gctgtccatt caggaagcga ccaacgcgtc tccggccaat ctggttaacg aagtggtcgt 30060 gacctaccac aacccgatca tggacgaaga ccagcaggtg cgtagccaca acctcgcgca 30120 gatccagaat cagggctgtc tgaacagcaa cacgattgaa tacctcggca tccctaccgg 30180 gaaactggcg atgcaggtgg cacagcgtga cttgcgcgcc gcgtctacca acgtccgccg 30240 tttcaccgtg atctgtgacc gccgggcatg gaacgtgcag ccgggcgacg tgctgaaagt 30300 gcgtgacccg aaacagcgcg gcctgaccga agttgttatc cgcgtcggta ccgttgaaga 30360 tggcacgctg ccggacggca agatcaagat tgtcgcgctg caagaccagt tcgccttcca 30420 gttaaacacg ttcaaccaag tcgagccgcc atccggatac gagcctgacc tgactcccgc 30480 gattgcccgc cgtatcgtgt acgaaatgcc gtacgtggat ctggtgcagc agctaccgga 30540 cggcgaactg aacgccgttg gcccgaacga tgcgttcatc aactcgcagg ccgagaagcc 30600 gactgacatg tccgccgcgt atgacatggg tatcatggcc gaaggtgaat cgggcttcga 30660 catccgtggc aacggtgact tcggtgcgtt cggatcgctg gccgccgaca tcgactacct 30720 gtcaacgcaa atcatcctgt ccgaaatgac cgacatgtgg gacgacgttc agccgggctt 30780 cgtggcgcgc atcgccaagc ccgttcttga cggccaacgc acgcagatgg aaatcgccga 30840 agagtttgtc cgcatcgacg ccatcaatgg caacgtgatt acagtggcgc gcggcgtact 30900 cgacaccatc ccattccgcc accagaaaag cgaaatgctt tgggtgacga cgtacgacgg 30960 cggtaccgac tggcagaaat acgcgggcaa cgagtccatc gacatcaaga tcctgccgtg 31020 gacattgggc ggtggccgct tcccgattga agatgcgccg attgatcatc tggacatgga 31080 cttccgccag atccgcccat acccgccggg caacgtgcag cactatctcg cttcgacgtc 31140 aacgctgcaa cgctggtatg tcccgtccgc gcttacctac acggccaacg ccggggaaac 31200 gcctgacacg tacaccctga cttgggcgca ccgcgaccgt atcctgcaag cggataagcc 31260 tgtgccgcac accgacggcg acatcgggcc agaaccgggt accacgtaca ccatccgcgt 31320 ctacaatcag gaaggcacgt tagttcgcac cgaatccggc atcaacggca cgtcgtggaa 31380 ttggccgtat gcaaccgcag cggcagacat gaatgtcgaa gccagtctgt tcgacccggt 31440 gctcgcgacg ctccgcctga cgtctgtccg cagtggccgc gaaagttggg aatactacga 31500 aatgaaggtc agcgtttata aaaagccgcc gcagttcgtg tatgacgcca gcctgatgca 31560 gttcgccgcg cagccgtaca acgccgccag tgatcctaac ccgccaacgc caccgatgga 31620 cggcccgtat gtttcgtccc tgatgcagtt cgccgcgcag ccggatggta gcggtgattt 31680 cggcgaagca gatagcatga gtggcccgaa cgtggcgctg ctgccgcacc aagttacgca 31740 gacgtcgtcc atcattacgc cgctcgatac gctgctgtac gaaaccccgt acatccagtt 31800 atcccgcgac ggccgtgacc tgaacacgtc gaaagtcagc gcatacgtgg cgcgttcgtc 31860 cgaccgcacc gtcgattcgt ataccctgtt caccaagcat gaagccgatg cggattacgc 31920 ttccgccggg actcagccgt ggacaccgtg gggcatgact accgtcggcc tcggcttctt 31980 ccggacgaa gtgacgattg acccgacgtc ggataaagac ggcgttccta tcgctggcgc 32040 gcaggtgggc gatctgctgc ttatcgatca ggaattgatg tccatcaccg ccatcaacgg 32100 ccgcacgtac aagattgggc gtggcgtggc tgacaccatt ccggcgcagc actacagccg 32160 ccgtccggtc tggctgatct ccgtgggctt cggctactcc gacatggcat tcggcgataa 32220 cgaaaaagcg atggtcatcg ttcgcccgga tacctacggc accgacatcc cgctgaacaa 32280 gctgtacccg ctccagttgc agatgcagta ccgtccgaaa cgtccatacc cgccgggtct 32340 gggatgatt ggcggccaac cgttcttcaa caccgcgagc ggactggcag acgatttcaa 32400 cccgtacaac aatctgaaag cgaaagacgt ggcctgcacg tacgcccacc gcaaccgcgt 32460 aactcaggca aacacggcgc gcgaccattt cgccgtcggc atcactccgg aaccgggcgt 32520 gaagtaccgc gtgcgcattg gttatgcgta ctggtcgcag tccggtagcg gcttctcgct 32580 gctgtctcag ttcgaaacgg aagacgcggg cttcattatg cgtgccgccg acctcgaacg 32640 ttggggccgt caggcgggct atgcgcagga cgctggtggc tattccacgt taaacgtgac 32700 cgtcaacgcg attcgtgatg gaatgctgaa ctggcaggga tatacgatga ccgtgcgcgt 32760 tccgtccttc ccgcttcctc cgggacagaa gccgggcggt ggcgatgggc cgtggaaccc 32820 gccgggcggt ggcaatactg gcggtactac tccgccgacg gagccgccgg atcgtccaga 32880 cccgagcgag ccgggcacca ctgacccatc cgagccgacg aagccggacg agccggaaga 32940 accgcagcca ccagtcgatc cggaagaccc ggatactgac ccgccggaac cgccggaacc 33000 gcccgttgac ccgaccaacg taccgggatg gtcattgagt tgggatcacg gttgggcgat 33060 gacactgcca gatttccgat atgttccacc agcagaagag gaacctgaat aatgccgaaa 33120 aagacagccc cgaatcaggg gctaccctac gggtggattc gcggggaaga ctattggggt 33180 ggcccgatgt ccgactcgca aatcttcctc gacaccacgc tattcctgac actggaatcg 33240 ctgaccttct ccgctccgcc gccggacgtg gtggacgggg ccacctacgt cgtcgcggct 33300 aacccgacgg gggcgtgggc cggacatgaa ggcgatgtgg ccgtgtacgt ggaaaaggtc 33360 tggacgtttt acacgcctcg ctacggatgg cgcgctcgcg ttcgttcgta cgagaaattc 33420 ttgtggttca atggcacgac gtggattgac gaggcgaccg gggaagaccc ggtcaaccct 33480 gaccctgacc caagcgttaa gccgaaatgg tacgacatcg gggtgacggt gtctgacagc 33540 atgtacgcga atgaacctgt cgtgcatctg ccgatccttg acccgatgta cctgcccgcc 33600 aacatggtag gatctcagtt tgacatggcc gacggtgcaa gcccggccta ttatcaattt 33660 cgcgtgcagc gcaacggcca gaacgtcgca acgatgaccg tggaaatcgg aaacttcaac 33720 gcgtcgttta ccacatccgg tggcaacccg atttccttcg cggccgggga tcgcatgacg 33780 gttcgcgcgc agcaggaagc cgtcgccggg atgaaaaact ttggcttcgt aattcgcatg 33840 ggactggtct aaggggatca catggcaagt tattttgatg gatatgagca gttccgttcg 33900 acggacacgt ctaacccggg aacctttatg cggctcgccg ggtacacggt tcgaggcgcg 33960 atttcggcgg gtcttggccg cctcccggcg agcatcggac tgtacacgct gaacagcgcc 34020 tacgagcgcg actggacgtg gggtggcgac accctcacca tcggctttgc gtgcaagcag 34080 caggcgcgtg gcgcgttatt cgggctaaag atcggtgacg ccactggccg cgacgtcccg 34140 catgtcgtcg tatacactga ccctgtgtct gcgctcgtca ccattcagac cgggccagac 34200 caactggaag tcggctatgt cacaccgatc ccgacgcgct ggtactacta cgaagtcgtg 34260 atgaaccgcg ccacgaaggt tatcgaggtg ttcgttaacg ggaagtccga cgtggcgttc 34320 acgctgccgg acgtcctcgc ggcgtcggcc gttgtgcgca tggtcttcaa cccgttcgac 34380 atgctcccgg cgttccctgc cgatgcgaac tacaaggaag atacgaaagt cttcgacgac 34440 atgtatgcgc aggacggagg ccgtttaggg gccatccaga ttagcggccg actgccggac 34500 ggcgaccgtg acaccgaatg gggcgtggcc gggcctaacc cgacggctgc acataaccag 34560 atggtcggcc gtcttccgcc tgacttggca aatctgttta tctatactgg cacgaatgac 34620 cgccatgact cgtttatctc gacggcgaag ttaccggata acggggccat tctggcgcag 34680 ggattgatcg cattagtccg gaaggccacg gccgacccgg tgtcgattat cgcgaacatt 34740 gacagtcaca ccgttaccct gtcgaatatt ggccgtaatt gggaatatcg ttacgcgcaa 34800 ttcaccacaa gcggttatga tcgagccgcc attgaagccg ccgaattcgg tgtgcgttca 34860 gtaatttaaa ggggttacta tgctgaaatt tatggacgga tttgaccagc tacgcggctg 34920 gacagacgtt atcgacgggc tgaccaagtg cgggtatacc gttgcaggta cgcccacgct 34980 ggaggaaggg cgcgtggcta cgcagatggc ggtgtcactg ccggatgcgg ccagcctgaa 35040 acgcgtgttc acctccgggg ccacgaaggt cgtgtttggc tttgccttcc gtgcgatcgg 35100 gaaacgtcac accctcgtca ccatcaaaga tgtcgcgacg gttacgtgga acgagacgga 35160 cgggaaaatc tcggcggcgg gcggtaccgg aacggcggtg ctgctgctgg acttgtggta 35220 ctacatcgaa gtggtgctgg acaagaccac cagcacgatc gaggtgtacg tgaacaacgg 35280 gctggacatc accgcgcctt cgccgtcgag tgctaacccg gtcactaact acgaagtcac 35340 atgggctggt gttgccacgg cgcaatacct tctcgacgac ttccagttca tcgacaatca 35400 gccgggcaag tacaccgacc gtatcgggcc tatccagatc accagccgcc tgccgatggt 35460 ggacgtggac aaagagtggt cgccgtcgtc cggtactgac cactacccgc tcgtctataa 35520 ccagccgccc gtcgaagggt cgtacatcca gtcgaacacg tccggcgcga tggacacgtt 35580 tttatctaac accgtcatcc ctgacacgca aaacattctg gccgtcggca tgaccgtact 35640 gaacaaaaag tctgacgtgg acaaccgtca gttaggcatg gtcatggggc caaaaggcag 35700 cacgcaaaaa gaagttatcg acgcggcgct atccacgact gaaaagtata gttatgctgt 35760 gtttgagacg aatccggccg gactggactg gaatgatgaa cgactcagcg aagcgccgtt 35820 cggcgttgcc gtaagaccat aaggagcata ccgaaatgtt taaattctgt gacgggtacg 35880 accactatgc cgaaatcggc gtgaagggta cggccctaca aagttatctg gaagccgccg 35940 ggtacgtggt gcgtaacgcc agcgatacca ccttttccgt cgtcgagggc cgccggactg 36000 gtgcgcgtgc gctgaaattc accgtggccg ccagttcgtc cgtcaacgcc tccctgtcat 36060 ggggcttcac cacgacggcc acgtcagtcg tcttcggctt cgccatgaag gccgggctgt 36120 cccgtatgcg tatcgcgcgt atcgagaaca tcgtggacat cgagtgggac accacgaccg 36180 ggaaaatccg cgtaggcgag cagctaggcg tcaacccgct gatcctgaac gcgtggtact 36240 actttgaaat cgagatcgat aagacggcca ataccgtcaa ggtctacgcg aataacgaat 36300 tgcagttaac cgtcgcgcca tcgagtgcgc cgaccaccac gtacaccatc gtttggggcc 36360 agacgggtac cgcgccgaat gcgggcgaac agacgctgga cgacttctac gtgatcgacg 36420 ggtcgggtag ccgtaacaac gcgcgcctga ctccggtcga agtgaccagc cggatgccaa 36480 cggccgacgt ttctacgcag tgggatgttg tgaacgcctc gtcgtccacg ccgcactatc 36540 agatcgtgtc gcagctttcc ccgggtggcg agtcgaagcc atacctgcaa tcgaacaccg 36600 caggagccac cgatatgtac cgctcgaacg tgacgctgcc gaacaataac cagatcttcg 36660 ccgtgtccgt catcgcgtat gcacgcaaag gcgaccttga tgatcgtaaa ctcgggctgg 36720 tggtcaatac ggaaggcgga acgccgactg aaattcaggt gccgcttacc gaatcgtaca 36780 agtattatca ggcatcgttc gagcaagcgc cgggcggaag tgactggaac cagaacaacg 36840 tcgaatcttt gcaattcggc atcatcacgc gataagggag ggacacacta tgcctgtagt 36900 atggcactgc ggttttaacg gcatcatcaa taacacgatg ggtcagggtc aatccaactt 36960 attcatcaat acgaacatga cgtctggcgc gtcgtatgcg cttgccgcgg ataacctgat 37020 taacaatgtt tttggacgag gggggaacgc cgaccagaac ggaggggcgt tacctttgat 37080 ccaatactac ggccgaaaat ggtcgtcaac ggcgggctac ggtaactcat atgtgtttgg 37140 ccgcgtgaaa ccatcgtatt tcgataactc cgtgccatgt ctcagttcag aaatgtctca 37200 accgactaac caatctgact ggtacggggg gtggagttta cgggtggata cgctattccc 37260 cacggactat cagactgcaa gttggaggat gggatttcga atcgggcgag caccccaccc 37320 gacgtataat atgggatacg ctgtttcgat ttgcccggca ggcgtcaatg cggccgaatc 37380 gctcgcgttt tataacatcg cggtaggcag cggtagcatt cccgaaaaat acgtggaagt 37440 tgaatggctc gcggacacta agtccctgaa catcttcgtg gatgatgaac tcgtttcggc 37500 cagaacgaat tacactgctg catcaatggc gttcgggatc atcctcatgt gcgagcatta 37560 tgtcggggga aatcttagcc aaacgccgtt cttcggtttt gagttaaggg atatgtatat 37620 ccagcgtatc ctttccgaag cagaccaacg cctcggatca tccacgaagg tatatgcgtt 37680 caatccagtg actgacgacg ccgtccagtt ctcacggccg gacgcttacg cgtcaaacgc 37740 ctccgtggta cagactccga tcggggcagg tgggatcact ccggccccgt cacaaacgcc 37800 tgcaatcctc ggggcggacg tggtcggcca gcaagacctg tataacatcg acatttccaa 37860 agtcagttcc cgcctcgcct cgatcgaagc agtcaacgtc cggacaatgg cccgtaaccc 37920 gttattgggg aaccgccaat ccgctgcgat cgctaagtcc ggggcgacac agacccaaag 37980 cgagaacact tctaaactcg aacagggcgg actatggcgt ttcaacacgt tgcagatggt 38040 atctgacccg gcggacggct cacgttggaa ccttacaaaa ctggccgggc ttaaagtcgg 38100 taccaaattc gtcgcgtaag ggggccatgt ggctgatgaa gacgtaggta tccgcgcggc 38160 caagttggtc ggcgaagcct tacagtggaa tggcgcgaag cccaaactca catacctgtc 38220 ccggtgtgta ggggaggcgt tagccatttc tgcgttaaac atcgacgcgg cgaaaaactt 38280 tggggaggtt ttacagcggg acggtaaatt gccaaccctc ttccgcgcgg ccgagttggt 38340 cggcgaagtg ctggcgattc agacgatgga gatccgcgcg tcaacgatgg tctgtgaagc 38400 gctggtcaaa gagccgcaat ttatggcggc cgaaatgatc tgcgaggcta tcggtcaggc 38460 ggccactgaa cggccaccgc tgtacgcgga ctcggtctac cagtatgtaa cgcagcacga 38520 cgtcgtaccg ccgttgtcgg atgtcatttc tacggaacgc ctgtactcgc tgtggaatta 38580 tgccatcgtt cagcagccga tcgatgaccc gatttcctac aacatcgtga agcaggcggt 38640 gacgctgtcc gttctggcgt cgcctatcgt gactatttcg tacgacaagg tgaagaccgc 38700 gtggtcgtat gccgtggtgt ccgcgacgct cggtgacttc ccgctgccgg atgaaatgtg 38760 gtcgaagatc agcacgaaac aggtgtggaa tctggtattg cagtcgatcg acatcccgta 38820 cgtcccgacg tctggtgtgt tcgttcgcca tgaatcgacg ctggtggttc aggcccatcc 38880 gctgccaatg tacacgacac ccgcatccgt taagtctaac gtggttctgg ccgcgcagaa 38940 gcgcccggtt gaacgcctgc cgcgctcgct tacccgcgtg gcgcaggaat tatcgcaggc 39000 cgtttccgcg ctgcctatcc cgatgccacg gtctaacgtg tttgccgcgc aggcgctgtc 39060 acaggctgtc atcccggcgg acttcgttga cccgactgtc ggtgtcgagc acgcggccgc 39120 agtgacgtcg tacgctgttt gcgagtatca ggtgtcgctt cctatcagtt acacgcgcgt 39180 gaagaccctg acgccgctgg tgctgcaatt tagcgataac caacgcacca tcccgctgtc 39240 cactatgcgc accaagcacg tctcgacgca gttcgtggtc aagtccgact tgttcccggt 39300 gctgtcgttc agccgggtga aacaggtgcg cacgtacgca gtgcaggaga cggcctacac 39360 gccgccgatt aacctgctgg tgtggtcgcg cgctgacgct gttcgcatgt cggtggtgca 39420 gcaggcaacg gactaccctg acccgggcat ccctacaacg tcggccatcg ttagtgcggc 39480 cacggcgtac cacgtcatga aagcgctgga ctacgaagac ccgaacgaca tgtgggaagc 39540 gagcaaggtt cagatcgtgc cgcagttctt agagaaggtg gtgaaagccc aaccgctgcg 39600 gctcccgatc tcgtacgctc cgctggcgca atttgttgaa ctggtgacgc ggtaccagta 39660 catgccgcct ccgggcgaag tcgccgagtc cggtatccat gttcgtcagg tgatgcagcc 39720 agtggcgttc cgcgcgacgt accctgaccc gactgtcccg gtatcccacc tgacggtcaa 39780 tcaggtgctg gagcacatcg cgatcccggc cgactatccg gacgtccacc tgccgacgtc 39840 ggacgcgatt gtaaatcagg tattggaaca cgttgcgcag attgacgatt tccctgatcc 39900 cggtaccatg ttccggccgc ttattgtcag ccagatcatt cagcaggcgg cacaggatgc 39960 gtcttacccg gattggagca cactgcataa accgcttgcg gttcagcagg taattcagca 40020 tgtatcaatg acggcggaat accctgacaa ggacgtcccg caatcatttg caaaagtgtc 40080 tcaagttttg cagcactatg ccgataaagc taattaccca gacaaagatg ctccccaatc 40140 cacgctacga gtccgtcagg tacttgagca ggtaatgatg cgcgacaagt cgatgtacga 40200 aatgccgcag ccaccacgga aacatcgggt gcaaatcact tgccgctttg tgtactgaca 40260 attgacgggt taatattctt agcgggcttg tcccgctaat tttttgtgag gaaaacaccc 40320 aatgagcgat atggaacggg aacggccgga cgataattcc gggggcatca agttagattt 40380 gtcggtcaat ctccccacga ttctgacgat ggcgtccatg ctggtagcgg cggtgctgta 40440 tgtcaataac cgttttgctg acctgtccaa ccagaatacc caagccgaga cgcgtttgac 40500 caacgtggag aaacggcagg acgcaactga tgctgcgttc acggtgctac gtgctgaaac 40560 cgtctcgcaa aactccgccc ttcgttcaga ccttcgcgcc gacatgcgcg acctgaaaca 40620 aagcgtggat accctatctt cccaaatgat gaggaaataa ccgatggcta aacagaagcc 40680 gcgcgggatt cgcaataaca acccgggcaa catcgagtgg gggtcgccgt ggcaagggct 40740 gatcccgcgc agtgaagcga cggacagccg cttcgcacag ttcaaagacc cggcatccgg 40800 catccgcgcc attgccgtca ctctgaccac ctactacgac aagcgcaaag cgaatgacgg 40860 tagcaaaatc gactcggtgc gagaagttat cgagcggtgg gcacccgccg ttgagaacaa 40920 cgtatcggcc tacgctaaac aggtggccgc cgttctggcc gttgacccga atagcgaaac 40980 gctgaacctc cacgactacg acaccatgcg cggtctggtg gaaggcatca tccgccacga 41040 aaacgggaac ccggaagcgt tcggcctgac gccgtacagc aacgcgaata cgtggtactc 41100 ggacgaagtg atcgaggaag gtctgcgccg ggcgggcatc gttaaggcgg ccaagcctgt 41160 taaccgtacc accgtggccg cgacgagcgt ggccggactc ggtgcggctc aactggtgga 41220 catggttcag cccgtcaagg ctgcgatgga cagcgcccac ggcgatattt cttccggtga 41280 ttgggtacgc atcgcattcg gcgcggccac aatcgcgatt ggcctgtaca tgggatgggt 41340 cgcgtaccgg aagcatcggg ccggggcggc tgcatgacag ggctattcgc ccggatcaag 41400 acgggcgtcc ttgcggcgct cgttttcgtt gtagccctct tcggcgtctg gcgggcagga 41460 cgcgcgaagg gcaaacagga tcagattaat aaccagaact atgacacgat gcgggaacag 41520 gcaaacgcgg ataaacatgt ggccgaggtg cataatgaaa tcaataaact ccctgatggt 41580 ggcgctaacg atctgcttcg ccgtaagtgg atgcgtcgaa aggattaacc ctgcgccgtc 41640 tttctgttct gtcgcgcgcg ccatctacat cggcgaggaa gacgtgctga cagatgaaac 41700 cgcacggcag atcgccaccc acgatgaaat cggcgaacgg ctatgcggct ggaaatagtt 41760 aaagagttgg gtgggtaata atcaccgtac gttcaccctc gattgtctgg tgggtctgta 41820 gcgaggacac cagatgatcg aggttattga agaccgggat attcagccgc ttcgcctctt 41880 ccacctccgc gcgggtacct gaactgcgca tcgcatcttc accaatcagc agaacagcgt 41940 cccaacggcg catcaattcc atcgtgcatt ccagccagta ttcgtccggt atctcgccct 42000 ctgtcttgta ctcgaagttg gccgtgttca ggtgcggcgt aaccgggaac atcttgtgga 42060 cgtagagttg atcaacaacc gacacggccg cctcttcggc tttcaggatg ttgaccttcg 42120 tcggaacggt gcagcccgaa ccgatgatgt acgggcggta tggcccggcc agatagatca 42180 tgacgggctt ggccgccggg ttacacatat tcacattcat acttcatcgt cctcgaattc 42240 ttcgtcaaaa aggtcggact ggaatggctc acaagatgat gtgcagccgt cgccatcgtc 42300 cgggttagtg cgggtcaggt acagcaggcg ctggcgatct tccagtccgg cgtcggccat 42360 aagctgatcc gtgtctcgtt tgccacggaa ccaagtatgg ccgtcttccg ctttaacgtg 42420 gccgtactgg atttccatgt cgcggttcca cgtaaaccat tcagggtgtt cgtgggctat 42480 caggtacagc tttgcattgc tctttttaaa gcaggtcagg cagttcccgt aatgcggcgg 42540 gatgttgagt ttgaacggca tggcatccca atagtcgttc acgtcctgct tatcgaaact 42600 tccccaatgg caaagggggt acaccagatt gaacttcttg gccgacgcct tattcgggtt 42660 agcccggtca ggctcgtcag accgcattcc aatggccgtc ttcgcactcc acccacggcg 42720 cgccagtccg ttgctgcgca tccagtggcg gatcgtctgc gttttcagat agtcgctgca 42780 cttctggcgc gatatgttcg ggatgccctc gacgctgatc aggctttcaa acggttcgcc 42840 cttgcgcgcg gccgtctcga aactgacaac ccggtggcgc atcccgacgc cgtgttccgg 42900 gctggcgtat ccttccagcc acaccagatt caggccgaag tgtttatcca ccttgtcggc 42960 gaagatcagt gtttcttcgt gctcccggcc cgtattggcg aacacgaaat gcaggtcata 43020 tttgtggccg tagttctcga tcaggtatgc acaaagaaag ccggacgact ggccgccgga 43080 aaagctgacg acaagcggct ctttgttagt tggcatcggt taaatcctcg cataatggta 43140 caaagtcctt cgggccatca cctgaacccg ggaagcgccg gaagaccatt tcatcttcga 43200 tggtcagttt atccttgctg ttcccgcgac ggtagaagtc ggccatgccg ctgccgtacg 43260 tgcggtcaat cccggcgcca tacgggtctt gcgggcataa ttccagcgcc agcacttcgc 43320 gccagtctgg ccgatcgata cggctgacga tgcgatggtt gttcgacacc cggcagtatc 43380 ccgaggcaat cagcgtttcc accagccggg cctgttcctt gccacgcatc gttttcgcgc 43440 gcttgacgtc ctctttcctt gcccgggcga aatactcccg cgagaagcgg ctaccgacgt 43500 tctccgcgaa gatgcgcagg tctttgatga ccatgcccca tttccggcgt ccgccctctt 43560 ccacgtagcg aatggctccg gcgtagcgca gcaacattgc gtcggtcagg cgggcattct 43620 tcgggtcttt ggctttccac cagccgtata cttccaccat gcgggccatc gccttgtcgc 43680 tgtaccatag ctcggcgtgg ctggtgggcg agtattccgg gtaacacttc gctccgccgt 43740 caaacgcccg gtagcagtac cagatcccgc cgctgtactg gaagtgcggt atcttgtgtc 43800 gctctttcat cgtagccatg tcagccctcg gccagttcag gattaatggt aaaccgcagc 43860 ccgtacttaa aatcgtttac ggacagcaga tggatgatgt cgtccgagtt ttctatcttg 43920 atgcccttcg cggccagcgc ttccctcggc ggaagccacg gatcgatagt caactggcgg 43980 tataccgggg ccatgtgtat cggcgagcct tcgcgcacga tgaatggcag cgggtacgga 44040 cggccagacc gggcgtggat acgacgtggc ccgggcgaac gggtaatacc gtagaacgat 44100 gccataacgt cggtcagccc tttgccgtac atcgagcgta cgacttcttc gtcgccgcgc 44160 aggagccgct taacgatatg ccggacgaac gggtacagat cccacttcgc gcgttggcgg 44220 atgaccgaag cctgtcggcg ggcctcctgc tggaataatg atccgcgttt cattttttat 44280 atttatcctc ttcgttgaaa ttacggacag gcacgccgtc ccaaatgcgt acgatgcccg 44340 ggtgcgcctc gcgccacgaa ccgagcaggt ggtcggccca ctctttcagg tcttcgctgt 44400 atgcggccag aagtcggtct tcaatggact ggcgttccgc tgcggacagc ggcccgacct 44460 gatggatgat gtcaaaccag atctcgtcgt ggacaccatt gacggccacc agatagcctt 44520 ttgcttccag atctgcgatg acggcgtggg tgcggtcggc tttcatgcgc atgtagttca 44580 tgcggcggcc ctcccccata agatttgtag ggtgcatggc ggctcgtccg ggtcggaata 44640 gatttcgaac ccggcgtcct tgatccagcc aatcagcatg gcgcgggatt tatcggacag 44700 cgggtggtcg ttgtgtaaca gcgcgaccgt agtggtggtc agcccggccg atgcggcccg 44760 gcggataaga cgctctacct cgccacgtac ttcaatcgct tcggcggtgt ggtgatcggt 44820 agcagcttta tacatttcat cggcagtgat agtcagcatg ttgcccccta aagtaaaaat 44880 ccccgttgtt gccttcgggg ataatatagg ttatacacgc gacgtatgtc aaacagtttc 44940 taagactttc ttttccgggc gtgctccagt cgatcaccgt ggcccttctc gaccatgaaa 45000 tcgcgggtct gtttgtgcag gttggcgatc gtctccttcg ttttccgcca gcgcgccact 45060 aacccttcgc gcacttcccg ttcgcggaac aggacggcca agtccgtcgc ggtgatgtcg 45120 tacccttcgc acgcggccaa aactccggcc ttgaattcgt agaacaggct ttggatctcc 45180 cggaattcgg cctcggttac gatctcgata ttgaccgttt tcccttcacc gatgtcgtcc 45240 tcggtaaaca ggcacccggc ttcgccaatg gcgcggccat cgtccttccg gtagcgcgta 45300 ttccctacga tgacctgcgt cggcgtggtg cgctctacgc gtcgtttcac ggcgaaggtt 45360 acttccgggc tgtacccgat acggatgctg ttacgctcca gtgtcatgtc gccagtttcg 45420 aggtgtccga atctcaattt tttatcggcc atcggtgtcc acctcgactt cgaacgtgcg 45480 cggaacggtc atggtcagca cgtcattctc ggtgaacggt gacaatggct cgtcgcccgg 45540 ctggaagcgc agcgtcacgc tgccgacgtt gtgatacgtg atatagcgaa ggcggaactg 45600 ctcgccaaag atggtcgcga gtacgcccgg cttaacgtct cctgcgggaa ttttcatggt 45660 catagtctcc agtctaacgg ctcccattgg ccgtctcgtt tgatgtagca gccgcttaac 45720 gggctgtatt tggttggtag ttcggtctgg acgctgacgt cgttacaccc gatcgccgcg 45780 cccacacggc caatgacaaa cagcacaccg cagataatca atacaaacag cgttagcgca 45840 gcgatcttct tcatggtggc ccctttattc cttcggtgga tttgattggt ggcgatttag 45900 ccagatgaca atcccggcag tataattgat cacgccgagc aactccctaa ccctttggtc 45960 atgcgttggc aggttgacgc cttctacgac cttcttgcac acctgataga ccaagccaag 46020 cgggctatcg atcaggtcag aaatggcgtt catcggctgc tggctgaacg gaaggtcatt 46080 ggcatgtctc tcgcgccctt taccatacgc ggcttgcgtc agcgcatcat gtagcacttt 46140 gactaattcg gaataatcgt tgctgtcttt gacgttccgg aattcgtact gccatgaccg 46200 ggcggccgac ggtgcaacgg cttccggagc gacgagggct tcgttaagcg gccatccctg 46260 cgcgaaggcg aacatgatgt aattcatcgc gtcgatgtac tgattctctt cgagacagcg 46320 gcggatggcg tcggaacatt gcttcggcca gtccggctgc atccatccca attccagtcc 46380 cgggtttttg cgctcgaact gcaccagacg atcgagcatg tagccgaaag tcttctcggc 46440 ctgttcaacc gttggctggc tcaacttagg acggcggagc acgatgaccg tctcggcttc 46500 ggcggatttc ggaatggcgg gctgattgtc catcatggaa ttgataaggc cgagaacgca 46560 gtcggtcgcg tcgtcgtccg tttcaaagtc atccagatcg aggttcagtg cgttcatcac 46620 tgcgttgcgg aagcgctgcg cggagggctt gccagcgtgc ttctgttttg atgcgtctac 46680 cataataaaa aaaactcccg ttggtggatt aaccttcggg agtaatatta attacacatg 46740 tcgcgtgtgt caaacgcttt ctttaaatct cgtcccacat tttgatccag cggttccgga 46800 tcttggccga tacgttagcc agtggtgcgc ctatttccag tagcgtttca cggtcgatgt 46860 actcgttggg gtggatgcgg tcactctccg gatcgaagat gtggatggcc gcgccgaacc 46920 cggccgacgt gatttcgctg ctgccttcct tcggccggaa ccagactggc ggttcgaacc 46980 cgatgcgccc tttgatgaag atggtgcggg tggctatggc gtccggccac cagctttccg 47040 atgtcgcaga cttggtcagc cagatggtgc ctgcaccttt ggcccgttcc tcgtcagcct 47100 tgcgcatgat gtgggtcatc ccggtcaggt gttctttccc ggcgcgcttc tggctgtacg 47160 gcgggttagc gtaggcccaa acgcgggtct tctggtcgaa cccgtcgctg gcgtggatct 47220 cggccagacg cttcgcccaa tcctgccgca gcgcgttatc ttcggccgta aagaatcgct 47280 ggcatttggc gttctggccg tcggtgaaca ggtcgagcac gattggcccg tacagcttgt 47340 taatcgcttg gaacagccat tccggcgtcc gccattggtc gccgatcaac ttcggttcgt 47400 gctcgtccat gcctttcagc gcgttcagcg catcaacgta ctggctcatt aaaacgtttc 47460 tccgttcagc aggtcatcgg cctcaacgta gaaggcttcg tatttgtccg gtgacatcgt 47520 gtacagcggc tggccgccag tatatttatg ggtcaggttg gtgtacgtgg atgcgctcat 47580 atcgagttga atacggttcg ccatgtcctt gaccatttgc aacgtgactg acttcggtcg 47640 cggtgacagc agcaggttat cgcacatagc atggatatac gtgacgatcg gtgcaggcag 47700 gtcggccacc agattcgcgt tcaggcggta atgcgggatg tcgaagaact ccgccgcgtg 47760 gtcagggtac gcccggaagc cggacaggat caccatgcgg gcggtggtca ggtcgatggc 47820 cgccagttct tcccacgcgg cacggacgga cgcggacttc cgggtgtaac gtacgtcatt 47880 ctgcatcgat ttgatcgtag cgcgcagctt agtcgtttcc cggtcgtgct cggtgttatc 47940 ttcggcgtga cggcggtggg cctcggataa ctggatttgc agactggtga cggcgctctc 48000 tgcggtatcg aggctgcggc tcttattatc cagcgcttca cgtagcgcgg ccagttcgct 48060 cggcgtgatg tcggcgaggc gcaacggctc gaagtctttc agggtaagcg ggcctttgct 48120 ggtcatctgc ccggtcaggc gctggatgtc ctgcatggcg aggcttccgg ccagttcgat 48180 gctggtggtg cctgcaatct tcgcgtattt agcgctggcg cgtaccgttt cgaggatggt 48240 gctcaggatc tgctggcact gctcgcgctc ccactcggtc acttctgcgg ccgttggcgt 48300 aaccgccgat gggcgtttaa ctcgcgtcca gtgaatgcgg atttcccggg gcattgcgtc 48360 gccgtccacc tcgacttcat cgactgactg gatgttccct tccggaacgc cgcagttatc 48420 gagaaggtat ttatgcgcca cgcacaggat atgatttggg gatatgccgc cttcctgctc 48480 gatagccatg acgaggatgt ctttcaggct tttgtcgtcg gtgatgtaag tgacttttgg 48540 ttcgctcatt tggatgcctt taagatgtta gagaattcaa cgatgttaac gattttgtgc 48600 tggcggccgc tgttcgatct ggcgaggctg gccgactcca tcacgcgagg attcatttca 48660 acgcgcggcg tgcgcccggt gccgtattga tccatgctgc tcatgcggtc aacgctgatg 48720 ctgtacatct ggctggccgt catccagtta cccatcgtat tgatgaaaaa ttccatcgcc 48780 tgcaatgcgt cgagcgcgta gaccatcgtg gctgcgttgt ccgtgtacag cacgtacgcg 48840 ccgacggtct gcacgacctt atcgacgccg ctcatgtact gctcgatcca ttctggcggc 48900 actggcggca cggccagacg ttccccggta gccgggctgc cgtacggata ggcgaaagcc 48960 gggctggacg acacgcgacg tccggtaggc gtggtcgtcg ccattgcttt cggcgcgtcg 49020 gattccggca cggtgcatgg gggcgggtcg atgtcattcg catcccatgt cttctgacag 49080 tcccggcaat agaattggtc actatgctgc acgactgata agtggcttcc gccagtggct 49140 tcgattctca tatctcgact tccccttcgg ttaaaatgcg aaacgcgccg ccgccgactt 49200 tccagccctg ccatacgtcg gccagaagcg ccgcccggta cttccggcga aacacgttgt 49260 agatcatgtc ccaattctcg gatggcaggt ctgacgcgat accgccgctc tcccaagccc 49320 aatcgcgagg cgactgcgtc agccatttca gcgggggccg ggggtagacg tacggattcg 49380 ccagaccgca ggccaccagc ccgcgtttaa tccatgcacg ttgccaccat aaatcgtgaa 49440 ccgtcttccc ggcgatgaat tcttcgtcca gcatcgcgca caggatcgtc atggcggcct 49500 tttctgcgtt catgggtacg ccaatgtcgt cccaatcttc cagcgcggcc ctgacgccgt 49560 cagcgctgcc gtcgtcgttg ctggaggctg aacggtcatg caggatgacg tcgaaaacca 49620 tgaacgggaa cacgacctgt tcgccgttcc gcgtgtattt cgggttaggc tcgctgaaca 49680 tagccgagta caggcggcgg atgtggtctt cgagatccgg gacgaagcca gtcaggtttt 49740 cggtggtgtc ggcgttaacg acttcaaggt cggttccgtt ggtgcggaca aggcagcgcg 49800 gcccgtacgg cagggggaac acgtacagca cttctttccc gatcaacgct tggttgaaag 49860 gtacgaattt cattacgtgg gtcttcatcg gcatgactcc tttgttatgg tacgggaata 49920 ttagtcgtac acgcgacgta cgtcaataac tttattcggg gcaaaagaaa gcccgcgtaa 49980 agggattgcg ggcttttgga aggtggcatc cgaagattac cactggctta aagcggacaa 50040 cgcaggagcc gataaggaag cactatgccg agtccgcagt taatcatatg tcggggttac 50100 tgtaaataag attccagtat ttgtactcct tcaagtccag ttcgatgacg catgggtcat 50160 ccagcgcgct caggcaggtg ccgtagtcct cgacaaagac gcaggaggat tccggatgat 50220 gcagcagtcg cacccggcgg tagttcaggt gccattccgg actgaacaca tggttctccg 50280 gctggcgtat cagcatgatc tggcctgtcg ttctgacctg ccagtattgg tctacttggg 50340 gcgtggcata ggctcccatt tcgttactcc tttttgtccg ctctgtcccg gtaaacgcgg 50400 gtacccatct tcccgtagat ctggtcttgc aacgtgaacc ggatggacag gcggatgtta 50460 agccgatagc ccattcgacg cagatagcca agatcctgcg gcttcacgcc ttcgataaag 50520 aacgaatcgc cgtccttcat cccgagcaga agtttttcgt gagtttcacg cacgcttttg 50580 tcggtagtcg cggccattgt taaacctcgt cgtcgtcttc ttcgtcgttc agcgttccgg 50640 tgtactcgac tacctgcccg gcctcgtcca gcacgctgat caccgcacgc agggacttat 50700 cgagttggaa caggttagcc atcatggtca ggaactggcc gccgtcggtg ccgtagcgtt 50760 cgagcgtggc gtcgtagttc gcatcaacgt cggcctcgcc gatgtggata ccggacagga 50820 tggctttgtc ggacaggcgg aagtacacgg tgtcgatgcc cgggcattgt gtatcgactt 50880 cgagatccag cgcgcggaca gtccagtgac gtgacagcag gtcaaccacc gactcgtctt 50940 tgaagatgtc cgtgtccttc atcttgacgt cgtcgccatc gtcgtgtttc aggtggataa 51000 acgcatcgcc cttgatgccg tcctgctggc cgagaaccgc gcgctgcatt aactggcgca 51060 gctcatactc gctcggcagg aggtgttcga cgggccatgt gccgagcgcg gtacggatca 51120 gt; acaggatgac gttgacgcgg gacgggcgga taggcgcgtg tttcagcatt tccgcttcca 51240 cgtcgtcctt catctgcgcc caatccttgc ggttcggctc ccaaccctcg atctctttct 51300 cccgcaacgc ggccatgcgc ttcacgactt catttcgtac gctaacgccg ggcagcactc 51360 gctcgcgctt ttcgatctgg aagacgatgg cgtttgttcc ggggatggcg agcgcaaact 51420 tgctgtcaac ctcttcgccg tacgctgcaa agcccattgt cttgtaagac gcctccggcg 51480 gatctgacac ttcatgcttt tcgagtgcat cgctcaacgc tccttttgcg aaagccgtta 51540 cggcctcttc gtcaacgttc ggtaggttta agatcaggcg gtgcagcggc ttgaccttct 51600 cgttaccgtc gttggccgcg tcgtacagcg cgccgaggtg caagataatg gctttctcat 51660 aacccttgat tttcatgctc tatggctcct acgttgatta aattggctgg tgctccgtgg 51720 tacatggatt gcgctgcatg gaacatcgcg gcgcgtagtg cttctttggc cgcccgaagg 51780 ctcgccattg attcagcgta ctggtcgctg accgtgttat atgccgcctc gtcgttgtcc 51840 agcgttgcga ggtcgagcat tttggtactg tgggtctgca cgtcttcttc catgtccttg 51900 aaaaacgtct caagggcaga cagtttgcag gtgatttggt ctaaccggac tttgcgcagt 51960 tcttggaact gcacgcgaac gaattcaatg ccttccacca gccacctcca cgacgacatt 52020 ctgcacttcc gggcatggca tattcgtcat ggcccaatcc agaaacttca cgatgtacat 52080 gcgcatggcc gtgcattcat ccgcgccgcg agcctcgccg tgcttcgcga tggccgggtc 52140 gccagactga tagaaggtca cgcggtgatt cgtagccgac gacaccttga tcccggtgtg 52200 cgtggcgtgg atggtcacgc tgtagatccg ttcgatagcg cggaacgccg ggacgacgtc 52260 gatgcggcct ttcgggtgca tggcgttgtt caccatcaac gcggtattag tcggctgcat 52320 tgagcggttc ctctttcaga cggtaggtga tggtgtccgg gcgagtcgcg tcgatttcca 52380 gttcaggcat ctgcccggtg atcagacggg acggcgtcag cacggcggtg cgcaggccat 52440 tctcgaacat gatgttggcc gcagtgacca gcgaaatgct ttcgatatgg tcttgggtct 52500 ggtcgaggcg gcgctgtagc agttctacct tctcggcgtt ctcgtccgga aggtcgaagg 52560 cggcacgcag tccggcgatc aatgcgatgg cgtccgcgcg ggtcaggggg gtggcgatct 52620 cggtgtcacc gttgtccttc tcgatgtaga tctcgccctc tttttggccg ggcttcttat 52680 agataaaggc cgtacgggca atgtcgcgta ccggactcag ttcaataggg tggtgcgttg 52740 ttttcatcgg tgtgcgtcct atagggtgtc aggtcaaaag ggtggtacgc catttttcag 52800 gcgtgccgtc ctacgttttt aaacttcgtc gtcgtccgaa acgtcggtgg ccgtacaggt 52860 tacgacggcc tgaatgaaca tgcggccaga cgggtaagtc accagtgcgg attccatgct 52920 gaaccgttgt ccgtgatgtt tcatggcgtt ccggatggaa gaattaaggc tggcccgcat 52980 cttcgctttg agggcattgg cctcgtcctg tacctgcgaa agcgtacggt caagcgggag 53040 ttcttgggac agggtgtaca cgccaccgac ttccagcgca atgaggttcc ccacgacgct 53100 gtacacgcgc acgcgtgacg gcagttcaac atccggctcg gtcgtcatca ttgcgccagt 53160 cagttcggcc gggcttagtt ctttgctcat tgggatggct ccagttggtt cgtgaatcat 53220 tatgttcgga gaacaatagt tctacacgcg gcgtatgtca attcatttca tccggcaatt 53280 atgatgaccg ggtggcactg cggttcagtt tcaaccggat gactttttga agtaaggctg 53340 gtggccgggc cgccgggctt caaccgggta actttttcaa gtaagcgcgg aacgtcgcaa 53400 cgacaaccgg acaggttttt caagtaagcc ggacggccgg gtgaaattgt cccgacgctg 53460 gtggataaaa acgtcagccg ggtaggtttt tgaagtaagg aatggcgcgg tcgttcaaaa 53520 tttttcaacc gggtaacttt ttcaagtaag cccgtcggga atttttatag agaaagtcag 53580 gcccggcgcg gcccggcgtc atggtgtgcg gccgtttttg gcgggatttc gtacaaatcg 53640 ttataaatca atgacttaac ctatgtgccg accgacccgc tttggcccgg ttggcgttgg 53700 tttttcgtat agaattcaaa ggtttaaggc gtggcgcggt gtgccgtttg gcggtgtgcc 53760 ggggcggcgg ccgtggggcg tggcggtgca atgcgtcccg gcgtggcgat cggtggcgct 53820 tttatgtgcg gttaatcaaa taatgtgaaa aaagatttga cgcacgccgc gtatatcaat 53880 tacatttaaa ttcccttcgg gggacggggc aaaacgtccc gacaaattaa caggagcgca 53940 caccatgaaa actaatctct ttttaatcgc gtttgccatc gcaatgatta gcggcggcat 54000 atggggcggg gaagacttcg ccgcgatcgt catcgttggc ggtttaggct tcgcggcgtt 54060 agggatcgcc gtcatggttg gcggcgcggc ggtgtgcgta ttacgcgaca tcgtgatcgg 54120 tgtgcgttac ctgttcaaac gttaatttaa agaggtggca tgatggaaaa ggtatataac 54180 gcgatcaagg gtcaaacgtt gcagcattgg ttaacgattg ccatcaagcg cgacgaaaac 54240 ggcggcgcta cggtcgaact acgcagcacg atccacgtta acaaggttag cccggtgtcg 54300 ttttcatggg gtaaggaata tagcgacgcg gatattctga atgatgccga tttattacgc 54360 gtcatttatc agaagtatgg acacgaaatt ttcaccgctg attttacacg taacatttaa 54420 tttaataagg ggatcactat gcaaactatt aacactaaat accataacgg gaaaatcgtt 54480 gctacttcgt ggaatggctg gacgcgttca gaatatgacg cggcgttatc cagtgaacaa 54540 aatcaccgcg ccgccgctga aaagctaatc attaaactga ataaaggccg ctcgatggaa 54600 tgggggatcg tgaatagcgc gccatccgtg ccgggcgtgc gtggtaaaga taacggatgg 54660 acgtttatta ttggctatgt gccggaagtc gcgccgctgg atatgtcgat cactgttaaa 54720 ttctgcccgg caacaaataa aggcccggca tacatgcgcg cttattcgtg gttagtcccg 54780 cgcgggatca aggttaatta ttctccgcgt gtcgccgacg ggtcagatat tcaggcaaac 54840 gcacgttatg cggcgggcat catgttagac cgtattaatg agatatgccg cgaaggtggc 54900 gatctcttag gctggaaaat tgcggactac gtgcaactgt acgacggcga tcgcctgttc 54960 acactgaaat ctaattaatc cgggggcgcg tcatgttaat ggctattatc ttgatcggtt 55020 attcaataat tgcggctgtt atatatcgtg acgcgcgggc ggtgtggcgt cgctgtaagg 55080 ttgagcgccg ttattctttc gggcgtaaaa cgttttattt acgcgagcgt aaaacaggcc 55140 gtttgatcgg ctgttcaaat aattttttca ctcttctgtt acaaggggct taatcatgat 55200 cctgaattcc atcaataaag agcaacgttt atatgtcatg aaagcgggcg cgggctttac 55260 ctgctacggc ttcgacgtgc tgaacgataa agcgcgccgg gcgttggaat ggttgaaccg 55320 tcacggccgc gcggctgaaa tgattttagg cgcgaagggg atcgacgtca ccgcgctgga 55380 tattccggcc cgcgtcggca ctaaaaagca ttttaccgcg tgcaataagg tattcgacgc 55440 cgcgcggatc tattccatcg caagcggcaa taaatgcgat gcggaatata cgccgcaatt 55500 agtcggcctt gagggccgcc gcgtggaagt ggtggacgcc tacggggaaa agcgccgttt 55560 catcgtgggg cggtcatccg gctggatgcc ttgccactta gagatcgccc gccgcgattc 55620 caacggcggc ccggcggtaa cgggcgcgcc ttttaaatcc gtatcagtca ttaactaacc 55680 gggggatcta tgggtcatat gtatggcgtt atcttgtcaa atggtacgca ttgcgacacg 55740 tcaaccacgg aaaagggcgc gaaaaattac gccacgcgcc acggttacaa ggaagtaagc 55800 cgccgcctga attgcggcta tagcgtcgaa atcgtagcac atcgcgccgg gtcgaaatgg 55860 atcgccgggc ctgctaacca ctcaaccaaa taaggggatc accatgaaac tgaatatcac 55920 cgcgccgggc cttttcttta atcgtagcgg cgaccgcgtc accgtggatc aaatcgtttt 55980 agaaaatagc gccggggccg tcgtgtcatt ccctgttaag ggccggatct ggaaaatgta 56040 ccggggcgca tatcgtccgc gtgagtttaa gatctggacg gcatcgggca agttttacgc 56100 gggcgaacaa tcccgatcgc cgtgggatat tgtcggcgag gttgtcgcgg acgatgtgcg 56160 cccggtctac tggcaagatc tgaccgatga ccagaaagcg acggcggcgg ccaaatatca 56220 caacggtgaa ggcacgccgg aagattattt tacctatacg ccggatcaat tcatcgtgat 56280 taatggtgac gtgtacgatc tggacgccga agcggccgtt tatgaggtcg aagggttaaa 56340 aggtcatgtg cgcatactgg aaagcacgaa ggcggacggg ggcgcgttgg cggtgcaatg 56400 cgtcgaaggt tgggaagtca taagcgttta ccgtgtacag gtgacatcat gaataaaacc 56460 ggattttcgg ggaacgccgc aagggccgcg atcaagttta ccgcgtgcgg cgcgatcgtg 56520 gcgtttatcg gggcggcggt tattttcggc gacaacgtcg ggccgtataa agtgtttttg 56580 tataccgtcg ccgggggcgt atgcgtgcgc ctgatttggc cgactcagga cgaaggcgac 56640 gcacttataa aacgggatga cgggccgtaa aggtgacgcg gccgacgatt aaagagaaaa 56700 gttattgacg cacgccgcgt atatcattta tatttaaatt cccttcgggg gccgggggca 56760 atggggccga cggttatcaa aacaggcgat acaaaatgac acatgatcaa atcgttgcaa 56820 tggttggaaa atacaacatc aaggccgatc gcgccgtgtc ttacaactat gatgtttacg 56880 gttcgacgca agtcgagatc cgcgaggttg acggcgggtc attaatctgg cgtgcgtggt 56940 cttatgaaac caacttcgaa aacgtgttag aaagcgcgct cgatcattcc ggcacactga 57000 aaaccgccga agaaatcgca gaacaggccg ccgccgcacg ctctcaaaag ctccgcaatg 57060 acgcgatgtt ttttgaggtt cacgggttcg aagatgattt cattctcaac ctttccgact 57120 ttacccgcgc tgaaatcgaa gcggagatca tcgcgatgaa tgcagagtta aacgcggcaa 57180 gcgccgacgc cgacgcggca acgtttgacg ccgccgatca cgttttacgt tcaaccaatc 57240 cggccgggga atatgtttat ttaaagcgcg gttactggat gttagacctt gctgacgcgg 57300 agatcctgaa cgcgcgcccg gatattcact ggataagcct ttacgacacg ccgatcgaat 57360 atgtgacgct tgaacaggcc cgcaaggatg acccggaaag cgatttcatc aaacaactga 57420 aagcgggcta tgtgaccgcc ccggccgact atgtcgatcg cgcttgcgcg gccgttgagc 57480 gtggcaacgt taacgcatcg gggatcgtgc tgcatacctt gcgtgaccag atcgacggcg 57540 aacactacga aatcacgaca cgtattagcc cggctgaata tcagctttat cgtgtgcgcg 57600 ttgtcgtgtc ctgctgcatt gatggcgcgt ttcataacgt cattgaactg gatcgcgatg 57660 tgagcacggt tgagcacgtc cgcgacgtgg tgcgcgtcgg ctgcatgaaa caatatatca 57720 agcaagcccg cgcgacgtat gccgcgaccg gggccgccac gctcaacaat gataagacgg 57780 cgacccgtat caagattaaa accgttgcgg gcatgacgtt gatcgctaaa accgaagacg 57840 tgattttttc agatagcggc ctgttttgcg ccgcgtggtt aatgcgtctt gatttgctgg 57900 aaaacaagac ttatgcgcaa gccgtcctaa cgtcgcagat caaaacaatc gaatttatca 57960 aataagggga tcgctatgtt ttacaccgtc ctgttaaacc ccgactacga aggcgcgaca 58020 aacgtcaaaa cgttttacgt cgaaggggaa acggtagaag acgcggtaac ggccgcgatc 58080 gatgacatca acgcgcacga agtcgagatc ggcggggaac cgttagcgcg tgatcaattc 58140 tcaatcattg cagtttttcc gggctatcag cctaatgtgt gggagggcta accgatggac 58200 atcaacgggg ttaaggtcgc aagcgtcgag cgtttcaaca aatatgcgga atggttagcg 58260 cgtcaaatgg tggcgggcgt gccgttggcg tcgcacgctt gcccgcattg cggatcaacg 58320 ttgcacgtta tcgcaaacgg tgacaagggc gatcaatggg attctacttg cgcttgtccg 58380 gtgtgcgcta aaatgtttca ccgctcaata ttgcacggcg aaggcgcgcc agcgattaac 58440 atcattaaac tggatcgggg ttggtaacta tggcgggctt atggtttttc gttgtgacga 58500 tgtgcggcgc ggttggcgct gataatctgg cggctgattg tcatgattat gtgattgacg 58560 cggggatctc ttatgacgat tgccgcgcgg ccgtggcggc ttatccgggc cgcattggct 58620 tatattccgc gcggtgtgac cggggcgaag cggtagaagg gggcaaccat gaataaaggc 58680 caataccgcg ccgcgcgtcg ccttatacgt gaaaacggga tctacgcgtt gcgatggctg 58740 gacgatgaca ccgcgccgat tatggacgtg ttagcaagcc agcccgatga ccagttagag 58800 acaagggcgg cgatcgtggc gtacagcgcc cgcgccgggc ttgcctgcaa tgttcgtaaa 58860 acggcgtcac ttgaactttt agcgcggtac aatgaccgca aggcggcgca caatggataa 58920 ggcgcaacgc ttaacggccg cacgtatggc ggcggatcgg tatgccggga tcgcaagggc 58980 aaaaggtttt aagcgtcatg ttgatggcgt gacttttacc cgggccgatg cggatctaac 59040 gtgggatgac cgggcgcgcg ctttccgtgt gacgctgtac aaaatggacg gggccgcgcg 59100 tcttgccgtt gccactgtac gcgctaacgc gatgcttaac gtgctgttaa aagcgtttat 59160 ctaaaaccgg attttcgggg aacgccgcac gatccctttt aacctgtttt gacatattcc 59220 ccatgtatca aaggccgcct atccgggcgg cttttttatt gcctttctga tatacgcggc 59280 gtgcgttgcg gtgccgtgta gcgggcctat ggtgtgccgt gtgcgatgat atacgcggcg 59340 tgcgtatggc ggcccgtcct gctttcttcc ttcctgatat acacgacgtg catcaatgcg 59400 gccttatgtg tgcgctatgc cgtatggcgg cccgctatgg tgctgcatga tatacgcggc 59460 gtgcgttggt gtgcggctgt atatggctat ctattcccct ttttatggta ttccctgcta 59520 tatggtgtgg cggggggtta ttgtatgccc ttatttcggc cgctatgggg gctagacggg 59580 gcgatcgtgg tgtaaggtat gactacctat tacgatctat cttgcgtggc gtggcgggcg 59640 gctgaggaag aattacggtc acgatttcaa aaaggtactt acccggcgat cggtcggccc 59700 gggggtgcgc agagc 59715

Claims (12)

Bacteriophage KFS-SE (KCTC13147BP), consisting of the nucleotide sequence of SEQ ID NO: 1, with the killing activity specific for Salmonella enteritidis. delete The bacteriophage KFS-SE (KCTC13147BP) according to claim 1, wherein said bacteriophage has a lytic activity in the range of pH 3.0 to pH 11.0. The bacteriophage KFS-SE (KCTC13147BP) according to claim 1, wherein said bacteriophage has lytic activity in the range of 4 ° C to 60 ° C. delete delete An antibiotic composition comprising the bacteriophage KFS-SE (KCTC13147BP) of claim 1. A composition for feed addition comprising the bacteriophage KFS-SE (KCTC13147BP) of claim 1. A feed comprising the feed additive composition of claim 8. A disinfectant comprising the bacteriophage KFS-SE of claim 1 (KCTC13147BP). A cleansing agent comprising the bacteriophage KFS-SE of claim 1 (KCTC13147BP). delete

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