KR20110134554A - Method for prevention and treatment of infection of salmonella choleraesuis or salmonella dublin - Google Patents

Abstract

PURPOSE: A composition for treating infectious diseases of Salmonella Dublin or Salmonella Choleraesuis is provided to ensure high specificity to the bacteria. CONSTITUTION: A composition contains bacteriophage with Salmonella Dublin or Salmonella Choleraesuis-specific apoptotic ability as an active ingredient. The composition is a feed additive, beverage additive, or disinfectant. The bacteriophage is a mixture of bacteriophage of deposit numbers KCTC 11173BP and KCTC 11174BP. The bacteriophage of deposit number KCTC 11173BP has a genome of sequence number 1. The bacteriophage of deposit number KCTC 11174BP has a genome of sequence numbers 2-22. A method for preventing or treating Salmonella Dublin or Salmonella Choleraesuis infection comprises a step of administering the composition to a mammal.

Description

Method for prevention and treatment of infection of Salmonella Choleraesuis or Salmonella Dublin}

The present invention relates to a composition comprising bacteriophages capable of infecting Salmonella cholerasus and Salmonella Dublin, and to a method for preventing and treating infection with Salmonella cholerasus or Salmonella dublin using the composition. will be. That is, the present invention relates to a composition that can be used for the purpose of preventing and post-infection treatment of Salmonella cholerasus bacteria and Salmonella dublin bacteria, and a method for preventing and treating the same.

Salmonella is morphologically and physiologically similar to E. coli but has become an independent genus by K. Kauffmann et al. For medical convenience. Salmonella has been isolated from patients with enteritis, gastroenteritis, and other diseases since Salmon and Smith first reported isolation of Salmonella cholerasus from pigs that died of don cholera in 1885. It has also been isolated from chickens, healthy animals such as cows, pigs, goats, dogs, and cats, and from the environment. More than 2,000 serotypes have been reported so far, and Salmonella genus can be divided into two types, one having host specificity and the other having no host specificity. Salmonella is a Gram-negative bacillus that does not form spores. Salmonella are all found as parasites in various animals. Salmonella infections occur in several forms, but enteritis is the most common form. Salmonella is also a major cause of food poisoning. Salmonella food poisoning is very common in Korea, and its severity continues to increase due to an increase in group meals. In 2000, a study on the causative agents of food poisoning patients found that more than 2,500 food poisonings were caused by Salmonella, compared with other major food poisoning agents such as Staphylococcus (about 1,000) and Vibrio (about 200). It is a high proportion. Salmonella infection has clinical symptoms such as coarse skin, lack of appetite, conjunctivitis, depression, thinning stools, increased spleen, and death. Salmonella can also be easily delivered to humans because they are present in the intestines of poultry and livestock such as chickens, which humans consume as food. In the United States, the economic damage caused by food poisoning is estimated to be $ 2-40 billion, which can be economically important.

Among Salmonella bacteria, Salmonella cholera suis is a bacterium that is well known as the pig cholera bacteria of the genus Salmonella, and infects both humans and animals. The incubation period of pig cholera is 6-8 days on average and shows symptoms such as body heat, anorexia, conjunctivitis, constipation, yellow-gray diarrhea, vomiting, posterior palsy and spasms of 40-41 ℃.

Salmonella Dublin is a bacterium belonging to the genus of Salmonella, which is generally known as 'a species that shows affinity for cattle' when it is divided into 'host specific group' and 'host non-specific group' according to its adaptability to the host. Bovine Salmonella is the main cause of infection. Salmonella infection caused by Salmonella Dublin bacteria occurs worldwide and is most common in the summer season, and carriers carrying Salmonella Dublin bacteria without contaminated feed, water or symptoms can be an important source of infection. Salmonella Dublin bacteria are contained in feces, milk, and vaginal secretions, and bacteria excreted in the outer world are known to survive for 1-6 months. If a cow is infected with Salmonella Dublin bacteria during pregnancy, it can infect the fetus by riding the bloodstream, which can cause the sepsis to die of sepsis, which can result in crude abortion. Salmonella infection caused by Salmonella Dublin bacteria is considered serious because of mortality rates of 50-100% in cows within 1 month of age. For the sake of simplicity, Salmonella Dublin is one of the many causes of cow mastitis.

The damage to the livestock industry due to the infection of Salmonella cholerasus and Salmonella dublin bacteria is considerable, and there is an urgent need to develop methods to prevent their infection and to effectively treat them. .

Although various drugs are used for the treatment of Salmonella cholerasus and Salmonella Dublin, infections are usually difficult to cure. Therefore, efforts to prevent infection rather than treatment may be more important and it is necessary to develop a method using a naturally-derived material for this purpose.

Typically, a variety of drugs can be used for the purpose of preventing infection by bacteria and further treating infectious diseases, but bacteriophages have attracted attention in recent years with the emphasis on nature-friendly methods.

Bacteriophages are a type of virus that infects bacteria, often called abbreviations. Bacteriophages have the ability to infect bacteria and multiply within them, and kill off the bacteria by destroying the bacteria when progeny bacteriophages come out of the bacteria. Bacteriophage bacterial infection is so specific that there are only a few types of bacteriophages that can infect certain bacteria. That is, certain bacteriophages can infect only certain classes of bacteria, thereby killing only certain bacteria.

의 세균학자 d'Herelle이 이질환자 변의 여과액 중에 적리균(Shigella disentriae) 을 녹이는 작용을 가진 것이 있다는 것을 발견하고 이에 대한 연구를 통해 독립적으로 박테리오파지를 발견하였으며, 세균을 잡아먹는다는 뜻에서 박테리오파지라고 명명하였다. 이후 이질균, 장티푸스균, 콜레라균 등 여러 병원균에 대한 박테리오파지가 계속적으로 발견되었다. 박테리아를 사멸시킬 수 있는 특별한 능력으로 인하여 박테리오파지는 발견 이후 박테리아 감염에 대항하는 방안으로 기대를 모았으며 관련한 많은 연구가 있었다. 그러나 1950년 Flemming에 의해 페니실린이 발견된 이후, 항생제의 보급이 일반화되면서 박테리오파지에 대한 연구는 일부 동유럽 국가들 및 구소련에 한정되어서만 명맥이 유지되었다. 그런데 2000년 이후에 항생제 내성균의 증가로 기존 항생제의 대체 물질로의 개발 가능성이 다시 부각되면서 다시 박테리오파지가 주목을 받고 있다.The bacteriophage was discovered in 1915 by a British bacteriologist Twort, in a study of the transparent melting of micrococcus colonies by something. In 1917, France

Of bacteriologist d'Herelle in Shigella disentriae ) They found that they had a function of dissolving the bacteria, and through this research, they independently discovered bacteriophages and named them as bacteriophages in order to eat bacteria. Since then, bacteriophages have been found for several pathogens such as dysentery, typhoid, and cholera. Because of its special ability to kill bacteria, bacteriophages have been expected to fight bacterial infections since their discovery and there has been much research. However, after the discovery of penicillin by Flemming in 1950, with the spread of antibiotics, research on bacteriophages remained confined to a limited number of Eastern European countries and the Soviet Union. However, since 2000, the growth of antibiotic resistant bacteria has highlighted the possibility of developing antibiotics as alternatives, and bacteriophages are receiving attention again.

Accordingly, the present inventors can be used to prevent or treat infections of Salmonella cholerasus and Salmonella Dublin by using bacteriophages capable of selectively killing Salmonella cholerasus and Salmonella dublin. And after endeavoring to develop a method using the composition, to secure suitable bacteriophage and to develop a composition containing these bacteriophage as an active ingredient, the composition is effective in preventing and treating infections of Salmonella cholerasus and Salmonella Dublin The present invention has been completed by confirming that it can be utilized.

Accordingly, an object of the present invention is to provide a composition that can be used to prevent infection of Salmonella cholerasus bacteria and a method for preventing infection of Salmonella cholerasus bacteria using the composition.

It is another object of the present invention to provide a composition that can be used to treat infections of Salmonella cholerasus bacteria and a method for treating infection of Salmonella cholerasus bacteria using the composition.

Another object of the present invention is to provide a composition that can be used to prevent infection of Salmonella Dublin bacteria and a method for preventing infection of Salmonella Dublin bacteria using the composition.

Another object of the present invention is to provide a composition that can be used to treat infections of Salmonella dublin bacteria and a method for treating infection of Salmonella dublin bacteria using the composition.

Another object of the present invention is to provide a disinfectant using the compositions.

In addition, another object of the present invention is to provide a negative additive using the compositions.

Another object of the present invention is to provide a feed additive using the compositions.

The present invention provides a composition comprising bacteriophages having an effective killing ability against Salmonella cholerasus and Salmonella dublin bacteria, and a method for preventing and treating infection with cholerasus and Salmonella dublin bacteria using the composition. do.

The bacteriophage of the present invention has a genome represented by SEQ ID NO: 1 or a genome represented by a partial gene sequence of SEQ ID NO: 2 to SEQ ID NO: 22. These bacteriophages have been isolated from nature by the inventors and confirmed killing ability against Salmonella gallinarum , Salmonella enteritidis , Salmonella pullorum , and Salmonella typhimuriun . There is a bar. By the present inventors, the bacteriophage having the genome represented by SEQ ID NO: 1 was named bacteriophage SEP-1 and deposited on August 21, 2007 at the Korea Institute of Biotechnology and Biotechnology Center (Accession No. KCTC 11173BP). The bacteriophage having the genome represented by the partial gene sequence of SEQ ID NO: 2 to SEQ ID NO: 22 was named bacteriophage SGP-1 and deposited on August 21, 2007 at the Korea Institute of Biotechnology and Biotechnology Center (Accession No. KCTC 11174BP). .

The present invention also provides disinfectants, negative additives, and feed additives that can be utilized to prevent or treat infections of Salmonella cholerasus bacteria and Salmonella dublin bacteria.

As described above, bacteriophages included in the composition of the present invention specifically kill Salmonella cholerasus and Salmonella dublin, thereby preventing various diseases caused by Salmonella cholerasus and Salmonella dublin. It is effective for infection prevention) and treatment (infection treatment).

Therefore, the composition of the present invention can be used for the purpose of preventing and treating swine cholera, which is a representative disease caused by Salmonella cholerasus bacteria, and also for the prevention of Salmonella infection caused by Salmonella Dublin bacteria and It can be used for treatment purposes.

The term "treatment" or "treatment" as used herein refers to (i) suppression of diseases caused by Salmonella cholerasus or Salmonella dublin; And (ii) alleviation of diseases caused by Salmonella cholerasus bacteria or Salmonella dublin bacteria.

Pharmaceutically acceptable carriers included in the compositions of the present invention are those commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate , Microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto. no. The composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like in addition to the above components.

The composition of the present invention may include bacteriophage SEP-1 or bacteriophage SGP-1 alone, and may also include both bacteriophage SEP-1 and bacteriophage SGP-1. The bacteriophages included at this time include 1 × 101 pfu / ml to 1 × 1030 pfu / ml or 1 × 101 pfu / g to 1 × 1030 pfu / g of bacteriophages, preferably 1 × 10 4 pfu / ml to 1 Bacteriophage from 10 × 10 15 pfu / ml or 1 × 10 4 pfu / g to 1 × 10 15 pfu / g.

The compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient, according to methods which can be easily carried out by those skilled in the art. It may also be prepared by incorporation into a multi-dose container. The formulations here may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of extracts, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.

The compositions of the present invention may be implemented with, but not limited to, disinfectants, negative additives, and feed additives, depending on the mode of application.

 Feed additives of the present invention may be in the form of a dry or liquid formulation, and one or more enzyme formulations may be added. Enzyme preparations can be added either in dry or liquid form. Enzyme preparations include lipolytic enzymes such as lipases, phytases that break down phytic acid to form phosphates and inositol phosphates, starches and Amylase, an enzyme that hydrolyzes alpha-1,4-glycoside bonds in glycogen, and phosphatase, an enzyme that hydrolyzes organic phosphate esters ), Carboxymethylcellulase that breaks down cellulose, xylanase that breaks down xylose, and maltase that hydrolyzes maltose with two molecules of glucose. (maltase) and sugar-producing enzymes such as invertase, which hydrolyzes saccharose to form a glucose-fructose mixture. Can be used.

In addition, other non-pathogenic microorganisms may be added to the feed additive in the present invention. Microorganisms that may be added include Lactobacillus, which has physiological activity and organic degradability under anaerobic conditions such as Bacillus subtilis , Bacillus subtilis , which can produce proteolytic enzymes, lipolytic enzymes and sugar converting enzymes, and bovine stomach. Strains ( Lactobacillus sp.), Filamentous fungi such as Aspergillus oryzae , which have been shown to increase body weight, increase milk yield and increase digestive absorption of feed (J Animal Sci 43: 910-926, 1976) and yeasts such as Saccharomyces cerevisiae (J Anim Sci 56: 735-739, 1983).

In addition, the feed additive in the present invention, raw materials such as peanuts, peas, sugar beets, pulp, grain by-products, animal intestine powder, fish meal powder, etc., including various grains and soy protein may be unprocessed or processed.

The composition of the present invention and the method for preventing and treating infection with Salmonella cholerasus or Salmonella Dublin using the composition are highly specific to Salmonella cholerasus and Salmonella dublin compared to conventional antibiotic-based methods. Can be provided. This means that it can be used for the purpose of preventing or treating infections of Salmonella cholerasus and Salmonella dublin without affecting other useful bacteria. Accordingly, the side effects of use are very small. When antibiotics are used, various types of common bacteria are also damaged, resulting in various side effects such as decreased immunity. On the other hand, the present invention also provides the advantage of being very natural because it uses a composition containing the bacteriophage already present in nature as an active ingredient. In addition, since the bacteriophages of the present invention have the ability to kill Salmonella gallinarum, Salmonella enteritidis, Salmonella Fulumum and Salmonella typhimurium, the composition of the present invention together with these bacteria Salmonella cholerasus bacteria and Salmonella The composition of the present invention may also be used for the purpose of simultaneously preventing infection by Dublin bacteria, and when the infection by Salmonella cholerasus or Salmonella Dublin bacteria occurs in combination with these bacteria, It can be effectively used for the purpose.

1 is a photograph of the entire culture dish as a result of lytic plaque analysis, which is a test for killing ability against Salmonella cholera suis bacteria of bacteriophage SEP-1 and bacteriophage SGP-1. 'SEP-1 spotting' is the result of dropping the bacteriophage solution of bacteriophage SEP-1, and 'SGP-1 spotting' is the result of dropping the bacteriophage solution of bacteriophage SGP-1. The clear, transparent circles are lysate plaques that result from the lysis of bacteria grown on the petri dish.
Figure 2 is a photograph of the culture plate as a result of the analysis of the killing ability for the Salmonella Dublin bacteria of bacteriophage SEP-1 and bacteriophage SGP-1. 'SE' is the result of dropping the bacteriophage solution of bacteriophage SEP-1, 'SG' is the result of dropping the bacteriophage solution of bacteriophage SGP-1. The clear, transparent circles are lysate plaques that result from the lysis of bacteria grown on the petri dish.

Hereinafter, although an Example demonstrates this invention more concretely, these Examples are only illustrations of this invention, The scope of the present invention is not limited to these Examples.

Example  1: bacteriophage SEP -1 separation

Bacteriophage SEP-1 of the present invention was isolated using Salmonella enteritidis. We collected a sample based on where Salmonella enteritidis is expected to isolate bacteriophages that specifically infect Salmonella enteritidis. After incubating the sample with Salmonella enteritidis, the culture solution was centrifuged to obtain a supernatant. The supernatant thus obtained was filtered, and then cultured Salmonella enteritidis was re-cultured together as bait for bacteriophage separation to determine whether Salmonella enteritidis was killed. Confirmation of death was finally determined by plaque assay.

In detail, 237 samples were collected from living streams in Seoul and elsewhere where bacteriophages are expected to separate bacteriophages capable of selectively killing Salmonella enteritidis, at 37 ° C with Salmonella enteritidis. After shaking for 3-4 hours, the supernatant was recovered by centrifugation at 8,000 rpm for 20 minutes. The recovered supernatant was filtered using a 0.45 μm filter, and then bacteriophage specific to Salmonella enteritidis was detected by lytic plate analysis using the filtrate thus obtained. The separated bacteriophages were named SEP-1 and deposited on August 21, 2007, at the Korea Institute of Biotechnology and Biotechnology Center (Accession No. KCTC 11173BP). A large amount of bacteriophage solution of bacteriophage SEP-1 was secured by repeating the method of growing Salmonella enteritidis and bacteriophage SEP-1 together, followed by centrifugation and filtration as described above. The bacteriophage solution of the finally obtained bacteriophage SEP-1 was used as it was, or it was used for further purification by adding a conventional polyethylene glycol precipitation process.

Example  2: bacteriophage SGP -1 separation

Bacteriophage SGP-1 of the present invention was isolated using Salmonella gallinarum. We collected samples based on where Salmonella gallinarum is expected to isolate bacteriophages that specifically infect Salmonella gallinarum. After incubating the sample with Salmonella gallinarum, the culture was centrifuged to obtain a supernatant. The supernatant thus obtained was filtered and then cultured with Salmonella gallinarum again as bait for bacteriophage separation to determine whether Salmonella gallinarum was killed. Confirmation of death was finally determined by lytic plaque analysis.

To explain this in detail, samples are collected from living streams in Seoul and elsewhere where bacteriophage is expected to separate bacteriophages that can selectively kill Salmonella gallinalum, and at 37 ° C for 3-4 hours at Salmonella gallinarum. After shaking for 2 hours, the supernatant was recovered by centrifugation at 8,000 rpm for 20 minutes. The recovered supernatant was filtered using a 0.45 μm filter, and then bacteriophage specific to Salmonella gallinarum was detected by lytic plate analysis using the filtrate thus obtained. This isolated bacteriophage was named SGP-1, and was deposited on August 21, 2007 at the Korea Institute of Biotechnology and Biotechnology Center (Accession No. KCTC 11174BP). A large amount of bacteriophage solution of bacteriophage SGP-1 was obtained by repeating the method of growing Salmonella gallinarum and bacteriophage SGP-1 together, followed by centrifugation and filtration as described above. The bacteriophage solution of the finally obtained bacteriophage SGP-1 was used as it is in bacteriophage SEP-1, or was further purified by adding a conventional polyethylene glycol precipitation process.

Example  3: bacteriophage SEP Gene Sequence Analysis of the -1 Genome

Gene sequencing of the obtained bacteriophage SEP-1 genome was performed. To this end, the genome of bacteriophage SEP-1 was extracted by conventional methods and used for gene sequencing. Specifically, first, a 1 ℓ flask TSB (T ryptic S oy B roth) medium (Casein Digest, 17 g / ℓ; Soy bean Digest, 3 g / ℓ; dextrose, 2.5 g / ℓ; NaCl, 5 g / ℓ Dipotassium phosphate, 2.5 g / L) was added to 200 ml of 50 ml of Salmonella enteritidis suspension having an absorbance of 1 at 600 nm and 1 ml of a bacteriophage solution filtered to 1 × 108 pfu / ml. 4 hours shaking culture. After incubation, it was confirmed whether Salmonella enteritidis was lysed, and when the lysate occurred, the culture was filtered with a 0.45 μm filter. Next, in order to remove DNA and RNA of Salmonella enteritidis remaining in the filtered culture, 200 U of DNase I and RNase A were respectively added to 10 ml of the filtered culture, and left at 37 ° C. for 30 minutes. . In order to remove the activity of DNase I and RNase A after 30 minutes, 500 μl of 0.5 M ethylenediaminetetraacetic acid (EDTA) was added and allowed to stand for 10 minutes. In the next step, 100 µl of proteinase K (20 mg / ml) and 500 µl of 10% sodium dodecyl sulfate (SDS) were added to disintegrate the bacteriophage outer wall, followed by reaction at 65 ° C for 1 hour. After the reaction for 1 hour, 10 ml of a mixture of phenol: chloroform: isoamylalcohol having a composition ratio of 25: 24: 1 was added to the reaction mixture, and the mixture was mixed well. Then, this was centrifuged at 18,000 rpm to separate the layers, and the upper layer was taken from the separated layers, and 2 vol. Ratio of cold 100% alcohol was added thereto to extract only pure dielectric material. To confirm whether the genome of the extracted bacteriophage was DNA or RNA, DNase I (10 U / μl) and RNase A (10 μg / μl) were added, respectively, and treated at 37 ° C. for 1 hour. In this case, Mung Bean nuclease (45 U / μl) was added to distinguish between single-stranded DNA and double-stranded DNA, and then treated at room temperature for 15 minutes. The samples thus treated were subjected to electrophoresis using 0.8% agarose gel to investigate the cleavage pattern by each enzyme. As a result, the obtained genome was sensitive only to DNase I and sensitive to DNase I means that the genome is DNA, and not sensitive to mung bean nuclease means that the genome is in the form of DNA double strand. The genome of the bacteriophage obtained from this result was confirmed to be DNA-type and double stranded.

The genome of the bacteriophage isolated as above is genomic DNA (gDNA). To first build a variety of restriction enzyme were treated with primary identify the cut pattern according to the restriction enzyme treated with (restriction enzyme), the restriction enzyme is a library (library) of gDNA When treated with Hpa II to analyze the gene sequence of the gDNA A gDNA library using a gene fragment prepared by treatment with Hpa II, which was determined to be the most suitable, was constructed according to a conventional method.

In detail, first, gDNA of bacteriophage SEP-1 was treated with Hpa II to obtain a gene fragment according to a conventional method, and pBluescript II SK (+) phagemid for preparation of a vector portion to be used later when inserting the gene fragment. A vector fragment of linear was prepared by treating phagemid vector (Stratagene) with Cla I. Gene fragments and vector fragments thus prepared were combined with each other according to a conventional method using T4 ligase. The recombinant vector into which the gene fragment of bacteriophage SEP-1 thus prepared was introduced was introduced into Top10F 'species (Invitrogen), a species of E. coli, through an electro-transformation method called electroporation. The transformants thus transformed were X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) and isopropyl beta-di-1-thiogalactopyranoside Ampicillin-containing agar plates containing (isopropyl β-D-1-thiogalactopyranoside; IPTG) were selected by conventional blue-white colony selection. Selected colonies (colony) were inoculated in the culture medium containing ampicillin and then shaken overnight, and the plasmid was extracted from the cultured cells using a plasmid purification kit (iNtRON). The extracted plasmid was screened for recombinant plasmid by checking its size through electrophoresis using 0.8% agarose gel.

The clones containing the selected plasmids were cultured again, and the plasmids were extracted from the cultured cells as described above, and sequencing using the extracted plasmids was performed in the forward direction of M13, a primer widely used in conventional sequencing. (forward) primers and M13 reverse primers were used. The sequence of each primer is as follows.

Thus, a partial gene sequence of the bacteriophage SEP-1 genome was secured, and a general whole gene sequence analysis was performed based on this. The total gene sequence of the finally identified bacteriophage SEP-1 is shown in SEQ ID NO: 1.

Based on the obtained nucleotide sequence of bacteriophage SEP-1, homology with the known bacteriophage gene was investigated using BLAST (http://www.ncbi.nlm.nih.gov/BLAST/) on the Web. As a result of the BLAST investigation, the nucleotide sequences of the bacteriophage SEP-1 were analyzed by the sequence of bacteriophage KS7 (GenBank Accession No. AY730274), the sequence of Salmonella phage SETP3 (Salmonella phage SETP3) and bacteriophage MB78. partly homologous to, but partially homologous to, the partially identified sequence of bacteriophage MB78 (GenBank Accession No. AY040866; AJ277754; AJ249347; Y19203; Y19202; AJ245858; AJ245537; Y18133; X87092; AF156970; AF349435; X86562). It was confirmed to be different. These results show that the genome of bacteriophage SEP-1 is characteristically different from bacteriophage KS7, Salmonella phage SETP3 and bacteriophage MB78. Therefore, bacteriophage SEP-1 may be referred to as a novel bacteriophage.

Example  4: bacteriophage SGP Gene Sequence Analysis of the -1 Genome

Gene sequencing of the obtained bacteriophage SGP-1 was performed. To this end, the genome of bacteriophage SGP-1 was extracted in the same manner as in Example 3 and used for gene sequencing. Whether the genome of the extracted bacteriophage was DNA or RNA was also performed in the same manner as in Example 3. As a result, it was confirmed that the genome of the bacteriophage SGP-1 was in the form of a DNA double strand.

Gene sequence analysis was performed using the genome of bacteriophage SGP-1 thus obtained in the same manner as in Example 3. Sequencing of the bacteriophage SGP-1 genome was confirmed only to partial sequences, and the identified partial sequences are shown in SEQ ID NOs: 2 to 22.

Based on the obtained nucleotide sequence of bacteriophage SGP-1, homology with the known bacteriophage gene was investigated using BLAST (http://www.ncbi.nlm.nih.gov/BLAST/) on the Web. BLAST results confirmed that the base sequence of the analyzed bacteriophage SGP-1 was partially homologous to bacteriophage phiYeO3-12 and bacteriophage T3. The homology between the obtained bacteriophage SGP-1 genome was compared with the sequence of the bacteriophage piYeO3-12 genome (GenBank Accession No. AJ251805) and the sequence of the bacteriophage T3 genome (GenBank Accession No. AJ318471). .

These results show that many of the subsequences of bacteriophage SGP-1 differ from bacteriophage pi Ye03-12 and bacteriophage T3. From this, it can be seen that the bacteriophage SGP-1 genome is distinctly different from bacteriophage pi Ye03-12 and bacteriophage T3. Therefore, bacteriophage SGP-1 may be referred to as a novel bacteriophage.

Example  5: bacteriophage SEP -1 and bacteriophage SGP Salmonella of -1 Cholera suis  Fungus and salmonella Dublin  Against fungi Death  Research

The killing ability of the bacteriophage SEP-1 and bacteriophage SGP-1 against Salmonella cholerasus and Salmonella dublin was investigated. The bacteriophage SEP-1 and bacteriophage SGP-1 were both bacteriophages isolated in a manner unrelated to Salmonella cholerasus and Salmonella dublin, and this was necessary. Irradiation method was different from each culture of Salmonella cholerasus (KCTC2932 strain) and Salmonella Dublin (BA584; strain isolated from the genome analysis of clinical isolates) of 1 ml at 600 nm TSA (T ryptic S oy a gar) plate medium (casein Digest, 15 g / L; Soy bean digest, 5 g / L; NaCl, 5 g / L; agar, 15 g / L) and then dried to spread on, carried out The bacteriophage solution of bacteriophage SEP-1 prepared in Example 1 and the bacteriophage solution of bacteriophage SGP-1 prepared in Example 2 were dropped. This was incubated in a 37 ℃ incubator overnight to determine the degree of lysis (lysis) of each bacteria. The results are shown in FIGS. 1 and 2. In this study, bacteriophage SEP-1 and bacteriophage SGP-1 were confirmed to be lytic (killing) activity against Salmonella cholerasus and Salmonella Dublin, from which the bacteriophage SEP-1 and bacteriophage SGP-1 of the present invention It could be concluded that both Salmonella cholerasus and Salmonella dublin had antimicrobial activity. From this, bacteriophage SEP-1 and bacteriophage SGP-1 of the present invention was confirmed that it can be effectively used for the prevention and treatment of infectious diseases caused by Salmonella cholera suis bacteria or Salmonella Dublin bacteria.

Example  6: obtained bacteriophage SEP -1, bacteriophage SGP -1, bacteriophage SEP-1 and bacteriophage SGP Salmonella with a Mixture of -1 Cholera suis  Fungus or salmonella Dublin  Application example for the prevention of bacterial infection

Two 9 ml nutritional broths (3 g / l beef extract and 5 g / l peptone) were charged with 100 μl of bacteriophage solution of bacteriophage SEP-1 at approximately 1 × 108 pfu / ml. In dogs, 100 μl of bacteriophage solution of bacteriophage SGP-1 at a level of about 1 × 108 pfu / ml, respectively, and 100 μl of bacteriophage solution of bacteriophage SGP-1 at about 0.5 × 108 pfu / ml, respectively. 100 μl of bacteriophage solution of bacteriophage SGP-1 at a level of 0.5 × 10 8 pfu / ml was added, and the last two samples were prepared without the bacteriophage solution. One sample from each of the two groups was added with a culture solution of Salmonella cholerasus bacteria so that the absorbance was about 0.5 at 600 nm, and the other sample was finally absorbed with 0.5 Salmonella at 600 nm. Blin bacteria were added and the growth of the bacteria was observed. As shown in the results of Table 3, in the medium without the bacteriophage solution, the bacteria grew so well that the absorbance at 600 nm was 1.3 at 60 nm after 60 minutes, whereas the bacteriophage was added in the nutrient medium to which the bacteriophage solution was added. Irrespective of the type of liquid, the absorbance at 600 nm after 15 minutes was reduced to about 0.1 level and after 0.05 minutes to 0.05 level.

From these results, bacteriophage SEP-1 and bacteriophage SGP-1 of the present invention are capable of inhibiting the growth and killing of Salmonella cholerasus bacteria and Salmonella dublin bacteria, and thus killing Salmonella cholerasus or Salmonella dublin bacteria. It can be confirmed that it can be used to prevent infection.

Example  7: Bacteriophage obtained SEP -1, bacteriophage SGP -1, bacteriophage SEP-1 and bacteriophage SGP Salmonella with a Mixture of -1 Cholera suis  Fungus or salmonella Dublin  To treat diseases caused by bacterial infection Application example

Four two-day-old chicks were fed with 1 × 10 7 cfu of Salmonella cholerasus bacteria to create an artificial infection (animals 1-1, 1-2, 1-3, 1-4) and another four 2 One-day-old chicks were fed with 1 × 10 7 cfu of Salmonella Dublin bacteria to create an artificial infection (animals 2-1, 2-2, 2-3, 2-4). Animals 1-1 and 2-1 were then fed by mixing bacteriophage SEP-1 to 1 × 109 pfu per g of feed, and animals 1 and 2-2 were fed 1 g of bacteriophage SGP-1. The mixture was fed to 1 × 109 pfu per animal, and in animals 1-3 and 2-3, bacteriophage SEP-1 became 0.5 × 109 pfu per g of feed and bacteriophage SGP-1 also 0.5 × 109 pfu per g of feed. They were fed together to feed, and animals 1-4 and 2-4 were fed only feeds of the same composition excluding bacteriophages. Two days after feeding, Salmonella bacteria counts in fecal and caecum contents were measured. In this example, a Salmonella bacteria selection medium (Rambach agar plate; Merck) was used to prevent interference by other contaminating bacteria. As a result, when the bacteriophage was fed with the feed, fecal irrespective of the type of bacteriophage solution was confirmed to reduce Salmonella bacteria more than 1,000 times compared to the control group, and confirmed that the contamination of Salmonella bacteria more than 200 times in the cecal contents. Could.

These results confirm that both bacteriophage SEP-1 and bacteriophage SGP-1 of the present invention are very effective in the treatment of infectious diseases caused by Salmonella cholerasus bacteria or Salmonella dublin bacteria.

Example  8: Preparation of feed additives and feed

The bacteriophage solution of bacteriophage SEP-1 prepared in Example 1 was prepared to include 1 × 10 9 pfu of bacteriophage SEP-1 per g of feed additive. The manufacturing method is spraying the bacteriophage solution prepared in Example 1 evenly on a powder medium consisting of 90% dextran, 5% silica, 5% calcium carbonate by weight and then dried at room temperature and finally pulverized to a fine powder form It was prepared by mixing well. In the drying process, reduced pressure drying, warming drying, and freeze drying may also be substituted.

A feed additive containing 1 × 10 9 pfu of bacteriophage SGP-1 per gram of feed additive was also prepared in the same manner using the bacteriophage solution of bacteriophage SGP-1 prepared in Example 2. In addition, a feed additive including 0.5 × 109 pfu of bacteriophage SEP-1 and 0.5 × 109 pfu of bacteriophage SEP-1 per gram of feed additive was also prepared in Example 2 and the bacteriophage solution of bacteriophage SEP-1 prepared in Example 1. It was prepared in the same manner using a mixture of the eastern blood of the prepared bacteriophage SGP-1 bacteriophage solution. For the control experiments, feed additives containing no bacteriophage were also prepared by spraying physiological saline containing no bacteriophage.

Each of the four feed additives thus prepared was mixed with 1,000 times the feed in a weight ratio to prepare the final four feeds. The feed thus prepared may be further processed through an additional feed manufacturing process, but in the present embodiment, this was used without being applied.

Example  9: Preparation of negative additives and disinfectants

Negative additives or disinfectants differed only in their application and the formulations were identical, so they were prepared in the same way. The bacteriophage solution of bacteriophage SEP-1 prepared in Example 1 was prepared to include 1 × 10 9 pfu of bacteriophage SEP-1 per ml of negative additive (or disinfectant). The preparation method is a bacteriophage solution of bacteriophage SEP-1 prepared in Example 1 so that 1 × 109 pfu of bacteriophage SEP-1 per 1 ml is contained in a sterile buffer commonly used in the art (in this example, using saline). It was prepared by adding and mixing well.

A negative additive (or disinfectant) containing 1 × 10 9 pfu of bacteriophage SGP-1 per ml of negative additive (or disinfectant) was also prepared in the same manner using the bacteriophage solution of bacteriophage SGP-1 prepared in Example 2. In addition, a negative additive (or disinfectant) containing 0.5 x 109 pfu of bacteriophage SGP-1 and 0.5 x 109 pfu of bacteriophage SEP-1 per ml of negative additive (or disinfectant) was also prepared bacteriophage SEP-1. It was prepared in the same manner using a mixed solution of the bacteriophage solution of the bacteriophage SGP-1 prepared in Example 2 and the eastern blood of the bacteriophage solution. For the control experiment, physiological saline itself without bacteriophage was selected as a negative additive (or disinfectant) without bacteriophage.

The four negative additives (or disinfectants) thus prepared were diluted with 1,000 times water by volume and used as final negative or disinfectants.

Example  10: Confirmation of effects in pig breeding

Salmonella cholera suis bacteria are the main pathogens in pigs, so the effect of the present invention on pig breeding was examined. By using the feed prepared in Example 8 and Example 9 and the negative and disinfectant was applied to the actual animal breeding was investigated whether the specification result when breeding the animal. In particular, the survey was conducted in terms of mortality. A total of 60 piglets were divided into three groups (group-A fed; group-B fed negatively; group-C sterilized) for 4 weeks. Each group is subdivided into five groups. Each subgroup is divided into small groups (small group-①) with bacteriophage SEP-1, small groups (small group-②) with bacteriophage SGP-1, and bacteriophage SEP-1 and bacteriophage SGP-. 1 is divided into a small group (small group-③) and a bacteriophage small group (small group ④). The piglets covered in this study were 20-day-old weaning piglets, and the piglets of each test subgroup were reared in groups of five. Each subgroup is divided and referred to as Table 4 below.

In the case of feed feeding, the feed prepared in Example 8 was fed according to the conventional feed feeding method according to the classification of Table 4, and in the case of negative feeding, the negative produced in Example 9 was classified in Table 4 According to the conventional drinking water supply method, and the disinfection treatment was carried out three times a week alternating with the existing disinfection. On the day of spraying the disinfectant of the present invention, disinfection using a conventional disinfectant was not performed. The test results are shown in Table 5.

As a result, it was confirmed that feeding and disinfection of feed and drinking water according to the present invention is effective in reducing mortality in animals. From this, it can be concluded that the present invention is effective for improving the specification results of animals.

Example  11: Identify the effects in cattle breeding

Salmonella Dublin bacteria are the main pathogens in cattle, so we examined the effect of the present invention on cattle breeding. By using the feed prepared in Example 8 and Example 9 and the negative and disinfectant was applied to the actual animal breeding was investigated whether the specification result when breeding the animal. In particular, this study was conducted for one month in terms of the incidence of diarrhea (yellow, malodorous odor) and mucosal stools. Jau was 1-3 months old. A total of 10 jaws were divided into two groups of five each, one group consisting of the feed prepared in Example 8 together with the bacteriophage SEP-1 and bacteriophage SGP-1 prepared in Example 8 and While the negative water containing bacteriophage SEP-1 and bacteriophage SGP-1 was also supplied, in addition, disinfection using bacteriophage SEP-1 and bacteriophage SGP-1 prepared in Example 9 was also performed. The other group did not perform any treatment related to bacteriophage as a control group.

In the case of feed and negative feeding, the feeding was carried out according to the conventional feeding method, and the disinfection treatment was performed three times a week alternately with the existing disinfection. Disinfection with the disinfectant according to the invention was carried out on both the barn and each animal individual. On the day of spraying the disinfectant of the present invention, disinfection using a conventional disinfectant was not performed. The test results are shown in Table 6.

As a result, it was confirmed that feeding and disinfection of feed and drinking water according to the present invention is effective in reducing diarrhea in animals. From this, it can be concluded that the present invention is effective for improving the specification results of animals.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

KCTC KCTC11173BP 20070821 KCTC KCTC11174BP 20070821

<110> iNtRON Biotechnology, Inc. <120> Method for prevention and treatment of infection of Salmonella          Choleraesuis or Salmonella Dublin <160> 22 <170> KopatentIn 1.71 <210> 1 <211> 43285 <212> DNA <213> SEP-1 Bateriophage gene <400> 1 aactcgaact gtgcggctgc gccgttgtcg ttagtaatgg tgaacgtgtc gataagttta 60 agtggcgcgg caccgtcata agcagataca tcaaccgatg caaacgggcg ggcactaaaa 120 ttagtcgtga agtcggaacc gctaggcggc gcctgtaaga tttcctgatt caagccgatg 180 aacgggaatg aaccggtcac cattgcgtta acagcctgtt cgatagtgaa cccggtaaac 240 tcgacaccac gggttacgat gtatgaatcc gggtttccgc atttaccttt caaccatgtg 300 aggattgaat aggtcttaca caagttaccg gtttccagtt tatctgcgat acgcaaatct 360 gcctggacgt cggattcggc ggtaagggta tgctggatgc ccgcaccggt tacgaccgta 420 gccgttactg cggtaacgag gaaagcttta tcgttattac cggacaaacc atcgaattgc 480 accaggtcgc ctacttcaac gccatcggtt acaaagctac cggaggcccg tgtgaaagtt 540 ttcgctaccg ggtcaaccgt aatgccgatt gccgatgcgg tggaacctgc tacccatgaa 600 ctggtcattg caccggctaa cagctcgtcc tggcttgtcg cgcttagttc gatagcgtat 660 tcaccagtta cctgacggtt accggtacgg atggatgatg tttcgcggct gccgtccagc 720 tcgttagaga tgagggcgtc gcgcgttacg gcggggatgc cacctgtgtt gcggagtggc 780 tgccataccg ggttagccgg cgtcacaccc ggtgttgtct caagtacgta aaattgcgcg 840 gtcatcgcgc ctttgtatgg ttgtaacgcc attatctaat cctcgctgta aaggctatga 900 aattaattga gagaggcctc ttcgcccatc cattctcaac aataagagga ccgaggctta 960 ctgattgtac ctcagcacag atttcgttac gactaaaaca gttaccggcg gcgaaggcgg 1020 cgtttagctt atcagctaat cggttaatag gagcgctacc caaggccgac ccgacgttaa 1080 tgtctacctg ataaacaccc gcccgttgtt cagtccaaaa caagtcggct tgttctgtat 1140 cagacaacag catataactc gcgatataag gtgcgtctgt agacgtcggc gcatcgatgt 1200 tctctaatgc gaccttgatg tcgttgtcca tgccgaacgt caccagcgcc gtgtcgaatg 1260 ctttcgttaa gtcctcaaaa tatgttgcca ttatttcacc ttagaggctt cttcatttaa 1320 tagttgctgg aacctgctaa cgttaactct aacgaatcct tgcggggcct gttgagacca 1380 accgtactcc agacgttgcg catacggcaa attgttagta agggtgaagg tgtgccagtc 1440 agcagcattt agaacgaaac ttgtaacttt accggttgct gtgttgccgg atttatcagt 1500 agcgtccgta gttccttcgg cgggggtgct tcctgaagcc atccagttca tacgaaatcg 1560 cccggtatct acgggactcg ctttgataat cgcagaaaac aactttatag aaacctgacg 1620 aattaccttt tcaggattct tcttagcctt ttccacgaac ttggacacat caagcgcaaa 1680 actcattttc tcacctgaat aaagtatgcc acaacgtcat cgttaaccat cttcttctca 1740 atagcaacta cagaccattg ttcaccgcta aaccgaacct tgtcatccat ctttggcact 1800 acgctgtaat ccgctttaac catcatgtcc ccggcctgaa tggtggtccc gtttacaaca 1860 ccggcattaa caggtacagg aaccgcttta agagggagta cgacgtcagg cccccacacg 1920 tactccccta aaacagggtc ccaatgtttt ttggcctttg cgcaccaaag ttaccgtgct 1980 gccgtatttt ggtagtaggc gagtacctac gccttgcatc cgtttactaa aagcggtgcc 2040 cattacacac cctccagtct tgagatgacc agcaatgcgg atggagctac accccaggaa 2100 gtaaccgttg ccgcttgtgg gtacagtcca ccgaagttag aaccggcggc gtcgcgcata 2160 atctttaccg taaaagtttg cccggctgcg gcattcacaa caacacgaga ttcaatagga 2220 acagtaacat cggtgctagc cagcttagcg acggcgggtg aaccgaattg agcgccggcc 2280 agcagaaccc gcgacaggag gatagaggtc ccgctcgccc cagtccgccc ggcctgtaac 2340 ttaacacgga tggcatagtt acccgcggca ttaaaacgta actacacccg acgcgttaat 2400 cattaccggg tcagcgcttg ttttctgtgc cccaccaaag gctacggtta atgctgtacc 2460 tgtggtggtc ggtgcctgaa tatctgtaga cgacgcgcgg aggacctcta cttccttagc 2520 cccgtacagc atagaatccg ccatctgagt ggtaacttcg cgcaactttt ccggtgtaat 2580 tagcccggac tggttatcag ggaagtttgc cccaattaag gcgaatattt ccgatttagt 2640 cttagccatg tttagcccct aaaaacatta aaggagtagg cattattgct accgcataag 2700 agaggtcgta gcgcatcatc ggctgccgtg atgcttactg tgccgcctga gtagccattc 2760 ttaaagtagg acaccgttac tgcaccctca acccgttcgg tctgaacctc tcgcccgtct 2820 gtagaccctc gaacgtcggt accagcgcca tactcgaccg cggccattac ctgagcttga 2880 ataactaacg atggaattac attggatggc tgggggaacc cgtgtaaagt aacaccggtt 2940 cgagggaatg ccagtgcctg gtttgcggat acgcgacgac cgcacatctg agactcaaaa 3000 aggccaacat ataccgcgcc attacggaga gaggcctccg ctgcgatgtc atcttctggc 3060 agctcaagac cgtacttaga ggccatggac ctcgcgtctt ccaggctgac gtaagagtca 3120 gcattcggga tgccttgacc ggtttccacg ataagcggca taaattattc ctctacgctt 3180 ttgcggcgac gacgctgctt aggttctccg ccgccgttgt aatgcggttc ttcggccact 3240 ggctgatttt cgattaatgc ttccgggttc gcttcttcgc gcatcgggac taactgcccg 3300 tcaacctcga ccacaccttt gtattgttcc cgtactacgt aattatctgt catgtctaat 3360 ccttaaagcg gcccggaggc cgcatgtatt taacttactg tcactacagt actatctgaa 3420 atgatattac cgtagccgtc gtgcgctacg actttgtaag taccggaatc cgccaccgag 3480 gagttggcct tagtgtaagt caaagcggtt gcaccagcga tagcgttatt atccttatac 3540 cattgaacgg ttttagggtc catgccgtcg gcgagtccca cggtcagtgt gatagcctga 3600 ccagtggtta ccgcggtagt atcattgaga ccggtagaga aacgcaacgg cattacgttt 3660 tccatatcaa tctcaacctg cccatcctgc ggactatcat cagacacacc aacaatacgg 3720 cgtttaatta catcaaccat ttttatcctc ctcaggatac agtaccagcg tttttaagag 3780 cggttagcag attagtaacg gcagtgcgca atgaagtcac atctgtacgc aacttattgt 3840 agttggtaac cagtgcatcg aactcttctt tcgtaggagc agccgcggcg gcggccccac 3900 tacccgcgct aatcgccggt ggtgccgcta cagtagcaga ttttttaacc cctccgattg 3960 ccgttgtggt ggcggcgggg atatcgaact ccgcgttaac caggctgcgg ggtagccctt 4020 tacctgtctt tgacattata ttgcctccgt ataaatgaaa agagggaccg aagtccctca 4080 agagtatacc ctacgtttaa gcaccgacgc cagttaccag gaacgcaatc ggtacgtgct 4140 tacggtcaac tacacggttc cagttggtag cgttagccag gtcttgccag ctagcggagc 4200 gggcgatagt ctcagtaccg ttgccggtga ttacggcgct ggtgaagctg taaccaaacg 4260 gatgcagcaa ccacgtttta cgcgtccata ggggtttcaa cgccgccgcc gttagcgcga 4320 gatgcttcac gctcgtattc caacggcatc tcaggattgc cttcgccata accgatagcg 4380 ccctggccga agatgacgga gatgaacttg cggctgcgca ccctggccta ctacggtcat 4440 gctgtcatcg acgataacac ggtagccctg gtaggtggcg aacatggtgt tgttctcagc 4500 gtcacggatg aagtcgataa gctgagcttt acgcgcctgc gcatatacga agctgtgcat 4560 cgcgatagca ccgagaacct caccaccatt acccatcaat gcgtcaccca tagtctgggt 4620 agcgtcaatg aatgcacctg cgtcgaagcc tgaagttgca gaaacgtcga ccaccatgtc 4680 gttctgcttg tgatatgcat cagtagcgga tacgttgtcg ttgtacagac cgagggcggt 4740 agcaatcaga cgacgctggg cctggcgctg ccagaagtta tccaggcggg aggctacgga 4800 ctgcaatgga ttctggctag tcagttcgac agtcaggtct gcctggccga aaccttcgtt 4860 cagatacgca acgcgggcca tcatttcacc ggtctgaata gcgcgcgggg tagcaatatc 4920 ctgatacaca tcgttcgaat agttaggttc gatagaggta tcgatggctt tccagaacgg 4980 caggttagcg atgttagacg ggccgcgggc aatctcggca gcatacggag ttggagtcaa 5040 aattccggag ttgaaaaacg cggttttctc taccgggtct tccgtcatat aggacgccag 5100 tacagggata ttgccagtta cgatgttgcc aatagtggta attgccatta tttatttcct 5160 cagggcttta agttgccgtt caaattcggc agggttagat ttatagagag ccaatcgctc 5220 gctttcactc atgtctttaa acgctggtgc ggccccgccg cctttattcc cggaagcccc 5280 gccaccggaa gctgcatttg ctttaatcaa atgtgaaaaa gctttatgtt cgcgcaggta 5340 tttgcggaat tgctcagggt cggtagtaac aacattgcca tccgcaccca tgaatttagt 5400 cactacgtct tcgccttcaa actcggtctt aacgaacggt gccaggattt ctaccgcttc 5460 tggggtgata aaatcaccag cgaaagaacc taaaacagcc ttacgttcgc taccaagaat 5520 acgcgccgct aaagatgcgt aacgctcttc tttctctttc agtacaggtt catactgact 5580 acgaatcgtc ttttcaaatt cgtccatttt accggcggcc ttcaaggctt cctggtgggc 5640 gcgttgccgt tcttcttcgg cttctttagc ccggcgcgca gcttcctttt tctcgttaag 5700 gagcgcttct tgattcgcct taagcccggc cacttctttc tcaatcagtg cccgcacttc 5760 ttcggctgtg tacatttttg gtgcgtcacc cgcacccaga tttatcttcc accccggctt 5820 cttcatgaag cgggtaacgt aaaaaacgat tcatagtcaa tatgtcccct ggactttagg 5880 acaccgggcc acccggtgtt tctgtaacaa gaataaatta ttccacttac taaggcaact 5940 attccagaat attccttatg tactcctgca accggaatac tttcaggcgc aattgccgtg 6000 tgcactcggc gttctgtacg tcgacggcca ggtcttcatc ggcatcacta cttggtagca 6060 ccattttgca cgggggctgc atcaacgtcg tatccgggga tggaattagc attctcagtg 6120 gcgcgtcgtt tgattgacac ccgttcagaa ggaaactcgc agacagtacg gccaggacgg 6180 ataacatatt tgatgacttc atgggtgata gcctccgaat tttctttgcc ttccgattcg 6240 gcggaggcgg ccttagtatc attctgctgc tgccgttgtg ttttcttcgt cagctcagct 6300 tgtgccttct tttgttgctg tgaaacgaga ttggcgcggc cttctaccca cccgcttctg 6360 tagttgtact ggccgtataa ccacaaggag aggataccgg ctgccgtggc taccgcggtt 6420 actttccaat tcatagtatc ccctttcttt atacgctaag cccgacgaag tcgggcgtta 6480 atctcgtata tctgtaaaca tcatacggcg aatcttagct aaatactgaa agcaatatag 6540 ctcggccctt ccgaatgcgt tcgcggttac gctagggcgt tgtgacgccc gcttcacgcg 6600 acccgcaccg cggtacgctg aaagcgtagt tagcgtagca tttaaaataa tagaaatcta 6660 ctttttgaat aaagtattcc tgaatttaca ttggtaagga ataactggaa taatcgctgg 6720 taaaattaag gccccgtcaa ggggcctttc gttacttagg ttctgagacc ttagccgctt 6780 tgctaaccac cgacacaccg tcggcatcta ctgccttgac gaaatattct ccggggtctt 6840 taacggttag ttccccttca acgtaaggaa cgtttattac ctctttacca tctttaaacc 6900 attgcaaatc gtagggcgct ttaccgcctt ttacaactac ggttaatttt gcagaaccgt 6960 ctttcagctc tacatctgcc ggttgcagat cgaagtacac atctcccgca ccatccagat 7020 acggtacttc gtacaggatg ccgtctgaga ttgtggtcag tccggtctta tccgcaaacg 7080 gcatttcatc taccgactcg ccgagaacag attcgtcttt aatataaaca acgccttcac 7140 ccgcaccaga cacacgtgcg tactgcacta ctcgacgtga cggcacatct ttaactttga 7200 aaaagcccat ctttattcct ctttcagata atcggcaaca cgtttatcga gctccgccat 7260 ctctttaaga gttaacgggc gcccgaatcc gtctaccgat attacacgaa attcttccgg 7320 tgttatccca ctattacgaa aaatcttacc ccttacgggg ccaagtgctt cgtcctggaa 7380 ccacgccggt tgttgtttaa ggaattcgta gtagctcgtg tccgcgctta cctgctgacc 7440 tccatctgca cccctggccg cccgttttgc acccttatcc aggaagtcga attctggact 7500 tattaccggt gctgtgctcg accgacagtt aggatgcgct ggcggtagcg gacctttacc 7560 tatttcgtat tgcatagaat cccgcgaccg acaaacggct gaggttctgg agtcaagggt 7620 agatacccac tcgtatttcg tgatgatgtc gccgttctga gcgtaaacct gttgccgtgc 7680 ttcgttggat acgtgcgcca gcgctgtccg gataacggtt gcagcgttac gctcggagat 7740 atcggccaga ccaccgggcc cgacaacgtt cttaactatc cgacgtgtgg tcatcccctg 7800 tacgaaccca gatttaacgc ccataacaag gcgatttacc tcggtttgcc gccagccgct 7860 cattagagaa acaaaatcaa ctggcttttc gcttagttca agaggcgcga aagttgcggc 7920 ggcccatact tgttcggcgg ccggtgttac gaaattagcg ttaacgttgg tcgacagggt 7980 ttttacgttc cagttaactt catagtctga aagctcacgg gcatcggcca gtagcttctc 8040 ataccaccca gacgtaatcc cgttcagcgc ttgctcaagt tcccgcagca tggtggtaag 8100 tctcgccgcc gttctgctgt catcaccaaa caacaacacc tgccgcttaa cctcatcgcg 8160 catctgcttg atgaatggcg ccaggtcttt cacctctccg gatgcggtgc gttgtaacca 8220 tacctggtgg cttattaatg atgtaagtaa gctcatagat aaaccctgtg atgctaaagt 8280 taatagcata ctataagaaa acccatccca aggctatggg cgaccctaat tagtagaaca 8340 acttaatcat agttctaaag ttaactagaa gtatttgacg aatactatat ctaggtggta 8400 gtattcattc aatacttcaa aaaggagata gaaacatgaa atcgcaaaca ctagaatttc 8460 cctatgggga tataactttc accccgagaa ggtgagctga acctgacggt attagtgaag 8520 cagatgaata agtggcgtga agaaaatggg atgggtgtga aaaacctatc gcagctgtta 8580 tctaccgagc aagccaggtc gttcgcgcgt gtgtgctgtg aggaattagg gatagaaagc 8640 gcgtggcgtg tggttagagg gaaaaacgcg gcaacatacg catgtctaca tatggctata 8700 tgggttgcgg agcaatattc tgactattac cacttcctcg ttatagaccg tttcttaact 8760 caacggcagg tagaactccg taacattggt gctgttagtt ttgtagaact taatgccgcc 8820 gttagccgga tgattgagcg tacggagggt aggattgggc gtatcggtca cttcatacat 8880 gtggcgaatg cgataaaaga gtctatagat atccgtgaag taaccggttt tgatacttgg 8940 gactctcagg acgccaaaac taatcaatta cggagtgata ttcaaaaggc tatggtgacg 9000 ttgttggata tggaagcagt caactcatgg gatgagctga aagagacgat accgcgggtt 9060 gtgcgtaaat gcgcggctaa tatccgctaa aactaaggcc ccgtaatggg gcctttttct 9120 ttactcctgt tgttgttgtg ccgcttgcgg aatctccccc gctacctgag taacagcacc 9180 cgacggcaaa ggtgcatctt caatagcgtt cagaatatcc tcgtcggtcc agtcagtcac 9240 ccccgcctta cgcaacgcag cgtaataagc agtggcgggc agtaatccgg cattaatgtc 9300 tgccatccac gcagccctgt cctgtgctgt cataggttgc aggaagaaat ccatgttaag 9360 ctggaactcg acttcagaat cttctggctt acccaacatc atagcaaccc atcgtaaagc 9420 atcggtatac gcctgactta cgttacgagc gattgtggcc ataacggatg tatcggcgcc 9480 gcgttggatg cgcgcggatt ctgcggtaat ttgctgagat ggtgtaataa gttgggcacc 9540 aatctggata gcctgctgtt ctttgtccag catattctgg cgggccaggt tgttttcgcc 9600 cgcctgaata agttgagcgc taccaccata accaaggtta tgcccgcacc gactgccaaa 9660 tttgatgccg ttggggttgg cttccttgaa cgactgtggt gtaaggttat ccccggggta 9720 gataaacagc gtaggctggc caactacaaa acttgattcc tcgttatcag cactgttccg 9780 gtagtgcccg atattaagct cggccaatgg caacaaagga gcgtcgtcaa tggtggcgtc 9840 gttattggta gctccgataa aggtaaacgg aattacgcca cgtaacgact cccctaaatc 9900 tgggtaaatc tccacaacct cttcctgagc tccgccttcc gcatcgaaac ggaacagtcg 9960 ctgacgataa ttaccatcgg tgtcaatgtc cagcacgcgg tactgctcgc cgtatttagt 10020 ttcgaactcg tttccaggtt cgtggtactc ccatgtctca cgcagcacaa ccatagttac 10080 ccggtttaca gaacctacgc gcgtgagtcg ccagttaacg atattctcgg tagtgtagaa 10140 agcgattgta gggttaagca agcccgcatt ttgttcggca gcggtggccg cggctgtttc 10200 cggagcatca acaagaagac caccacgacc tactgagtca atctccatga gtgtatcttg 10260 cgcgtgctgt attaggccta caccagaccc atctgcattt ttaagcaggt attccagctc 10320 cttcggaata ttgatttctg gttctttgcg catgacgcta ccgaccattc ccgacagcgt 10380 tcgccgggtg aagttgtaga cgataccccc ggcctcgtat tctgcctgac gcgcttcgcc 10440 gtatgcttta tctggttcat taaggccgac gttgcgtaag tagccaacca ggtctccagc 10500 aagcgcatgg cgcaccttct gccatttcgg cgcgtagtgc agccattcgc ggtgcttggt 10560 ttttacgcca gaaccctggc cgtttgctgt taacatttaa taatcctctt aaagtgcaaa 10620 agtaaccggg atatgggaaa caggtttaac caacggcatt tcgtaggcga tagggtatcc 10680 tgtcgcgtcg ttctggtggt cgttgccgct tgtcttatcc ggtataccgt ttttatcgta 10740 agcctgttgc tccaggcaac gtgctgtaac cgggcaagcc tgctcgttga ccattaatct 10800 accagactcc agcgctttat tcaccgatgc tacacggtct ttaaccgcag ggttaactga 10860 tttcgcacgt atctcgaaac ccgcgttctg taattgggca atatctgacg tactggcatc 10920 agtcgattta cggttcttgc cgctggcatc cggatacatg acgatgtggt ggccctgtcg 10980 cccccatcgc tcggtaattt ccctgaccac atccggggtg tcgaacatat ccaccagctc 11040 ggctaccgcg tgccagacat actcacgctg aacgtatacg gtgctagcca tatgcccgac 11100 gttgaagtcc tgaccgatgt acagcgtctc tccaggttgt atagtctccc gactgctgtt 11160 cttacgtcgg tcgtaagcat aatacacgct gccggatgtc aggttgacaa attcgccgtc 11220 gatgtaggca tcaattaact ggcctggata cgtatcccgc aacgactgca cataatcttc 11280 cggtagaaag ggattggatg ttgtcgaggc ctgaatcatt tcatagcctg gtttcttttt 11340 tacagcccat ctgtcgtgca caaaacggaa gccttctggc gtcgtaaata ccgaaactgt 11400 attagcgggc ttcggggtaa ttggacgata tgtacgcggc aactgacggt tacgggcgat 11460 aactttgttc caggcgtgct cggcgtggtc tttatttaac gtgtccaact cgtcgatttt 11520 tgcgcggaac gattcgtagc caacaattcg tgctggatta tccaatgtgc gaaggacaaa 11580 atcctcgaat tgcccgaatt gcccggatga ggtataaatg atgttgtcgg atttattata 11640 cttgtagcga ataccccaat cagacaactt ctcttccata cgcggagcga ggataaggcg 11700 caccaggtcg tatgtcggtt catacatggc gataagtgaa tcactaccgc cctccatgct 11760 gtcgagtagg gcggaattgc acatgacctc tgatttacct gtgccgaagc ccgcaacgaa 11820 ggccgggaac ttacaatgca gattaaggaa cgcgccttgc ggttccgtag ctgtgatatc 11880 aacgttcacc ggacaccacc tttaatagtt acttcgctaa ttggttcgtc gcgcgcttct 11940 tccacaacag ttttattaag ccccagtttc gccgcggcaa acgtagcgga tatcccggct 12000 gcaccggtct cagtgaaata cgcctcttcc agggcttgtg ccgtctcata tgcttcggcg 12060 aatgcaggaa cctctcggag ccatagcttg ataatcggga tggttacgcc gatgtgtaat 12120 gcgaagcggg ccagcgacgg cggtttgtcc tggataagcg ggcgttcgtc gcctttagac 12180 gtgggtacga gttcccatga cgcgcggtcg aagaacctga ttaactcgtc gcaatagtcc 12240 ggatcccata gttcggcgga atgtctggat gactgataca ggctttgttt accacgcggg 12300 cgtttacggc gacggtttgc actaaccgct tcttcgtgcg cggcggccac tacttcggcg 12360 tcaggctgtt ttaatttgag tttcatgcaa tcctctctta aatgcgtatg accatcatag 12420 cgcatcagac gccccctgag cgtctattaa ggattgtaca ggggaatgaa ggcctgtgta 12480 cagaaaagcc cgcagaagcg ggcttgtgtc agttatatag gccgtgtcgt agtgctcttc 12540 taccatcgcc gccgtaatgg tcgttagctg cggcgataac ccaggcgttc gggatgatta 12600 gcaatctcat ttgaaaaacc acagcaggat gccggccgct cctgatacac ccaagcctaa 12660 aaacataggc caccacagat atcagggaag actgttaaac acagtcggtt gttttattag 12720 cgcactgaga aaggcaaacc aaaacatcag gtcgccagtg gcatcggtag ccttcaaaat 12780 ctcaatcatt ctccactctc ctctttataa acccaccatt tacctacggc ttcccatcgc 12840 cacccgcgac tatgtatggc ttcgaaccta cgaccaaata ctgtttcccc gtaccacatg 12900 acaaatagct tcaccgcggc accagcacca gctttatgag cattcaaacg gcgatacgct 12960 agtgacatcc tcgtaagact taccgcctca acaaatggcc aaagaaacaa aatccacatc 13020 aggtacatag caccggcaag cagtaatgtt gtgaatccta tacccatgag gatgagaaaa 13080 ctatccatca cccctccagc ttcatacggt cgttgcgcga taatccacgt ttcgtgtaga 13140 tgaccggtga cgttttcgta tgggtcatca gtcggttgtt atagatgaca tgttgctcgc 13200 attttgtgtc atggcggtac ttggacaatg tcatctcatt aagtcccgtc tcccggcacg 13260 tatcagccag cgtgccgtgg gtctctatta gttttgggat actagtaatc attcctgttc 13320 gctctccttc tcaagtcgtt ccagcagcat cttgtatgct tctaactgga attcttcgtt 13380 cattgacagg cccagcttac ttacagactc caaatacgtg atgcgttggc gcaacgtttc 13440 tttgcatact cgtttcatcg tttaatcgcc cccgcactta acacactgtc cgcacatatc 13500 tttaggattg ttaacaaaat cgtgctcgca cttattcagg tcataacggt tgataagctc 13560 ccggcgatac tcattaaggt agtcaagcgc agcctggtgt gcctcgattt gcgcgtctac 13620 tgcttcaatc ttaaacctgc cgttggcggc ttctaccatc atcttttcat attcgttgtt 13680 agtcatttct tcttacccca cattcgattg agatacttgt tcttgtccgg tcctgggaaa 13740 ctgttacgtt taatcagttc ctcgcgtgtc gggaagggcg tgttacttac cttaccgccc 13800 accttaaacg ttaccgttac ttgcccgttg tcgctcattc ctgcacctcc catccgagta 13860 catttacgcc gtcatagtaa tgtgaggtgc gctccagctc ggcggcggct tcggcctttt 13920 tttcgctgag ataaacatca agaacgtaag tgtccccgtc gtgatgccat cccggatacg 13980 atatatactt tcatttttgt tctcctgcat aagctgcttt cagtgtctgc atcgcggcga 14040 accagaacgc atccgctgga atccgtgtac ttaggttatt aacggcaatg cgtgccatca 14100 gttgtgcgtc tttgaatgct tcggtgtcca gtagcggttt gcaatggtaa aaaactttat 14160 cgctttgtaa gttcatcttc tctactccgt tctcgttgtc gatgaactaa atataatagg 14220 gttaatgtta ctagtcaact attatttctg aatacagata aaaaaaaatc ccggtaaggg 14280 tcagttaccg ggataaagga gctagaggga tgaataagca ggagcgacac cagtatctta 14340 ggaatgctcc tatgtgtcaa cctactttgc ttcgcggccg atggcttccg cctgttgcca 14400 cggcataccg tcgaatagcg caacacgacc cgcggcacgc cgacgcaatc caaggagtga 14460 tttgccgttc tgataatgga actgagatag cttattccgc agtgtggcaa cgtcgccctt 14520 acgcagggcc tgccctgttc cggtagaagc cgcaatcaca ccggcgccag cgttatacac 14580 caggtcgcac atcgcatcga actgtgactg attgagcgac ggatgcgcta cagcgtctac 14640 cgcagctacg gccttagcca tatccttgtg cagcagcaga agaccctgtc cctcggtaat 14700 cttctggcct tctttcacat ctgcgccgta gtggccatag ccgatagtaa gtacttctca 14760 ctcttcgttg ccttgtacgc ggttccgcgg aaaccttcga acgcggcggt gaatttgatg 14820 ccgttgttac taatgtttcg gtttgacata catcctccgg tatttgtagt agcgagcgcc 14880 tttcacgcaa atgtgacgca ccgatttaag cgcgaatata gcgacgagag tcgtaattat 14940 gaacggtgga atggcaccgg taaagagatg ggcgagacct ccggcggcgg tcaccaccga 15000 cgccatgtac agcaggcggc cgaaaagacc gtcttcaacc cagtcggcgt agatattaat 15060 taacgaggtg gcgatgatgg caaccaggca aatgacgcca attgatgagt tgagcatgtt 15120 atctccccca cggtaactta aagttgctga gggatgagtc agcagcttcg aataacttaa 15180 gccagaacat acctaccccg aaaggaacca ggtactgacc ctcagactgt gataatttca 15240 tagtaactta taatgatggg ggatgcgtac actgcacaac aggcggaagc ggtgagatgg 15300 cacaatcgca accatggcga aatagcctca cgcttcttaa tctgagatac ggcacccccg 15360 gcgaggcccg ccacactaag ccaaaggtat ttatcgtcca caataaaaaa tctccgaaag 15420 ttggtttatc cttcggagat tagcatttat acgacaattt tagtaacaga taattcggat 15480 ttatcctagt cgatgtgtta tcacgcttcc atcggtacca gggtcaaacc tcttcgcaac 15540 ctctaccgct tccgttgccg tcgcgcccat gaacatggct gtaagtgcat acggagcccc 15600 ggagcctacg gcagcagcgt tacctgcaat tggtataacc gagcttaacg gtgcgtccca 15660 cgccctacag aattctaccg cgaaaatacg gttatcctct ttaacaaaca ccagcgccga 15720 aaagtcgaag tcgtagtgct taggcgtcag tatctcatca tcggcaacca acaacatacc 15780 caccccggcg tccccggata cccctataac aaagtggtcg ttctcgtaaa tcttggtatc 15840 cgtgtggtat ttgtaattgc cggtgacaca ggtatcacag gccatggttt tgccgtcgaa 15900 agctatcgta gtcatataaa ttacctgaac gtataaagtc gaatagatgg cttgtgcggg 15960 atttcggaat acccacgttt acgccacagt ttgtacagtt taaccgccat atgcgccctt 16020 gcctggctac tgaactcccc caggtttacc actttcttcc cgtcgtaaga gtgcgcacgc 16080 cacagacctt tgcctacaca atacttaggc tggtgcaccc cagggatgcg cggatacacg 16140 tccggcatct cattaactgg ctttgcgcgc ggtttaggtt tacgcttaac cttttcttta 16200 acgcgagtaa ccttctttgt taccgccgat gaaggaggtg ccaggttgat gccggcgtgt 16260 ttcagtatat cggttgcgag actcatttct ccccctgctt agataaccgt ttcagtgtgc 16320 aaataccgcg tacctgagcg tcaacaggac gctctacgaa agttaacggc gcgttctccg 16380 cagcgcgtgc ggctacctgg cacgcctcta acgtataaaa cgtttctgtt ccggcaagtt 16440 gtagctgacc cgcggataca acccatatga aaagtatact tgtcatttga ttatcctgta 16500 cttaactata tcccagttgt tgttatcaat cccccaccgg tagtcctggg ctgcgccgac 16560 ggctatagag ccattgcgga attcaacctc aacctcacaa tcgctaggca ccgggcacgc 16620 cttacgtcct ggattttcta tccaaccatc gtacagctca tgttgtgggt tcaatactgc 16680 cccgcattta gtacactgtt ttaagtcgct actccacaga accaaacggt agtagtgctc 16740 acattcttcg cgtttagtca ctcggtaaaa gtcggcattc ataagttcga tagtatcccg 16800 tagttcctta accttatcta cctcccggcg taacctgtct ttcagcatgc gtatgtcgcc 16860 gtctttacgg gcgaggtgtt gttgtaagtt atggatgatt tgtttgctgt cagaatcatg 16920 gcctatgtag gcgtcgataa ttgcgttctc ctcgtccgag taccctgagc ggcctgttat 16980 cgaccggtct atgctcatct tgtgtatttc tgctacaacg tcgaacttat tttgcgcgct 17040 gacgtagttg gtcaatcagg tattcaaata cttcgtcttt gctactcatc tccacactcc 17100 tcgcatacga tagctttcag gtccccgtca caaactacat cttctccggc gcaccatata 17160 tacactgcat cttccaggct aatatcctcg tacattcctc ccgcccctct cgtttcgata 17220 cagtaatagt acactattat gtttggttat gccaatacta ttttacgatt aatttatccc 17280 gacgcaccgt aagataacgg ggtttcttcg catgagcacg ccaccactca acacgggcgt 17340 agatgccctg aagcgatgtt actttgccgt gtctgatttc tccgttccgc ccggtccagg 17400 ttacaaggtc atttggtttc attcgttagt ctccgttatc gattcgatga aggtcatgag 17460 ttccttgtaa caagcatccc acctcatagc cgctgcttca ttactcctcg tcgtcgaccc 17520 tgattgtata ggacatatcc gccgatgcat cgtctatacc gtccaccagt tccatcagat 17580 atacagcctg ttgtttagta atcattgttg cgtatcctgc aacaagttgc gttgattgtt 17640 acatttcgtg aaatagaaaa atattaacaa aaatggtacg ctgctactca gcagcgtaag 17700 tcactcagta aaatcctaaa ctactgttac aagcttcctg cggttccgat aacagcgccc 17760 aggtgcggcc ccgccaccac gataccccag gaactatcca cccatcatca tcggcgtccg 17820 gatgcggccc cggggaccac taaaacccta gccataaacg gcaacctcct ttaagacccg 17880 agaatcaccg gcgcggactc tttagaatct tttgccgtgg ggttatttac agggaggtcc 17940 ctgccccatt agagtgcctt tattctcgtc cctcgtcagg taagcgcttg ccgcaaggca 18000 agtgcgttaa tccctgagta gtagactaag aatctttcgg catttcagtt acgttgccgt 18060 tgtccagtgt tgtgccttta ttcttattac ttggtaacta aagggcaggc cttaagcctg 18120 cccactaatc gttacatttt tacggttgcg aatttacggc acatcaagga attcagatac 18180 ttaagagata tgctgccgat gagaatatag ctcgcccagg gcagttcaga tactaaggaa 18240 gggaagttac tgagttttaa gtttgcgggt ttcagtaagc cttaacgcgt atccgcttgg 18300 ggcggacccg ctaagctctc acaaggtgaa acagaaactt aagtaacttg aagttacaag 18360 agacaactta caacgaaaaa aatgaaaatg tcaataggtg tgtttatagt tgatttccca 18420 ttatgccgtg ggtatacttg ctttactaac tgagatggag gatttgagaa tgacacagaa 18480 tgaagtagct gagcttattg gggttacccg ccgcacgctg aataactggt taagagacgg 18540 caagttcccg gattgttgtg tccggattat gggtcgccgg atgccgggta cgtttgaccg 18600 ggagaaagtg gaagcgtgga ttagggagaa tgtaaagtga ccgatttcca gaaaagggta 18660 ttcagaactg tagtcgcgct tacgcgtaaa aaagggtcgt gtagtgttct tgacttgcgc 18720 cgcgcttatt ttcagtacta tagttcctct ctcatcgaga ggtcgctgga agttttggtt 18780 agagacggag cagttaaaag taaggccgga aaatattcgg cagtagctga agttaaaggc 18840 tcccaggcga ccccagaaga ccttgaataa aagaaaagcc ccaacgcgcg aacgtcaggg 18900 ccttaaacac tataaagaaa aagtaaatga aaacaggcca tttaccatac tcatggccgc 18960 gacccactcc ccgtgaggaa cgcgctgaca ttaacgttta caccacgacg tccggcacct 19020 gggatgagtt tgtcgaactt atgcagccgc ttaaacgttc gcgccggaac cccaagacag 19080 accccggcta cattaccgcc gcttgtaccg ccacggtaag ctctaccggt aaagaagccg 19140 ccgaaggtat gttctatcgc tgcaatgcgt ctgttacgtc ttcatccctg gcctatgccg 19200 acgtggacag cgcgacgccg gaagagttcg caaccgactg tgagatggtg cgcgagtcac 19260 gttacgcaat gatgctctac accacggcat cccacaccga agaagcgccg cgctaccgcg 19320 tcgttatgcc tgtacgcact ccggtaaccg gtggcgacat catccgcatc cggtacggcc 19380 tgttggcaca cttccttaaa ggtcgtgacg tggatagcgc cgggttcacc ctgtcgcagc 19440 ctatgtaccg cccgccggta ggaagccagg tcatcgtgtc tgaaagtagc cgcatgatta 19500 cggcgagcaa gcttatggag gaagttcctg aaattaacgt taccggtgct tctgattata 19560 aagttccgga gggtgaacaa tccgaattaa ccgacctgtt tgaagagttc gcttttgaat 19620 tcggcggacg tatgaccgac cgcggcctgc aaatgcccgc cacgccggag cacgccgccc 19680 aatacactac cggcgaacct aagcaggacg acttcctgtt ctgttggccg cgcgacggct 19740 tcgagcgacc caacgttacc ctgtaccacg ataccgacct ggtagctacg ggcgggatga 19800 agcctggcgg acgggatatg tgggcttacg cttgtgcggc caccggctta ccttttgacc 19860 gtgtggaggt ggcgcttggg tgggcggagg gggtcacttg cgacgaagaa gacctggacg 19920 acgaagaacc accagcacca caagcggact tcatcgtcga gggctatatg ccgtctaatt 19980 gcatttggga tatcgtcggt gaatccggga cgtataaatc gttctacaca ctcggcatga 20040 tgtatatcag cgccgccggg tatcggttcg cgggggcaga cacccgtaaa gcccatcatt 20100 tctatattga tggtgagggt ggggagttca cgcacacccg tatcgctgcg ttggccgcta 20160 aatacggcga tgaaggtatg cgatatatgc atgtgcttga cgctggtgaa ttcgcggata 20220 cgaagaaact ggtgcgtaaa atgcgccaga tagcgggcag tgaacctgtc gggatggtcg 20280 ctttcgacac tcttaaccag accttcggga actggataga caagttcaat gagaactcag 20340 ccggacagga cggtatgggc cgtgtcgtgg ctatgctcaa agaagtgcgc gacggtacta 20400 aaggcgctgt aggcgttgtc caccacaccc cgaaaggtgg aagtaaggcg cgtggaagtg 20460 gtgcgctgta tgccggtgtc gatgtggaac taacgcttgt ccgcgctacc gagaaacaaa 20520 taaacgttgc gcacaccaag aacaaaaacg gtatgcagca gaaaacaata ggcatggtgc 20580 ttgaaccagt acagtttcgc gaggccccgc cgccgaaaga gttccaggcg gtagaattcg 20640 ctgggggtga gggatacggg gagatagtca acctcgacct tccggagccg cataaagctc 20700 tcgtacttat gccgtggggc ttccagccgt tcgaaacgga cgaggaaaaa gagcgaaacg 20760 aaggattgga tggtaaaggt aaagattccg taaaggatac ggttaaacgg tctaaagacg 20820 cttcggcccg tgagtcggtt atgtccgctc tcgaggattt acagcaagcc gatgatacgg 20880 gacgagggtt cacacaacgg caaatagtcg cgagagctgg agaccacagt atcacaaacc 20940 tggtgcttga gaagatgcta cgcgagggtg aattgattct tggttgcgac gaaaacggtg 21000 aagttgtaac gaatacttac aggttgccga tgggtataga cgaccgtaag cgaccgaaga 21060 accggtatga acctaacgac aacataaaga caacagaagg ggatttagag taaaaagaaa 21120 ggggccatca ggcccctgta tcaccaaagc ttaaccatcg agggtcaaca ccgagagcat 21180 cggcaatgtg gaagatttta aaacagtcta ccttacacac ccggccggtt gccaggtggg 21240 taatcaggct ttgggatacg ccagctttga aagcaagctc agtttgaccc atgccggatt 21300 tctttacggc ggcgctctat gcgagcgcct agctcagatt gctgcatatg tttactcctt 21360 agttagatat gcagtaatag tacagtaata aaatacacga cgcaatttac gaaaaagttc 21420 ttgcataaat aatagagtac gattatagtc tttatcacca actaaacagg agaagaaata 21480 atgttagacc aattcttaaa aattacttga acgtttcgta gtcgcccatg aactgattgc 21540 tgcgaatagt gtgaaacaga cgtctaacaa atccgtaagc gaaatgactg ttgacgtctc 21600 cgttgccggt gtcgagacag ttaaaaaaga actggctaaa gcactggaag aagtcgagaa 21660 agcaaaagaa attccggtcg aaggtgagga catcgtcgac actaaaccgg cagaagagga 21720 gaaaccaaaa cgtaagccgc gtaaggcaaa agtagaggaa ccagtacaag aagagaagga 21780 agaaattgat taccaatctc ttcgcgacca gattcaggct atcgacgatg caattaacga 21840 aggccctagt gatgctgcgt gcgatgattc cgatgaactg ctggaagagt tcaccggtaa 21900 gaaaatgaag attgccgcga ttaaagacga agaccttgct gaatacctgg aacgtctgac 21960 agcaatcaag aacaagtatt tcgaagaaga ataactactt aacttatggt gccttcgcgt 22020 aagagggcac ataactgagg gtaaagaaat gatcttagac atcagcatta aatgtgaaaa 22080 cttagcggtt tctactgtac gtggtggctt atacgtagac attaaaaacg cagaattagc 22140 ataaaacgtc ccggtagaag atataggccg tcacgtgtcg cagattaagc gcacgacttt 22200 cgccaagcct tacttttatg tctttgagtc gatgcatgga tgggctatgc attgcgagca 22260 taagacaaga catttaattg accagtggga gaacagagca tgactgataa atggtggctt 22320 aatttgcgca gcggcgacaa agtttacacc gtcggcgacc gcagaaagaa cagcggtgta 22380 gctactgtgc tacaaaacgg caaaaagtac atacacattg cgcatgaagc tcgggcgctc 22440 cgcgtgaata aatcgtctgg taggcttgag gattacccga aaacattaat atacaaaagt 22500 gaaaagcatc acatggtcgc tgttgacgca agacgtcagt tcgcgggaat gctgcaacag 22560 ttaagcgact tacatcgcga tgattctttc attcccacag aagaacagtt acaggcctta 22620 aacttatatt tggagagttt acgatgattc taaaagaacg cggcggcaat aacgatgttc 22680 acgccttact gtcgccgtca ggtgctaaaa aatggctgtc atgcgctgca tcactggcct 22740 gtgaaaaaga tattcctaac acgtcaggta aagctgcggt attaggcaca gctatgcaca 22800 ccatagcgga gatgcacctt aacgcctata tacgcggcac tgcactgccg ttagaacgtg 22860 aagtcggcgc ttatgttctg gatgaaggta aaggccagat taaggcgcta atcagcccga 22920 tgaaaggcgc ggtactgatt acggcgggca tgattgagca agtgcgcaag tacaccgact 22980 actgcaaacc gattattgat gtagcgactt acgccaagct ggaaatgcgc gtcaatctta 23040 ctgaggtatt gcatccaggg tacatccata gccctgacgg acggcaagaa gaactacaga 23100 cattcggaac ggctgacctt gtcgccattc aggaactggc taacaccaac gagcacatgc 23160 tcattattgg cgacctgaaa acaggacggc atcgtgtcga agcgaaagaa aacaaacagc 23220 ttatgcttta cgctcttggt gtttatcgcc gactcaagag acgttatagc ataacaactg 23280 ttcgtctggt catattccag ccgtatgctg gcggcgcgtc ggaatgggac atttcggttg 23340 aaggtctgga actgttcgct aagttcgcgc agaaacgcgc ggtagctgcg cttgacgcgt 23400 atcggcgtgg taagaaaaac cttaaaccat cagacttcaa gccgtcggtc gacggctgtc 23460 agtggtgtcg gttctccgaa cagtgcgccg cgcgtacaaa aaccgttaac tctgtactgg 23520 cggaagaact ggaggacgac tttgcgctgg aacttacacc cggagcaact cgtagctgag 23580 tatgagaagt tgccactgct gcgccagcac atcgacaagg tcgagaaagc gatggctgcc 23640 gcgttgcatt ccggtaagaa agtgccgggg tacaagctgg ttgaaggccg cccgggtaat 23700 cgtgcgtgga aagataccga tgcgctgctg gaaacgctat cacattttga gttgggcgct 23760 gaaatgcttc ataaagaagt actaatgacc ccgaccgagg ccgagaagca gcacaaaggt 23820 tctgagctat gggcggcgct ggagaaacac gtaacccgta agccaggagc gccgtgtgtg 23880 acaacggtcg aagacaaacg gccagaatgg aaaaatgtta ctgaagacga tttagaggcc 23940 ttgacatctt aatcgtactc tattatatta ctaatcactg accgggcagc tccccggata 24000 aactctaaaa cgcgagaaaa ctaaaatggg aattaaactt aatcttcgta aagtacaaac 24060 cgcatggctg aacgtattcg aacgcgctaa agaccgtgaa aataatgacg gttccgttac 24120 taaaggtacg tataacggta cttttatcct gactccggaa cacccacaaa ttgaagagct 24180 tcgcgatacg gtattcgcag tagtgtcgga agcattggga gaggccgccg ctgagaaatg 24240 gatgaagcag aacttacggc gaaggtaagc acatggacaa gtgcgctgtg cgcgacatcg 24300 ctgagcgcga taatccgttc gaagacttcc cggaaggttt ttacttccag gccaagaaca 24360 agcaacaacc attaatcctg acttcggtta agggtgaaaa gcaggtagaa cctgacttca 24420 atattgatgg tgagcagatt gaaggtgagc aggtatatag cggttgtgtg gctaatatct 24 480 caatcgaaat ctggttctcc gagcaatata aagttttagg cgcaaaacta aacggcatca 24540 aatttgccgg tgaagggaag gcattcggtg gttccgcggt ttctgccagt gtcgacgacc 24600 tggaagatga tgaagacgaa acaccgcgtc gcgaacgccg ccgtaaccgt taatatattt 24660 tcttttcaat taaggcggct tcggtcgcct tttctataag ggtcaaataa atgaatctgc 24720 tttatctcga tactgaaaca ttttcagaag ccgatttaaa aaaagtcggt tcctatgctt 24780 acgccgaaca tccgactacc gaaattgtta tctgcaccta cgctttcgat gaaggccctg 24840 tgcaagtatg ggacgccacc gacggcagcg atatgccgcg tgatttgcgt cgggcgatgc 24900 taaagctgca aaaaccagac agcaatctca aactggtagg ccaaaacttc cttatgttcg 24960 accgaccagt tattaagcat tgctggggat tcgaactcct ggtagaaaac attatagaca 25020 ctatgatagt cgcgttccga catgccctcc cgggttcact ggccgcgctg tgtgaggttt 25080 taaacattga cgcaagcatg gctaaggata aacgtggtaa ggcgctgata cagcgattca 25140 gtaagcctac gcccaagaac tataagattc gacgttatac tgccgatacc cacccaaaag 25200 agtgggcaga atttattgca tacgcaaaaa gcgacattac gtccatgcgt gaagtgtata 25260 agaaaatgcc taagtggggg aattctgagt tcgaagaccg cgtgctgcac ttagaccagg 25320 tgattaatga ccgaggattt aaggttgatg tggcattggc ggaagcagcg attgaagcag 25380 tcacgcgcca taaagaagag ttacaggaag aagcccaacg taaatatggc ggttcactaa 25440 ccggcaaaga cttcttgcct attttacagg agctagcgcc agcgcaccgc atacacaacg 25500 cacagaagtc aacactaaac gatttgctgg cggatgagga tttaccggac gacgcccgca 25560 ctattatcga gatgcgtctc ggggctgctt ctaccgcatc gacgaaatac gcgccgttgc 25620 tgttaggccg ttcttcagat gaccgccgcc gtggttgcct gcaatacgga ggagcgaagc 25680 gaacattgcg gtgggcgggg aaaggttttc agccgcaaaa cctggcgcgc gggtattatc 25740 acgacgatga actggatagg ggtattgccg cgttacttaa aggccgtgca caccgccgtt 25800 ttgatgtggc caagctaaca gcgtctaccg tccgaagctg cattatcccg gaagccgggc 25860 gtaagtttgt tgttgcggat tactctaacg tcgaaggccg tgggcttgca tggctggcgg 25920 gcgaagaaac cgcgcttgat acgttccgcg ccgggctgga tatttactgc gtaaccgcag 25980 gtaagatgtt tggcatggac cccgacgata tcaagaaaga acgtaaagac ttacgccaga 26040 taggtaaggc ttgccttcac cgtcataccc aagttttgac cgattggtgg atttaaggat 26100 attatggcgg ttacatcagc atataaagta tggaatggcg aaaaatgggt gaatacaaaa 26160 ggcacgcacc ttatggggtg gaaaccggta ataaatgtgg acggggttct aatgaccgaa 26220 gaccacaaaa tcttgacgca ttcctggaag gcggcaaagc aactcgtttc aaacaaatat 26280 atgatgggcc tcgccctggg gagaggtggg gacgcctggt tatcctacgt gagctaccaa 26340 aacgacaagg ccaaagacca actactcgtc caatgtgatt gcggagagat gccggggcgg 26400 gtctattatg acaatgtccg agcgggtaaa accactcaat gcaacatctg cgcccttgaa 26460 gcaacaaaaa aatatcgtaa aaaatatttc tgctacgaag acgcaatgcc ggatgatgcg 26520 cacagaacgc gattgctcaa ccggttgtcc gcggcaatcg tgcggacaac aagccccgga 26580 aacaagagtt acaaaaacta tggggctagg ggtatcacgg tatttgacca gtggagggcc 26640 gataaaaggt cgtttcttag atatgttcaa actttggaag gctgggatga cccaaacctc 26700 gaaatggacc gaatcgacac agacggtaac tacgagcctg gcaacattcg gtttgtcagc 26760 cgttctgaaa actgccgtaa cagacgtcga atacctgagc ttcaacgaaa gtatgatgca 26820 gccgttgccc gcattgctga gcttgaacgg gaaattaacc tactgcgagc cagtttacga 26880 cctgattgat gtggaagacg gcaaccgttt tcttatagcc tcagactcgg gctttcttgt 26940 ggcgcataac tgcgaactgg gcctcggcta cgagggaggt gtcggagcgt tcgttacgtt 27000 tgctaaaaac ctgggtcttg accttattga gatggcaaaa acaatggacg ggactttccc 27060 cgaccacatc tgggctgcta ctgcacgtgg gtatgagtgg gcgcgtatcc aggaagccaa 27120 gagaccaccg catcccggtg aaaaggatga caggccgtcg tatatacttg acaagaaagt 27180 atggcgcaca tgtgacgcga tcaagcgtat gtggcgtgag tcacaccctg aaacagtagc 27240 gttctggcgc gaccttaaag acggaatttt agccgctgtt cgtaatcctg gtcgtgaatt 27300 ttgggctggg gcacacttgc gccggaatgg tgaaagggct atccgcatat ggcgtaccgt 27360 agaatttgat tcgtcgggca ggaaggttcc tggctggtgg ttgtgtatgg agttgccgtc 27420 aggccgtatc ctatcgtatc cgggaatcgg cgttagtgtg acaaaggaaa cagacgaaga 27480 cggacggata aacaccaatg taagaattaa gtaccagggt gagaaccagt taacacggca 27540 atggaccacc ctgtacacac acggcggaga aggcttgtga aaacattgtt caggcgttgt 27600 gccgtgactt attggcctat gcgatgctta atgtagaagc cggtgggtat ccaatagttc 27660 tttctgttca cgatgaactg gtatgcgaga ctccggatac atcagattac acggtagctg 27720 aactggaaaa actaatgtgt gcattgccag aatgggctga tggttttcct cttgtagcgg 27780 aaggtgcgga gttaaaacgg tatgctaagt aaactgatta tttattattt cagggaagaa 27840 gactgtcgaa tctgtccgcg ctgtggggtt aagcacacga aacgtgaggg gtgcaataga 27900 tgagtacgcc tgagggccgc gtccagaaat atgcaaaaga gcgattcgag gccctggggg 27960 gcctcgtacg caaattgtcg tacgaaaata gagtgggcgc tcctgacctg ctggtgattc 28020 tccccggtgg catcatctgg ttcgtcgagg tgaagaaaga cgaaaacacg aagccagacc 28080 cgcaccagtt gcgcgagcac gagaggatgc gtaaacgcgg cgcaaatgtt tttgttgttg 28140 ggtcgaagaa acaggttgat aaattaatag aacactatta tatttagtta acaccaaaca 28200 gaaataaggg aattgagaaa tgaaatatga atatgaccgc aaaccagcac gtgacatcgt 28260 accaggcgat atgattttca acgttaaaac ccgccagccg gttgccgttg atactgtgtt 28320 cgtcgagtca aacggtaaac tggttatcga agatgtaact ggcaatgtta cagcgttcgg 28380 gcgtaaagag ctggtgctgg tgctgaaatg agtaaattca atagaagacc cgatattgag 28440 aaggcgaaac gtcttcttaa ctatgacccc gaaacaggta tatttacacg aaaagtaagc 28500 ataaaaggga gaaatgcggg ggaggtagcc ggagggccga acgacaaagg atacatagtt 28560 attacagtct ccggtgtaag aataaaagcc caccatctgg catgggcttt cgtctacggg 28620 gaataccaca atggtgagct tgaccataaa gaccgtaaca gggccaataa cgccataaat 28680 aatataaggc ccgcaacaag gagccagcag attcagaacc gcgactgctc atcgcataac 28740 actagcggcg cgataggggt ttaccaaata ccatctggca ggtggcggcg ccagaatagg 28800 cgtcaataac aaatacatcc accttggcta ttttgacact atagaggaag cgtcgcgcgt 28860 ataccaaaga gctgcggaaa tatacttcgg ggagttcaag gcatgagtaa gtttaggcgc 28920 agggaatacc agaaaataat gacgtcgttt atgctacagc acccacgttg caatatatgg 28980 tgcggtatgg gtggcggcaa gacctcgtcg acaatgtggg tgcttagccg cctgttccgt 29040 aatgggcaac ttaatgacga cgaccgagtg ttaattctgg cccctttacg tgttgcgtca 29100 ggtacgtggc cagcagaaca agagaaatgg aacttcccgt gtctgagtgt agtagatgca 29160 actggttctg agaagcgacg catcgcggcg ctggagtcag acgctaacgt ggtttgcaca 29220 aattacgaag ttatagaatg gcttattgac tactacggca aagacgactg gccttttacc 29280 gttatcgttg ccgatgagag cacgaaactg aaatctttcc gcagccgttc aggcggtagc 29340 aagcgggcaa aggcgcttag taaggtggcg ttcggtaaag ttaagcgttt cattaacctg 29400 accggtacac catcaccaaa cggcctcaaa gacttgtggg gtcagaactg gttcatcgac 29460 gcgggtgaac gccttgggtc ttcatacacg gcctttaccg atagatggtt taactcggta 29520 cagaaaggca aatctgcgat ggcgcgggag taccatgctc gcccaggcgc ggataacgag 29580 attcaccaga agatgaagga tatcagcctt accattgatg ccgccgagtg gttcggttgt 29640 gaagcaccgg ttattgtacc ggttgagatt gacctgccga agaaagcgcg tcaagcctac 29700 atcgatatgg aggagaagtt attcgcggaa ctggagagcg gagaagttga agcggctaac 29760 gccgccgcta aaacggctaa gtgcttgcag attgcttccg gtgccgtgta tgtgtcgggg 29820 ccggatggtg aagcaacgaa agactgggag aaagtgcacg acgcgaaact cgatgcgtta 29880 gagtccattg tcgaggagtt gcagggtgcg ccgctgctgg tggcctatca gttcaagcac 29940 gaacttgagc gcattcttag gcgattcccc caggcgcagg cgtttgcgaa aggtgctaag 30000 ggtaataagc agatggaatc ttggaaccgc ggggaaatcg agattttgtg cgtgcaccct 30060 gcatcggcgg gccatggttt gaatttacag gacggcgggc atcatctggc gtttatttcg 30120 caaggctgga acctggagca ctatttgcag gttgtcgagc gtataggtcc tgtacgccag 30180 aaacaggctg gccacgagcg tccagtgttc ctgtatcaca tagtcgctaa agacacgctg 30240 gatgaggtcg ttgccgcgcg tacggacgag aaaaaatctg tccaggaaga gttgcttaat 30300 tatatgaaga gacgaggtaa gagatgaata atgagtttga tatcgacgct tgcaacgaat 30360 tgataaaaga cgccattaat tcccgcgagc aacttctagc tatgcaatta aagcgagaaa 30420 taaaacgtat cagggaactc gaagaagagg ttttacggct acggcagcaa agagacgctg 30 480 ctaacgcgca acttgactgg ctactagaac aacaggaagt tgacaaagag aggggcgcta 30540 aatgaaatac tcaattggtg cgaaatgctt tttaataggc tacggattgg ctgcggtcgc 30600 tttacttatc gctgctgtgg ttctgttatc ataaattaag gccccttacg gggcctttac 30660 tatttatgcc aaagttaatc ttgtatagct tccatcggcc cgcttaacta acgctttaag 30720 gctatcgccc tccagataca gtgatataga cccgttatcc ctgacagcgt tgtctggtag 30780 ggcggacatc gctaccggta tctcaacagc acgataatcg cccctattta tttttaatga 30840 cacggcatcg ggcaagctgc ccgaaatagc tgtgacttct gtccacgctg aaccagaacc 30900 tgggccgccg tttatgtggg agtacacgga accactatca agggtctttg acagtttatg 30960 tatccgtaac cgaagcgccg cagaatcatt attgaaactg tttatgcgag agttaccaag 31020 accttcctcc atcaaattgg ctacgttaat ccttgagggg tctacgaagc cggttatacc 31080 acttaccgtg gagttaggcg cgtctatcgt caacccctgt ccactagtgg agcgtatgcc 31140 taccaaacga agaccgttca cacggcaggc cccggagatg tagatctgat tcgctggaaa 31200 atctttagta tttgtgtcga ttatagcaat attagtgaat accgtttcgt gagtaagaag 31260 atatgcaccc gaaccagcgc aatcctcgac ggttatgtta gagacataca gacctttccc 31320 atccatgccg aaacctacgc ccaaagaacc cctaaccagt agattgtcga tcaaatggtt 31380 tagggggagc atatgtaccg ggtattgaga atacgggaaa tcccccgggc ggtcatcttc 31440 tgggttcatg tcggtatctg cgcctaaatc gaagccgtcc catacagggt atagcaccgc 31500 cgaatcccgg aattgcaggt tgtagttacg agaggtcgta gaacctaccg tgccttgcca 31560 tgttttaaca ccactttcgc cggcgcgata cgaggtaaac ccgatgaccc cgccatcgcg 31620 cgcgaaaccg ccattgtttc gtaagaattg agcgctactt accgaaccgt aacttgtgcg 31680 cccgccaata acatagttac ccttgcccca atcgccgctc aggttttcaa aggtaattac 31740 gccatctttt ccgccgctcg ggttgtcagc gtctaccatc ttacagaaat ggcatccgcg 31800 gaacaggaaa cacgccataa gaccactcgc ccggtgaacc tcgacgcctg tacattcccg 31860 aatttccagg gtagaagata tgctttgccc tttagcttcc ggagggagaa gggattctat 31920 accaggaaac ttggcgtaat cgtttaccgt cggctggtat ccatctgttt tagactgttt 31980 aagtgtggcc acgattgccg cggggtcggt tatccactga ttatcgtcgg tccacggttt 32040 taatcaccca cggcgttgta gcactctcca taaagggggc tattacaatg gaacctttac 32100 ctaattgcgt aaataccagg tttccgtcgc cgataaactt cgctttacag tcgatggtca 32160 gggtcttccc gctgaagtta acggtctcgt tattactgaa agtgtaatcg cggtcgataa 32220 gaacgctatc aacggcggca tccgccaatt gttgtaacgt agtaaagtcg gacagcttaa 32280 cggagtattt aaatttctta tccgcttccg cgcgaaaggt aatatctccg aacgcgaccc 32340 aagcggaatc cgaaatgccg cctgtcgtaa gaggtgtcga cgccgcaggg ataactttcg 32400 gcagggggcc acgccatgcg taatagttac cgtccccgcc atcttctttc ggccaaagaa 32460 ccgctttatc cgcgtcgttt acgccgagag tacccccggt cgtaaagtta aaagatgccg 32520 gggagaaacc agcatcacgt aagactgcgg gaagcgtctt ctgcgtctgc ccggttacct 32580 ggttagtggc gtagtcgata tccgcaccac cagctacacc gccttgttta ccggtgatca 32640 cctcggcttc gaaaatctgg tgttttttag ctatttgtaa atcattaagt gacaatacat 32700 caccgcaacc gctggacata tggctatcct cttagttaaa accgttatcg aatccgctgg 32760 agaacgcacg accgaaagga ggcacgttgt caaacttata gaaatccttg tcatagttaa 32820 acccggttat tttgactgtt ctgtcatcgc cagggtctac cgtagaaaca agaatcatct 32880 gagcattatg ccttgcttcg ttgccgaatg aaaattcagt tttcaaagcg ctattaccag 32940 tgtaaatggc ttcttgcggc acagacgtca taattacttg tcggtcatgt gcgccaggga 33000 caacacgaac actttggact ccgccatcac gtaacttcag aaccaaataa tggtcatctc 33060 cggatgtgaa cttaaccggc tgggatagct ctacggtaag accgttaacc gcggttacgt 33120 agccgtcgta agtagacata cgcgaaccct taaccacact aacggcacga ttcggaaggg 33180 caaaaatacc ctcttcagtg gcggtaaacg acaccgaaat tttctttaaa atgttcttct 33240 ggtggcgacg gtttgccgcc cagaaagcct gcttatagtt gcggattcct tttgagtcgt 33300 aagtctccgt tttaaggccc ccggtttcgg gtatggttat agtctctttg acgttcgtct 33360 tagggtctat gtacgagaat ttaaggctgt caaaaacctg agaatcgtta aatgttctgg 33420 tccacttttc agaagtaccg gccttgcttc ggtgggtgaa caccatctcg gggcccatgc 33480 gagggcgctc aaaatcgaga aggatatccg ccccacgccg atatggggtg cagaatatgg 33540 cgtctgctat agtactaact atgtcctgca tggtggtttt atagtcatca aacgtgtagc 33600 aaaattcccc cgcctgtttg tcaccgaaat acgcttcgac ctcgttctgc acggcaagta 33660 acttatccat gttgcgtacc gtcaggttta aaccacccac gtccgggtca cgtgccaggc 33720 ggatgagaga ttgcacggcc tgggtattgg gagtcatcgt atcctcaaaa acgccattac 33780 caaggtattt gtataccatt tcagtagcaa tcatacgcaa ctgcggttgc ttaacttctg 33840 cagcacgcgg tgtctgcttg cgcatcgagt gtacagtagt tctgttgccg tagtgcggag 33900 tgttatcgcg tgtttgcccg tacaagttaa cgtacgttat ctcatcaact acagacccct 33960 cgaaattaaa gtcgaggtct gtcacccttc tggcgcgcac ccgcaccctc gacacaaccg 34020 gcaggtcggc gtaaatagtg acaccgttgt agtctggggt tcgcccggac acggttcctt 34080 gtgcagtgta tatcggccca taaggggtgc tattttcatc aagcagctgg tactgtatct 34140 ccgcggtaac cgacgctagt gttttagatt tgccgttgtc tttatacata ccattggcgg 34200 cggcgatatt ggcaactatg cgctcaacct cggtacggtt tattgacacc caatcggtta 34260 aggttttctc gtatgtgtta tccggttgca gcgacgcctc tccgtcacct cgaaaaaatg 34320 aacctggtct tatttcttgc caccgaccga taaggccgcc aggtacaata aatgtaattg 34380 tcccttcgga tacgcttaac acttggtacc cacccccgga aaggtcaaca tctgtatttg 34440 tccacgctga caccaacgaa aacgtgtcat taggcaccaa cactccagaa aaatccgaac 34500 cacctgttgg gtcatatatt gtgccaatgt tcccggacaa agacgggtac ccggacatgt 34560 aagagaaaga aatacccagg ccgttgggag ctttaagaac cacgccatct acttcgttag 34620 attctacggt aatgtacagg ccctgctcta tagggtcgcc gaccatgacc tggggtgccg 34680 atgtgttatt gggcgatgta tatggtgagt acacggcaac cgacgtgcct gttatgtctg 34740 atacgcgggt gtccccgtca gttatacctt cagggtgtat gtctaggtaa ccacgcccgg 34800 cgtcgtaata gccgtactct acaattttac cagcggcgtt aaacacctta taagtagaca 34860 taaggttatt tgggatagtt tgcaccgttc cgcagatgtc gtagctgcgc tcgtatgggc 34920 gggccttgtt gttacggtcg gtgagactgt tgttcgggga atccgcctgc gagttcgcaa 34980 ggttagattg acgccccttt cactgatggt gataatagtt tcgctagcgg tttaaggata 35040 acgctaaaaa tcttcattac gcctttaatg gcaccaccac cggcagattc cacaatgtgg 35100 aaagtcgcgt tttctttcag tgcttcgaag tcttctgtta cgtcgttatc ttctccgatt 35160 tcgtcaatga aaacacgcac cggcaccccg tccggaacat ggttcacgac gaagttcatc 35220 ggattatcgc gataacgttt aacgtcaaac gtgccgtctt cgttgcgggt atagtggatt 35280 actagcgcca aaattcaatc tccgaataag tatctttaag gtcttccaga ctatccagtc 35340 taacctgtct ggacgccagc tcacagtggc ttaccattcc ttcgtaatat accccagcat 35400 gccacactat tcgcccccta tgtttcactc ctagtagaac cgcgtcaaaa ttctgcggtg 35460 tgaccgctcg taccagacct ttcggattag agtggccatc gtcgaaggcg gcgcctattg 35520 ccgttgggct tgttacatcg aacatcggtg tcgataaccc cgcgtccgcg cgaacgttgc 35580 ggacgtgatg ccagcaatta cggcggcgaa agtcatacgg caagccagtg taatcattaa 35640 tattcacgat gtcagtatcc cgcgaagtaa gggaatctct tccggtgtca tcagaatccc 35700 tgttgcccgt tggttaagca tcggcgtacc ggtttccgct gtgaatacgc ccttttcttg 35760 cgtgagtgtt tgtaactcgt acaccaccgg accgtcgcat gggtatgtca ggtcggtact 35820 tacataacgg cggaagatga atttaggtaa ctccttatta tcatagggga tgcggtccat 35880 ttcctcgtct agtatgttaa gcacatccgg cagcgaaaag gatgcggtct ggtccatatc 35940 gttattgttg gccgcgttct tggcctccat tggcgtacct tcgaatgtca cgacctcccc 36000 ggtctcaaga gtagctgtta aatcgtcagc cccgcgcaca agtaaccacc gtttcgagag 36060 aagtggatgg tatatctcaa gagttataaa gtccatctca ccatcgggat tagacgctag 36 120 tttacgtcgg tatgctgctt ctactgattc ctgactcatt gcatagggtc ccagattcgg 36180 gggaatgtag tctgatacac tccgtatgtt ttaagaaatt cgcccaggca atcaccatag 36240 cacccgtaca aatcgggtag gttctggttc aggcaagcat tgttggtttc ctggaatggt 36300 gatttctcgg cggtggccgt gaaggtgatt acccaattct ttccgtcatc tgtggactcg 36360 ttccacgtgg atgttaacgt tacctggtaa tcctcaatac cctgacccaa gtcgtgcttc 36420 atccagaaac tggacgcacc accgtctacc ttttccatga aactgaggaa tgcctgccgc 36 480 cccaacggcg atacgaccag ggtaacgtta atcggaaaca cgtcaaaata ggtgtcgcgt 36540 ccctggcgaa ccccgccacc ggccaggtct acccgccaca cgttgttacc gcgggtcatc 36600 gagtagcctt tcgatactat aggcctcaaa gaggccggga aatggtaatc gctcatatca 36660 atatcctggt tggccgcgtg tagcgcgacg tgacttagaa atggcgctgt tactatcttg 36720 taacgctgaa cttacggttt cactgattat aatacgtaaa cgaccttcgt catcgcgttc 36780 tgttgcagcc gaatcaattc ttccagtggt gttgttcacg atagtaacat tatctccacc 36840 ggatttagcg ccattctcgc ccataatctg tcgcatctgc tccgccgttc ggacacggga 36900 ggcgttggca ggcataatca cttctggttt accgcgttcg gcaatagtag acatctggcc 36960 tgcggccagg ttgccgccct gctcgcgcgc ggagcgaatc ttacccacgt tggcgagacc 37020 tgcggctacg gcagcagcag cagcaactgg cgctaaaaat ggccctacta ctgggatggc 37080 cgccgttgat ttatacgctt ctactgctga cgtgtacgta gcgattgtcg cctgtgcgat 37140 ggcaaatgct ttatatgccg ttgatgcttc ccctaacgcg gaccctatgt tgctcgccat 37200 attgccgaaa gccgcgccgg tcgccttcgc tcggtcaagg gagtaagatt catccatggc 37260 gcttaacgtt tgttggtaag tttcttcggc aatcaaaccc tgcgcatgaa aatcgttaag 37320 tttttgtagc ttgacctcgt actgccggtc taattccgca aattctcctt cagcaagggc 37380 ctgcatctga gcgacatacg catccgcgga gaactgtttt tccaggcgct ctttctcgcg 37440 tttatctaac tcagcctggc gggccgtatc cgcctcgagc ataatctgcg tcttagcggt 37500 ttcgtattgc tggtcactaa ggagacggtt gctgtagaac tcctgtagct tagtaagttg 37560 ctcttcttgg ttagcttgta tcttctggaa tacgtcgtcg ttaaggtgga ttgactgata 37620 atagaaatcc tcagcggctt tcttctgacg ttcgaatgcg tcgacggatt tcttagcagc 37680 attgtccgcg gaatcgccta cagaaccgtt gtctttaccc gacactttaa acttaccaag 37740 accctcggcc ttggccgctt ctttatcgat gtcgtaagcc gcgcgcttca atgcgataag 37800 gtcttcctga gcttctacct gttcacggaa acttttacgt gtcaggtcag tcacagctct 37860 ttgggtatcc atagcaagcg ctaactggaa gttaaggttc ttggcctcga ccaccgactc 37920 accgggggtc agcgccgcct ttaccagacg accggcgtct tgtgcgccct taaccatatc 37980 ttcaaaaaac cgctccatcg ccgtgaattc ctgggcgata aagtccaggg cgtccgccgc 38040 ggattcggct atcgtagaag cggtttcttc tccgctcccg gtgatgcttt cgctggtata 38100 gttccaagcg gagtttaaag cgtttagggc gtctccaacc atcccgaagg cggcgtcgaa 38160 tgaggcctct atagcacctc cgacttcgga cgccatatca ccccacatct tcatctcagc 38220 gacaaactca cctgacgcta cccgagaatt tatctcctgt atcaggtcat caacatagcg 38280 taaaggttcc gccagtgacc ctacatctat actagtggca agggtcatct tcaactgaga 38340 ccatgagtct tctgcgctag caatagcgcc attgagcgtg ttcgcctggt ctgccatagc 38400 gcccgcgaag ttcacgttac ctatgttaag aagatactgc tcgatatcgg cggcattttt 38460 cttaactaca gtggttgttc cctggaaggt gaattcgata tccttttggt tctgcttggc 38520 cttgataccg aattctttca gacgttcgaa ttcaaatgta ctcgcatccg ccaccgcttc 38580 aatcatctgg ctaaggtctt tacccatcgc cgacgcggta ttgccgtaag agcggagcgc 38640 ttcttccgat ggagttaggc ctagagctac cagcttacgg aatccttcga ccgcttgctc 38700 aagtccgtaa ggggtgtcgc gggcgaagtc ctgtaagata ctcagggcct gctttgcgcc 38760 ttgcgtgctt cctgttagtg ttttcaggct ggcggacatc ttgtcgagtt gccgttgtga 38820 ctcgactagt tcctgcgccg ccttataggc tgtagcggct acaacggcgg ccaggccggt 38880 gacagcggcg cccgcaactt ttgcggattt agagaggtta tcaagacggt ctgatgtggc 38940 cttcgcgcct tgttcggtta cctttactac taagctagct acatcagcca tcgtttctac 39000 cctcaaaaat ggcgtctaag cccatgataa tttcagattc aaacatccct atattctgac 39060 ctgaaatagc gctatacgcc actaaatctg accaactaag ttgctctctt gggtatagct 39 120 ttacgccgtc gtcatcaacg cggcgggtga atttaacatc acgatacttc tcaaaagtgg 39180 tgagtagttc aggggggcat tcaggcccgg cgtcctgcgt tgtttcttcc gcgtcttcta 39240 ttacccccat agcgataagc gcggctttgt gcccgtcggc tatactgtca aacttgcgcc 39300 tttcgtggcg ggttatgaag ttccaacggg cgaactgaaa caacgcgtct acttttcctg 39360 caattggcgc aactgctcat ggtggaatac gactacatgt tctgccaggc gcttatattg 39420 ggcgagaaga gtctcaagat tttccttagt gaactcgtca tcaagactcc agccattcac 39480 caatttcgag ggccaattgc cggtttaggt ctcccgccgc gtcttccatt tttaaattgt 39540 attctgagaa gtctttttgc tcttcgcatt tatcccgaag cggttttaat ttacctaccg 39600 ccgctcggta cgccagggtg aaggcccgca tagctttaac cgcaacatca gcctcaggag 39660 agacaacgtt caaccattcc cctgaatctg taccatcttt taaaggaatc ggcatacgcg 39720 cgccttttct cggcttctgc ttcgtagtaa aaatcgctaa gtttcatagt aatacccttt 39780 ggttagtcgg ttactggttg gtagttgcgc caggcgggaa ccacccgctt ttccggtgcg 39840 accgtaggcg ccaaataaga ataacaaaat agcttgcata accgttaata gtggcctatt 39900 atccttacgt caactaaaca atatgaggtt aagaacatga ggcgtttcta tcggtgttat 39960 cgcgtttatc tatttcatcc cgttcctggt agccttactg cgtaatcata aagctaaatt 40020 aggaatcttt gtatgcaatc ttctacttgg gtggattctt ctgccgtggg taggcgctct 40080 catctgggcc tgtaactcta acgttaatgg taaataatct ggcttaatta aaattacatt 40140 attagaatca gatttgtctt aaacaatcag gggaaataag catggtcact cgcaaaataa 40200 cagatgaaca gttgcagcaa gagctaaacg ctggtctcgg cccgaccgaa atagccaaaa 40260 agtacaacat gtcccgccgt aatgttcagc ttagttctgc gcgtctggca aagaaaggag 40320 ttggtcacgg gcgtgacgta agccacctgg tgccggatgg gtataagatt aagggtacgt 40380 cgtcactggt ggatgagttc ggcaatacga aacttcagtg ggttaagact gacgccgacg 40440 ccgagcgtca ggtcgagttg atgcgagccg taatagatgg gatgaagtcc gatattacgc 40500 cagtttcttc ggtccctcgg cctaaaaagc gactaaacga aaagttgcta aatctttaca 40560 cggtttctga cttccattta ggtatgttgg cctgggcgga tgagagtggc gacgactggg 40620 atatgaagat tgcagaagac ctgttctcga gatggttcga cgcggcgttt caaaagcgct 40680 gatgcggtgt gggtgttatt aacctattag gggactttgc gcattttgac agccttgatg 40740 ccgttacgcc tgccagtggc catgtactgg acgcggacac gcgttaccag aaactggtgc 40800 gctacatgat tcgtatggtc aggcgcgtgg ttaatatggc gcttgttaag cataaaaatg 40860 ttcatctcct aattgtgcaa ggaaaccatg acgagtcagg catgatttgg ttggctgaaa 40920 tgtttaacac tctttacgat aatgaacctc gtgtttttgt agatacatcg gctgatgtct 40980 acaagatggt ccaacacggt aagacgaccc ttttctttca ccatgggcat aaggcgagat 41040 tcgatgctat cgaaccggtt atgatcgcca agttccgtaa ggcgttcgga gagagcgttt 41100 acagttacgc ccatgtgggc caccttcacc atcagaagat tgtggaaagc cgtaacatga 41160 ttgttgaaca acaccgcact ctcgcggcga aagatgctta cgcatcacgt ggtggatgga 41220 tgtcaggccg cagcgcgaat gttattactt acagtgccga atacggcgaa gtcgcgcgtt 41280 taactatttc accggagatg ctgggatgac taacaaatac aatcgcacaa tgacaaatac 41340 tgacggagat agcattacct gtgatgtgta cgacgttctg agggcatttg atatccgcga 41400 cccagcgcta cagcatgcgt tgaagaaact gttgtgcatg ggcttgcggg ggcacaagga 41460 cacaggaacc gacttagcag aagcaattga aagtctggag aagttacgga aataccgtag 41520 taatattgat gagtgaggca tgatttggtt ggctgaaatg tttaacactc tttacgataa 41580 tgaacctcgt gtttttgtag atacatcggc tgatgtctac aagatggtcc aacacggtaa 41640 gacgaccctt ttctttcacc atgggcataa ggcgagattc gatgctatcg aaccggttat 41700 gatcgccaag ttccgtaagg cgttcggaga gagcgtttac agttacgccc atgtgggtca 41760 ccttcaccat cagaagattg tggaaagccg taacatgatt gttgaacaac accgcactct 41820 cgcggcgaaa gatgcttacg catcacgtgg tggatggatg tcaggccgca gcgcgaatgt 41880 tattacttac agtgccgaat acggcgaagt cgcgcgttta actatttcac cggagatgct 41940 gggatgacta acaaatacaa tcgcacaatg acaaatactg acggagatag tattacctgt 42000 gatgtgtacg acgttctgag ggcatttgac atccgcgacc cggcgctaca gcatgcgttg 42060 aagaaactgt tgtgcatggg cttgcggggg cacaaggaca caggaaccga cttagcagaa 42120 gcaattgaaa gtctggagaa gttacggaaa taccgtagta atattgatga gtgagaaaaa 42180 ggcccctttc ggggcctttt ttttatcgtc acaataaaga atctgatttg aacgataagc 42240 atgttcgaag ttaaatataa ccataccaac aagaggagat gaaaccatga acgaattaat 42300 gaatgttagt gaagcacaaa ccatgtccag ccgtgagatt gcggaactga caggtaagga 42360 gcataaaaat gttctggctg atatccgcaa aatgttgtct gagattcaat cggctgaaaa 42420 gtcagccgat tacaaagaca gcaaggggcg cacgtataaa atgcttcttt tggataaaga 42480 ggaaacgtta attctgattt cagggtacag catcaaaatg cgcgctgcca tcatccgccg 42540 ttggcaggaa ctggagtcac aagcgagcaa accatcctta ccggtgccca agacaatggg 42600 ggaggcttta aggttggctg cggacttgtg ggaggagaaa gaacgccttg cgcttgagaa 42660 caaggaaatg gcaccaaaag ctgatgtcta cgaccgcatc atcgacagaa ataacttgta 42720 caacgcgacg caggtggctc agaagttcgg ccaatctgcc gtgtggatga ataaacaact 42780 tgaacaattt ggcgtataca atcgctccgt aaaacgcggt cgtgtcttcc agcaatggtt 42840 tgtggataaa ggctacggca tcatgaggga aacagaaact gggcattcac aggctatgtt 42900 cttcgctgaa ggagagatgt gggttattgg taagttaacc gaagaaggct tgatttaaaa 42960 ctaaggcccc tttcggggcc tttttcttag gcgtacttaa tgcgctgaat aacaattgaa 43020 gactggaact ggttaccggt tgcctgtccc tcaagagaaa gggtaataga ctcagggccg 43080 ccgatttcag gagttgcgga cgtcagtgaa gcacgcttga gtgtaaagct catagcgcca 43140 tccggaccat caagaaccga agatacctcc atttctgttt cattccagga atttattcaa 43200 caacgtcagg tcgtacagtt tacccgccaa cgagaaggta ttagcggcgc gaccgcgctc 43260 aacaaatgcc acgctgttat tcccc 43285 <210> 2 <211> 311 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 2 tttgaatagg gtacccgggc cccccctcga ggtcgacggt atcgataagc ttgatatttt 60 tttttcgttt gggtaggcaa taagtcacac attcgtttcc taaagaccat aggcgcggta 120 ttccataatg agtttaccgc tgatggtcaa ttccaattat tcactatcag taggaggtaa 180 ctatgtgctg gatggaagcg attcctattg ccatgatggg tgcccaagct ctaagtagcc 240 agaacaagtg ctgacaagcg ccgagtggga caagacccga actgggacgc aaacagggca 300 atggagatgg g 311 <210> 3 <211> 227 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 3 cagccagggc atatgtacgc aggcattcac agacaacgtg caccatcagg gagagcaccg 60 ccatgtcact gtcgaacatg ttacgggctt ccaagaagtt ggtgttctcg atacccacca 120 gaccctcaat gttatcctca atgacagagg aggcagagaa ctggtcagtg tgcgaaatac 180 catacgcctt atccatcgcg tgcttctgca ccatctcagg cgtaact 227 <210> 4 <211> 449 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 4 cctttgaacg ccctaaccct attaaaggga acaaaagctg gagctccacc gcggtggcgg 60 ccgctctaga actagttttt cccccgggct gcaggaattc ccatcattgt taacgccaat 120 agtatcagcg gtaaggtcac gtgcttcctc agcaacgtga agggtttgga cctgagagat 180 gttcaggtcg tatgcccgaa ggattgaacc atcggtgaag tcaaccagcc tatcggtagc 240 ggaggtaaat cgacggattt caatcatgtt ataaccgtca tctggcccca atgctctcgt 300 tgtggagatt gtggtcttag tcgcaaaacg gtagtcctga ttcaagatga gttcctttcg 360 gtcaacaccg ataagggtca ctctgacgaa cttacgcgct agatactcga acggaatatt 420 aaagtccgta gtggagccat ccagcgggt 449 <210> 5 <211> 440 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 5 ttgaataggg taccgggccc cccctcgagg tcgacggtat cgataagctt gatatttttt 60 ttctttgagt aaggagcgca cactatgagc aagtcaatca cccacgcaaa caccatccgt 120 ctgccagaca ccgctgacca gttctcccgc cgagtacaca tcaatgtgcg tggtgagaag 180 gtcacgatgg tctaccgctg gaaagaccac aagtctccga aagcacacac tcaacgaatg 240 accttggatg acaagcaggt cggtcgcttg atgggtgccc tgactatggc tgctgacaag 300 gtggtcggtg ataaccgtga gcgtctggta gagttcggcg ctggtatgca agagattatc 360 gagaagtgat aaactcaagg ggcgccaagg gtggccttta tgattatcat ttacacgaaa 420 ccaaaggagg gcattatgct 440 <210> 6 <211> 478 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 6 agctctttaa acgacccact aagtttgaaa aagggaccgg gccccccctc gaggtcgacg 60 gtatcgataa gctcgatatt tttttctgcc tgcatacgac cgacgagtag atggtgatgt 120 tgcaatcatg ggcggttccg ttaagactac caaagacctg aaagatgaaa tcatggcgct 180 ggataagcag tcagagggta acggcacact gaaaggtgaa gtggaagctc tcaaggacac 240 cgtgaagatt ctgactggtc gtgctcgtag gaacccggaa ggcgcttggg gtacagccct 300 gcgctccgta aatgacctca cgttctttgc taagaacgcc tatatgggcg cacagaacgt 360 tactgaaatc gctggtatgc tggcgaaggg taaccgtctc tgcgataact catggcatcc 420 ctatgattaa cgattgggtc aaccggggta aacccgcttc gtgcctctga gataaagg 478 <210> 7 <211> 430 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 7 aacccttttt aaaaggaaca ccaagctgga gctccaccgc ggaggttggc cgctctagaa 60 ctagtttttc ccccgggctg caggaattca ttaacgcttt atgtaagctg gtagttaaac 120 tgtacttcat cgaagccaag aagctggaca agaaggccaa agctgattca cagcgagcca 180 ttgagttggc gaaacagtcc cgtgagaaat ccgatgcggc tgttagtggc atccacaagt 240 cggcagcgat tgcagccaaa gcacaatcca tgagcaaatt cccatcattg ttaacgccaa 300 tagtatcagc ggtaaggtca cgtgcttcct cagcaacgtg aagggtttgg acctgagaga 360 tgttcaggtc gtatgcccga aggattgaac catcggtgaa gtcaaccagc ctatcggtag 420 cggaggtaaa 430 <210> 8 <211> 403 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 8 gatatttttt tctgcctgca tacgaccgac gagtagatgg ggatgttgca atcatgggcg 60 gttccggtaa gactaccaaa gacctgaaag atgaaatcat ggcgctggat aagcagtcag 120 agggtaaccg gcacactgaa aggtgaagtg gaagctctca aggacaccgt gaagattctg 180 actggtcgtg ctcgtaggaa cccggaaggc gcttggggta cagccctgcg ctccgtaaat 240 gacctcacgt tctttgctaa gaacgcctat atgggcgcac agaacgttac tgaaatcgct 300 ggtatgctgg cgaagggtaa cgtctctgcg ataactcatg gcatccctat gattaacgat 360 tgggtcaacc gtggtaaacc gcttcgtgcc tctgagatta agg 403 <210> 9 <211> 452 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 9 aaccagcaac tcttttgaca cgcccaaccc ttttaaaagg aacacaagct ggagctccac 60 cgcggtggtt ggccgctcta gaactagttt ttcccccggg ctgcaggaat tcccatcatt 120 gctaacgcca atagtatcag cggtaaggtc acgtgcttcc tcagcaacgt gaagggtttg 180 gacctgagag atgttcaggt cgtatgcccg aaggattgaa ccatcggtga agtcaaccag 240 cctatcggta gcggaggtaa atcgacggat ttcaatcatg ttataaccgt catctggccc 300 caatgctctc gttggggaga ttgtggtctt agtcgcaaaa cggtagtcct gattcaagat 360 gagttccttt cggtcaacac cgataagggt cactctgacg aacttacgcg ctagatactc 420 gaacggaata ttaaagtccg tagtggagcc at 452 <210> 10 <211> 442 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 10 cttttgaaac gcccccaacc ctttttaaaa ggaacacaag ctggagctcc accgcggtgg 60 tggccgctct agaactagtt tttcccccgg gctgcaggaa ttcccatcat tgttaacgcc 120 aatagtatca gcggtaaggt cacgtgcttc ctcagcaacg tgaagggttt ggacctgaga 180 gatgttcagg tcgtatgccc gaaggattga accatcggtg aagtcaacca gcctatcggt 240 agcggaggta aatcgacgga tttcaatcat gttataaccg tcatctggcc ccaatgctct 300 cgttgtggag attgtggtct tagtcgcaaa acggtagtcc tgattcaaga tgagttcctt 360 tcggtcaaca ccgataaggg tcactctgac gaacttacgc gctagatact cgaacggaat 420 attaaagtcc gtagtggagc ca 442 <210> 11 <211> 488 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 11 cccccccccc taatcaaccc tttgaaacga cccaccaaag ttttgaaaag ggtaccgggc 60 cccccctcga ggtcgacggt atcgataagc acgaaatttt tttctgcctg catacgaccg 120 acgagtagag ggtgatgttg caatcatggg cggctccggt aagactacca aagacctgaa 180 agatgaaatc atggcgctgg ataagcagac agagggtaac ggcacactga aaggtgaagt 240 ggaagctctc aaggacaccg gaagattctg actggtcgtg ctcgtaggaa cccggaaggc 300 gcttggggta cagccctgcg ctccgtaaat gacctcacgt tctttgctaa gaacgcctat 360 atgggcgcac agaacgttac tgaaatcgct ggtatgctgg cgaagggtaa cgtctctgcg 420 ataactcatg gcatccctat gattaacgat tgggtcaacc cgcggtaaac cgcttcgtgc 480 ctctgaga 488 <210> 12 <211> 472 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 12 tacacccctt tgaacgaccc accacagttt gaaaagggta ccgggccccc cctcgaggtc 60 gacggtatcg ataagctcga tatttttttt cgtttgggta ggcaataagt cacacattcg 120 tttcctaaag accataggcg cggtattcca taatgagttt accgctgatg gtcaattcca 180 attattcact atcagtagga ggtaactatg tgctggatgg aagcgattcc tattgccatg 240 atgggtgccc aagctctaag tagccagaac agtgctgaca aggcgcgagt ggcacagacc 300 gaagctggac gcagacaggc aatggagatg gtcaaagaga tgaatatcca aaatgccaac 360 gcctcgctgg aacaacggga cgcccttgaa gctgcatcct ctgagttgac tacaccgtaa 420 catgcagaag gtacaggcta tgggaaccat ccgtgcagct attggcgagg ga 472 <210> 13 <211> 417 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 13 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atattttttt 60 tgctgcgctt ctgaaaccgc tgccgacttc tcgcgttatc taccttggta cacctcaaac 120 cgaaatgacc ttgtacaagg aactcgaaga taaccgtggg tacaccacaa tcatctggcc 180 tgcgctctat ccgcgtagcc gtgaggaaga cttgtactat ggcgaccgtc tggccccgat 240 gctccgcgaa gagttcaacg atgggttcga gatgctccaa ggtcaaccga ctgaccccgt 300 gcgcttcgat atggaagacc tgcgtgagcg tgagttggaa tacggtaagg ctggcttcac 360 tttgcagttc atgctcaacc cgaacctgag tgatgccgag aagtacccct tacgcct 417 <210> 14 <211> 407 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 14 tttggaaaac gacccccggg agaactgcag aaaatcttca agtccggggg gggtttttgg 60 ttcaccaacc tctggaaacc ggcggccggg ggaaccgtac tgtggacact gccagactga 120 ctgcgggccc aaccgctggc gcaccgctga gtgggttcgg catggggaga agggagcaac 180 ttacttaata gctccggcgg acttaaaggg gatccgcaac ggcttggggg tggctttacc 240 ggtggaggaa tcgaaagcct ctgacccaaa cgcccaaggg cgggaataat aaaggcccaa 300 agggataaat gggccagaaa tcggggggaa tgaacaaacc gggcacccaa acttcttggg 360 ggggaaaaaa accttccccc gctggccacc ctaaaaacct tttggcg 407 <210> 15 <211> 321 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 15 gcggtattct gatctcctaa tagacgctga gcttttacgt agcatcagca gcttgggtag 60 ggcgatatta cgctaagcat ctgactcgcg tagccgctcg accattaaac tcgcatctac 120 cgttatcttc tggaagtccc ggtaaggggt ctttgatgcg gacttgacag cccgcttatt 180 aataactacc gcaagggtca attaaggcat ccctatcaga ggctgccttt caatggcggc 240 aaatcttctt ctttgggaac ctcacgggga aaaccccgga ctggaccttc tggccctgga 300 aatttggctg gccggctggc t 321 <210> 16 <211> 803 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 16 cggactagga tgttatttgg tgcatctatg taaaatctga aaattgatgg gaggtgttat 60 gctaaaactt ttacgcagcg cactcccttg ggtactcgcc gggacactct ttatgggtgg 120 ctggcactta gggtcaaccc atgagagagc aaactggaag gaggtaatcc aaaatgaata 180 cattgcgaaa acaaaagcaa ccgcagcaac tcaggcagag gtcagtcggg tatcccgtga 240 gtaccaagaa gagattgcag ccattgaagg cagcactgat aggatgctta atgacctgcg 300 tagtaataat aagcggctgt cagtccgcat caagaccctt accggactac cagaagataa 360 cggtagatgc gagtttaatg gtcgagccga actacacgag tcagatgcta agcgtattat 420 cggaataacc caagctgctg atgctcacgt aagggcgctc cagcgtacta ttaaggagat 480 gcagaatgag cgacacccaa gcaaaccgta atgcgctaat catcgcgcag cttaagggcg 540 actttgtggc cttcctgttc gtattgtgga aggctttggc tctcccgcca ccgactaagt 600 gtcagattga tatggcccgg tgtctggcta acggagacaa caagaagttt atcctacagg 660 ctttccgtgg tatcggtaag tcattcatca cctgtgcgtt cgttgtgtgg acgttatggc 720 gtgaccctca gttgaagata ctgattgtct ctgcatccaa agaacgtgcg gacgctaact 780 ccatcttcat caagaacatc agt 803 <210> 17 <211> 281 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 17 ttcttctgta ctccgggata cccgactgac ctctgcctga gttgctgcgg ttgcttttgt 60 tttcgcaatg tattcatttt ggattacctc cttccagttt gctctctcat gggttgaccc 120 taagtgccag ccacccataa agagtgtccc ggcgagtacc caagggagtg cgctgcgtaa 180 aagttttagc ataacacctc ccatcaattt tcagatttta catagatgca ccaaataaca 240 tccataaagg ctccccgcgt ggagaacctt gagtatgtaa a 281 <210> 18 <211> 232 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 18 agtggcgctc tggagaactg gcagaaaaac ttcaagtccg ttgttgagaa cggttcacca 60 acctctggaa acggcgggcc ttgggaacgt actgtgtaca ctgcccagac tgacttcggt 120 ctcaaccgct ggcgcacgct gagtggttcg tcatggggag aagtagtcaa cttacttaat 180 agctccggcg tacttaaagg gtatccgcaa cgtcttgttg ttgacttcac gg 232 <210> 19 <211> 281 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 19 gtccaatgat tatggccaga atcgtgtatc acaacggcac caactctggt gatatacttc 60 ccgctgccac ctcaagctct gcgcacgcga caaacgggtg gaacattaag atggctacac 120 gattgttgta tggtactgat ggcggtcaac cagtagttga gattggtgcc ttcaaggatg 180 gttctgacca gcagacggca aacgtgccag ttaaagtagg cttgcgcctc gttcacgtgt 240 tctaaggagg acataatgtt atcacttgat ttcaacaacg a 281 <210> 20 <211> 186 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 20 tttcaccaac ctctggaaac ggcgggcctt gggaacgtac tgtgtacact gcccagactg 60 acttcggtct caaccgctgg cgcacgctga gtggttcgtc atggggagaa gtagtcaact 120 tacttaatag ctccggcgta cttaaagggt atccgcaacg tcttgttgtt gacttcacgg 180 ttgagg 186 <210> 21 <211> 222 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 21 gggcgtcgct cattctgcat ctccttaata gtacgctgga gcgccctttg gtgagcatca 60 gcagctcggg ttattccgat aatacgctta gcatctgact cgtgaagttc gcgtcgacca 120 ttaaactcgc atctaccgta atcttctgga agccggaaag ggtcttgatg cggactgaca 180 gccgcttata attactacgc aggacattaa gcatcctatc ag 222 <210> 22 <211> 358 <212> DNA <213> SGP-1 Bateriophage partial gene <400> 22 cttattacta cgcacgtcat taagcatcct atcagtgctg ccttcaatgg ctgcaatctc 60 ttcttggtac tcacgggata cccgactgac ctctgcctga gttgctgcgg ttgcttttgt 120 tttcgcaatg tattcatttt ggattacctc cttccagttt gctctctcat gggttgaccc 180 taagtgccag ccacccataa agagtgtccc ggcgagtacc caagggagtg cgctgcgtaa 240 aagttttacc ataacacctc ccatcaattt tcacatttta catacatgca ccaaataaca 300 tccataaagg ctccccgcgt ggagaacctt gagtatgtca cttactgtac cgtgattg 358

Claims (10)

A composition comprising a bacteriophage having a specific killing ability against Salmonella Choleraesuis or Salmonella Dublin, as an active ingredient. The composition of claim 1, wherein the bacteriophage is selected from bacteriophages of Accession No. KCTC 11173BP, bacteriophage of Accession No. KCTC 11174BP, or a mixture of the two. The composition of claim 2, wherein the bacteriophage of Accession No. KCTC 11173BP has a genome represented by SEQ ID NO: 1. The composition according to claim 2, wherein the bacteriophage of Accession No. KCTC 11174BP has a genome represented by a gene sequence comprising SEQ ID NO: 2 to SEQ ID NO: 22. The composition of claim 1 wherein the composition is a feed additive, a negative additive, or a disinfectant. Salmonella Choleraesuis or Salmonella Dublin bacteriophage bacteriophage, which has a specific killing ability, has a composition containing as an active ingredient Salmonella Choleraesuis or Salmonella dublin. ( Salmonella Dublin) A method for preventing or treating infection with Salmonella Choleraesuis or Salmonella Dublin, which comprises administering to a mammal that may or may be infected. The method of claim 6 wherein said bacteriophage is a deposit number KCTC 11173BP bacteriophage, the bacteriophage of the accession number KCTC 11174BP, or Salmonella choleraesuis, characterized in that which is selected from a mixture of the two devices (Salmonella Choleraesuis) bacteria or Salmonella two Dublin ( Salmonella Dublin) A method for preventing or treating an infection caused by a bacterium. The method according to claim 7, wherein the bacteriophage of Accession No. KCTC 11173BP has a genome represented by SEQ ID NO: 1 Salmonella cholera suis ( Salmonella) Choleraesuis or Salmonella Dublin bacteria to prevent or treat infection. The bacteriophage of Accession No. KCTC 11174BP has a genome represented by a gene sequence comprising SEQ ID NO: 2 to SEQ ID NO: 22, Salmonella Choleraesuis or Salmonella Dublin ( Salmonella Dublin) A method for preventing or treating an infection caused by a bacterium. 10. The method according to any one of claims 6 to 9, wherein the composition is administered to the mammal as a feed additive, a negative additive, or a disinfectant, or Salmonella Choleraesuis or Salmonella Dublin. Salmonella Dublin) A method for preventing or treating an infection caused by a bacterium.

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