KR101993121B1 - Novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the same - Google Patents

Novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the same Download PDF

Info

Publication number
KR101993121B1
KR101993121B1 KR1020170172207A KR20170172207A KR101993121B1 KR 101993121 B1 KR101993121 B1 KR 101993121B1 KR 1020170172207 A KR1020170172207 A KR 1020170172207A KR 20170172207 A KR20170172207 A KR 20170172207A KR 101993121 B1 KR101993121 B1 KR 101993121B1
Authority
KR
South Korea
Prior art keywords
bacteriophage
clostridium perfringens
present
feed
bacteria
Prior art date
Application number
KR1020170172207A
Other languages
Korean (ko)
Other versions
KR20180074583A (en
Inventor
김영주
연영은
한범구
김현일
박철세
Original Assignee
주식회사 옵티팜
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 주식회사 옵티팜 filed Critical 주식회사 옵티팜
Publication of KR20180074583A publication Critical patent/KR20180074583A/en
Application granted granted Critical
Publication of KR101993121B1 publication Critical patent/KR101993121B1/en

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/195—Antibiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3571—Microorganisms; Enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/381—Microorganisms
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/48—Medical, disinfecting agents, disinfecting, antibacterial, germicidal or antimicrobial compositions
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2795/00—Bacteriophages
    • C12N2795/00011—Details
    • C12N2795/00031—Uses of virus other than therapeutic or vaccine, e.g. disinfectant
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2795/00—Bacteriophages
    • C12N2795/00011—Details
    • C12N2795/00032—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Virology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Food Science & Technology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Husbandry (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Pest Control & Pesticides (AREA)
  • Environmental Sciences (AREA)
  • Dentistry (AREA)
  • Plant Pathology (AREA)
  • Agronomy & Crop Science (AREA)
  • Molecular Biology (AREA)
  • Physiology (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nutrition Science (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)

Abstract

본 발명은 클로스트리디움 퍼프린젠스 (Clostridium perfringens)에 특이적인 사멸능을 갖는 신규 박테리오파지 CP3, 상기 박테리오파지를 포함하는 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제 및 상기 박테리오파지를 개체에 투여하는 단계를 포함하는 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법에 관한 것이다. 본 발명의 박테리오파지 CP3을 이용한 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법은 기존의 항생제 등의 화학물질에 비해 클로스트리디움 퍼프린젠스에 대한 특이성이 매우 높고, 상기 박테리오파지 CP3은 상기 균에 감염하여 균 내에서 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있다. 또한 본 발명의 박테리오파지 CP3은 사람, 동물, 식물 등 세균 외의 다른 숙주에는 감염되지 않으므로, 항생제의 오남용으로 인한 항생제 내성균의 문제점, 식품 내의 항생제의 잔류 문제, 광범위한 숙주범위의 문제점을 해결할 수 있는 장점이 있다. 따라서, 본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.The present invention relates to a novel bacteriophage CP3 having specific killing activity against Clostridium perfringens , an antibiotic composition containing the bacteriophage, a composition for feed addition, a feed, a disinfectant or a cleansing agent, and the bacteriophage The present invention relates to a method for preventing or treating infectious diseases caused by Clostridium perfringens. The method for preventing or treating infectious diseases caused by Clostridium perfringens using the bacteriophage CP3 of the present invention has a very high specificity for Clostridium perfringens compared to the chemical substances such as existing antibiotics, It has an advantage that it is highly resistant to physicochemical stimulation and has excellent lytic activity by proliferation in bacteria by infection with bacteria. In addition, the bacteriophage CP3 of the present invention is not infected with any host other than bacteria such as human, animal, or plant. Therefore, the bacteriophage CP3 of the present invention has an advantage of being able to solve problems of antibiotic resistant bacteria caused by abuse of antibiotics, have. Therefore, the bacteriophage CP3 of the present invention can be used variously in the fields of prevention or treatment of infectious diseases caused by Clostridium perfringens, antibiotic compositions, composition for feed addition, feed, disinfectant, or detergent.

Description

신규한 클로스트리디움 퍼프린젠스 특이 박테리오파지 CP3 및 이를 포함하는 항균 조성물{Novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the same}[0001] The present invention relates to novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition,

본 발명은 클로스트리디움 퍼프린젠스 (Clostridium perfringens)에 특이적인 사멸능을 갖는 신규 박테리오파지 CP3, 상기 박테리오파지를 포함하는 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제 및 상기 박테리오파지를 개체에 투여하는 단계를 포함하는 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법에 관한 것이다. The present invention relates to a novel bacteriophage CP3 having specific killing activity against Clostridium perfringens , an antibiotic composition containing the bacteriophage, a composition for feed addition, a feed, a disinfectant or a cleansing agent, and the bacteriophage The present invention relates to a method for preventing or treating infectious diseases caused by Clostridium perfringens.

클로스트리디움 퍼프린젠스 (Clostridium perfringens)는 1890년대 처음 확인되어 영국, 독일, 프랑스에서 각각 개별적으로 발견되었고, 과거에는 클로스트리디움 웰치(Clostridium welchii)라 불리었다. 클로스트리디움 퍼프린젠스는 임상 재료에서 분리되는 클로스트리디움속 세균 중에서 약 50%이상의 비율로 가장 높은 빈도로 분리되는 세균이다. 클로스트리디움 퍼프린젠스의 특징으로는 그람 양성, 운동성이 없는 혐기성의 아포 형성 간균으로, 건강한 사람이나 동물의 장관 내, 토양, 하수 및 먼지 등 자연계의 다양한 환경에 상재하고 있는 세균이다. 이 세균은 독소의 종류와 양의 비율에 따라 A, B, C, D, E 등의 독소형이 존재한다. 이 중 A형은 다양한 곳에서 분리 및 보고되며, 특히 사람에게 있어서는 식중독을 일으킬 수 있는 대표적인 원인균이라 할 수 있다. 이 세균의 최적 생장 온도는 일반세균보다 높은 43~45℃이나 보통 37℃에서도 잘 생육한다. 특히 클로스트리디움 퍼프린젠스의 세대시간은 10~12분으로 대단히 짧아 소량의 세균이 감염되더라도 단시간 내에 질병을 일으킬 수 있는 수로 증가될 가능성이 높다. 또한 이 세균이 생성하는 아포는 내열성이 높아 100℃에서 1~4시간 가열하여도 쉽게 사멸되지 않으며, 다양한 소독제나 약품에도 견딜 수 있다. Clostridium perfringens was first identified in the 1890s and was found in England, Germany and France, respectively, and in the past was called Clostridium welchii . Clostridium perfringens is a bacterium that is isolated with the highest frequency in a ratio of about 50% or more among Clostridium bacteria isolated from clinical materials. The characteristic features of Clostridium perfringens are anaerobic apo-forming bacilli which are gram-positive and non-motile, and are found in various environments of nature such as intestinal tract, soil, sewage and dust of healthy persons and animals. These bacteria have toxins such as A, B, C, D, E depending on the type and amount of toxin. Among them, type A is separated and reported in various places, and it is a typical causative microorganism that can cause food poisoning especially in humans. The optimum growth temperature of this bacterium grows well at 43 ~ 45 ℃, which is higher than normal bacteria, but usually at 37 ℃. In particular, the generation time of Clostridium perfringens is extremely short as 10 to 12 minutes, and even if a small amount of bacteria is infected, it is highly likely to increase to a number capable of causing disease within a short time. In addition, the bacterium-produced apo has high heat resistance and can not easily be killed by heating at 100 ° C for 1 to 4 hours, and can withstand various disinfectants and medicines.

클로스트리디움 퍼프린젠스에 오염된 식품이나 물 등을 통하여 사람이 섭취할 경우, 식중독이 흔히 발생하며, 급작스런 복통을 일으키는 괴사성 장염을 유발하고, 피부나 연조직에 감염될 경우 괴사, 봉와직염, 절단면 감염, 농양 등을 발병시킬 수 있다. 뿐만 아니라 클로스트리디움 퍼프린젠스는 사람 외에도 다양한 온혈동물에 감염되어 질병을 유발하며 그 중에서도 특히 육계, 산란계, 종계 등 가금에서 많은 장내 질병을 일으켜 생산성 감소, 폐사율 증가, 닭의 동물복지 저하 등의 원인이 되고 있다. 그 중 괴사성 장염(NE, Necrotic Enteritis)은 가금산업의 발전에도 불구하고 현재까지도 계군에 지속적으로 영향을 미치며 3주령 즈음하여 성장저해를 유발하는 만성적인 질병 세균이다. 최근의 조사를 보면 괴사성 장염은 전 세계적으로 80% 이상에서 나타나고 있으며, 유럽은 94%로써 89%의 미국, 75%의 남미, 65%의 아시아에 비해 가장 높은 발생률을 보이고 있다. 또한, 알파톡신을 생산하는 클로스트리디움 퍼프린젠스(Clostridium perfringens type A)는 이와 가장 관련이 큰 세균으로서 대부분 무증상(잠복성) 감염과 연관되어 있다. 이 세균에 감염되면 장 점막이 손상 받아 영양소의 소화흡수가 저해됨으로써 사료 효율이 떨어지게 되고, 생산성의 저하를 초래한다. When food is contaminated by Clostridium perfringens, food poisoning occurs frequently and causes necrotizing enteritis that causes sudden abdominal pain. In case of infection with skin or soft tissues, necrosis, Infection, and abscess. In addition, Clostridium perfringens is infected with various warm-blooded animals in addition to humans and causes diseases. In particular, it causes many intestinal diseases in broilers such as broiler, laying hens, and poultry farms, resulting in decreased productivity, increased mortality rate, It is becoming a cause. Among them, necrotic enteritis (NE) is a chronic disease bacterium that continues to affect the martial arts even in the development of the poultry industry and causes growth inhibition around 3 weeks of age. Recent studies have shown that necrotizing enterocolitis is present in more than 80% of the world, with 94% in Europe, 89% in the US, 75% in South America, and 65% in Asia. In addition, Clostridium perfringens type A, which produces the alpha toxin, is the most relevant bacterial strain and is mostly associated with asymptomatic (latent) infection. Infection with this bacterium damages the intestinal mucosa and inhibits digestion and absorption of nutrients, resulting in lower feed efficiency and lowering of productivity.

과거에는 원인균 제거를 위해 항생제를 사료첨가제로 많이 사용하여 왔으나, 항생제를 사용함으로 인해 농장에서 항생제 내성 세균의 빈도율이 높아지게 되고, 축산물을 통해 잔류하는 항생제로 인해 사람에게까지도 비의도적으로 항생제가 전달될 수 있는 부작용이 나타나고 있다. 특히 EU에서의 성장촉진용 항생물질(AGPs) 사용금지 조치와 국내에서의 사료 내에 항생제 사용 금지는 가금의 장 질병 제어를 어렵게 하였다. 따라서 양계업계에서는 괴사성 장염의 예방과 제어를 위한 새로운 전략이 필요하게 되었으며, 이에 클로스트리디움 퍼프린젠스에 대한 백신, 프리바이오틱스, 미생물제 사용 같은 방법들이 해결책으로 등장하고 있다. In the past, antibiotics have been widely used as feed additives for the elimination of causative organisms. However, the use of antibiotics has increased the frequency of antibiotic resistant bacteria in farms, and the antibiotics remaining through livestock products have unintentionally transmitted antibiotics There are side effects that can be seen. In particular, the ban on the use of antibiotics (AGPs) for growth promotion in the EU and the ban on antibiotics in domestic feeds made it difficult to control the intestinal diseases of poultry. Therefore, a new strategy for prevention and control of necrotizing enterocolitis (NEC) has become necessary in the poultry industry, and methods such as vaccines, prebiotics and microbial agents against Clostridium perfringens have emerged as a solution.

한편, 박테리오파지(bacteriophage)는 특정 세균을 감염시켜 감염된 세균의 성장을 억제하고 저해하는 세균 특이적 바이러스를 의미한다. 박테리오파지는 박테리아에 감염(infection)한 후 박테리아 세포 내부에서 증식을 하고, 증식 후 자손 박테리오파지들이 박테리아 밖으로 나올 때 숙주인 박테리아의 세포벽을 파괴하는 방식으로 박테리아를 사멸시키는 능력을 갖고 있다. 박테리오파지의 박테리아 감염 방식은 매우 특이성이 높아서 특정 박테리아에 감염할 수 있는 박테리오파지의 종류는 일부로 한정된다. 즉, 특정 박테리오파지는 특정 범주의 박테리아에만 감염할 수 있고 이로 인하여 특정 박테리오파지는 특정 박테리아만을 사멸시키며 다른 박테리아에는 영향을 주지 않는다. 따라서 최근 세균성 질환의 대처 방안으로 박테리오파지(bacteriophage)의 활용이 크게 주목을 받고 있다. 특히 2000년 이후에 항생제 내성균의 증가로 인하여 기존 항생제의 한계성이 나타나고, 기존 항생제의 대체 물질로의 개발 가능성이 부각되면서 다시 박테리오파지가 항-박테리아제로 주목을 받고 있다.On the other hand, bacteriophage refers to a bacterium-specific virus that infects a specific bacterium to inhibit and inhibit the growth of the infected bacterium. Bacteriophages have the ability to kill bacteria by infecting bacteria, multiplying inside bacterial cells, and destroying the cell walls of the host bacteria when the progeny bacteriophages come out of the bacteria after proliferation. The bacteriophage infection method of bacteriophage is highly specific, so the type of bacteriophage that can infect a specific bacterium is limited to some. That is, certain bacteriophages can infect only a specific category of bacteria, which causes certain bacteriophages to kill only certain bacteria and not other bacteria. Recently, the use of bacteriophage has attracted much attention as a countermeasure against bacterial diseases. Especially, since 2000, the limit of existing antibiotics has appeared due to the increase of antibiotic resistant bacteria, and bacteriophage is attracting attention as an anti - bacterial agent again with the possibility of development as a substitute for existing antibiotics.

그러나 현재까지 클로스트리디움 퍼프린젠스에 의한 감염증을 예방 또는 치료하기 위한 신규한 박테리오파지에 대한 연구는 여전히 부족한 실정이며, 클로스트리디움 퍼프린젠스에 특이적인 용균활성을 갖는 박테리오파지 및 이의 응용에 대한 기술 개발이 필요하다.However, research on novel bacteriophages for preventing or treating infectious diseases caused by Clostridium perfringens is still in the present day. Bacteriophages having lytic activity specific to Clostridium perfringens and their application Development is needed.

이에 본 발명자들은 클로스트리디움 퍼프린젠스에 특이적인 용균활성을 갖는 박테리오파지를 분리하고 분리된 박테리오파지의 형태적 및 유전적 특성을 분석하여 상기 박테리오파지가 클로스트리디움 퍼프린젠스를 선택적으로 사멸시킬 수 있음을 확인함으로써 본 발명을 완성하였다.Accordingly, the present inventors isolated bacteriophage having lytic activity specific to Clostridium perfringens and analyzed the morphological and genetic characteristics of the isolated bacteriophage to selectively kill Clostridium perfringens from the bacteriophage Thereby completing the present invention.

본 발명의 목적은 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 CP3을 제공하는 것이다.It is an object of the present invention to provide a bacteriophage CP3 comprising the nucleotide sequence shown in SEQ ID NO: 1, which has a killing activity specific to Clostridium perfringens.

본 발명의 또 다른 목적은 상기 박테리오파지 CP3을 포함하는, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제를 제공하는 것이다.It is still another object of the present invention to provide an antibiotic composition, composition for feed addition, feed, disinfectant or detergent comprising the bacteriophage CP3.

본 발명의 또 다른 목적은 상기 박테리오파지 CP3을 개체에 투여하는 단계를 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법을 제공하는 것이다.It is still another object of the present invention to provide a method of preventing or treating infectious diseases caused by Clostridium perfringens, which comprises the step of administering the bacteriophage CP3 to a subject.

상기 목적을 달성하기 위하여, 본 발명은 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 CP3을 제공한다.In order to achieve the above object, the present invention provides bacteriophage CP3 comprising the nucleotide sequence shown in SEQ ID NO: 1, which has a killing activity specific to Clostridium perfringens.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 항생용 조성물을 제공한다.The present invention also provides an antibiotic composition comprising the bacteriophage CP3.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 사료 첨가용 조성물을 제공한다.In addition, the present invention provides a composition for feed addition comprising the bacteriophage CP3.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 소독제를 제공한다.The present invention also provides a disinfectant comprising the bacteriophage CP3.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 세척제를 제공한다.The present invention also provides a detergent comprising the bacteriophage CP3.

또한, 본 발명은 상기 박테리오파지 CP3을 개체에 투여하는 단계를 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.The present invention also provides a method of preventing or treating infectious diseases caused by Clostridium perfringens, comprising the step of administering the bacteriophage CP3 to a subject.

본 발명의 박테리오파지 CP3을 이용한 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법은 기존의 항생제 등의 화학물질에 비해 클로스트리디움 퍼프린젠스에 대한 특이성이 매우 높고, 상기 박테리오파지 CP3은 상기 균에 감염하여 균 내에서 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있다. 또한 본 발명의 박테리오파지 CP3은 사람, 동물, 식물 등 세균 외의 다른 숙주에는 감염되지 않으므로, 항생제의 오남용으로 인한 항생제 내성균의 문제점, 식품 내의 항생제의 잔류 문제, 광범위한 숙주범위의 문제점을 해결할 수 있는 장점이 있다. 따라서, 본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.The method for preventing or treating infectious diseases caused by Clostridium perfringens using the bacteriophage CP3 of the present invention has a very high specificity for Clostridium perfringens compared to the chemical substances such as existing antibiotics, It has an advantage that it is highly resistant to physicochemical stimulation and has excellent lytic activity by proliferation in bacteria by infection with bacteria. In addition, the bacteriophage CP3 of the present invention is not infected with any host other than bacteria such as human, animal, or plant. Therefore, the bacteriophage CP3 of the present invention has an advantage of being able to solve problems of antibiotic resistant bacteria caused by abuse of antibiotics, have. Therefore, the bacteriophage CP3 of the present invention can be used variously in the fields of prevention or treatment of infectious diseases caused by Clostridium perfringens, antibiotic compositions, composition for feed addition, feed, disinfectant, or detergent.

도 1은 클로스트리디움 퍼프린젠스에 대해 특이적인 박테리오파지의 스크리닝 결과를 나타낸 도이다.
도 2는 박테리오파지 CP3의 클로스트리디움 퍼프린젠스에 대한 용균반 형성 활성을 확인한 결과를 나타낸 도이다.
도 3은 전자 현미경을 통해 박테리오파지 CP3의 형태학적 특성을 확인한 결과를 나타낸 도이다.
도 4는 박테리오파지 CP3의 단백질 패턴을 SDS-PAGE를 통해 확인한 결과를 나타낸 도이다.
도 5는 박테리오파지 CP3의 pH 안정성을 확인한 결과를 나타낸 도이다.
도 6은 박테리오파지 CP3의 열 안정성을 확인한 결과를 나타낸 도이다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing a screening result of bacteriophage specific for Clostridium perfringens. FIG.
FIG. 2 is a graph showing the result of confirming the activity of the bacterial papilloma CP3 to produce a bactericidal effect against Clostridium perfringens. FIG.
FIG. 3 is a graph showing the morphological characteristics of bacteriophage CP3 through an electron microscope.
Fig. 4 is a diagram showing the results of SDS-PAGE of the protein pattern of bacteriophage CP3.
5 is a graph showing the results of confirming the pH stability of bacteriophage CP3.
6 is a graph showing the result of checking the thermal stability of bacteriophage CP3.

이하, 본 발명에 대해 상세히 설명한다.Hereinafter, the present invention will be described in detail.

본 발명은 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 CP3을 제공한다.The present invention provides a bacteriophage CP3 comprising the nucleotide sequence shown in SEQ ID NO: 1, which has a killing activity specific to Clostridium perfringens.

본 발명에서, 용어 박테리오파지(bacteriophage)는 특정 세균에 감염하여 당해 세균의 성장을 억제하고 저해하는 세균 특이적 바이러스로, 단일 혹은 이중 사슬의 DNA 또는 RNA를 유전 물질로 포함하는 바이러스를 의미한다.In the present invention, the term bacteriophage refers to a bacterium-specific virus that infects a specific bacterium and inhibits and inhibits the growth of the bacterium, and includes a single or double-stranded DNA or RNA as a genetic material.

본 발명에서, 상기 클로스트리디움 퍼프린젠스는 생산하는 외독소와 효소에 의하여 A, B, C, D, E형 등으로 나뉠 수 있으며, 보다 구체적으로, A형 균은 알파 톡신, B형 균은 베타 및 엡실론 톡신, C형 균은 알파 및 베타 톡신, D형 균은 알파 및 엡실론 톡신, E형 균은 알파 및 아이오타(iota) 톡신을 각각 생산하고, 본 발명의 범위에 제한 없이 포함된다. In the present invention, the Clostridium perfringens can be divided into A, B, C, D and E type by an exotoxin and an enzyme to be produced. More specifically, the A type bacterium is an alphathoxine, Beta-and epsilon toxin, C-type bacteria produce alpha and beta-toxin, D-type bacteria produce alpha and epsilon toxin, E-type bacteria produce alpha and iota toxin, respectively, and are not limited to the scope of the present invention.

본 발명의 박테리오파지 CP3은 약 200-220 nm의 크기로 정이십면체의 머리와 수축성 꼬리를 가지는 마이오비리대(Myoviridae)에 속한다. 상기 박테리오파지 CP3의 단백질 패턴 분석 결과 51 kDa, 49 kDa, 42 kDa, 그리고 34 kDa 크기의 단백질을 포함함을 확인하였으며, 박테리오파지 CP3은 열 및 pH에 대한 안정성이 우수한 특성을 가지고 있다.The bacteriophage CP3 of the present invention belongs to the Myoviridae having a size of about 200-220 nm and having a truncated sided head and a contractile tail. Protein pattern analysis of the bacteriophage CP3 revealed that it contained 51 kDa, 49 kDa, 42 kDa, and 34 kDa proteins. Bacteriophage CP3 has excellent stability against heat and pH.

본 발명의 박테리오파지 CP3은 서열번호 1로 표시되는 염기서열을 전체 유전자의 전체 또는 일부로서 포함할 수 있다. 또한, 본 발명의 박테리오파지 CP3은 서열번호 1로 표시되는 염기서열, 및 상기 염기서열의 기능적 동등물로 이루어질 수 있다. 상기 기능적 동등물이란 염기서열의 변형, 치환의 결과, 상기 서열번호 1로 표시되는 염기서열과 적어도 70% 이상, 바람직하게는 80% 이상, 더욱 바람직하게는 90% 이상, 더더욱 바람직하게는 95% 이상의 서열 상동성을 갖는 것으로, 서열번호 1로 표시되는 염기서열과 실질적으로 동질의 생리활성을 나타내는 서열을 의미한다.The bacteriophage CP3 of the present invention may include the nucleotide sequence shown in SEQ ID NO: 1 as whole or a part of the entire gene. In addition, the bacteriophage CP3 of the present invention may comprise the nucleotide sequence shown in SEQ ID NO: 1 and the functional equivalent of the nucleotide sequence. The functional equivalent means at least 70% or more, preferably 80% or more, more preferably 90% or more, still more preferably 95% or more of the nucleotide sequence shown in SEQ ID NO: 1 as a result of modification or substitution of the nucleotide sequence. Or more and having substantially the same physiological activity as the nucleotide sequence shown in SEQ ID NO: 1.

상기와 같은 클로스트리디움 퍼프린젠스 특이적 용균활성, 내산성, 내염기성 및 내열성은 본 발명의 박테리오파지 CP3을 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 및 치료용 조성물, 및 상기 박테리오파지 CP3을 유효성분으로 포함하는 다양한 제품에 적용함에 있어, 다양한 온도 및 pH 범위의 적용이 가능하게 한다.The bacteriophage CP3 of the present invention is useful as a composition for the prevention and treatment of infectious diseases caused by Clostridium perfringens, and a composition for preventing or treating bacteriophage CP3 It is possible to apply various temperature and pH ranges when applied to various products including as a component.

본 발명의 박테리오파지 CP3은 도축장 및 양돈 농장 부근의 하수로부터 시료를 채취하여 분리한 박테리오파지로서, 본 발명자는 이를 박테리오파지 CP3로 명명하고 2016년 10월 21일에 한국미생물보존센터에 수탁번호 KFCC11699P로 기탁하였다. The bacteriophage CP3 of the present invention is a bacteriophage obtained by collecting samples from sewage near a slaughterhouse and a pig farm, and named the bacteriophage CP3 as bacteriophage CP3. The bacteriophage CP3 was deposited on October 21, 2016 with the deposit number KFCC11699P .

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating infectious diseases caused by Clostridium perfringens, which comprises the above bacteriophage CP3.

본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 가지므로, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물에 사용될 수 있다. The bacteriophage CP3 of the present invention has a killing activity specific to Clostridium perfringens, and thus can be used in a pharmaceutical composition for the prevention or treatment of infectious diseases caused by Clostridium perfringens.

본 발명에서, 클로스트리디움 퍼프린젠스에 의한 감염성 질병은 식중독, 괴사성 감염, 가스괴저병, 연조직 감염 및 화농성 근육염 등을 포함하나, 이에 제한되는 것은 아니며, 클로스트리디움 퍼프린젠스에 의한 2차 감염을 포함한다. In the present invention, infectious diseases caused by Clostridium perfringens include, but are not limited to, food poisoning, necrotizing infections, gaseous germosis, soft tissue infections and pyogenic myositis, Infection.

본 발명의 약학적 조성물은 1 × 103 내지 1 × 1010 PFU/mL의 박테리오파지를 포함하며, 바람직하게는 1 × 106 내지 1 × 109 PFU/mL의 박테리오파지를 포함한다. 본 발명에 사용된 용어, PFU(plaque forming unit)는 박테리오파지가 플라크를 형성하는 것을 수치화한 단위이다.The pharmaceutical composition of the present invention contains bacteriophage of 1 × 10 3 to 1 × 10 10 PFU / mL, preferably 1 × 10 6 to 1 × 10 9 PFU / mL of bacteriophage. The term " PFU (plaque forming unit) " used in the present invention is a unit in which bacteriophages form a plaque.

본 발명의 용어 예방이란 조성물의 투여로 질병을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.The term " prophylactic " of the present invention means all actions that inhibit disease or delay the onset of the disease by administration of the composition.

본 발명의 용어 치료란 조성물의 투여로 상기 질병의 증세가 호전되거나 상기 질병의 억제 또는 경감 및 이롭게 변경되는 모든 행위를 의미한다.The term " treatment " of the present invention refers to any action that improves or alleviates or alleviates the symptoms of the disease upon administration of the composition.

본 발명의 약학적 조성물은 약학적으로 허용 가능한 담체를 추가로 포함할 수 있다.The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier.

본 발명에서 용어, 약학적으로 허용 가능한 담체란 생물체를 자극하지 않고 투여 화합물의 생물학적 활성 및 특성을 저해하지 않는 담체 또는 희석제를 의미한다. 액상 용액으로 제제화되는 조성물에 있어서 허용되는 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로오스 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다.The term pharmaceutically acceptable carrier, as used herein, means a carrier or diluent that does not irritate the organism and does not interfere with the biological activity and properties of the administered compound. Examples of the pharmaceutical carrier which is acceptable for the composition to be formulated into a liquid solution include sterilized and sterile water, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

본 발명의 약학적 조성물은 질환 부위에의 도포 또는 분무하는 방법으로 이용할 수 있으며, 그 밖에 비강 분무, 경구 투여 또는 비경구 투여를 통해 투여할 수도 있으며, 비경구 투여의 경우 정맥 내 투여, 복강 내 투여, 근육 내 투여, 피하 투여 또는 국부 투여를 이용하여 투여할 수 도 있다.The pharmaceutical composition of the present invention can be used as a method of applying or spraying on a diseased site, or may be administered through nasal spray, oral administration or parenteral administration. In the case of parenteral administration, intravenous administration, intraperitoneal administration Administration, intramuscular administration, subcutaneous administration or local administration.

상기 비강 분무의 경우 일정량의 박테리오파지 CP3을 물에 희석하여 분무기나 분무 시스템을 통해 비강으로 흡수 되도록 할 수 있다. 상기 비강 분무를 위한 비강 분무제 또는 호흡기용 제형으로는 에어로졸제 등을 들 수 있다.In the case of the nasal spray, a certain amount of bacteriophage CP3 can be diluted in water and absorbed into the nasal cavity through a sprayer or spray system. Examples of the nasal spray agent for the nasal spray or the aerosol agent for the respiratory system are mentioned.

상기 비강 분무제 또는 호흡기용 제형은, 수분산 된 농축물 또는 습윤 분말을 분산시키기 위한 추진제 및/또는 기타 첨가제를 추가로 포함할 수 있다.The nasal spray or respiratory formulation may further comprise a propellant and / or other additives for dispersing the water-dispersed concentrate or wet powder.

본 발명의 약학적 조성물의 적합한 도포, 분무 및 투여량은 제제화 방법, 투여 방식, 대상이 되는 동물 및 환자의 연령, 체중, 성, 질병 증상의 정도, 음식, 투여시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사나 수의사는 목적하는 치료에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다.The appropriate application, spray and dose of the pharmaceutical composition of the present invention may vary depending on factors such as formulation method, administration method, age, body weight, sex, disease symptom level, food, administration time, administration route, Response sensitivities, and the ordinarily skilled physician or veterinarian can readily determine and prescribe dosages effective for the desired treatment.

본 발명의 약학적 조성물을 유효성분으로 포함하는 경구 투여용 제형으로는, 예를 들어 정제, 트로키제, 로렌지, 수용성 또는 유성현탁액, 조제분말 또는 과립, 에멀젼, 하드 또는 소프트 캡슐, 시럽 또는 엘릭시르제로 제제화할 수 있다. 정제 및 캡슐 등의 제형으로 제제화하기 위해, 락토오스, 사카로오스, 솔비톨, 만니톨, 전분, 아밀로펙틴, 셀룰로오스 또는 젤라틴과 같은 결합제, 디칼슘 포스페이트와 같은 부형제, 옥수수 전분 또는 고구마 전분과 같은 붕괴제, 스테아르산 마그네슘, 스테아르산 칼슘, 스테아릴푸마르산 나트륨 또는 폴리에틸렌글리콜 왁스와 같은 윤활유를 포함할 수 있으며, 캡슐 제형의 경우 상기 언급한 물질 외에도 지방유와 같은 액체 담체를 더 함유할 수 있다.Examples of formulations for oral administration comprising the pharmaceutical composition of the present invention as an active ingredient include tablets, troches, lozenges, aqueous or oily suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or elixirs It can be formulated into a zero. A binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin, an excipient such as dicalcium phosphate, a disintegrating agent such as corn starch or sweet potato starch, a disintegrant such as magnesium stearate , Calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. In the case of a capsule formulation, in addition to the above-mentioned materials, a liquid carrier such as a fatty oil may be further contained.

본 발명의 약학적 조성물을 유효성분으로 포함하는 비경구 투여용 제형으로는, 피하주사, 정맥주사 또는 근육 내 주사 등의 주사용 형태, 좌제 주입방식 또는 호흡기를 통하여 흡입이 가능하도록 하는 에어로졸제 등 스프레이용으로 제제화할 수 있다. 주사용 제형으로 제제화하기 위해서는 본 발명의 조성물을 안정제 또는 완충제와 함께 물에서 혼합하여 용액 또는 현탁액으로 제조하고, 이를 앰플 또는 바이알의 단위 투여용으로 제제화 할 수 있다. 에어로졸제 등의 스프레이용으로 제형화하는 경우, 수분산 된 농축물 또는 습윤 분말이 분산되도록 추진제 등이 첨가제와 함께 배합될 수 있다.Examples of the formulations for parenteral administration containing the pharmaceutical composition of the present invention as an active ingredient include injection forms such as subcutaneous injection, intravenous injection or intramuscular injection, aerosol formulations such as a suppository injection method or a respiratory apparatus It can be formulated for spraying. For formulation into injectable formulations, the compositions of the present invention may be formulated as solutions or suspensions in water with stabilizers or buffers in water, and formulated for unitary administration of ampoules or vials. When formulated for spraying, such as an aerosol formulation, a propellant or the like may be formulated with the additive such that the water-dispersed concentrate or wet powder is dispersed.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 항생용 조성물을 제공한다. The present invention also provides an antibiotic composition comprising the bacteriophage CP3.

본 발명에서, 용어 항생용 조성물은 약제 형태로 개체에 제공되어 균을 사멸시킬 수 있는 제제를 의미하며, 방부제, 살균제, 항생제 및 항균제를 총칭하는 것이다. In the present invention, the term antibiotic composition refers to a preparation which can be given to an individual in the form of a drug to kill the bacterium, and collectively refers to preservatives, bactericides, antibiotics and antibacterial agents.

본 발명의 박테리오파지 CP3은 기존 항생제에 비하여 클로스트리디움 퍼프린젠스에 대한 특이성이 매우 높으므로, 유익균은 죽이지 않고, 특정 병원균만 사멸시킬 수 있고, 약물 내성 내지 저항성을 유도하지 않아, 기존의 항생물질에 비하여 제품수명(life cycling)이 긴 신규 항생제로서 이용될 수 있다.Since the bacteriophage CP3 of the present invention has a high specificity to Clostridium perfringens compared to existing antibiotics, it does not kill useful bacteria, can kill only specific pathogens, does not induce drug resistance or resistance, (Life cycling) is longer than that of the conventional antibiotics.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 사료 첨가용 조성물을 제공한다.In addition, the present invention provides a composition for feed addition comprising the bacteriophage CP3.

축산, 수산업에서 사용되는 사료 첨가용 항생제는 질병의 예방 목적으로 사용되고 있는데, 예방 목적의 항생제 투여는 내성균 발생 가능성을 높이고 가축에 잔류하는 항생제가 사람에게 전달될 수 있어서 문제이다. 항생제가 육류를 통해 인체에 흡수되면 항생제 내성을 유발해 질병의 확산을 부를 수도 있다. 또한, 사료에 섞여 먹이는 항생제의 종류가 많고 이는 다제 내성균 발생 확률이 높아지는 문제점이 있기 때문에 좀 더 자연 친화적이면서도 기존의 항생제의 사용에서 발생한 문제를 해결할 새로운 사료 첨가용 항생물질로서 본 발명의 상기 박테리오파지 CP3을 이용할 수 있다.Antibiotics for feed additive used in animal husbandry and fisheries are used for the prevention of diseases. The administration of antibiotics for prevention is problematic because it increases the possibility of producing resistant bacteria and antibiotics remaining in the livestock can be delivered to humans. If antibiotics are absorbed into the body through meat, they may cause antibiotic resistance and spread the disease. In addition, since there are many kinds of antibiotics mixed with feeds and there is a problem that the probability of occurrence of multidrug-resistant bacteria increases, it is possible to solve the problems caused by the use of existing antibiotics while being more environmentally friendly, Can be used.

또한, 본 발명은 상기 사료 첨가용 조성물을 포함하는 사료를 제공할 수 있으며, 본 발명의 사료는 박테리오파지를 사료 첨가제 형태로 따로 제조하여 사료에 혼합시키거나, 사료 제조 시 직접 첨가시켜 제조할 수 있다. 본 발명의 사료 내 박테리오파지는 액상 또는 건조 상태일 수 있으며, 바람직하게는 건조된 분말형태이다. 건조방법은 통풍건조, 자연건조, 분무건조 및 동결건조가 가능하지만, 이에 제한되는 것은 아니다. 본 발명의 박테리오파지는 분말형태로 사료 중량의 0.05 내지 10 중량%, 바람직하게는 0.1 내지 2 중량%의 성분비로 혼합될 수 있다. 또한, 상기 사료는 본 발명의 박테리오파지 외에 사료의 보존성을 높일 수 있는 통상의 첨가제들을 추가로 포함할 수 있다.In addition, the present invention can provide a feed comprising the feed composition, and the feed of the present invention can be prepared by separately preparing bacteriophage as a feed additive and mixing the feed into the feed, . The bacteriophage in the feed of the present invention may be in a liquid or dry state, preferably in the form of a dried powder. Drying methods include, but are not limited to, air drying, natural drying, spray drying and freeze drying. The bacteriophage of the present invention may be mixed in powder form in a composition ratio of 0.05 to 10% by weight, preferably 0.1 to 2% by weight of the feed weight. In addition to the bacteriophages of the present invention, the feed may further include conventional additives that can enhance the preservability of the feed.

본 발명의 사료 첨가용 조성물에는 비병원성의 다른 미생물이 추가로 첨가될 수 있다. 첨가될 수 있는 미생물로는 단백질 분해 효소, 지질 분해효소 및 당 전환 효소를 생산할 수 있는 바실러스 서브틸리스(Bacillus subtilis)와 같은 고초균, 소의 위와 같은 혐기적 조건에서 생리적 활성 및 유기물 분해능이 있는 락토바실러스 균주(Lactobacillus sp.), 가축의 체중을 증가시키며 우유의 산유량을 늘리고 사료의 소화 흡수율을 높이는 효과를 보여주는 아스퍼질러스 오리자에(Aspergillus oryzae)와 같은 사상균 및 사카로미세스 세레비지에(Saccharomyces cerevisiae)와 같은 효모로 구성된 군으로부터 선택될 수 있다.In the composition for feed addition of the present invention, microorganisms other than non-pathogenic microorganisms may be further added. Examples of microorganisms that can be added include lactobacilli such as Bacillus subtilis capable of producing proteolytic enzymes, lipolytic enzymes and glycosyltransferases, lactobacilli having physiological activity and organic resolution at anaerobic conditions such as bovine, ( Lactobacillus sp.), Filamentous fungi such as Aspergillus oryzae , which increase the body weight of livestock and increase the milk yield of milk and increase digestion rate of feed, and Saccharomyces cerevisiae ). ≪ / RTI >

본 발명의 박테리오파지 CP3을 포함하는 사료에는 식물성으로는 곡물류, 근과류, 식품가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류, 곡물부산물류 등이 있으며, 동물성으로는 단백질류, 무기물류, 유지류, 광물성류, 단세포 단백질, 동물성 플랑크톤류, 남은 음식물 등이 있으며 이에 제한되는 것은 아니다.Examples of the feedstuffs containing bacteriophage CP3 of the present invention include cereals, rootstocks, food processing, logistics, algae, fibers, pharmaceuticals, buses, oils, fats, But are not limited to, proteins, mineralogy, oils, mineralogy, unicellular proteins, zooplankton, and remnants.

본 발명의 사료 첨가용 조성물에는 품질 저하를 방지하기 위하여 첨가하는 결착제, 유화제, 보존제 등이 포함될 수 있고, 효용 증대를 위하여 사료에 첨가하는 아미노산제, 비타민제, 효소제, 생균제, 향미제, 비단백태질소화합물, 규산염제, 완충제, 착색제, 추출제, 올리고당 등이 포함될 수 있으며, 그 외에도 사료 혼합제 등을 추가로 포함할 수 있다.The composition for feed addition of the present invention may contain a binder, an emulsifying agent, a preservative, etc. added to prevent deterioration of quality. An amino acid, a vitamin, an enzyme, a probiotic, a flavoring agent, A nitrogen compound, a silicate, a buffer, a coloring agent, an extracting agent, an oligosaccharide, etc., and may further include a feed mixture.

또한, 본 발명은 박테리오파지 CP3을 포함하는, 음용수 첨가제를 제공한다.The present invention also provides a drinking water additive comprising bacteriophage CP3.

본 발명의 음용수 첨가제는 상기 박테리오파지 CP3 또는 이를 포함하는 조성물을 음용수 첨가제 형태로 따로 제조하여 사료 또는 음용수에 혼합하는 방식으로 사용되거나, 음용수 제조 시 직접 첨가하는 방식으로 사용할 수 있다. 상기와 같이 음용수에 혼합하여 공급함으로써 지속적으로 클로스트리디움 퍼프린젠스의 숫자를 감소시킬 수 있는 효과가 있다.The additive for drinking water of the present invention can be used either by preparing the bacteriophage CP3 or a composition containing the bacteriophage CP3 separately in the form of a drinkable water additive, mixing the mixture with feed or drinking water, or adding the bacteriophage CP3 directly in the production of drinking water. As a result, the number of Clostridium perfringens can be continuously reduced by mixing and supplying them to the drinking water as described above.

본 발명에서 음용수는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 음용수를 사용할 수 있다.In the present invention, the drinking water is not particularly limited, and drinking water commonly used in the art can be used.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 소독제를 제공한다.The present invention also provides a disinfectant comprising the bacteriophage CP3.

클로스트리디움 퍼프린젠스에 특이적 사멸능을 갖는 본 발명의 박테리오파지 CP3을 포함하는 소독제는 병원감염을 막기 위한 병원 및 보건용의 소독제로 유용하게 사용될 수 있고 일반 생활 소독제, 식품 및 조리 장소 및 설비의 소독제, 양계장, 축사 등의 건물, 축체, 음수, 깔짚, 난좌, 운반차량, 식기 등의 각종 생육 용품의 소독 등에 사용될 수 있다.Disinfectants containing the bacteriophage CP3 of the present invention having a specific killing activity against Clostridium perfringens can be usefully used as a disinfectant for hospitals and healthcare to prevent infection by hospitals, and can be used as general disinfectants, foods, cooking places and facilities Disinfectants, poultry farms, buildings such as farmhouses, shafts, water bottles, litter, stagnation, transportation vehicles, dishes, etc.

또한, 본 발명은 상기 박테리오파지 CP3을 포함하는, 세척제를 제공한다.The present invention also provides a detergent comprising the bacteriophage CP3.

본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 특이적 사멸능을 가지므로, 클로스트리디움 퍼프린젠스에 노출되었거나 노출될 가능성이 있는 개체의 피부 표면 또는 신체 각 부위 등을 세척하는 용도로도 사용될 수 있다.Since the bacteriophage CP3 of the present invention has a specific killing activity against Clostridium perfringens, it can be used for the purpose of washing the surface of the skin or the body parts of a subject who is exposed to Clostridium perfringens or is likely to be exposed Can be used.

또한, 본 발명은 상기 박테리오파지 CP3을 개체에 투여하는 단계를 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.The present invention also provides a method of preventing or treating infectious diseases caused by Clostridium perfringens, comprising the step of administering the bacteriophage CP3 to a subject.

본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 가지므로, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료에 이용될 수 있다. The bacteriophage CP3 of the present invention has a killing activity specific to Clostridium perfringens, and thus can be used for the prevention or treatment of infectious diseases caused by Clostridium perfringens.

본 발명에서, 클로스트리디움 퍼프린젠스에 의한 감염성 질병은 식중독, 괴사성 감염, 가스괴저병, 연조직 감염 및 화농성 근육염 등을 포함하나, 이에 제한되는 것은 아니며, 클로스트리디움 퍼프린젠스에 의한 2차 감염을 포함한다. In the present invention, infectious diseases caused by Clostridium perfringens include, but are not limited to, food poisoning, necrotizing infections, gaseous germosis, soft tissue infections and pyogenic myositis, Infection.

본 발명에서, 용어 "개체"는 클로스트리디움 퍼프린젠스에 의한 감염성 질병이 발병될 수 있는 모든 개체를 의미하는 것으로 인간을 포함한 포유동물, 가축류 또는 가금류를 포함하나, 이에 제한되는 것은 아니다. In the present invention, the term "individual" refers to all individuals to whom infectious diseases caused by Clostridium perfringens can be developed, including, but not limited to, mammals including humans, livestock or poultry.

상기 용어 "가축류"는 인간에 의하여 순화, 개량되어 사람과 함께 공동생활을 하는 유용한 동물을 지칭하는 개념으로, 예를 들어, 돼지, 소, 닭, 말, 오리 또는 개 등을 포함하나, 이에 제한되는 것은 아니다.The term " livestock animal "is intended to refer to useful animals that are refined and improved by humans to co-live with humans, including, for example, pigs, cows, chickens, horses, ducks or dogs, It is not.

상기 용어 "가금류"는, 가축 중 조류에 속하는 동물을 총칭하여 이르는 개념으로, 예를 들어, 닭, 오리, 꿩, 메추리, 타조, 거위 또는 칠면조 등을 포함하나, 이에 제한되는 것은 아니다.The term "poultry" includes, but is not limited to, chicken, duck, pheasant, queen, ostrich, goose or turkey.

본 발명의 방법에서 상기 박테리오파지 CP3 또는 조성물은 약학적 제제의 형태로 동물에게 투여되거나, 가축의 사료 또는 음용수에 혼합하여 이를 섭식시키는 방법을 통해 투여될 수 있다.In the method of the present invention, the bacteriophage CP3 or composition may be administered to an animal in the form of a pharmaceutical preparation, or mixed with feed or drinking water of a livestock and fed to the animal.

본 발명의 방법에서 상기 박테리오파지 CP3 또는 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 경구 또는 비경구의 다양한 경로를 통하여 투여될 수 있으며, 구체적으로, 구강, 직장, 국소, 정맥 내, 복강 내, 근육 내, 동맥 내, 경피, 비측 내, 흡입 등을 통해 통상적인 방식으로 투여될 수 있다.In the method of the present invention, the administration route of the bacteriophage CP3 or composition can be administered through various routes of oral or parenteral administration as long as it can reach the target tissues. Specifically, the oral route, rectal, topical, intravenous, Intravenous, intramuscular, intraarterial, transdermal, intranasal, inhalation, and the like.

본 발명의 방법에 투여되는 상기 박테리오파지 CP3의 적합한 총 1일 사용량은 올바른 의학적 판단범위 내에서 처치의에 의해 결정될 수 있다는 것은 당업자에게 자명한 일이다. 특정 개체에 대한 구체적인 치료적 유효량은 달성하고자 하는 반응의 종류와 정도, 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여시간, 투여 경로 및 조성물의 분비율, 치료기간, 구체적 조성물과 함께 사용되거나 동시 사용되는 약물을 비롯한 다양한 인자와 의약 분야에 잘 알려진 유사 인자에 따라 다르게 적용하는 것이 바람직하다.It will be apparent to those skilled in the art that the appropriate total daily dose of the bacteriophage CP3 administered in the methods of the present invention can be determined by treatment within the scope of sound medical judgment. The specific therapeutically effective amount for a particular individual will depend upon the type and extent of the response to be achieved, the age, weight, general health, sex and diet of the patient, the time of administration, the route of administration and the fraction of the route of administration, It is preferable to apply differently depending on various factors including drugs used or co-used and similar factors well known in the medical field.

본 명세서에서 달리 정의되지 않은 용어들은 본 발명이 속하는 기술분야에서 통상적으로 사용되는 의미를 갖는 것이다.Terms not otherwise defined herein have meanings as commonly used in the art to which the present invention belongs.

이하, 본 발명을 실시예, 제제예에 의해 상세히 설명한다. 단, 하기 실시예, 제제예는 본 발명을 예시하는 것일 뿐 본 발명의 내용이 하기 실시예, 제제예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail with reference to Examples and Preparation Examples. However, the following examples and preparative examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples and preparative examples.

실시예Example 1:  One: 신규한New 박테리오파지의 분리 및 배양 Isolation and culture of bacteriophage

1.1 박테리오파지의 스크리닝 및 분리1.1 Screening and separation of bacteriophages

클로스트리디움 퍼프린젠스에 특이적인 박테리오파지를 스크리닝 및 분리하기 위해서 용균반 분석을 수행하였다. 시료는 박테리오파지가 존재할 것으로 예상되는 도축장 및 양돈 농장 부근의 하수로부터 채취하였으며, 박테리오파지 분리에 사용된 클로스트리디움 퍼프린젠스는 (주)옵티팜에서 채취한 야외 시료로부터 분리 동정된 것을 사용하였다.Sputum analysis was performed to screen and isolate bacteriophages specific for Clostridium perfringens. Samples were taken from the slaughterhouse where the bacteriophage was expected to exist and the sewage near the pig farm, and Clostridium perfringens used for bacteriophage isolation were those isolated and isolated from the outdoor samples collected from Opti Pharm.

구체적으로, 채취한 시료를 균질화 한 후 클로스트리디움 퍼프린젠스와 함께 35℃에서 하룻밤 동안 진탕배양 하였다. 이때 클로스트리디움 퍼프린젠스는 산소가 있으면 생육이 저해되는 혐기성 세균이므로 배양은 모두 혐기 조건에서 배양하였다. 상기 배양액을 4,000 rpm으로 20분간 원심분리한 후, 상등액을 공극(pore) 크기가 0.45 μm인 필터를 이용하여 여과하여 시료 여과액을 얻었다. 상기 여과액을 이용하여 스팟 테스트를 통해 대상 시료에 클로스트리디움 퍼프린젠스를 용균할 수 있는 박테리오파지가 존재하는지 스크리닝을 수행하였다. 그 결과를 도 1에 나타내었다. Specifically, the collected samples were homogenized and cultured with Clostridium perfringens at 35 DEG C overnight with shaking. At this time, Clostridium perfringens was an anaerobic bacterium in which growth was inhibited when oxygen was present. Therefore, cultivation was carried out under anaerobic conditions. The culture solution was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was filtered using a filter having a pore size of 0.45 μm to obtain a sample filtrate. Using the filtrate, a spot test was conducted to examine whether a bacteriophage capable of lysing Clostridium perfringens was present in the sample. The results are shown in Fig.

다음으로 전배양된 클로스트리디움 퍼프린젠스 150 μL를 취해서 멸균된 튜브에 넣고 3 mL의 BHI(Brain Heart Infusion) 탑 아가(top agar)((Brain infusion solids 12.5 g, Beef heart infusion solids 5.0 g, Proteose peptone 10.0 g, Glucose 2.0 g, Sodium chloride 5.0 g, Disodium phosphate 2.5 g/L) +0.6% agar)를 첨가하여 잘 혼합한 후 미리 만들어 둔 2% BHI 아가 플레이트에 중층하였다. 실온에서 30분간 방치한 후 상기 시료 여과액을 각각 10 μL씩 점적하고, 다시 실온에서 30분간 방치한 후 35℃에서 24시간 동안 배양하여 박테리오파지의 유무를 확인하였다. Next, 150 μL of pre-cultured Clostridium perfringens was placed in a sterile tube, and 3 mL of BHI (Brain Heart Infusion) top agar (12.5 g of Brain infusion solids, 5.0 g of Beef heart infusion solids, (10.0 g of proteose peptone, 2.0 g of glucose, 5.0 g of sodium chloride, and 2.5 g / L of disodium phosphate) + 0.6% agar), and the mixture was layered on 2% BHI agar plates. After allowing to stand at room temperature for 30 minutes, the sample filtrate was dropped by 10 μL each. Each sample was left at room temperature for 30 minutes and then cultured at 35 ° C. for 24 hours to confirm presence or absence of bacteriophages.

박테리오파지의 존재를 확인한 시료로부터 박테리오파지를 순수 분리하기 위해서 소프트 아가 오버레이(soft agar overlay)법을 이용하였다. 박테리오파지가 존재하는 것으로 확인된 시료를 식염수로 적절히 희석하여 각 희석단의 시료 희석액 100 μL를 취하고, 대상 세균(클로스트리디움 퍼프린젠스) 배양액 150 μL에 혼합한 후 0.6% BHI 탑 아가 3 mL을 첨가하여 잘 혼합한 뒤, 미리 만들어 둔 2% BHI 아가 플레이트에 중층하여 방치한 후 35℃에서 24시간 배양하였다. 배양한 각 플레이트에서 형성된 용균반을 확인한 후, 이를 선택하여 400 μL의 SM 완충액 (NaCl 5.8 g/L; MgSO47H2O 2 g/L; 1M Tris-HCl (pH 7.5) 50 mL)에 현탁하고, 실온에서 4시간 동안 방치하였다. 상기 박테리오파지액 100 μL를 0.6% BHI 탑 아가가 포함된 세균 배양액 500 μL와 혼합한 뒤 150 mm 지름의 BHI 플레이트에 분주하여 중층한 후 완전히 용균이 일어날 때까지 35℃에서 배양하였다. 배양이 종료된 후, BHI 플레이트에 15 mL의 SM 완충액을 가하고 실온에서 4시간 동안 천천히 교반하면서 박테리오파지 용액을 회수한 후 4,000 rpm으로 20분간 원심분리하여 상등액만 취하고 이를 0.45 μm 필터로 여과하여 최종 박테리오파지 용액을 회수하였다. 이렇게 얻어진 최종 박테리오파지의 용균반을 확인한 결과를 도 2에 나타내었다.A soft agar overlay method was used to isolate the bacteriophage from the sample which confirmed the existence of the bacteriophage. A sample of which the bacteriophage is confirmed to be present is appropriately diluted with saline, 100 μL of sample dilution of each dilution stage is taken, mixed with 150 μL of the bacterial (Clostridium perfringens) culture, and 3 mL of 0.6% BHI topa Well, mixed, and allowed to stand in a 2% BHI agar plate that had been previously prepared, followed by incubation at 35 ° C for 24 hours. After incubation, the supernatant was added to 400 μL of SM buffer (NaCl 5.8 g / L; MgSO 4 7H 2 O 2 g / L; 1M Tris-HCl (pH 7.5) And allowed to stand at room temperature for 4 hours. 100 μL of the bacteriophage solution was mixed with 500 μL of bacterial culture containing 0.6% BHI topagase, and the mixture was divided into 150-mm-diameter BHI plates and cultured at 35 ° C. until complete lysis. After incubation, 15 mL of SM buffer was added to the BHI plate, and the bacteriophage solution was recovered while slowly stirring at room temperature for 4 hours. The supernatant was centrifuged at 4,000 rpm for 20 minutes. The supernatant was collected by filtration through a 0.45 μm filter, The solution was recovered. FIG. 2 shows the result of checking the thus obtained final bacteria bacteriophage.

도 2에 나타낸 바와 같이, 약 2 mm의 크기를 가지는 용균반을 형성하는 박테리오파지를 확인하였고, 상기 박테리오파지는 클로스트리디움 퍼프린젠스에 특이적인 용균능을 갖는 박테리오파지임을 확인하였다.As shown in Fig. 2, a bacteriophage forming a fungus having a size of about 2 mm was confirmed, and it was confirmed that the bacteriophage was a bacteriophage having a specific ability to Clostridium perfringens.

1.2 박테리오파지의 대량 배양 및 정제1.2 Bulk culture and purification of bacteriophages

상기 실시예　1.1에서 확인한 클로스트리디움 퍼프린젠스 특이적 박테리오파지를 대량 배양하기 위하여, 1.1에서 회수한 최종 박테리오파지 배양액을 1.0×108 PFU/mL로 희석한 후 전배양된 클로스트리디움 퍼프린젠스 1.0×109 CFU/mL과 혼합하여 MOI (Multiplicity of infection)가 0.1이 되도록 하여 35℃에서 120 rpm으로 9시간 동안 배양하였다. 배양이 종료된 후 배양액 전량을 6,000 rpm으로 30분간 원심분리하여 상등액을 수득하였고, 이를 0.2 μm 필터로 여과하여 최종 박테리오파지 배양액을 회수하였다. 분리된 박테리오파지를 박테리오파지 CP3(Bacteriophage CP3)로 명명하고, 2016년 10월 21일 한국미생물보존센터 (Korean Culture Center of Microorganisms, 서울, 한국)에 기탁하고 수탁번호 KFCC11699P를 부여받았다.In order to mass-culture the Clostridium perfringens-specific bacteriophage identified in Example 1.1 above, the culture of the final bacteriophage recovered at 1.1 was diluted with 1.0 × 10 8 After dilution with PFU / mL, the cells were mixed with 1.0 × 10 9 CFU / mL of pre-cultured Clostridium perfringens and cultured at 35 ° C. and 120 rpm for 9 hours in order to obtain MOI (multiplicity of infection) of 0.1. After the culture was completed, the whole culture was centrifuged at 6,000 rpm for 30 minutes to obtain a supernatant, which was then filtered with a 0.2 μm filter to recover the final bacteriophage culture. Separated bacteriophages were named bacteriophage CP3 (Bacteriophage CP3) and deposited with the Korean Culture Center of Microorganisms (Seoul, Korea) on October 21, 2016, and received the accession number KFCC11699P.

실시예Example 2: 박테리오파지 CP3의 전체 유전자 분석 2: Whole gene analysis of bacteriophage CP3

박테리오파지 CP3의 전체 유전자를 분석하기 위해, 박테리오파지 CP3 배양액에 20 mM EDTA, 50 μg/mL의 프로테나아제 K, 0.5%(w/v) SDS를 가한 후, 50℃에서 1시간 동안 정치하였다. 동량의 페놀-클로로포름-이소프로필 알코올 (25:24:1)과 혼합한 후 실온에서 12,000 rpm으로 10분간 원심분리하여 상등액을 회수하였다. 회수된 상등액을 동일 부피의 페놀-클로로포름-이소프로필 알코올 (25:24:1)과 혼합하여 실온에서 12,000 rpm으로 10분간 원심분리한 후 상등액을 회수하고, 상기 상등액에 3M 소듐 아세테이트를 전체 부피의 10%(v/v)가 되도록 가한 후 혼합하였다. 여기에 2배 부피의 차가운 95% 에탄올을 가하여 혼합한 뒤 -20℃에서 1시간 동안 정치하였다. 반응이 끝난 후 0℃에서 10분간 12,000 rpm으로 원심분리하여 침전물을 획득하였고, 차가운 70% 에탄올로 2회 세척한 뒤 에탄올을 완전히 제거하여 DNA 펠렛을 회수하였다. 회수한 펠렛을 최종 부피가 50 μL가 되도록 TE 완충액(Tris-EDTA, pH 8.0)에 용해시킨 후 DNA 농도를 측정하였다. 전체 유전자 분석을 위해 PacBioRSII를 이용하여 유전자 시퀀싱을 시행하였다(마크로젠, 한국).In order to analyze the entire gene of bacteriophage CP3, 20 mM EDTA, 50 μg / mL proteinase K and 0.5% (w / v) SDS were added to the bacteriophage CP3 culture medium and left at 50 ° C. for 1 hour. The mixture was mixed with an equal amount of phenol-chloroform-isopropyl alcohol (25: 24: 1) and centrifuged at 12,000 rpm for 10 minutes at room temperature to recover the supernatant. The recovered supernatant was mixed with an equal volume of phenol-chloroform-isopropyl alcohol (25: 24: 1) and centrifuged at 12,000 rpm for 10 minutes at room temperature. The supernatant was recovered and 3M sodium acetate was added to the supernatant in a total volume 10% (v / v). The mixture was mixed with 2 volumes of cold 95% ethanol, and the mixture was allowed to stand at -20 DEG C for 1 hour. After the reaction was completed, the precipitate was collected by centrifugation at 12,000 rpm for 10 minutes at 0 ° C., washed twice with cold 70% ethanol, and then the ethanol was completely removed to recover the DNA pellet. The recovered pellet was dissolved in TE buffer (Tris-EDTA, pH 8.0) to a final volume of 50 μL, and the DNA concentration was measured. Gene sequencing was performed using PacBioRSII for whole gene analysis (Macrogen, Korea).

시퀀싱 결과, 박테리오파지 CP3의 전체 유전자의 크기는 52,068 bp이며, GC함량이 34%, 전체 ORFs의 수는 74개임을 확인하였다. 분석한 박테리오파지 CP3의 전체 유전자 서열을 서열번호 1로 나타내었다. As a result of sequencing, it was confirmed that the total gene of bacteriophage CP3 was 52,068 bp, the GC content was 34%, and the total ORFs were 74. The entire gene sequence of the bacteriophage CP3 analyzed is shown in SEQ ID NO: 1.

실시예Example 3: 박테리오파지 CP3의 형태학적 특성 3: Morphological characteristics of bacteriophage CP3

박테리오파지 CP3의 형태학적 특성을 알아보기 위해, 배양액을 0.01% 젤라틴 용액에 희석하여 2.5% 글루타르알데하이드(glutaraldehyde) 용액으로 고정하였다. 이를 carbon-coated mica plate(ca. 2.5 x 2.5 mm)에 적가한 후, 10분간 적응시켜 멸균증류수로 세척하였다. 이를 구리 격자(copper grid)에 끼워 2% 우라닐 아세테이트(Uranyl acetate)에서 30-60초간 염색하고 건조시킨 후, 투과전자현미경 (Tecnai G2 Spirit Twin, Bio-Transmission electron microscope, 120 kV, 120,000~400,000)으로 관찰하였다. 그 결과를 도 3에 나타내었다.To determine the morphological characteristics of bacteriophage CP3, the culture was diluted with 0.01% gelatin solution and fixed with 2.5% glutaraldehyde solution. The mixture was added dropwise to a carbon-coated mica plate (ca. 2.5 x 2.5 mm), and the mixture was adapted for 10 minutes and washed with sterile distilled water. The resultant was stained with 2% Uranyl acetate for 30-60 seconds, dried, and then observed with a transmission electron microscope (Tecnai G2 Spirit Twin, Bio-Transmission electron microscope, 120 kV, 120,000-400,000 ). The results are shown in Fig.

도 3에 나타낸 바와 같이, 박테리오파지 CP3은 약 200-220 nm의 크기로 정이십면체의 머리와 수축성 꼬리를 가지는 마이오비리대(Myoviridae)에 속함을 확인하였다. As shown in Fig. 3, bacteriophage CP3 was found to belong to Myoviridae having a size of about 200-220 nm and having a head of trilobes and a contractile tail.

실시예Example 4: 박테리오파지 CP3의 단백질 패턴 분석 4: Protein pattern analysis of bacteriophage CP3

박테리오파지 CP3의 단백질 패턴을 분석하기 위해, 1.0×1010 PFU/mL로 준비된 박테리오파지 CP3 40 μL를 10 μL의 5X 로딩 완충액과 혼합하고, 10분간 끓인 다음 12.5% SDS-PAGE를 수행하였다. SDS-PAGE 수행 후 단백질 전기영동 결과를 도 4에 나타내었다.To analyze the protein pattern of bacteriophage CP3, 40 μL of bacteriophage CP3 prepared at 1.0 × 10 10 PFU / mL was mixed with 10 μL of 5X loading buffer, boiled for 10 minutes, and subjected to 12.5% SDS-PAGE. The results of SDS-PAGE and protein electrophoresis are shown in FIG.

도 4에 나타낸 바와 같이, 박테리오파지 CP3은 51 kDa, 49 kDa, 42 kDa, 그리고 34 kDa 크기의 주요 단백질을 가지는 것을 확인하였다.As shown in FIG. 4, it was confirmed that bacteriophage CP3 has major proteins of 51 kDa, 49 kDa, 42 kDa, and 34 kDa in size.

실시예Example 5: 박테리오파지 CP3의 다양한 세균에 대한 감염성 조사 5: Investigation of infectivity of bacteriophage CP3 against various bacteria

박테리오파지 CP3의 다양한 세균에 대한 감염성을 조사하였다. The infectivity of bacteriophage CP3 against various bacteria was investigated.

우선, 박테리오파지 CP3의 클로스트리디움 퍼프린젠스에 대한 용균 활성을 확인하기 위하여, ㈜옵티팜에서 분리하여 보유하고 있는 클로스트리디움 퍼프린젠스 균주를 대상으로 소프트 아가 오버레이법을 통해 확인하였다.First, in order to confirm the lytic activity of Clostridium perfringens of bacteriophage CP3, Clostridium perfringens strain isolated from OptiParm Co., Ltd. was confirmed by soft agar overlay method.

클로스트리디움 퍼프린젠스에 대한 용균활성 확인 결과, 본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스 39균주에 대해 감수성이 있는 것을 확인하였고, 이는 박테리오파지 CP3이 클로스트리디움 퍼프린젠스에 대한 용균활성이 우수함을 의미한다.As a result of confirming the lytic activity against Clostridium perfringens, it was confirmed that the bacteriophage CP3 of the present invention was susceptible to 39 strains of Clostridium perfringens, indicating that the bacteriophage CP3 has a lytic activity against Clostridium perfringens .

다음으로, 박테리오파지 CP3이 다른 세균에 대해서 용균활성을 나타내는지 확인하기 위하여 ㈜옵티팜에서 분리하여 보유하고 있는 균주와 미생물 자원센터 등에서 분양받은 균주를 대상으로 소프트 아가 오버레이법을 통해서 감염성을 확인하였다. 그 결과를 표 1에 나타내었다.Next, in order to confirm whether bacteriophage CP3 exhibits lytic activity against other bacteria, infectivity was confirmed through a soft agar overlay method for strains isolated from OptiParm Co., Ltd., which were purchased from strains and microorganism resource centers. The results are shown in Table 1.

Figure 112017124753071-pat00001
Figure 112017124753071-pat00001

표 1에 나타낸 바와 같이, 본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스 외에 대장균, 살모넬라 등의 다른 속 균주에 대해서는 용균활성이 없는 것을 확인하였다. As shown in Table 1, it was confirmed that the bacteriophage CP3 of the present invention had no lytic activity against other strains such as Escherichia coli and Salmonella in addition to Clostridium perfringens.

따라서, 본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 특이적으로 용균활성을 나타내는 박테리오파지임을 확인하였다.Therefore, it was confirmed that the bacteriophage CP3 of the present invention is a bacteriophage exhibiting a lytic activity specifically on Clostridium perfringens.

실시예Example 6: 박테리오파지 CP3의 pH 안정성 조사 6: Investigation of pH stability of bacteriophage CP3

박테리오파지 CP3의 pH 안정성을 확인하기 위하여, 다양한 pH 범위 (pH2, 3, 4, 5, 6, 7, 8, 9, 10, 11)에서 박테리오파지의 수를 측정하였다. 소듐 아세테이트 완충액, 소듐 포스페이트 완충액, Tris-HCl을 이용하여 각각의 pH에 맞게 조제한 후 990 μL로 분주하여 준비하였다. 박테리오파지 CP3 배양액을 5.0×108 PFU/mL로 준비한 다음, 상기 배양액을 10 μL씩 취하여 pH에 맞게 조제한 각각의 완충액에 첨가하여 2시간 동안 상온에서 방치하였다. 방치 후 각 반응액을 단계희석하여 소프트 아가 오버레이법에 의해 타이터(titer) 측정을 수행한 결과를 도 5에 나타내었다.The number of bacteriophages was measured in various pH ranges (pH 2, 3, 4, 5, 6, 7, 8, 9, 10, 11) to confirm pH stability of bacteriophage CP3. Sodium acetate buffer, sodium phosphate buffer, and Tris-HCl, and the mixture was divided into 990 μL. The bacteriophage CP3 culture was prepared at 5.0 × 10 8 PFU / mL. Then, 10 μL of the culture solution was added to each buffer prepared for pH, and the mixture was allowed to stand at room temperature for 2 hours. After allowing to stand, each reaction solution was diluted stepwise, and a titer measurement was performed by soft agar overlay method. The result is shown in FIG.

도 5에 나타낸 바와 같이, 본 발명의 박테리오파지 CP3은 pH3-11의 범위에서 2시간 동안 안정한 것을 확인하였다. 이를 통해 박테리오파지 CP3은 pH 안정성이 우수한 박테리오파지임을 알 수 있다.As shown in FIG. 5, it was confirmed that the bacteriophage CP3 of the present invention was stable for 2 hours in the range of pH 3-11. As a result, bacteriophage CP3 is a bacteriophage having excellent pH stability.

실시예Example 7: 박테리오파지 CP3의 열 안정성 조사 7: Investigation of thermal stability of bacteriophage CP3

박테리오파지 CP3의 열 안정성을 확인하기 위하여, 40℃, 50℃, 60℃, 및 70℃ 등 다양한 온도 범위에서 일정 시간 방치 후 박테리오파지의 수를 측정하였다. 보다 구체적으로, 박테리오파지 CP3 배양액을 5.0×106 PFU/mL가 되도록 희석한 후 배양액 5 mL을 멸균된 튜브에 분주하고 30분, 1시간 및 2시간 동안 각 온도에서 방치하였다. 방치 후 각 반응액을 단계희석하여 소프트 아가 오버레이법에 의한 타이터 측정을 수행한 결과를 도 6에 나타내었다.In order to confirm the thermal stability of bacteriophage CP3, the number of bacteriophages was measured after standing at various temperature ranges of 40 ° C, 50 ° C, 60 ° C, and 70 ° C for a certain time. More specifically, the bacteriophage CP3 culture was diluted to 5.0 × 10 6 PFU / mL, and 5 mL of the culture was dispensed into sterilized tubes and left at each temperature for 30 minutes, 1 hour, and 2 hours. After allowing to stand, each reaction solution was diluted stepwise, and the result of performing a tester measurement by soft agar overlay method is shown in FIG.

도 6에 나타낸 바와 같이, 본 발명의 박테리오파지 CP3은 40-70℃의 범위에서 약 60분 이상 안정한 것을 확인하였다. 이를 통해 박테리오파지 CP3은 내열성이 우수한 박테리오파지임을 알 수 있다. As shown in Fig. 6, it was confirmed that the bacteriophage CP3 of the present invention was stable for about 60 minutes or more in the range of 40-70 占 폚. As a result, bacteriophage CP3 is a thermophilic bacteriophage.

상기 실험으로부터 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 대한 특이성이 매우 높고, 균 내에서 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있음을 확인하였다. 또한 본 발명의 박테리오파지 CP3은 사람, 동물, 식물 등 세균 외의 다른 숙주에는 감염되지 않으므로, 항생제의 오남용으로 인한 항생제 내성균의 문제점, 식품 내의 항생제의 잔류 문제, 광범위한 숙주범위의 문제점을 해결할 수 있는 장점이 있다. 따라서, 본 발명의 박테리오파지 CP3은 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.From the above experiments, it was confirmed that bacteriophage CP3 has a high specificity to Clostridium perfringens and has an advantage of being excellent in lytic activity and strong against physicochemical stimulation by proliferation in bacteria. In addition, the bacteriophage CP3 of the present invention is not infected with any host other than bacteria such as human, animal, or plant. Therefore, the bacteriophage CP3 of the present invention has an advantage of being able to solve problems of antibiotic resistant bacteria caused by abuse of antibiotics, have. Therefore, the bacteriophage CP3 of the present invention can be used variously in the fields of prevention or treatment of infectious diseases caused by Clostridium perfringens, antibiotic compositions, composition for feed addition, feed, disinfectant, or detergent.

비록 본 발명이 상기에 언급된 바람직한 실시예로서 설명되었으나, 발명의 요지와 범위로부터 벗어남이 없이 다양한 수정이나 변형을 하는 것이 가능하다. 또한 첨부된 청구 범위는 본 발명의 요지에 속하는 이러한 수정이나 변형을 포함한다.Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

이하 본 발명의 박테리오파지 CP3을 포함하는 감염성 질병의 예방 또는 치료용 약학적 조성물의 제제예를 설명하나, 본 발명을 한정하고자 함이 아니고 단지 이를 구체적으로 설명하고자 함이다.Hereinafter, formulation examples of pharmaceutical compositions for the prevention or treatment of infectious diseases including bacteriophage CP3 of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically explained.

제제예Formulation example 1: 의약품의 제조 1: Manufacture of medicines

1.1 주사제의 제조1.1 Manufacture of Injection

박테리오파지 CP3 1 × 106 PFU/mLBacteriophage CP3 1 x 10 6 PFU / mL

주사용 멸균 증류수 적량Sterile sterilized water for injection

pH 조절제 적량pH adjuster

통상의 주사제의 제조방법에 따라 1 앰플당(2 mL) 상기의 성분 함량으로 제조한다.(2 mL) per 1 ampoule in accordance with the usual injection preparation method.

1.2 1.2 액제의Liquid 제조 Produce

박테리오파지 CP3 1 × 106 PFU/mLBacteriophage CP3 1 x 10 6 PFU / mL

설탕 20 g20 g of sugar

이성화당 20 g20 g per isomer

레몬향 적량Lemon incense quantity

정제수를 가하여 전체 1,000 mL로 맞추었다. 통상의 액제의 제조방법에 따라 상기의 성분을 혼합한 다음, 갈색병에 충전하고 멸균시켜 액제를 제조한다.Purified water was added and the total volume was adjusted to 1,000 mL. The above components are mixed according to a usual method for producing a liquid agent, and then filled in a brown bottle and sterilized to prepare a liquid agent.

1.3 정제의 제조1.3 Manufacture of tablets

박테리오파지 CP3 1 × 106 PFU/mLBacteriophage CP3 1 x 10 6 PFU / mL

옥수수전분 100 mgCorn starch 100 mg

유당 100 mgLactose 100 mg

스테아린산 마그네슘 2 mgMagnesium stearate 2 mg

상기의 성분들을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조한다.After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

한국미생물보존센터(국내)Korea Microorganism Conservation Center (Domestic) KFCC11699PKFCC11699P 2016102120161021

<110> Optipharm.CO.,LTD <120> Novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the same <130> 1.37P-1 <150> KR 10-2016-0178172 <151> 2016-12-23 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 52068 <212> DNA <213> Unknown <220> <223> Bacteriophage CP3 <400> 1 aagtccttgt tttcaatgat aatttctttt tcattagata atataacttt catcatggtt 60 atccctcctt tactgtcatt ataagagaag taagtacaac tgtcaacgtt cgggagaact 120 ctactactgc tctctctctt actctctctt tccaaccaaa caaaatcaag agctcgatcg 180 ctgaccgtct gggtttttct atataagcta aatcttggac tacagaaggt gaactccaac 240 gttgcggaaa aatgcttccc aagctgtaag ctactccaag gtactttctt gagatctgta 300 agctatacct ttctctctcc aagctatacc ttcctaacct tggttgctta gaagctccaa 360 gccttgggca tggatatgcc tgccgtagtg taaacgaagg caaggcaata gcctgccaat 420 tatgtaatca ataaaacata gttatatata gcctttggct atatactata tataataata 480 tgtaattatt tatatatagt atatattgta ataatatggg gtatttctga tatccctcta 540 cgctagtgat accaacaaat ttgattacaa aaattgtaaa ctctatatgt gatatcaggt 600 gtaactatgc tgatatgcta cccatatcag aaaaaatggt actgatatgc ctatgtggac 660 tttgacatat ttatcttata tgtgtataat gtaagcatag gaggtgctat aaatgataga 720 taattgggga gaattaagaa agctaatcca gaatagagaa ggtagatctt tgaatgagca 780 tgagcttacc agagctgtta ctcatctctc tttcattgtt tctcaactaa aaaatgagaa 840 taatatgcta aagaaccaga taagagcagt gtctggatac ccactaatta actttaaaga 900 tcctaaagct aaggtagatg agttctcttt atatatgtat ttacaggaat tttggcctga 960 agtattccat gactataagg ataagatcca gagtagtaag aagttcaagg ctaaaaagtt 1020 tactaatcta aagcaagcta atgagtttgt atggactgaa ttgataaagc caaaggataa 1080 tgagagccta tttgtattaa gtaaatgggt agaaggtatg ggtatatctc atcacttcag 1140 aactaaacag gagtttatta ctgcttgtct gtgctatatg gactataaag gatatctgga 1200 caaggaaggt aagaccttta ataagctagg agacatcaag agaggaagct tcagatttaa 1260 ttctgatgag gaaacctatg caatactaac caatactttt gatactgtgg gtaaatataa 1320 atctggaatt gagaaattca aattaaaaat aaaggagtta aaagagaatg attaaaagaa 1380 ttatgtcatt actaacaata attgtgatag gagggctact gttaaccata cagagcctaa 1440 acaatcattg gggaaatgct gtccaagaat atattgagtc gaataaacaa ttaactgagc 1500 agactgaaat actgatggat aatgtagaag gattaaagta tcagaatgat aaactgagaa 1560 ctgataaaca aaatctaatt gagatgataa attcatttaa aggaatggag ttcaaagttg 1620 aagttactta ctattcatca gatgaaccta cagatgtggc aggatatgag accaagaata 1680 gtataggagg agatcttaag gttggtgact gggcagctcc agactttatt cctctacact 1740 cattacttca agtaacatat gcagatggga ctcaagagat tgcttacatc attgacaggg 1800 gtaaccctga tgtcattaag gtttttgata gagaagacaa aagaattata caggactcaa 1860 caatgagact tgataagtat gtccatgatg tgaatggcaa gggagttgat ggagcaacaa 1920 taaaaatatt aagatgggga gaataacatt ctctcttttc tttcgggatt tctattgaca 1980 ctctctggac tctctcttat aattaaagca taaacaaact gaaggagatg ttaaatatga 2040 attataataa aatgactaac tgggaaatat tatttgaact tgaggagaac ctattatgct 2100 ttggatataa attattaaga agagaacatg aagaagtaat acaagatata agagatacac 2160 aacctaagga tctatacaaa tatgctcaat cacttattaa ccaaactctt gaagaagaat 2220 atatctgtgt atactcaata agataatgtg ggaggtattt attacctctc ttttttattt 2280 gacagagtat gacagtatca cttataatag tgttgaggag gtgatactat ggataatcat 2340 tcacttgagg ttatagaact tcaatctgaa tacagagaac ttgaaagaag atggcaccaa 2400 ttcctggatg gccatgctca ttgcgaagaa gaagctatcc aagttaagga agagaatgga 2460 gaagatcaag agaactttac gaaacttgaa ctaattaagg atttttccag aactcaaaaa 2520 attataaaaa ggagggttaa tccctccctt atttttagaa aacctattgc agtctcttta 2580 gaactgatac aatatgtctt ttctctcttg gtgtaaacat ccaataataa ttgtgatttg 2640 ctttattctc atgtgatctt aatagctttc tggctttcct ttcatttatt ccaacctctt 2700 tggcaatatc tcttaatggt attaagttat tatctttcat ttattccaac tccttgattt 2760 tgtttttata gtctatatta tgctcagccc ggaagatatt atacgaattt cttcgagaaa 2820 atctttgaca aacttccgag gctctcttat aataaaatca acaaataaat acaaaaagga 2880 gaactgatat gaaaagaaga tatatcgttt tacaagtcac agttgatgtg gattggaacc 2940 ctgatgttag tgtgttagca cataacaaat atttaggagt acactgggta tctgttaagg 3000 atgaacacaa ggaagatata atctattttg atactgagga agaagctaag attgaagcta 3060 agcatagacc tggagctgta gttcaggcca catggaggta gataatatga aagtaagatt 3120 agtgattgta acatcaaaaa ggatcttgac cagatatttt gaagttgata acaaaagaca 3180 ggctatagga tatattccta aagtattaca ggaggaaggt atatctgata aggttagaag 3240 atatgaagtg attgaggttg atgagaacaa cagaagaata aaaagatatt aaggggagag 3300 atgatctccc ctttaacata gaaaggagaa ttaacatgaa agagtatact tatgtattat 3360 gtaagaatga ccaaggacaa cagtataaaa ggtatgtgga agtattatgt gagtttgatt 3420 atatgggcag gaccttattc attcatagac ctgtaggagc ccctcataat acagtatgtt 3480 atactatatc agataaggag acaggatgct gtcttgctcc taataactgc tctataaaga 3540 caccttcaaa gagatccgca gtaagagtat tgaatgagta ttttagagcc aatgaaatac 3600 atgagtggga tatgaagaga gtcagaaagt tgaaaaagga tttagccaga gctccaaaat 3660 atcataaagt caagaaaacc ccagagccta aaaaagtaaa accaaagcca gcccaatgta 3720 ttgatctgga ttatggtgat atacctttct aaagtttctc ttgacatatg atagggaata 3780 gtatataata aaattgaggt gataaactat ggaaaataat aacaacagaa taagcagaga 3840 aacactttac atggaagttg ctaaacttat gacaggaaga tctacttgcc taagaaaagc 3900 agttggttgt gtattagtta gagataacag gatagttgct acttcgtata atggagtacc 3960 ttcaggagta gaacattgta aaacttgttt aggtcctgga tgtgatattg tgatccatgc 4020 agaagctggc ttaataagtt actgtgctaa acatgggatc tctactgaag gaactactat 4080 gtatgtaacc ttatccccat gcgaaagctg tgctaattta ataatcaatg caggaattaa 4140 gaaagtggta tatcttgaga aatacagatt aagtggagga atagctaaat taatttgtaa 4200 tgatgtggag gtagaacaat atgagtaaag aggtatctaa tatattagag gatgagaaat 4260 taagagagtt tttgaatgag gaacaactag aagctattct aaatatagct aaagctggta 4320 ataatgataa agaagaaaat cttgagaaat ttgaaaagct attggaagag aaagaaaata 4380 tattaggagg tattagccat gagtaaatta atatgtaaaa gaagatgtta tagaagggat 4440 ccagcaggta actgcttagc atatacaatt ccatatgggc caggagaaaa atgccctagc 4500 caatgtaata atccacatca attaaagaaa gagtgtcaag acatcatttc taaaaatcaa 4560 cctaacaacc caatagtaaa agaatgtctt tacttaatag agaagatgga tctaataatt 4620 ggggttgaga ttaataaagc ttatgaagaa gataagaata gaggttcagg tggaggatct 4680 aagaactgta aacctaatac agcagctcta aagcaaaaga tgaaagataa tagacctatt 4740 gagtgtaagc aaactaaaga agagaataag gaatatcagg aagaattaaa acattggcaa 4800 gaagaccata atgaaaaact agaagttcat aaatcagttg ggtatggtat gaacaggggc 4860 aaggtagact catatactgg tatagaactt gatactaaag gtaatccagt atattcccca 4920 gaagaaatac aaattgcagc tgataacctg ctacaggaag gaagaataga tgtggatatg 4980 tatgaaaaaa tattgacagg tgaaataaaa ttataaaacg ttgacataat taatccagat 5040 gtggtataat agtggtataa atccatagtt aattttgcca tttggttggt attactagca 5100 ttaaggttag tgtgccaacc atttttcacg tagtaaagtc tggattttaa gggggtgata 5160 ttgtattatg agctataagt catacactga accagagtac tgtactcctg aacatttgaa 5220 gaaagccaaa aaattgatat ataatgccct gaaggctgat ccctttacac atcctaaaca 5280 attaagaaac cttgctaagg tatcacagac ctttgttagg cagtgtctaa gagagtttaa 5340 agagcagggc ataacatttt tagaagaaga aggtgatatt gtggcattga gtagcaaaac 5400 aaaaaagact attaagacct tggctgagaa gtatgatctt gatgagaaag aagagttgtt 5460 tgtatttcat tacatgaagt gctttaatgc aactacagca gctgctaggg caggctatcc 5520 tgtgaataat tctcatgcac atgggtctaa actattaaag aagaagaaaa tacaggaagc 5580 tctaaaggct atcagagccg atagagatga agaattacta tgcgatggta tggatgtaat 5640 aaggcagtac atgaaaatag cctttgcaga tataactgat gtgtgccaat tcaaggatga 5700 tctagtccta ttaaggtcta gtaaggctgt agatggtact ttaatcaatg agattaaaca 5760 aagtaaagat ggcgtatcag ttaaaatgga agatagactt caggcactaa aaagacttga 5820 ggtatacttc ggattaagta ctgatgtgat gaaagttgaa cttgaaaaag ttaaattact 5880 tttacaaata caggaaaatc agaataatgc tggtaaagga aaagatcttg aggttcttag 5940 agccaaaatg gctgatagaa agaaaaagaa tgcaggtaaa ggaaaatcta atactactaa 6000 gtaaaaaggt attacaggac taaataaagc actttactaa ggaggtgtat ataattgttt 6060 gatttagata gaatgattga gaaattagag ctagatgctt tggttgataa gttccacatt 6120 aacaaggaag ccctagaaga aatatccttt gaagaattag tagatctata ctatgacgat 6180 ccttgtggat tctcagaaga tatactagga gtaaacttag atccacatca aaaggatatt 6240 ttaataagtg taagagacaa taagagaacc tctgtaaggt caggacaggg ggtaggtaag 6300 actatggcct gtgcttgtct tataatttgg tatatgacct ttaggcctga atgtaaggta 6360 attgctacag ctcctacatt ttcacagtta acgtctgtac tatggcctga aatatcaaaa 6420 tggctacaag gaactattgt ggaagattat tttacagtaa ataaaaccac tatgactaga 6480 aattgtagtc ctatgacttg gtttgctatg tgtaaaacta gtaatagaaa agagaatatg 6540 cttggtcttc atagtgagtt catgctaatt atatgtgatg aggctcctgg aatagatgat 6600 gagatactag aagcattact tggtactata tcagatgagg aaaataagat aattttgatt 6660 ggtaaccctg ttaaaaatac aggtatcttc tttgattctc accacagtct taaggaaaat 6720 tttaaatgta ttcatattga cgctacaaaa gttgatagaa ctgataaaga aaaccacaaa 6780 atgttaagga aaaagtatgg ggaagagtcc aatgtctaca gatgtctggt aaaaggggac 6840 ttccccctag aagaagatga catatttata caattatcca cagtatatag aggtatcaac 6900 agtgaaataa ccagacctac agaaccagtt accatagata tgggtgtaga cgtagctaga 6960 tttggttcag ataagacctc aatcatgact aaaattgatg gttatgtgga agaacctatc 7020 tattaccaag gtaaggacac tatgaaaaca gtaggagaag tgctaagaaa ggctaaatct 7080 ctacatggtg agttccctaa actagcagta gttgtgaaaa tagacgacac tggtgtaggt 7140 ggaggagtaa ctgatagatt aaaagagata aaattcgaag aaaaactgga ctggttaaag 7200 gtagtaccat gtaagtttgg tatgccacta aagagagggt cccaggctta tgccttttat 7260 tatgatacaa ccacatatat agcctctgta gttaaggaat tactggagga tgatgagtta 7320 agactaccag ataatacaga tcttgtggga gaattgacta ccagaaagtt taaagtattc 7380 aactccaaag gtcaaataaa gctggagagt aaggatgaaa tgaagaaaag aggactacat 7440 tctccagact tagcagactc tctattctta gcttgccttc ctgttggatc taaattaaga 7500 aaaatctagt aactgatatg cctattaaag ccaataattt tttattctga tatgtgagat 7560 atcagttttg aaattgacta ggctctatat tagtaataat gtgatataga aggcatatca 7620 gaattaccac ctttatatat atgtaatata agtatactat atactatata atttatttca 7680 taaattaata tagtaatata gtatacatta ataattatat aattttatat ggggaatata 7740 tgcggaaaat cttccggaaa ataatataaa aataaaacgt tgacaaattt ttgtgagtta 7800 tgttataata aacgtatcaa attatagtta tttttaaatc atgtgaatgg ccgtatttga 7860 cactatatgt ggagttgatg tagtcctccc tgaaatatga ggccatctgg attttataat 7920 ttgatatata atgtaagaaa ggaggtaaag taactttgaa aaagatatcc gccagaagta 7980 tatgtggaga ccctatccaa aagtctttgg atccagtgga taaatccaat caacaaacca 8040 gtgtacttgg aattgaagtt gatgagatat cccctgaata tgacccccaa ctattagaag 8100 tgattataag ggagtctaca ataatacccc aatgcaaaaa ggcctaccga aataatattg 8160 ctggattcgg aataggtatt aggtatattg agaaggtaga tgaacctact ccagaaatga 8220 aagctgaata taaacagctg gaagaattaa ttgataccct gtcttttgat aaaccaacca 8280 agaatgtctt tgaagagata gttgattgga gagaaggact tgggtattct tttttagaga 8340 tcgtagaaga tcctaaagac tcagctctcg taaaagagat aaaagaaatt gaagacccta 8400 agacaattaa aattcttaag ctagatgatg agagtacaga gtacactgta acccgatatg 8460 gtaaacagta cactagaaga aaaaggttca gaagatttaa gcagactgta aataatcaat 8520 caatcttctt taaggaatgg ggagatccta gagatctaga tatgaatact ggaaagtatt 8580 atgaaaaggg agagatacct cttaaagatc gagctaacag tatacttcat tttaaaacag 8640 gttatggtgt ttatggtaca ccaagatggg aaggtcctat gttagccgct cagggttcaa 8700 gaaaggccga attattaaac tggagatatt ttactaatgg taaacatact ccattaatgt 8760 tactaattca aggtggtact cttaatgata aatcttggaa tgagttaact gagtacatga 8820 atagtgtaaa aggtgaaaat tcccagcatg gtttcttagt attagaaact gaagctgcag 8880 agaaagacac tgcccttgat gattctaagc cagttactgt tgaggtcaag cccctatctg 8940 aaatattaca gaaggatgag ttattccaag actacatgga taatactagg aaaaagatac 9000 aatcttcttt cttactacca gacatatatg tagggtactc tcaagattac aatgtggcta 9060 cagccagaga ggctacaaga aaaactgaag agcaagtatt tgttcctgaa agagaaaacc 9120 taacatggat actaaataat gttattctat ctggctatgg ttttaaatat gtggaagctt 9180 actttaagag tcctaatatt gagaactctg actatcagaa aggaatgttt gctatagcct 9240 ctagagctat gactcctaat cagattaatg aggagtacta ttcatcaatc aataaggact 9300 atactcctat tgatgaagaa tgggcaaata tccctactta ctcagttgcc tcttcaatga 9360 gtaataactt aacaagtata attgagaagg ctcaaaactc taatgggagt gatgaggtga 9420 tagctgtact taaagaggct aagaaacagt tattaaacta tcaattaaaa ggaggtaagt 9480 aatagtgggt actgatgagt tgatcgctaa aattgataag atcttaaaga atgaagatga 9540 tagccttaag aaagaattga aagaagctgg ctatgaggag attgattata caatagaaag 9600 gatctctgaa ttagaagaca ctatttctga tctccttaca gaagaattaa aacatcttat 9660 aaaagatgga gactttgaaa ccattgaaga atttatggaa tatgtggagt ctcaaatggg 9720 tacagataat atcgatactt ctttagcaga aactcttaag agttgttatc aagaagctat 9780 agaaaattta ggagatactt atttacaagg tattgataag gacttatccc taaccaaagt 9840 cttttctcca tatactacta cctttataga tgagtggtct tctaaactag gggaaatgat 9900 gaaattaact tcccatgaaa aactacagaa tattctggat aaggctcttg aagatggaga 9960 gtctgtagaa gatgtagttg gtaaattatc tgagacttat ggctttagta gaactagagc 10020 tagagctact gctattactg aaatgttaag agcccactca tatgctaagg atgaggcttt 10080 taatcaatgc cccgcagtaa caggaataca gtggaaacac tcagggggaa ggggtattca 10140 cccaaggcaa caccatgtcg atctggatgg tactgtcatt aataaaggtg agtacttcac 10200 tgtaggaatt tatgaagcta gatttcctag ggatatcaac ttaccagctt ctgagactgt 10260 gaattgtcat tgtactcatg ggcctgtaat tgataaggat atactaggat tatctaaaga 10320 agaaaaagaa gctatcagag atcaagtgat agcagaatat aatttgtaaa ggagggagag 10380 taaatgccag taaaatcagt taagaaagct attgagatct cagatgctaa aattgattac 10440 ctttcattag ttgataaagc agctaataaa tggcaatgga ttgtggctaa agaacttgga 10500 gctcctaata gatttagata tgataagcca atcttaaagt ctgagtctga aggagataat 10560 cattttgtta tcggtgtagt atatgagcca atgactgctg actaccatga taattttatg 10620 actgaggaag agatcaaaaa agcttgtgag tggtttaatg aaaatggcct tggtagtgat 10680 ttacaacaca atgagaacac tcttgagggt gtcaagattg taaaatcatg ggtaactgag 10740 gaagatacca caatagaagg tacagtagtt aaagctggaa cttggttagc taaagctgag 10800 attatcgatg atgatctatg gagcaagatt gagaaaggcg aaattactgg atggtctatg 10860 ggtggtactg gtaaatattc tacacaagat gtagatattg atgagattga gaagtctata 10920 ggtcaatcta tacttgataa aattggaaag gccctagggt taactccaaa gcatatacag 10980 aaaggtgcat actctgaaaa atataaagag actgcaaaaa gaaataattt ctggaatgca 11040 gttagtacct tagaagatat tctatatcca tattctagtt gggattacac tagatctttt 11100 gaaagtgatg tagataagat aaaggaggct cttcaagaat tttctgaaac tatggaggga 11160 atattattgg gaactccaga agaagtagct aaatcttttg agttaccagt caataaggca 11220 ggtaagaaaa tatcttcaaa aaatgtggaa aagttaaaaa cagcacaaag tgctattaat 11280 gaattattag actcagtgga gggggaagaa gagatgaatg aaactcaagt acaaaaaatg 11340 atagacgaag ctatcaaaaa ggcattagat ggaactaaac ctgttgaggg tgaagttaat 11400 aagaatgaag gtcaaccagc tactactgaa gatatagttg ctaaggctgt ggaaaaagct 11460 atgttagagg ctggcttaat ccagaaacaa gaagaaaaaa ctgaggaaga aaaaatagct 11520 gaggcagtag ctaaagccat ggctccatat gtaggaagta aacaaattgc taaaaacgga 11580 gaagaaggac aagaagttca aaaatcagta tttaatggat tatttggtgg aaaccaagaa 11640 taataacaat taggagggaa agttatgtca aaagtaaata aagctgtatt taacgctgcc 11700 gtattaaatg gtggaggtaa attaaatcca gaacaagcta acgcttttat taaaatgata 11760 attgatgagc caacaatatt aaaagaggct agaactgtac caatgaaagg agactctaaa 11820 aagattgagt caataggatt tggtcaaaga gtattaaatc ctgcagctga aggaacagat 11880 ccaggacaat ctaaatattc agctccaact actggaacag ttactttaca agctaaagag 11940 tttatagctg tggttaagtt aactgatgat actatagaaa caaatataga aggaaaatca 12000 atagaagaca ctgtaatgca attattagct gagagagttg ctcttgacat ggaagaaata 12060 attgtaaatg gtgatactgg acatagtagc gatactttct tacaacaatt agatggggtt 12120 agaaaacaat taactgctca tgtagtagat gctaacaaag ctgctttaag taaagaccta 12180 ttaaaacaag ctaagaaagc tgtaccagct aagtatttaa gaaacccagg tgaatggaga 12240 ttctactgct caaatggttt agaaactgat tgggtagact tagtaggaga aagattaact 12300 gctaatgggg atgactcatt tataaatggt actgttaaat ctgcttatgg agtacctgtt 12360 aaaggtatag ctatgctaca agattatgac gactcttcta ctcaagtatc agacataatc 12420 ttaactcacc ctaagaatat agttataggt atctcaagag atatcagaat agagttggaa 12480 agagctccta gagaaagatc tacttatgta gttttaactc ttaagatgga tgttaagttt 12540 gaagatccta aagctgctgc taagatagtt aaagtaaaag cttaatcatg aaggcccact 12600 gtaaatgtgg gctctttttc attagaaagg agagatatta tggctaaggt caaattagca 12660 aagggtaaat tacaaactct ggttattaaa ggtcatagaa ttacccaaga ccatgtagaa 12720 gaagttaaag atagtctagt aaattattta aaacagaact atgatgtgat tgtcatagat 12780 gaacctgttg aggatgaacc tctagaagat tttactgaag aagaagtacc ttctacagat 12840 gacaataaag aagaacctgc tagaagattt acaaacaaga ataaaagaaa cagatagggg 12900 tggctcatat gaatattact ccggatgaaa taattgactt ctcaagtaat cctgagatta 12960 ggaaaagaga ccccaaaaat attgagatgg atatattaag agctaaagcc aagatctcag 13020 gaatttttaa aaattcaagt aaagactatg atactattgt ggagaaaaat cccaatagta 13080 aagaattaag gttagcccat atactgtatg ctgagtatta tggactacaa tccttggtca 13140 ttagcaaggc tgatattcaa tccgagacct atgatgacta ctcatataca agaagaactg 13200 ataaggctgt tatagagcct gatattatgc ctctaattaa acctttctta gaagatgata 13260 gtccatctat cagaacaact actatgagat taagggtact gtaatatgat tagaaattca 13320 tataacaagc tcttaaatca tagatgtaat atataccacc tattagaaga tcctgaagat 13380 cccggatatg ggctagccca tacccctagc tttagttatc ctgatgagcc agacatgatt 13440 gaggtaccct gtcattttaa atcaggaact ctagaaaaac ttgagtacaa agaacctagg 13500 tttaagatta cccaggagta ctcagtagat tttgctattg gaactgatgt tagacaaaat 13560 gataaaattg tggaccttga gacgggtatc acatatatag ctaaagctcc tagggatata 13620 agaggtctaa aaattaaggt tggtatagag gctaaggatt ttagatagtt atggcttcaa 13680 attctggggg catagaaata gatgctaaac aaatagaaag gtttataaag aagctaagag 13740 atgccgctaa tagtgatttc aaaaagcaat tagcagtatg gtttgaagct gcagggatag 13800 acttccttga tgtggttcaa gatgaaataa ttagactaca agtagtagac actagattat 13860 tacttaactc ctttcaaaaa gggggtaaga atggtatatg ggtaagtagt gatggaggac 13920 taactttaga agttggtact aacctaaaat atgctaagtg ggtaaacgat ggtcactata 13980 ctaatccaca aggacaatca tttagattcg ttcctggcat atggtcagga gataaattta 14040 tttatcaacc aggagctaag actggtatgg tactaagaca aaagaaagta cctggtaaac 14100 cctattggga caatgcagta aaaatatatg aaaaaatatt tgaacaggct ttccaaagaa 14160 aactagaaga ttggttagcc gaagttttag ggggtagtta ttaatgacag cagatatagg 14220 gagtattgct aaattcatat ctgatctaac tggaattact accctctatt ttgagcgaat 14280 acctgaaaag ttcaagtatc cctcattata tttcaagaag gtatttaatc ctcagaatga 14340 tactctttgt agttatagaa ctgagaatac tttaagagtt aaggtatttg aacctaagga 14400 ctctcttgaa attgctcaga atatagctaa tgctatatac ttgagaaata gaaaaattcc 14460 tatacttaat agggatggta gtaagaccaa caaaaatctc aaaataaaga atgtggaagt 14520 atttgaaggt gaaactgaag aagattctca ggtagagatc aactatgata ttgtggttag 14580 tctttataga gatccaactt acttcgaaaa ctttgagata acaaaggggg taaaatagtg 14640 aaagtacaaa aattttccat acctcaatta agagcccatt gtcaacaact tttccaggtt 14700 gaggagtttg tcttcaacgc tgcagttcaa tcccttccac ctgatgaaga gttaacaata 14760 aaacaaacta gtcagattat aaaagactgg ctaaagaagg aggtaaaata gatggcagga 14820 ggaactttta ttccaggaca aaataaagta agacccggta tgtatactag atttgtaagt 14880 ggtggactta atgagcctaa gcttaataat ctaggagtta tgatcttacc attacaactt 14940 aagtggggta atgctggaga tgttatcaag gtaaccaatg aaggtggcct agaagcatta 15000 tatcaacttg ggtatgacat aactcaccct actgtattac tggttaaaga ggctttaaaa 15060 gaagctgcaa ctgttttagt atataaccta acaggtggaa ctaaagccaa agctgtagta 15120 gggtctctaa catttacagc tactaaagct ggagaaaggg gtaataaatt atctctaact 15180 attaagacca atcctcttgg aggaaaggat atattattat atcttgaagg atctaaaatc 15240 tttgaggcta caggagtaac tgaaatatca acagttaagt ctcaatggtt tacagtaagt 15300 ggaagtggtt ctatagatga gactgctaag actgagttta gtactggtac ttctactcag 15360 atgaataact ctgatgttat taacttctta gataactgtg agttatacaa ctttaaaact 15420 atggccttcc catttactga gagtgaatta cacactgctc tattatcaaa acttaaatac 15480 ttcagagatg aggttggtaa gaatattgtg gcagttactt ctgattttgc tacttgtgat 15540 tacccatata tcatcaatgt taagaatggg gttgtattag atggaggcac taagctaaca 15600 gcttctcaag caactgcatg ggtagctggt gctagagctg cagctggata tactgaagac 15660 cttacttata aagcatatcc tggagcagta gatgctaatc caagactaaa acactctcaa 15720 atagttgatg gattaaaaac aggtcaattt atattctcaa ctcaaactga ctcaaattct 15780 gaaagagtaa aggtagtagt tgaggaagat ataaatacac accatacttt tacagaagaa 15840 tgggatgaaa gctttacaga taacaaagtt gtggctgtat tagatgctgt tatagatcta 15900 ggatctcaga tcttagttcc taatgttttc cctaaccaca aagatggtat aaagctagct 15960 caggatagac taataagttt attaactaca ctacaagctc aaggagcatt aaagaatgtg 16020 gaccctgact ctgatgtagt ggtagatgaa gaagctagta aaggaaagac tatgtatgct 16080 actatactta cacaacctgt aagaactttc aagaaatact ttgtaactgt taagaataat 16140 taaggaggta tagcccatga gtgaagttaa tagaatgtct gctagggagg gtagagcttt 16200 tgtatcaggt aagcaagtaa ctagcttaat aaaactagaa gctctattta ctcctgaggt 16260 aactaagaaa agggttatgg gtcagaaagg attaacctca aaagttatag gttatgatat 16320 aactggaact ataactcaat tcaaagctac tccaatgata agagaagcta ttaaagaata 16380 cttaaaaact ggtatattcc ctaagatgga tatacaagct gtatgtgatg acccagattc 16440 tgactatgtt aagaattatg gtgaagacag agtacaatta ctaggagtac aattagaagg 16500 agatttaccc ctaattggac tagatgctga gggagaagaa attcaagaag aaatatcatt 16560 ctcagcttca gaagtaagat atttataaaa gattaagaat attgtggagg tagattatta 16620 tgtcaaaatt tagtatggaa tttttcatgg aaaatgataa gggtaaacaa tcacccgaaa 16680 ttgaagttga aggggtagct agatttagtt tccctgaggg tcatgaatta gcgggccaac 16740 ctataccttg gatattaaag ccaatcacta ctaagcaaat ggaggatatc caaaagagaa 16800 atacagtaat tactagaaac aaaggtataa ctaactcaat aacagaccaa agaagagcta 16860 ctagggaaat gatactagaa acaatagtat tccctgactt taaagatcca aaatggttag 16920 agtctactaa gtttgtggat ccagtagatt tacttatgga tgtactaagc ttaccagggg 16980 atttcactag aataactagt gaggttatgg atataaatgg attaggcgat aataaggatg 17040 atctaatcaa agaagcaaaa aactaataaa ggccgacact agtgcatttt gggctcatgt 17100 gttattccag aagcataata tactaccctg ggaatttgat aaactatctc ccaataggaa 17160 agcttttctt atagcttcat gtgaactggc tttagaagaa gataaaaaag ctagggataa 17220 attaaagaaa ggctcttaat cataaaggag ggatataatg gcaagtcaag ctgcatttac 17280 tgccatattc tctgcccaag ataagatctc tggtgcattt aaccagatgg ctgatagtgg 17340 taattccttc atgggtactc ttgccaaaat tggtggagta gctgctggag tattctcagc 17400 taaggctgtc tgggatttta ctaaggacag tgttaaagcc tttggggatt ttgagcaagg 17460 tttaaatgag gttatgactc tattacccaa ctctactcaa gagacctttg ataagatgtc 17520 cggccaggta aaaaagttaa gttctgatat agggatattg cctgataaag tagtacctgc 17580 tctctatcaa gccatatcct catcagttcc agaggataac gtttttagtt tcttagaaac 17640 agctggtaaa gcttctatag gtggtatcac atcaatagaa actgcggttg atggactaac 17700 tactgttatt aactcatatg gagctaagaa tatggatgtt caaagggcct cagaccttat 17760 gtttactaca gtaagacttg gtaagactaa ctttgagcaa ctatcacaat ccctattcaa 17820 tgtgctacct tcagcctcag ctgctggggt atcttttgaa gaagtatctg cctctatagc 17880 tgcattaaca gctcaggggg ttccaacatc tgtagctact actaagataa gagctgctat 17940 agatgagcta tctaaatcgg gaactaagac tgataaggtc ttccgggaaa tagctggtaa 18000 gggtttcaaa gacttcatta aaggtggagg taatatccaa gaggcccttc agatgttaga 18060 aaaacacgcc cagaaaagta atctgggtat caatgacctc tttggttctg tagaagctgg 18120 ttcagctgca ctagccttga ctggtgaagg tacggaggcc tttactaatg ctcttaatga 18180 gatgaaggat agtgctgggg ccactgaagc tgcctatgaa aaaatggata agggctttaa 18240 taggaccatg gagagaataa aaaccaaagc tgctgtagtt atgttagatg tgggagatgc 18300 cctagcccct gccgttgagg ttatagctaa tgctgtccta aatggtatca catttataca 18360 agaaggcttt gaaaaactag gacctatatt ctcaatggtt gtttctgctt tccaaggttt 18420 aggagagact atatattaca tcatgaatgg ttgggagttt ggggatgcta tgcaaccctt 18480 tatagatcta accaacgata tattatataa gttcttacct gccgatgtgg cagatcaaat 18540 taatgagaag atatggagtc ttggggaaga catatataat aattttggag agatcttagt 18600 aggtctacca gattatttat cagaggctaa caataatata caagaagcct ttagtggttt 18660 agctgagcct atcatgggag tattagctcc tataggtcaa gtacttcttg atatgtttgg 18720 aactttagct tctatattta tgaataacct aattccagcc atagacttag tagttcttta 18780 tgttcaagga tttactcaag tatggcaaac tgtattggaa gtgttaggtc ctatattaga 18840 agtattaggt tctatgttct ctgctacttt tacagctctt ggagaattga taaccaatgt 18900 tttcatgcca attgctactt ctttagctga ggcctttgct acttatatag ctcctgcaat 18960 ttctcaagtt gtggaagcta taggtaactt cctaatgcca atattacaag gactagcaga 19020 ttggtggaat aatacactac tacctatgtt cttaagcttt gctgagaatg tagttccata 19080 tattcaatca gctattgaga gttttggtat gatattccaa gggttggtgg atatgctaac 19140 tccactactt aaccttttag ccgcatcttt ccaaacagta tggcctgtta tatcctcagt 19200 tgttactggg gccgttcaaa ctatatcagg agtactacag ggattaatgc agatgttgag 19260 tggtttaata gcttttgtgg ctggagtatt cacaggtaat tggtctaaag cttgggaagg 19320 tgttaagaac atctttggag gtattttcca aggtatgtta tccctagcaa aagctcctat 19380 taatagtatt attggtatga ttaataactt aattggtgct cttaatacta ttaagatacc 19440 tgactgggta cctattattg ggggtaaggg tattaatatt ccccagatac caatgcttgc 19500 tgaaggtact gacttcttta cagataaagc tgccatggtt ggtgaagctg gtcctgaatt 19560 agttatgggt cctacattct caaaaatgcc tagaggctct caagtattac ctgcagataa 19620 aactcaggat ttactaagac aatctggagc aggaagtgat gaaggatcta aagttatgaa 19680 cattaacaac tcatttaatt tcacagtaca ggctgaagat ggaacaatct cagaaagtga 19740 attaatgaga gtatacagat ttattgtgga aatgttaaat caggaaatgt tcgagggagg 19800 tgaactggac cttgcatagt aaatatcaat tctggttaat acagggagat actaggttta 19860 ggttaccagt aaatcccact gatataaata tctctgatag tagagataat agtacagttt 19920 ccattgaagg cttaggggaa ctctctttca taggagaccc taagctaaaa tctttttctt 19980 ttaattttga gttaccaagc tcctactacg atggatgtga gtatactgat attgcagacc 20040 cccgagtaag ttatgctaaa tttattgaaa tgggtaaaga tccagtaaga tttactatta 20100 ctagtactga tattaactta ttagttacta ttgataatat ctcatatact gagaaaggtg 20160 gagatattgg tactcttcaa gtaaaaataa aactaactga gtataagaag cctgaggtta 20220 aatcagttag tttcacaatt ccaaaaccta aactagccta tagtggtaat acaggaggta 20280 gttcttctgg aggtactcaa ccttgggcag ctactactac taatgtttac tcatggctat 20340 gggtaagaac tggaccaggt atcaataatc ccactaaggg taagatatgg cctaatgaac 20400 actttacagt attaaggtct caaaatggat ggcattatgt aaagtataaa ttagataatg 20460 gttcaggata taaagaaggt tggagttctg gtgattatat aaggaggtta taatatgcaa 20520 ttaattgtgg tacaaggaga taaatccttt gatattacaa atacctccag tggtaagagt 20580 tgggcagggt cttcttcatc tgcccctcgt accctatcat tccaatataa tgctcaagat 20640 atacatccaa tttatgatgg agatcaggta gtactagtcc atgagggtaa gaatatcttt 20700 aatggtatta ttatgaatat ctcagataat gagaagaagc atttacttaa ggttaaagct 20760 tatgacccta tgatttattg ggctaataat aaagatagtt ttgtatttga aaaccaaact 20820 ggcgaagcta tatttgagtc attatgtaaa gcctttgggg ttgcccctgg agtagtagct 20880 aaaactggat atactttctc tagtttagta tgtgagaaac aaactcccta tgatattatg 20940 aagaaggtaa taaaagaggt ttatgaacaa actggagaac agttttatat ttactataat 21000 tctattacag gtaagtttga ttttataaac cgaagaacca acaatgaatt gtggaaattc 21060 gaatatggta aaaatattgg aactttttca agaagtagaa actttgatag tattgtcaac 21120 caggtaaagc tagtatgtaa atataaacag ggtaagggta aagatgatcc cgagattact 21180 ttatttgcct ctgctaaaga tgatagcagc caaaagaaat atggggtatt tcaattatac 21240 gatactatca gtgatgaact aaaccaagca caattaaatg agaaggctag aataaagcta 21300 gaaacaggta agagacctaa aactgatatc tctataactg ttataggtgt acctgactgt 21360 attagtggta aagccattgt ggttagtata ccacatttaa aaattaataa ggctttctac 21420 attgagtctg atacccataa atttgatact gactacacca tgaatttaaa acttaaatgt 21480 actgaggagg gtgatattta atggctaata atttattaga ttgtgtaaaa caagtatata 21540 gtaatatgtc caccccccta aatatttata gagctaaggt acttagtact agccctttaa 21600 aatgtgagtg tcttcacaat agaaaattag tactatctgg aaatattata gatgttccaa 21660 aacatattga gtctattagt gagggcgata ctgtatatct tatgccctat gatgatggac 21720 aaagatattt tatgttaggg atcttaggag gttgatataa tgctacctaa tggctttaaa 21780 gataccctta tagttgatga gaaaacaatg gagaatacca agacctataa actagatcta 21840 aatcttaata gaatttatga taatgtggat ggtcttgagg ccttaaaaca atctttatat 21900 aaaagattaa taactcctaa aaatgcctat atgatttatc aagatacttc ttatggttgt 21960 gatattagag ctttgattaa tgactctcta gtaactcctg aacttcttga aatggagatt 22020 aatagggcta tccctgaatg tctcttagaa gacgatagag tgactaaagt aactgatcta 22080 aagtataatg tgcaaggcaa gaatgtatat attgaagtat ggataaatac catatatgga 22140 gatacaaaac tggaaggggt gatataatgg gattttctga agatttattc tcccgaacac 22200 ataaagaatt attagaagag atgataggcc tatcccctga tgatattgac actagaactg 22260 gaagtttcac ctatgatatg gctatggttt ttgctgttaa gctagctgag ttttatgctg 22320 ttaatgttaa tcagatatta gactcagtat ttctaagtac tgctacaggt aagttcttag 22380 atgagagagc tgccaatgta gggatatcca gaataattcc aactaagtgt agaaggtcag 22440 ctaagtttgt tggctatgaa cccgatgttg gttcaagatt ccaaggagga gattatttct 22500 ggcaagtgat agagcctaat attattgaat gtgagacctt aggttcagga cccaataagg 22560 tatcaacagg tacacatcta atccctatca atgctatagc agaacttaag agtgctacta 22620 tgggagatat acttgttatt ggtagagatc aagagacaga tgatgagtta agagaaagag 22680 ctaccaaagc cactatacaa ccatcgggag atggaaatgt tcaacagtta actgattggt 22740 gtaaaagtgt agctggagta ggtaaagctg tggttcttcc ctgttggaat ggtataaata 22800 ctgctaaagg tattataacc acagttgaga ataagcctgc caatcctgaa ttagttaata 22860 aagttcaaga atacatggat ccaggcaaga aaggtttagg tgaaggtaaa gctcctatag 22920 gatgtgtttt tactgttgag tcagctcaaa cctttaatat cactgtaaga gctcaagtta 22980 agaaattgag ttcagttaca gaagaacaaa taaaattaaa cttcaagaaa gaactagaag 23040 aatttctaaa tagtcaagta tttaaaatta ctagcctatc tataaataga ataggggcta 23100 tgcttttaaa tattgaggga gtagaagatt atacaaccat tacacttaat gggagtagtc 23160 aatctttaac tattcctgct aatcaaatac caaccattac tgaggtggtg gtaacttatg 23220 tatagtctag aacaggttca aaaagaatta ctattgtacg ttcctgaatt ttatgataat 23280 ataaaagagg ctgataactt agtaaacact attatggcta ctgaacttca gaagctatat 23340 actaagggac aggaattaca agactatgtg gatcccaaga ccactcctaa ggaaggtcta 23400 gaaagatttt tcactgacct tgaattatat aaatactttg attacccacc aactgaccaa 23460 gagaaaagag ctattattac agtagttcta tctgcttgga aaaagtttaa tagtacatct 23520 atcaaaaatc ttactaaggc ctatacgggt ggaaaggtaa aggtagagtt catagctgaa 23580 gagagtaaga tcaaaattag ctttactgac ctaccagggg taccacctaa cctaaaggct 23640 ctagaaagag tattcagaga aagaatgcct gctcatttaa tatatcaaat tatattactt 23700 tatattactt gggagatgtt cgataggtat gattatatgt ggagcaagtg ggacagtcta 23760 gaccttacct gggaaaaatg ggaagaatat gctataactg aataaaggag gtatgcaaaa 23820 tgccaagtac tggtaaaact cctactctag gtttaaataa gtgggagtta acagataagc 23880 caaaaatgaa tgattttaac aaggacaatg agaacattga gaacttcgct gccgaacata 23940 aacagaaaat gactgaggta gaaaagaagt ctaaatctaa tgagaatagg ctagatgctt 24000 taagggttgg tactattaat aggcttgagt ggtataattc tgagtcttat actggtcctg 24060 gtaatatcac atctattaca gatgctccaa ctaagagacg aggttttaaa tggtctggta 24120 tgtcaacagg tccacaaact cttgaaatta aaactctgcc tatagaacca aatagtgatt 24180 atgtattagc ctttctagaa gatattgaag gtagttttac tagctatgaa attaaactgg 24240 cttacactaa tgggggtcaa caatataaga gtattgtaaa acaaggtagt acaccacaga 24300 gtaatgggta caacttaaaa actcataaat ttagtattcc ttctggagct actgatacta 24360 aaattcaatt tgtggtacag ggtgcaacct caagcacatc attaaaggta ttaaatatct 24420 tattagctag gggtaatgta gtacccgact ataacctagc tcctgaggat gtgctaaaaa 24480 gattagacct attatttgag aaagacacta gtttacaaaa tcaagtaaac caaaaagctg 24540 ctaaaactga tgtactttta aaggttagtg ctcccctgga tagagattgt aattcattta 24600 aggatattaa tactttttgt acctttgata caggagatgg taattttaag aatacccctg 24660 atggtactct atcccaagga agtgctcagg tatttatggt agttaataga ggtcatcata 24720 gtactagatt acaacaggag tttatatacc tatatcccaa ggatagtgta gctcgctata 24780 agagaaactt tgacggaact cgctggggta actggtataa ggaatatgat agtgctaata 24840 agcccactgc agaagaacta ggagtagctg gggtgaatat gcccatggct gatacagatc 24900 taaataatca taagaatcct ggtttctatt ggggttatca aaatatggtc aatgcccctg 24960 tacccaatgg tgtagcaatt atggaggtag ttaagtataa taatgattgt gtattacaaa 25020 gatttacctc agttagagca gatagtaacc atactactta tattagaacc tgctattcaa 25080 ataaatggtc tagctggaag aagatttatg atgagggtaa taagcctacc ccagcagata 25140 taggggccat accccagata cccattagaa taactcagga tcttaacaca ttcactacac 25200 ctgggttata tattgtggat agagcaactc atgctccaca atcatatgga agattattag 25260 ttctaggttg ggacaacgac cagaagtggg taactcaggt attctatgct gatattacta 25320 ataaagtata tgttagatgt agcactaaca acactgctac tgaatggact tcttgggctg 25380 aacagtactc aactagtttt aaaccaactc cccaagacat tggggcttct cctagtaacc 25440 ataatcataa taaggtatat taccctatac ctaaacaggc tattactgat tttaattccg 25500 gtatagctat aactgagggt caacatctag tatcttctag tacagctata cctaattcac 25560 ctcctttata tgctcaagat ggtagtagta tatatggtat acttcatgta tatgttagtg 25620 ttggtgattc ttataatggt gtgaataact ggatatggca agactttcat gatactaggg 25680 gtaatcacta ctggaggtat aaaattaata atgctgactg gagtccttgg agagctgtat 25740 ataattctgt taataaacca accccccagg ctataggggc tgtagctact gaaggtgatg 25800 gagtaataaa tggaactcta gaacttagaa aagccgataa gctaataagg tttgctactg 25860 agagaggctg gattttaaaa cagggaggaa ctgcagcaga aacctccttg gatctagttt 25920 ctgaaactga tggaaaggcc tttagggtat tagaaccttc gggaaaaaag ggcatctata 25980 tgtgggtatc atcaggggga acacaattta atgtagatgg caattctgtg taccatcaag 26040 gatttaaacc tactccatct gatattggag cagcctcttc tagccataac catgatagtg 26100 cttacctggg taaaactgct acagctgcta acagtaataa acttaagaac tgggacttag 26160 ctgaagggag taataacttt gtgggtatac catttatacc ccatgatgga gtcatggagg 26220 taggtagaat ggttgacttc catgaacctg ggacaaataa agactttaat actagattag 26280 agtctattaa tggagctcta aaatgctggc aagatttctc agctgaaaac ttatgggcca 26340 gaaaatatat gagagtaaat gactggtatg gtggatctga agatggccga ttctactata 26400 aacaagatgg aaaaggacta tatactgaga atgtggaaaa cctctatgta aaaggtagtc 26460 ctgtggctaa aggagactat tgggatgtag gggataatac cagagtagtt agtctggggg 26520 gaggactaag attagttcgt caaatagtta agacaggcta tgctaactca ggtggaggtg 26580 taaattatac tatacatttc cctaaggctt ggaattgggt attacctata tctctagtat 26640 cacataacta taatgacgct tctaataata cctcaggtta ttgtactatt gactattgga 26700 ataactcata tttgaatggt ggatgttatc aaatggtagc tggtaagcca actcaaataa 26760 tactaaccta tttagctact gagtaaggga gttgataaaa catgagaata gtaggattta 26820 attattctgt ggaaactaat aaggttatag accttatatt ctatgatgat gaaaaagaaa 26880 tgctggtaaa tcaggcagaa gactcagaag atgtaagatc atttatagat aaagaccatg 26940 aagaagttac taaaatatta gacctaatta atagacctga acctctatac tatagagagg 27000 atagtgaaat attaaataat atagaagata ttgtatgtct ccatatacct cataattcac 27060 aagggcattt agcctttgtg gataatactg aggttattga tctagacaat cttcctgtta 27120 tacagatgac tataagtcta tttgaccaca taaagggtgg cccaactaaa tttgatatca 27180 gtaaaaagag atcagattta tacagtatca aagataagga cctatttact tcatatgagg 27240 ctatagatcc taatcctcta gatattaata aattaattgg ttatcaagcc caggctcttg 27300 ctaatactaa attagagcta atgcaaacta aagccataat gaaagtatta acacaagaac 27360 tagctgatat aaaaattaaa ttgatgaata aataaggagg tagagattat gaatttctgg 27420 gaaatggctt atagaattgg agcaataggt aaggacaaat tacatcaggc agtaaaatgc 27480 tcttctaatc cagaaggaga aattacacca gaggactata aaagaatatg tggtgaagac 27540 tttaatgaaa gggtgtaatc tatggaaaaa gaactactaa ctcaagttct tagccaaggt 27600 atcttttgta gtctatttgt atggctatta tatgacacta gaaaagaggc tagagaaaga 27660 gagaataaat taaatcaagt gatagataag ctagttgata aattcaacgt agttgaggat 27720 attaaagagg atgtagatac tataaaagat cacctattta gtaacaaagg ttaaattata 27780 agcctagcag gtcatttaag gcctgttagg ttttatataa gaaaatatat taaggaggtg 27840 gttaaaatgc tgtagcgagc ctctcaggcc aaataaaaca cttataatat ttaaaggaga 27900 tgtttattat gataatcaat aaaagattaa gtactactaa tgttacctta aacgctaata 27960 atccagccta tataattatg cacgaaactg ataacactga tagaggagca ggggctgaaa 28020 gacactgtag ggctcaagct aatggaaatt tagaagatgc tagtgttcac tattacgttg 28080 atgacaccgg agtataccaa gctgctgagc ataaacacgc tacttggaat tgtggagatg 28140 gccataatag atatggtata aataatagaa atacaatatc tatagaaata tgtgttaatc 28200 ctgactctga ttataataag gcagttgata atgctgtaga gctagttaga tacctaaaaa 28260 atggctacta ttctaattgt aaagtagtaa gacactatga tgctagtaga aaaaattgtc 28320 ctagaagaat gatagctaat ggttactgga atacattcct agaaagagta aattcaggag 28380 agagctcaag ctcaccagta attaataatc ctaagggatt ctatgagtct aatgagacaa 28440 gaactaatgc tactttagtg ggggaaggtt caatagaagt tctagatgag gattgtaagc 28500 cagtccctgg tagatttatt gatagcctag ataatttatt tgttcttgga atatatcctt 28560 ctagaaattt catagaagtt gtatatcctg gaaaggataa gaaataccat gcttacattg 28620 acataaaaca ttatagcaga ttaagttttg actaccacat gaaatatcaa aatgataatg 28680 gaataactta tgtgtggtgg aaccctgaag acattaatgt caaagatcat aatgaagaat 28740 tacagcctgg tcaaaaggct agcccaatgt atagaactaa gggctggcta agaataacat 28800 tctatagaga ggatggaact ccatctgatg gatacgttcg ctatgagggt gagcaaagcc 28860 agaagttcta tgaggatatt aaacaaggaa tagttaaggt taacacttct cttaatgtaa 28920 gagatgatgt gaatggtaat ataataggct cagtatttaa taatgagaga gttactatat 28980 taggaagtaa gaatggttgg tatcatatag aatataatac tagccatggt aaaaagcaag 29040 gttatgtaag ttcaaaatat gtagagatag tttagggggt atttatcatg gaagaaaatt 29100 tagtaaactt aatattcaat tacattgatc cacagctttt aataatcatt gtatcatgtt 29160 actgtttagg tctgtttatc aaggctaagc cttatatacc agattggtct ataccattaa 29220 tacttttggt attcagtata tcattgtcaa tcttatatat ggcaattcaa ctagaattag 29280 gttttacagc taagaccttt ttgaatggct ttatacaagg tttaatatgt gcagctgtag 29340 cttcctttgg taatcaagta tggaagcagt taatggataa gagaaaatca gatcctcctg 29400 taaatcaata ggggatagag gttgggccta ctttatttag aaagtaggtc cttaaccttt 29460 ttaaattctt cttcattatc ccattcccat tgctttcctg gttttacaac ttctttgttt 29520 cttaagattc ttcttgcctt tttaccattc atatttaatt cttcaagtat catctttaag 29580 gtgaacttac cttccttagt ttttggttca tccttttcct ctggttcctt aacatctgtt 29640 agatcaagag taattttagt ttgttcagcc ttttcttcgg gttcagctag ttgttgtacc 29700 tcttcaactt tttcttcttg tggttgttgg atttgttgga tagcagctat tagatcagcc 29760 tttttcatag tccaccagtt tttaactttt aatttctttg ctaattcttt aatttcaaca 29820 gctttcatag tatttaaatc tttcatatca tttacctctt ctttcttatt tgttttatct 29880 acattttcat tataagagat ttcctctggt ttgtcaggga tttcttttaa taattttgga 29940 cctccccagt tacattcttc tccatcagca ttatcaaaga cctttcttcc ttcttctgtg 30000 taacctatta cctctccatc agcaaatact aaaccctttt taacaaggga tgagatcaca 30060 cctctagcct tagttccttc tatacctgag ttttctatta ctgagaatac ccatacacag 30120 tcaacatcta gtccatcatc aaattcattc tttctcatag catttaatat ctttatttcc 30180 attgcagtta agttcatatt taacatctcc tctttgtttt tgtttacaag tttattataa 30240 gagttattgc cagagatgtc aaaggttttc tccacagatt tataaaattt ctttgagatc 30300 ttagttggta tcacatattt aaagccgtta catttaaagc aagtagtgtt accatctgag 30360 gttcttgagt aatggccagt gcctccacat ctagggcaag ttttaagtaa acctttctta 30420 gcaacctttg acatagtgct agcccatatc ttaccaaagc catatttgat tttagcttta 30480 tctgaaagat cagggtacac catattaatt atatcaccct ttaatggatc atccttgata 30540 taactattat accaatccaa tttaacttgt gtatgtgatc tcctacttat taccttagta 30600 acctcatact cctttaacca tccatctatg gcctcattgg ataatttatc agcctcttca 30660 aaagttatgc tattaacagg tatactaaat gctcctgttg ctacccagct tccatcttct 30720 ttttggcaac caaagaagtt ttcagcttct acttgaatac cataatatat atctatcatg 30780 tgatttacct cctaagtttt ttgttaatat aattatacca gcactctcga gatatgtcaa 30840 cgaaaaaaga gaagaagttt taccttcttc ccctaacctt taagttgatt aatagatcat 30900 atatatcttt aggcatacct tgctcccact cccactgttt acctggtttc tcaatgttat 30960 tctttcttaa tatacttctt gcctttcttg gatctagttc tatcaattca cataaatctt 31020 ttaaggtgta tatatcactt ctagtttgtg ctacagctga gttttcaccc tttctagcct 31080 gttgcacttt gttgtaataa agactagtta attttttatg taaaggacta tgtgaccaat 31140 aagttaagtg agctaattca tggcatatag tttcaagaac acagtcctta atatcccagc 31200 caaatttttt gattttatca taagcttttt ctccttcctg tgcaaacctg aaagggaaaa 31260 ggttaattgt aatcccagta aatttattac ctgatctatg gcatagacca tagtagttag 31320 catccacatc agattctata taaagatcaa cagcacttac accaaatgct tttcctaaga 31380 tcattgtggc ctcattgtat aactccttta cttcatattc agctaattga ttttctatca 31440 tatctctcat tttgtttacc tccaatagtt aatttatgat ttaattataa gagagaactc 31500 tggatctgtc aacgaaaaac ttcgaagaaa aaaaggatgg ggttttacca tcctagatca 31560 tctcttcttc ctatatcata tattgcacag atgcaagcct caagatgttt ttcaatctca 31620 gcttttttat gatgtttaca tacataatat ttagttactc ccatttgttg tagtagttct 31680 atcaccttat ccaagaattg tttttggaag ttcttttcaa aaaccacctt atttattttt 31740 tctatctcat ttaacatttc taatttagtc atttttcatt cctcctaaga attaattaat 31800 attgattaca atattattat accagagggc ccagactatg tcaacgaggt tctggagatt 31860 tttttcgaaa aaaaatagat ggccttttgg ccacccatat taattggatt ttttcatttt 31920 taggataacc tcctcatagg caaccatgca gcaagaaagg tagtctccta attcactttc 31980 cctaactttg ttgcttagtc caaaggattt tacctgttta tctagttctt ttctaaattg 32040 tggatagatt ttattatcca tagccttttg agcaattcct tttattacat taagtttttc 32100 tgatcttgtt agttccatag ttaacatctc cttcaattaa ttttgattac aatattatta 32160 tactagatag tcccgatcct gtcaacgatt aaatccagat tttttcgaaa aaataaaaaa 32220 ggaggttttt acacctcctt agttttatag atccttgaac atttcagggt agttacataa 32280 agtatcttcc acatattgtt ctgggggtaa tcatattcta tgatgtattc aacagcagct 32340 attaagtcat tctcatcatt aaattctacc ccttgtattt ctagttcctc ctttaattgt 32400 tctaaagttt cttgaaattt tttgttattc ataattaaca actcctttga attaattttg 32460 tttacaagat tattatacca gactctggag tctatgtcaa agaaaatccc gaatttcttt 32520 ttggcatgaa agttgctagg tatcggatcg ctttaaaaag aagcaactat ttagttgctc 32580 ccactggggt taatgaaaac cctgatagtt ttttattctt tccccaccta gcacaagatt 32640 ccattgtggg ctcaataccc aagatattgc atatatccaa atattctttt aaccataata 32700 acctcataat aactaatcct cctttgagta tcttaacttc ttagattttt tattcttata 32760 catcattgag tcggcctgta ctatagcttc ttcaatagtt ttacctttac catagccaca 32820 tgatagctca gggtatatat tcaaggtatt gcacagatta taagctatat cactgataac 32880 tacaaactca tcaccacccc atctaatcaa tatatccttc tcaggatgta aatgggcctt 32940 aataagattt acagtgtatt gtatatacac atctccttct agatgaccta atttatcatt 33000 gaactctttt agatcattaa gatcaatata tacaatctca tactcatcaa ggtcaatctt 33060 atagtttcta ttataggctc ctgtccaatc aatagataac tcctttacaa gttcaatcaa 33120 ttcttccctt gtaagttttt caagatcttt catattcagt tctctccttt ttgtttgatt 33180 acaatattta tattaccata tactgtggat attgtcaacg aaaaagaaga ggtttttaaa 33240 cctccccctt attttaacat tttcttaata tcctcaagtt cagatcctga ccatttatag 33300 ggtttgttta ccccttgctt tcttagcttg gctctggctt tacttggatc taatcctaat 33360 gactcacata aagattttaa atcaactgtt cccataggtg cattacttcc aggagtagta 33420 gggtaatctc tcttggcctc aactttattc agatcaacct tctttatagg tttgtccttt 33480 ttctttcttg gcttcttggg ctcattagga ttaaattctt tcattaattc tctctccttc 33540 ttcttttcct catttaggag tcttaacctt tcaagtccct tttgtttttt cttttcctcc 33600 ttgactttaa tcttggttag ggctttaata gttttaccag cctcaattaa agctttagct 33660 atttcagtat gattacttat tctaggttgt tcttttctca ttacttatca cctcaattct 33720 atattaccac attacttatc aatagtcaac agtttttctg ctctatccct tttaagctgt 33780 attgatttta atattttctc atcaatagta ttctcagtta ttaagtgtat tatttgccaa 33840 ggctcctttt gaccattcct tttgattctg gcaatagctt gcacataatc atcatattta 33900 tagttccaac tatagaatat taaccttgag gccctgaaga ggtttagacc aacactacct 33960 gaacttattt gtagtattaa tatgtcaatc ttgccctcct gaaaatcctt attgatctca 34020 gctctattct tagtagatcc tgttaagcat tctactctgt gggtcttttt aaaagtatca 34080 tgtataagtt ttatttcctc aagaaaatta catattacaa caacaggctt tttatatcct 34140 tctagatagt cctgtaataa gcctaatttg cctttatcaa tattaatact atttcctaca 34200 ttgtttgtaa taaatcctcc tgagatctgc tgtagcctca aatattgggt aatggtaaga 34260 tcagctgtta attggttgat aaacatatta gcccttaaga ataaatcctc atatggctca 34320 ttactctcat agggtatacc attagacctt aataaggcct ttaattgctt cctaggaatg 34380 ttttcctgta ctatatctag ttgtgctatc atttcctttc taaggtcatt ataggccttt 34440 ctggctttat tatttaattc acaggttaaa tacagatcag gaagttgttc tggtaaatca 34500 atacaatctt ctattcttac tatatagcaa ttctggtgca taaggttctt taatatttcc 34560 tcattcttat accccacaat ttcatagttc atataacctc ctttgattat aaagtcatct 34620 tcaaactgtt tccaattaac tcctagaata tttgagttca taattttata ttgcataaat 34680 aaatcttcat aaccattagg agtaggagtc cctgtcatta tcaatctata tctacattta 34740 gtagcaattt tatatagagc cttagatatc tgggcatttc tattctttac cttatgggat 34800 tcgtcaacaa caataaactg tggctttaag ttcataaggt atttatccat caacctagcc 34860 ttctcaaaat tggcaataat aatctttaat cctgagtagg attttatagc tctataggtt 34920 tccataacct tggttttatt accctgtagt ttaaatattt tatactcctt aagtttagta 34980 aatttatgga tctcagactc ccagttaaat acaatagatt taggtgagaa tattagtaca 35040 gtatctagac cttcaagttt ctcaagggcc acaataggaa ctaaggtctt acccgtaccc 35100 atgtccataa atagggcaaa cttttcttta tttaaaaata gttgtaatgc ttctttctga 35160 tgtttaaata attttctctt catactaaaa aagaagaggc attaaagcct cctccttata 35220 cctccttagc cagttttaca agttcatctc ttgaggcctt actatggaat ttaaccccat 35280 tatcttttag gtattgcttt aaggattctt tattcatact atcaaaatct aattcttctt 35340 cgtcgtaaac attgtcatca tcatctaagt cctcatcgtc attatcatct atgtcttctt 35400 catcatattc ttctttacta agatcatgta tttctgagtt atcatcacta gttaattcca 35460 tagcagccat aatagcatag ttagctaagt ctagtaaagt atcaaccaca gattcttttt 35520 gagcagcata tgaagggaag tcatcttgat ctccgagtaa taaactttta gctcttccta 35580 gcttatcttc aagtcttata catagaacag ggtttccata ttcaagataa gtcttgtgga 35640 aattatcccc atatagatta ttctttaatt catacagatc tcctagttct cctaggaatt 35700 gttggtgtct ttctaaattt ttcatgtaat ctcctccttg tatattcagc tgaggtagcc 35760 ataaaagcta ttcctcctgc tagatttatt ttatttataa tgtctttttg tttttgagtt 35820 aggttctttt cttttccagg aagtttaact tcaataccaa taaacttacc tttgtgacaa 35880 cctagtatat caggtaagcc agctatttga aagggtcccc catgagtttt aaaccaaaaa 35940 ccagggtatt cttttttaag catattcatt atggatttaa ccacatctct ttcaagcata 36000 atagtagcct ccaagggtaa ataaaggagg accttaaagg ccctcatatt ttaataatta 36060 aattttattc ttcatcatct tcttctagca tttctatgat gtcatcctta gacattcctt 36120 ttttaacctt taagcctctg tccttagctt cagctttaag ttcttttaaa gtcattgact 36180 catagtcatt ttcttcttca tcttcctctt cttcatcctc atcttcttcg tcatcttcat 36240 cagagtctcc gaataggtca ttatattgtt cagctatttc ttcttcgtca gcttcttctt 36300 caattagatc aattagatct tcttcagttt tagccttctt gattttctta gctgggattt 36360 ttaaagcctt agctaattct tttagctcat ccttgtctaa ttcttcaaga tcaacttcag 36420 actcatcgtc ttcatcatct tcttcgtcat catcagagtc agcttcgtca aggtttataa 36480 aatcagttat tcttgatttc ttcttacctt catatatttc atgtgatact tctaccccac 36540 attgtaaacc aactagagct tctgtatcta attcgaactc ctcatcaggg atatcaaatc 36600 ctaaagcaac taacacacct tttaagttaa ataaagcttg tggttgaagt gagcaaatgt 36660 ggtataactt ttgtcctttc atctttcctt ctgctattac aaaagtaaat gataaataat 36720 tgcttccact attttcagaa accttaacct ctacatcttc tacttccaca atgtaatttc 36780 cttctgggat agtaactgag gtttcaacgt ttgaaagatc aatctttaat cctcctttgt 36840 ttttgttacc tctcttagta gtctttttag ccatgttatt gtcctcctta aattttaaaa 36900 tatcattatg agataaatat ttgtcaagat attgtacctc tttgtttaca agtttatatt 36960 atcagatttt taattggttg tcaaaggtat tttttagaaa atttttattt tttcttttta 37020 ccagatttct tactggtttt tttaccagat ttagctggct tattattcca ttcaccagcc 37080 agaatagttt caatatcctc ccagatagtt gttggtgatt ttaaatcatg taccagataa 37140 gctggacatt cagatccttg aggtctagta aatttagtaa tataataagg gttaggtcct 37200 agtcttagtc tatactggat ttctttccta accttcatac cctctttttc tgagtgttca 37260 tataaataag tatgacctat tactcttgat acagcacata aatatttaga tacagatggc 37320 attaatcctg gtcctacctc aggcattaat tggtcttctc cttcaccatc accagattct 37380 aacctatctt gtactataaa gcatggtagt ataccttcct catttaattc tttataaagg 37440 tttataacct ctttcatata attagccaca tttccaaaca ttctttggct catctggtcc 37500 ttaccttcct cagctttgac cttttcatta cctaattctt gtaaagcagt taaatggtct 37560 ataactactg ttttaaattt atcagtgttt tctgataggt aatcataagc ctcgtatatg 37620 tcatcaaaac tctttagact aaatacttgt atatctcctc ttttaactct aagcttgttt 37680 ctagctgata gagtaccttt gtcctttaca tcaataaaga atattggttt gggtagtgtt 37740 cctgagatag tagttttacc tgaaccagat cttccataaa gtgttaatat tgtgggctca 37800 tctatatcat atagatccac taatctgtcc tcaaaactat ttgattttga tttttgcttt 37860 tttgccattg tcactttcct ccttttgttt atagtcagcc ttaataataa attctttatc 37920 aagatttaaa agttcagctt gacataaaga cttatattca cagaaagagc aaccttttcc 37980 aaggttcttt tcctttcttt tgtggccctc atctctaata agtttagcta tctcaatagt 38040 atcatccatt acactgttaa gtatattctt tgataatatt attgggtgtc ttacaaagaa 38100 gtcttcccat ctagcagaat taatataaga ctcataatct ttaggatcta atcctaattc 38160 tcttattcct ctttctaact ctaaaggtgt ggatttaatc ctcttttgag ataatacccc 38220 tttttgagtt aattctggtc tacctggttg ctgagctaat attatatccc atatagtacc 38280 acttactttt gggtattcat gtagaagagc ccaggtatat atagcagatt gtctattgaa 38340 gatcaggaaa tttctatcag gcatcttttt gtaggtttta gtttccttag cccatatctt 38400 tccatccaca tctaagacac tatcaatata gccttctaga gttacaccct tacatagttt 38460 taattcgaag tgatgttcat tttccacata ggttacatta tcttcttttt catcataata 38520 gtggaaatag ttatcacata aagaataaac catttttgga atatctccaa attcaaggat 38580 ctcttcttta aatgtatttt tgtaaaactc tttggaaaat ttatcaacag attttttcca 38640 actcttccca ttatggtagt cttctatagc ctggtggata gcagagcccc tttgtaaggc 38700 agctccactt cttttaggta ccagaccttc atagtattta acataatggg cataaggaca 38760 agacaagaat ttgttaattc tggaattact tatttttaac atccttatac ctccttttca 38820 ttacaatatt tatattatca gataattatt catctgtcaa cacttttctt atctggcttt 38880 tccaatcacc tttaaaatcc agttcaacac atcctcccca agaataacca actgatatat 38940 ccatgattat tgggactgtt ggggtaaatt taaatacttt ctttaatact ttattatttt 39000 caactattga ttttactttc caagctaatt cttccacata gtctttatct acttcaaata 39060 atattgagtc atgtactgag cctaatacat caaacttaga cttatctaat acatattcag 39120 gattaacata tttagcatag cctgtaatct cacataggcc taataaagtg atatctgaac 39180 caaaaccttg aactggtgag ttaatacatt gtctttcggc ttcagcagct ttagatctat 39240 cagatgaata tatatctggt aatcttctaa ttcttcctat tggagatctt acttcaccat 39300 tggctttaac aatctttctt tgtttatcat gccatgttaa tagtttagga tatttattaa 39360 agaagttcct tctatatgtt ttggcttcct catctgttaa tttaagacca tagttgtctc 39420 ttccataatc cttgaatttt ttccattgca taccatataa gaaaccaaag tttacagcct 39480 tagccttttt ccttttctcc ttcttaactg ctggatcttt ctcatcattg atatcttccc 39540 cagatataat attatatgtg gagtcatgta tatctccacc agtctggtaa ataaatttca 39600 tattatcctc atcggccatc atagcagcta ttctaagttc ggcctgggat agatcggctt 39660 caatgaatac tcttccagga gatggtccta ataaactcct gattttctta tctctaggaa 39720 cttgctgtag attaggatta ttacttgagg ttcttcctgt tacagtccca tgaagtttaa 39780 agtttggata taatctatga ttgtgcatcc tatttagcca accatctata aagtgagaaa 39840 tttgtatatg aactccccta tgctgtaata gtagttctac tgctggatgt ttatctctta 39900 gtcttaataa tacactttct ccagtagctg gtgctccaga gtcagtaacc tcaatcacag 39960 gaagaccaag tgtatcatat aaaaattctc caacctgttt aggtgagctc cagttaatat 40020 cctggccatt agtatacttc ttaagtccag tctcaatctt atctagttca gatcttaagt 40080 atttttcaac ctctttgaat ttctcctgat ttacatatat acccttgaac tgaacatcct 40140 cataagcttt actagctggg atgtataagt ggtggaatag tttatataag ctctcatccc 40200 ttttaagtct tttattaaat attttatata gagcaaatgt atagtatata tcatacccca 40260 gatacctaac ataatcctca aagtctgatt tagtctccac atttccagtc tttaatttct 40320 tatttatatc ccagtcttta gcattacaat gtaataaagc attctcctta agaccattag 40380 gagtattctc atctaaacaa tgtgaggcta gtaaagtatc aaaggttatg attggcttaa 40440 taccatattt ctcctttaag aaattatcat caaacttacc attctgggct attctttcct 40500 tcatctcaga attaagtctt ttaattaagg atttagctaa ccttctttgt gctatgggct 40560 taccccttaa tgggctatat ttgacttcta aaggcagtat atattgtact tgagtattac 40620 caaaaccaaa tagagtaatg tcattattaa acctatctaa gcctctagtt tctatatcat 40680 atgatactct attataacct tcctctctta aatatttgaa agcctggttt aactctttca 40740 tactggtaat aagttttatg ttaagctctg gtaattcatg ctctttacct tttaccatgg 40800 ctttaaagtt attaaaggcc tgatctacaa attgtgcttt agttggatca tatgcaacta 40860 cactgggtga atagctggca ataaacttaa agccatattt ctcatgcttc ttgggtactc 40920 cacatattga ggttatacct tcattactta gcatctttaa agcttgagat cctaagatca 40980 taacatattc aggtttatat ttttcaattt ccttaagtaa atgttcttgg catttcttaa 41040 tctctgatac tttgatttta gtacccttag gagttctaca cttaatagca ttggtataat 41100 aaaagtctag accttccatt ttatcaagta atgatttatc aggagtagct attctattgg 41160 cctcatccct atcattagct aaactattta tgaccattat tcttggcttc tttttctttc 41220 ctacacaagt tccttctata catgagggag ctgagtgagt atataggtta cagttgttac 41280 atctcattta aaatattacc tcctaaatag aaaggacccc taaaggccct tatctttatt 41340 ttatgattga gttacatact ggacagacct tatgtttttt gaagggtatg gttactaccc 41400 ataggcctcc agttaagaaa atcatgatcc aaaacatagc tccatatttt ctttttaatg 41460 ttactgagtt tttacacata ggacaaatta tatttttcat caggatagcc ccctttaaat 41520 tagaatatca tatgatatat tatattggca atatcaaaaa ttattaccca taatcctata 41580 gctgctaaga ttataattaa acaaccaaga cagccattgt tattgttctt aggtctgtta 41640 tcatcactat aattatacat attacttact ccacagctca ttcatataat ttatatagtt 41700 attaagttca ttcctagtca tgtccccaag gttagctggc tccaaatcaa attccttttc 41760 agtattccta ataaattgct ttagggtttc cttcttgtta accttattat taagatcaga 41820 atgtttatat ctttcttttc tcatataact catttattag aataccccct taatatcttt 41880 gttgtaaaca tgaagagaga aaattgtatg agagaaactt cctggttcat agccagtttg 41940 ttcagctacc caatgtaata gtttcatagc aagataaaca tcattagcaa agtgagtagc 42000 aaaatcagaa cttctcataa cataatggat atttagctta ccttctctta cttggaagtt 42060 atatcccaat gaacatggta ctctagttac tcctcctata ttgaatggat caatagctgg 42120 attccacaga gataaccata attgtcttga gcctggatct tctttaattc tattgatgat 42180 atcctcaatt tgtctgtatt gccaaattct ttcattgtaa gaatatccaa attttccatc 42240 tcttagatat tcattccata cctcaggtct taattgataa gctttaccag gatttatgaa 42300 agcctcccta gtatgatgtg gatgtatacc aaataattct tctctttctt caagagagta 42360 ttgattgatg acaccttgtg gatcgtagat cctttctcta aattcagcat ctgcccaagg 42420 ttgagataca cctggaactt cttcaggttt actttctaga attgtgtaac tatagttttg 42480 taactccttt gtgaaataat ctggattacc ttctataact ttatcttgca tagttttggg 42540 tcttacctca atccccattt cacataaatc tctttttact tcttcaaatg cttcttgagc 42600 attaatataa attctcatag tttaattcct ccatttattt aattttgtta agttaattat 42660 actacttatt aaagtacttg tcaatagata atttattgat atctatcttt ggaaatttct 42720 ccagaccaaa atgtaataac tgcatcttct ggagagcttt ataagagtgt atctggtcct 42780 cagatttaaa gaacctgtta tagttactat tcaaggattt atgccaagga tgtttactat 42840 tagcaatact ctttcttggt accttaaaat aattgaagta tccatttata aacattcctg 42900 acaaataagc ttgagacata tgaaaagtaa tattatctat ctcacatact tcttggggta 42960 gttcttcaat gaacctgttt attagtacta gatcagctgc gaatttcttt tgtaattcac 43020 atactctata cataattttt aagtgtttcc aaggtccttt tctttttggt ctagttacta 43080 ctgcagctat caagcaagaa ccattagtag cctttttcct attgaaataa taagtatagc 43140 tcataccact tatcttggta agcttttctc taaattcaat taattgtgga taatcaatgt 43200 atgatcttaa taggtgtccc cacttacctt ctttatatcc aaagtcccct agatttaaag 43260 tacatttatg ggacttacag gagagaacta agtcctcctg aaagccctgt atgttctgag 43320 tatacatcat agtttcttct ggattggtaa tcatttccct attaagattt aagaataagt 43380 catcaaaatc gttatagtct cttattagca tattaccttc tcctaagtat ttattttagt 43440 aattagatct ttggcggaat ttatttacct gtgatttctt aaggtatata tttactatgt 43500 catctacctc taagccagat acagctaata atttaatata acatacccat acattaaata 43560 gtctttggtt aaaagcattt acatctgtta gcattgaagt ttgtttccaa ggtttattct 43620 taaggcaatt acagcacata cccatttctc ttaccaggtc agttacagct tcttctacta 43680 acatttcagg aaattctggt gatgtagtag ccttatgata taagttctct aggctatcaa 43740 ctagtatagc attatcatct atagtattat aatccttacc agctaggatt gtaaattcag 43800 ttaagaagtg taatccatca attaattctt cttggtaatg gtcttcatga ttataagcat 43860 ctaatgcttc tcccacttct tcagtcattc tccaagcaaa atctttaagt ctagcttgac 43920 ctcttttatc atttaagtta actggacagt cttcagtttg gcataatcct gatctagctt 43980 ctatatgatg atatttctcc atcaattcct tttgtctatt aaagatagct tgtaatctat 44040 ctccttctat tttttcttct actgcatggt ttatattcat aacaatctct cctcttatct 44100 tgatatattt ataaattggt ttatagcttc tttagcactt gagtcatcta ttaactggga 44160 tagcttctcc acattggaag tagtataatc atatactagt acattccatc catcacagaa 44220 taacttccat attagttcat catatctagc cagtagatgt tcagctcttt caattactcc 44280 tggcatctgt tctctatcac caaaattgaa gatcacttcc ctaggtggtc tagcatatac 44340 aatagttggc ttacatagat gttttaagat ctttagctcc ttactgtcaa ggttgaaatt 44400 actactatct ctaatgatag gtccatacac catttcttca aagcaagtaa atctatcatg 44460 tatagcttcc ctttcacctt ttctcacaat ttgagttaat acccattctc tttgttcttc 44520 ataagaacca ggtcccattg attggatata ttctcctcct ctgtgtttag ttaggttctt 44580 agctgttgtg gtcttacctg agttatccat tcctaaaatg ataatcatta aagctctcct 44640 ccattcttta gatcttctac tattaagttt agtatatatt gtactccttt atcattgtat 44700 tcaaataaag gatttataag ggacttactt ggagttacat aataaccaag ttctttagca 44760 aggattgggt ctattatttc aagtgtatgt tcattatacc ttctggcaaa tccctttctt 44820 actaaatacc tatttagttt tatcacacta actcctaatg ctcctgctat ctgagtagca 44880 gtggttaaac ttcccttcca tagttggggt ttgtcatcat taaggtcttt caagatgtaa 44940 tactttttat tgaccattaa attctcctcc ttctacattt ttatattacc ataattaatt 45000 tattatgtca aaggttattt tagaattttt aggtatatct ataacagcca gaagattttc 45060 tggtctatta ctatcataaa caagtatagt ataattctga gctcctaagt tattgaccat 45120 aacctcatta gctaacttag ataaattgtt aggggtaagg tccttaggac tattaagatt 45180 tagtttgatg ataagatcct ccccatctag tccagatgga gaagtctcaa taacattata 45240 gtctaaagca ttatgaagct tttctactaa cttttcttga ctttttcttt ggtgatgact 45300 ttctattagt tttgccccct gagttagcat tatccccatt gacaatgata tccctagaat 45360 tagtactact tccttcaagt ttcttcttcg cattagccat agcctcccta tactttatat 45420 tcatacattt tacataaaag tcttggtcac ttataccaca gatagggcaa ttcttgtaac 45480 ctccttggct tctcttaaat tcttgaatag tttgacaata tggacaccac atcttctttt 45540 taggtatttc ttcgccttca gggatcttat gagcttcatg gacatccacc caataaaatt 45600 cagcccaggg acattcttcc ataactttgt ctataaataa tttagctgat ttataggtat 45660 agaaatattt caatcctgaa tgataggtga ttttattctt cttaccccta gcaataatta 45720 ctactacaaa ggcctttaca tgattatgac ctgctggcct cttacatttt attatcatgc 45780 ctttttcctc ccaatattta gtttcttact tatattataa cctttatccg gagaatatac 45840 aatagtatct tttaaagtat ctagaactag attaaccttc tgtgaattta atcctgatat 45900 tgacactatc tcctccttag tatgagctcc tgattttaca gcctctataa ctttggcttt 45960 gtcctcaggt ttcacaatta cttcttttcc aggtattgct acttctacat gataatatgg 46020 atctccaaat tctcccattg ataaattcaa ttcaatttga gggaattgtc ctgcaagacg 46080 gaactctcta tccataatga ttttacttgg taaatggctt tgtttatcaa gattatctat 46140 atctacttct atttcctcat catcatcttc taagtcatca ggtcttttta gatatagagc 46200 agattctacc caaccatgta atatgaatga accagccatc ttttgaccac ctctagtttg 46260 agtagcattt ccacctttat tatagtggtg tattagcata actcctgtat gcttctcatt 46320 cttaagttta agacaccatt gtagtacagg attaagatca gctgcattat taagatctcc 46380 actaaacata agatataatg ggtcgaatat tactagtaca ggttttatct catcaattaa 46440 ttcttctatc tgttttctat gggattcttc atctaacata aatccctgtt gattaataaa 46500 agtgatagga agatctggag caaacctaac tttaagtctt ctcttacctt ttatatccac 46560 attaccaact actcctctgt ggctaattat cttttcagtt ctatctctca taatccagtc 46620 agcattctca ttttgaacta caattactgg gccaggttct agaacgggat attgacctag 46680 gaatggtctt ccactagcaa ctgatatagc taagtcctgt gtaaatgtgg atttaaatac 46740 ctttggttgt ccagccacaa ttccatgaga tcttctaccc cagaaacctt gtactaacca 46800 accagggaag gctccattat tacccataac atcctgataa ctatctattc tcagtttagt 46860 actttcagcc ttttcaatat cagcctctat ttctcctcct atgattttat ccaattcttt 46920 tcttaatctt gtttcttcat cttttcttcc cttatattta ttaaaagctg agcccttaac 46980 taataatata atctcattag gctctaatcc tatttcatgt aatttattct caatatacca 47040 tatagtagag cttctatcta aagaagtaat atcatctaaa gccagtaagt ctctaacctt 47100 ctttggtata ttatatttag catatatctt tctttcagat tgtgaggcct cagatcctcc 47160 tatattgacc ttaacactat cattagattt acttacagcc ttaacagcct tagcgatcac 47220 cttaggctta tatatctcct tagtgtgtat aggtcttttt atttttggtt tattcttgta 47280 tttgtagtta atagttcctg gtattctata tacatgagta acatcaaagc aggagtcaca 47340 gcctatatgt ttagctaatg ctggatttat ttcgtcatac tgattagcct ctatatacct 47400 atccatttcc catagcccct gatattttcc aggagaactt tcccacagat aacttggttt 47460 aggtttaatc cccaaggggt cagtatgctc atctatatct tgtattaagt atttagtctc 47520 tatgaagtta actatccttc tttgaggatt actatagggc attgggctcc aatataaatc 47580 ataaccctta tactgttgtt caaagtcctt tagttttttg tcaatgttat tcttattata 47640 tttgattggt acatctttcc attctttacc ttttctagct gctaggataa tgtaatctcc 47700 attttcacat tgtttagaga atatatcctt caagaacttt atgtccatat attatacccc 47760 ctattataag tctatattat cccaacctaa ttctttcaag tatggaacta cttctcctct 47820 tggctttaat tttctagtgc cttcctctat ctcagcgaat actaatcccc ctatatgtgg 47880 tttagtcccg ttaactactc tcattatact aacagccgat actcctaatt gtttagctgc 47940 atatctgaag gatctatact ccctattatt gttgtaacaa tatactgctt tagtataagt 48000 tgaatttcca cctcttgcca tttatacatc cccctctaac atctgtttct aaaataagta 48060 aaaaatttga ggagcaagtt tgctcccctt attatttgat gagaattatt cttcatcatc 48120 agcttctaat aattcaatta tctcagctgg tttcattttc ttattaactt ttattcctct 48180 ttctttagca agagcaatta agtctttctt agtcatttca gagtagtcag gagtttcatc 48240 ctcttcttct tcctcttcgt cttcttcttc accttgagct tctttgatta aagcgattaa 48300 ctcatcttta tcttttttct tagctccttt gatacctaag tctttagcca tagcagctaa 48360 ttccttttta gataatgatt ctaaatcatt ttcatcttct tcttctgctt cgtcttcatc 48420 aacttcttcg aagtcagcat ctactacatc ttcatcttcc acagcatcta tttctgcttg 48480 gtatgcagcc tttacagttc ctacagccat ttctaatgat tctaatgctt cctctacagt 48540 tatttgtcca tctgtgtcag gagttctttc atctaattcc acaatagctt ttaaagcctt 48600 tttaacttct ttctcatctc cagaatttaa agcagattgt actattacaa ttaactcctt 48660 attatttgct tcctctaacc attttgcttt tccttttgcc ataataatca ttcctccata 48720 tttttaattt ttaaataaca aatttagtat agtttgaaat taactaatta taaagttttg 48780 gttggtgttt gtaatttgat tacaatacta taatatcata tcctatatag cctgtcaacg 48840 agttatttag aaaaaattaa tttattataa cattcattac aaacggcttc atattcaaca 48900 ctttcgggct ggttatcaat ttgtatctgg ttaccctggt cagtatattt accttgatat 48960 ttcaatattg agaatgtggc ttttttacca cattgacaaa tgttctttaa ttcttcaatg 49020 ttatgggcca cttgtaataa tcgggtactt ccttcaaatc cttccctttt aaagtcagtt 49080 ctcaatccat aacatatgat aggataatgt aatgataaat ttaacagatc atcaacctgt 49140 tctctggata agaattgtac ctcatcaatt aatataacat ctactgccca ataattagac 49200 tcaaagtcag aaagtatttt tgataattta acgttaggag gtagaagaac atctaccttt 49260 cttgctaccc ctattctact gattatacat ggtccgcctt tagtatcaat tccgggctta 49320 actactaagg gcttcattcc tctttcttca tagttatgtg ctacctggat taactgagta 49380 gtctttccag agttcatagc actataccta aaatataact tattgctcac aactgatcgc 49440 ctccattata caccatacta ttacaagtat gcctaatatt ctaaatgccc gtaataaata 49500 tcccaagatt atcactcctt tgactcctcc tctaaatcaa atatacctgg tggtactaaa 49560 tccttaatct ctcccattat ggtttcatag ttaagtgttc ccttctcaat ttcctctgtg 49620 tgtttactca tctcttccac aaacctagtc attctagtct taccataacc ttccttcttt 49680 aaggtatagc atgataaggc taacatgata tccacagctt taagaacatt aagatttttg 49740 ttctctcttt cagctcttct tcttgcagct cttgctcttg acataataat ctactccttt 49800 atatatttag tttctaaaaa tattgatact attatagcta gtatcgcaaa acctagttcc 49860 atttcttcac ctcaaaacct ataaacactc ctagtaaaaa tattgatact agggttatta 49920 cccctagcac catttacttc tcctcccata acaggtcaga ataatcagtt ccttggtcat 49980 atgagaatat taatcctttc tgtgaactga tataatcagc caagtgtact attctctttt 50040 gataagtatt gattgggtac tctccccatt gaccatgatg agactcaatt agttctatta 50100 ccttttcata ggttaatgga aactcttcct ttagtggtaa tagtgtctct ctaaccatat 50160 atggatgctc cttagcaggt ttattaatat catgtagtat cagagcagcc tgtatcagat 50220 ccttatcttc ctgacatatc atctcagctt cgcataatgc tttacctata taacaagcag 50280 ctttagtatg tcttaacaaa cctcctaccc ctgcactata agttggatga tatttcccag 50340 tactacttgc aggtattata aagaacttct catcaactat ctcatttagg gcctttctta 50400 caaaattttg tatatcccta tctcttatat catgtatttc gggtattagc attctcatgc 50460 ccatttctct tatcataact attcctccta tcccatgtag tagcctctat attttatctt 50520 ctcccactca tcaacagtaa atactaattt agttctttgt ggtatgttag agtaacaaag 50580 ataatcaagg tctacttcta catattgttt acctttagcc caaggtttat tatcaaattc 50640 atggtactct cttacactgt gtaccttttt tatataatgc tcaagtaggt tcatttcttt 50700 ctcctccttt tctttttctt actctttcca tcaagtcctc gtttaacatg aaagtcaagt 50760 aagttttgag cccaccttag catcttaata tcttgcatac atgaaagatt aaagccagtg 50820 gatccatcaa aaactttaag gtcattccta aatggcatac tgtttactaa acctggattc 50880 tcattcttaa gtttaactac ctctctggca gtctcatccc attgttggtc tgtccagata 50940 ttgttgtcat acctataata tattattgaa tgaagacaaa cctgtaacca cttctttcgt 51000 aattcaattc tttctggagt ttcttcgcag tcatacttga gaattaacat agcacttctt 51060 ctccccaata ttatcacctc cttttctatt ataccatatt cttttacctt ctacaataga 51120 aaaagtttga ccaatatgtt ctggttcaat ttcctcaatg gtaagtgttc ccccatgctt 51180 acatagtatc ttagctaata cttgtcccat gtctgtatct gctaaagata ctccactatt 51240 ttttctggtt attacatacg acttcataac ttagccccac tctcaaattt attaactgat 51300 tatttatctc ttgtatttct tttaacttct tctcagtcaa tcctaagaac tgtcctaatt 51360 ttatcttctg ttcttttgtt aaatgacaat actgtttctc aatcaaatat ctggcatagt 51420 ttaggttctt tcttatctca tcagtatttt gcttatgctt ttcctctatc ccttccagtc 51480 tctttaataa ggatgtgttt atcatgtgat acctccctaa taagatattt tgttatcttt 51540 ggctttctgt cactaaagtc cattactgtg gctacatccc catcgatgaa tactactgat 51600 ttaccaccaa acttatcttt taagtcattt ttattcttga ctattttacc attacataaa 51660 tacatgatat acctccttta aaggcctcta gcagctcctg taaggtgtct ctaccacatt 51720 ataagtaaaa tcacattacc acatatctaa aaggccttag aattgattgt gggctcttat 51780 atactgtatc ttatgatagc tctattgcca ataacctcat ataaatgagt attatcctta 51840 tataagtatc ttggtatgga tttatcctca tatataagaa tacagtactt aggtacttga 51900 gatactaatt ctccgcaatt cttatctatt gctctagata gaataaatct gtcctcttcc 51960 attattatga agaagtcctt gttttcaatg ataatttctt tttcattaga taatataact 52020 ttcatcatgg ttatccctcc tttactgtca ttataagaga agtaagta 52068 <110> Optipharm.CO., LTD <120> Novel Clostridium perfringens specific bacteriophage CP3 and          antibacterial composition comprising the same <130> 1.37P-1 <150> KR 10-2016-0178172 <151> 2016-12-23 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 52068 <212> DNA <213> Unknown <220> <223> Bacteriophage CP3 <400> 1 aagtccttgt tttcaatgat aatttctttt tcattagata atataacttt catcatggtt 60 atccctcctt tactgtcatt ataagagaag taagtacaac tgtcaacgtt cgggagaact 120 ctactactgc tctctctctt actctctctt tccaaccaaa caaaatcaag agctcgatcg 180 ctgaccgtct gggtttttct atataagcta aatcttggac tacagaaggt gaactccaac 240 gttgcggaaa aatgcttccc aagctgtaag ctactccaag gtactttctt gagatctgta 300 agctatacct ttctctctcc aagctatacc ttcctaacct tggttgctta gaagctccaa 360 gccttgggca tggatatgcc tgccgtagtg taaacgaagg caaggcaata gcctgccaat 420 tatgtaatca ataaaacata gttatatata gcctttggct atatactata tataataata 480 tgtaattatt tatatatagt atatattgta ataatatggg gtatttctga tatccctcta 540 cgctagtgat accaacaaat ttgattacaa aaattgtaaa ctctatatgt gatatcaggt 600 gtaactatgc tgatatgcta cccatatcag aaaaaatggt actgatatgc ctatgtggac 660 tttgacatat ttatcttata tgtgtataat gtaagcatag gaggtgctat aaatgataga 720 taattgggga gaattaagaa agctaatcca gaatagagaa ggtagatctt tgaatgagca 780 tgagcttacc agagctgtta ctcatctctc tttcattgtt tctcaactaa aaaatgagaa 840 taatatgcta aagaaccaga taagagcagt gtctggatac ccactaatta actttaaaga 900 tcctaaagct aaggtagatg agttctcttt atatatgtat ttacaggaat tttggcctga 960 agtattccat gactataagg ataagatcca gagtagtaag aagttcaagg ctaaaaagtt 1020 tactaatcta aagcaagcta atgagtttgt atggactgaa ttgataaagc caaaggataa 1080 tgagagccta tttgtattaa gtaaatgggt agaaggtatg ggtatatctc atcacttcag 1140 aactaaacag gagtttatta ctgcttgtct gtgctatatg gactataaag gatatctgga 1200 caaggaaggt aagaccttta ataagctagg agacatcaag agaggaagct tcagatttaa 1260 ttctgatgag gaaacctatg caatactaac caatactttt gatactgtgg gtaaatataa 1320 atctggaatt gagaaattca aattaaaaat aaaggagtta aaagagaatg attaaaagaa 1380 ttatgtcatt actaacaata attgtgatag gagggctact gttaaccata cagagcctaa 1440 acaatcattg gggaaatgct gtccaagaat atattgagtc gaataaacaa ttaactgagc 1500 agactgaaat actgatggat aatgtagaag gattaaagta tcagaatgat aaactgagaa 1560 ctgataaaca aaatctaatt gagatgataa attcatttaa aggaatggag ttcaaagttg 1620 aagttactta ctattcatca gatgaaccta cagatgtggc aggatatgag accaagaata 1680 gtataggagg agatcttaag gttggtgact gggcagctcc agactttatt cctctacact 1740 cattacttca agtaacatat gcagatggga ctcaagagat tgcttacatc attgacaggg 1800 gtaaccctga tgtcattaag gtttttgata gagaagacaa aagaattata caggactcaa 1860 caatgagact tgataagtat gtccatgatg tgaatggcaa gggagttgat ggagcaacaa 1920 taaaaatatt aagatgggga gaataacatt ctctcttttc tttcgggatt tctattgaca 1980 ctctctggac tctctcttat aattaaagca taaacaaact gaaggagatg ttaaatatga 2040 attataataa aatgactaac tgggaaatat tatttgaact tgaggagaac ctattatgct 2100 ttggatataa attattaaga agagaacatg aagaagtaat acaagatata agagatacac 2160 aacctaagga tctatacaaa tatgctcaat cacttattaa ccaaactctt gaagaagaat 2220 atatctgtgt atactcaata agataatgtg ggaggtattt attacctctc ttttttattt 2280 gacagagtat gacagtatca cttataatag tgttgaggag gtgatactat ggataatcat 2340 tcacttgagg ttatagaact tcaatctgaa tacagagaac ttgaaagaag atggcaccaa 2400 ttcctggatg gccatgctca ttgcgaagaa gaagctatcc aagttaagga agagaatgga 2460 gaagatcaag agaactttac gaaacttgaa ctaattaagg atttttccag aactcaaaaa 2520 attataaaaa ggagggttaa tccctccctt atttttagaa aacctattgc agtctcttta 2580 gaactgatac aatatgtctt ttctctcttg gtgtaaacat ccaataataa ttgtgatttg 2640 ctttattctc atgtgatctt aatagctttc tggctttcct ttcatttatt ccaacctctt 2700 tggcaatatc tcttaatggt attaagttat tatctttcat ttattccaac tccttgattt 2760 tgtttttata gtctatatta tgctcagccc ggaagatatt atacgaattt cttcgagaaa 2820 atctttgaca aacttccgag gctctcttat aataaaatca acaaataaat acaaaaagga 2880 gaactgatat gaaaagaaga tatatcgttt tacaagtcac agttgatgtg gattggaacc 2940 ctgatgttag tgtgttagca cataacaaat atttaggagt acactgggta tctgttaagg 3000 atgaacacaa ggaagatata atctattttg atactgagga agaagctaag attgaagcta 3060 agcatagacc tggagctgta gttcaggcca catggaggta gataatatga aagtaagatt 3120 agtgattgta acatcaaaaa ggatcttgac cagatatttt gaagttgata acaaaagaca 3180 ggctatagga tatattccta aagtattaca ggaggaaggt atatctgata aggttagaag 3240 atatgaagtg attgaggttg atgagaacaa cagaagaata aaaagatatt aaggggagag 3300 atgatctccc ctttaacata gaaaggagaa ttaacatgaa agagtatact tatgtattat 3360 gtaagaatga ccaaggacaa cagtataaaa ggtatgtgga agtattatgt gagtttgatt 3420 atatgggcag gaccttattc attcatagac ctgtaggagc ccctcataat acagtatgtt 3480 atactatatc agataaggag acaggatgct gtcttgctcc taataactgc tctataaaga 3540 caccttcaaa gagatccgca gtaagagtat tgaatgagta ttttagagcc aatgaaatac 3600 atgagtggga tatgaagaga gtcagaaagt tgaaaaagga tttagccaga gctccaaaat 3660 atcataaagt caagaaaacc ccagagccta aaaaagtaaa accaaagcca gcccaatgta 3720 ttgatctgga ttatggtgat atacctttct aaagtttctc ttgacatatg atagggaata 3780 gtatataata aaattgaggt gataaactat ggaaaataat aacaacagaa taagcagaga 3840 aacactttac atggaagttg ctaaacttat gacaggaaga tctacttgcc taagaaaagc 3900 agttggttgt gtattagtta gagataacag gatagttgct acttcgtata atggagtacc 3960 ttcaggagta gaacattgta aaacttgttt aggtcctgga tgtgatattg tgatccatgc 4020 agaagctggc ttaataagtt actgtgctaa acatgggatc tctactgaag gaactactat 4080 gtatgtaacc ttatccccat gcgaaagctg tgctaattta ataatcaatg caggaattaa 4140 gaaagtggta tatcttgaga aatacagatt aagtggagga atagctaaat taatttgtaa 4200 tgatgtggag gtagaacaat atgagtaaag aggtatctaa tatattagag gatgagaaat 4260 taagagagtt tttgaatgag gaacaactag aagctattct aaatatagct aaagctggta 4320 ataatgataa agaagaaaat cttgagaaat ttgaaaagct attggaagag aaagaaaata 4380 tattaggagg tattagccat gagtaaatta atatgtaaaa gaagatgtta tagaagggat 4440 ccagcaggta actgcttagc atatacaatt ccatatgggc caggagaaaa atgccctagc 4500 caatgtaata atccacatca attaaagaaa gagtgtcaag acatcatttc taaaaatcaa 4560 cctaacaacc caatagtaaa agaatgtctt tacttaatag agaagatgga tctaataatt 4620 ggggttgaga ttaataaagc ttatgaagaa gataagaata gaggttcagg tggaggatct 4680 aagaactgta aacctaatac agcagctcta aagcaaaaga tgaaagataa tagacctatt 4740 gagtgtaagc aaactaaaga agagaataag gaatatcagg aagaattaaa acattggcaa 4800 gaagaccata atgaaaaact agaagttcat aaatcagttg ggtatggtat gaacaggggc 4860 aaggtagact catatactgg tatagaactt gatactaaag gtaatccagt atattcccca 4920 gaagaaatac aaattgcagc tgataacctg ctacaggaag gaagaataga tgtggatatg 4980 tatgaaaaaa tattgacagg tgaaataaaa ttataaaacg ttgacataat taatccagat 5040 gtggtataat agtggtataa atccatagtt aattttgcca tttggttggt attactagca 5100 ttaaggttag tgtgccaacc atttttcacg tagtaaagtc tggattttaa gggggtgata 5160 ttgtattatg agctataagt catacactga accagagtac tgtactcctg aacatttgaa 5220 gaaagccaaa aaattgatat ataatgccct gaaggctgat ccctttacac atcctaaaca 5280 cattgaga agagcagggc ataacatttt tagaagaaga aggtgatatt gtggcattga gtagcaaaac 5400 aaaaaagact attaagacct tggctgagaa gtatgatctt gatgagaaag aagagttgtt 5460 tgtatttcat tacatgaagt gctttaatgc aactacagca gctgctaggg caggctatcc 5520 tgtgaataat tctcatgcac atgggtctaa actattaaag aagaagaaaa tacaggaagc 5580 tctaaaggct atcagagccg atagagatga agaattacta tgcgatggta tggatgtaat 5640 aaggcagtac atgaaaatag cctttgcaga tataactgat gtgtgccaat tcaaggatga 5700 tctagtccta ttaaggtcta gtaaggctgt agatggtact ttaatcaatg agattaaaca 5760 aagtaaagat ggcgtatcag ttaaaatgga agatagactt caggcactaa aaagacttga 5820 ggtatacttc ggattaagta ctgatgtgat gaaagttgaa cttgaaaaag ttaaattact 5880 tttacaaata caggaaaatc agaataatgc tggtaaagga aaagatcttg aggttcttag 5940 agccaaaatg gctgatagaa agaaaaagaa tgcaggtaaa ggaaaatcta atactactaa 6000 gtaaaaaggt attacaggac taaataaagc actttactaa ggaggtgtat ataattgttt 6060 gatttagata gaatgattga gaaattagag ctagatgctt tggttgataa gttccacatt 6120 aacaaggaag ccctagaaga aatatccttt gaagaattag tagatctata ctatgacgat 6180 ccttgtggat tctcagaaga tatactagga gtaaacttag atccacatca aaaggatatt 6240 ttaataagtg taagagacaa taagagaacc tctgtaaggt caggacaggg ggtaggtaag 6300 actatggcct gtgcttgtct tataatttgg tatatgacct ttaggcctga atgtaaggta 6360 attgctacag ctcctacatt ttcacagtta acgtctgtac tatggcctga aatatcaaaa 6420 tggctacaag gaactattgt ggaagattat tttacagtaa ataaaaccac tatgactaga 6480 gt; cttggtcttc atagtgagtt catgctaatt atatgtgatg aggctcctgg aatagatgat 6600 gagatactag aagcattact tggtactata tcagatgagg aaaataagat aattttgatt 6660 ggtaaccctg ttaaaaatac aggtatcttc tttgattctc accacagtct taaggaaaat 6720 tttaaatgta ttcatattga cgctacaaaa gttgatagaa ctgataaaga aaaccacaaa 6780 atgttaagga aaaagtatgg ggaagagtcc aatgtctaca gatgtctggt aaaaggggac 6840 ttccccctag aagaagatga catatttata caattatcca cagtatatag aggtatcaac 6900 agtgaaataa ccagacctac agaaccagtt accatagata tgggtgtaga cgtagctaga 6960 tttggttcag ataagacctc aatcatgact aaaattgatg gttatgtgga agaacctatc 7020 tattaccaag gtaaggacac tatgaaaaca gtaggagaag tgctaagaaa ggctaaatct 7080 ctacatggtg agttccctaa actagcagta gttgtgaaaa tagacgacac tggtgtaggt 7140 ggaggagtaa ctgatagatt aaaagagata aaattcgaag aaaaactgga ctggttaaag 7200 gtagtaccat gtaagtttgg tatgccacta aagagagggt cccaggctta tgccttttat 7260 tatgatacaa ccacatatat agcctctgta gttaaggaat tactggagga tgatgagtta 7320 agactaccag ataatacaga tcttgtggga gaattgacta ccagaaagtt taaagtattc 7380 aactccaaag gtcaaataaa gctggagagt aaggatgaaa tgaagaaaag aggactacat 7440 tctccagact tagcagactc tctattctta gcttgccttc ctgttggatc taaattaaga 7500 aaaatctagt aactgatatg cctattaaag ccaataattt tttattctga tatgtgagat 7560 atcagttttg aaattgacta ggctctatat tagtaataat gtgatataga aggcatatca 7620 gaattaccac ctttatatat atgataatata agtatactat atactatata atttatttca 7680 gt; tgcggaaaat cttccggaaa ataatataaa aataaaacgt tgacaaattt ttgtgagtta 7800 tgttataata aacgtatcaa attatagtta tttttaaatc atgtgaatgg ccgtatttga 7860 cactatatgt ggagttgatg tagtcctccc tgaaatatga ggccatctgg attttataat 7920 ttgatatata atgtaagaaa ggaggtaaag taactttgaa aaagatatcc gccagaagta 7980 tatgtggaga ccctatccaa aagtctttgg atccagtgga taaatccaat caacaaacca 8040 gtgtacttgg aattgaagtt gatgagatat cccctgaata tgacccccaa ctattagaag 8100 tgattataag ggagtctaca ataatacccc aatgcaaaaa ggcctaccga aataatattg 8160 ctggattcgg aataggtatt aggtatattg agaaggtaga tgaacctact ccagaaatga 8220 aagctgaata taaacagctg gaagaattaa ttgataccct gtcttttgat aaaccaacca 8280 agaatgtctt tgaagagata gttgattgga gagaaggact tgggtattct tttttagaga 8340 tcgtagaaga tcctaaagac tcagctctcg taaaagagat aaaagaaatt gaagacccta 8400 agacaattaa aattcttaag ctagatgatg agagtacaga gtacactgta acccgatatg 8460 gtaaacagta cactagaaga aaaaggttca gaagatttaa gcagactgta aataatcaat 8520 caatcttctt taaggaatgg ggagatccta gagatctaga tatgaatact ggaaagtatt 8580 atgaaaaggg agagatacct cttaaagatc gagctaacag tatacttcat tttaaaacag 8640 gttatggtgt ttatggtaca ccaagatggg aaggtcctat gttagccgct cagggttcaa 8700 gaaaggccga attattaaac tggagatatt ttactaatgg taaacatact ccattaatgt 8760 tactaattca aggtggtact cttaatgata aatcttggaa tgagttaact gagtacatga 8820 atagtgtaaa aggtgaaaat tcccagcatg gtttcttagt attagaaact gaagctgcag 8880 agaaagacac tgcccttgat gattctaagc cagttactgt tgaggtcaag cccctatctg 8940 aaatattaca gaaggatgag ttattccaag actacatgga taatactagg aaaaagatac 9000 aatcttcttt cttactacca gacatatatg tagggtactc tcaagattac aatgtggcta 9060 cagccagaga ggctacaaga aaaactgaag agcaagtatt tgttcctgaa agagaaaacc 9120 taacatggat actaaataat gttattctat ctggctatgg ttttaaatat gtggaagctt 9180 actttaagag tcctaatatt gagaactctg actatcagaa aggaatgttt gctatagcct 9240 ctagagctat gactcctaat cagattaatg aggagtacta ttcatcaatc aataaggact 9300 atactcctat tgatgaagaa tgggcaaata tccctactta ctcagttgcc tcttcaatga 9360 gtaataactt aacaagtata attgagaagg ctcaaaactc taatgggagt gatgaggtga 9420 tagctgtact taaagaggct aagaaacagt tattaaacta tcaattaaaa ggaggtaagt 9480 aatagtgggt actgatgagt tgatcgctaa aattgataag atcttaaaga atgaagatga 9540 tagccttaag aaagaattga aagaagctgg ctatgaggag attgattata caatagaaag 9600 gatctctgaa ttagaagaca ctatttctga tctccttaca gaagaattaa aacatcttat 9660 aaaagatgga gactttgaaa ccattgaaga atttatggaa tatgtggagt ctcaaatggg 9720 tacagataat atcgatactt ctttagcaga aactcttaag agttgttatc aagaagctat 9780 agaaaattta ggagatactt atttacaagg tattgataag gacttatccc taaccaaagt 9840 cttttctcca tatactacta cctttataga tgagtggtct tctaaactag gggaaatgat 9900 gaaattaact tcccatgaaa aactacagaa tattctggat aaggctcttg aagatggaga 9960 gtctgtagaa gatgtagttg gtaaattatc tgagacttat ggctttagta gaactagagc 10020 tagagctact gctattactg aaatgttaag agcccactca tatgctaagg atgaggcttt 10080 taatcaatgc cccgcagtaa caggaataca gtggaaacac tcagggggaa ggggtattca 10140 cccaaggcaa caccatgtcg atctggatgg tactgtcatt aataaaggtg agtacttcac 10200 tgtaggaatt tatgaagcta gatttcctag ggatatcaac ttaccagctt ctgagactgt 10260 gaattgtcat tgtactcatg ggcctgtaat tgataaggat atactaggat tatctaaaga 10320 agaaaaagaa gctatcagag atcaagtgat agcagaatat aatttgtaaa ggagggagag 10380 taaatgccag taaaatcagt taagaaagct attgagatct cagatgctaa aattgattac 10440 ctttcattag ttgataaagc agctaataaa tggcaatgga ttgtggctaa agaacttgga 10500 gctcctaata gatttagata tgataagcca atcttaaagt ctgagtctga aggagataat 10560 cattttgtta tcggtgtagt atatgagcca atgactgctg actaccatga taattttatg 10620 actgaggaag agatcaaaaa agcttgtgag tggtttaatg aaaatggcct tggtagtgat 10680 ttacaacaca atgagaacac tcttgagggt gtcaagattg taaaatcatg ggtaactgag 10740 gaagatacca caatagaagg tacagtagtt aaagctggaa cttggttagc taaagctgag 10800 attatcgatg atgatctatg gagcaagatt gagaaaggcg aaattactgg atggtctatg 10860 ggtggtactg gtaaatattc tacacaagat gtagatattg atgagattga gaagtctata 10920 ggtcaatcta tacttgataa aattggaaag gccctagggt taactccaaa gcatatacag 10980 aaaggtgcat actctgaaaa atataaagag actgcaaaaa gaaataattt ctggaatgca 11040 gttagtacct tagaagatat tctatatcca tattctagtt gggattacac tagatctttt 11100 gaaagtgatg tagataagat aaaggaggct cttcaagaat tttctgaaac tatggaggga 11160 atattattgg gaactccaga agaagtagct aaatcttttg agttaccagt caataaggca 11220 ggtaagaaaa tatcttcaaa aaatgtggaa aagttaaaaa cagcacaaag tgctattaat 11280 gaattattag actcagtgga gggggaagaa gagatgaatg aaactcaagt acaaaaaatg 11340 atagacgaag ctatcaaaaa ggcattagat ggaactaaac ctgttgaggg tgaagttaat 11400 aagaatgaag gtcaaccagc tactactgaa gatatagttg ctaaggctgt ggaaaaagct 11460 atgttagagg ctggcttaat ccagaaacaa gaagaaaaaa ctgaggaaga aaaaatagct 11520 gaggcagtag ctaaagccat ggctccatat gtaggaagta aacaaattgc taaaaacgga 11580 gaagaaggac aagaagttca aaaatcagta tttaatggat tatttggtgg aaaccaagaa 11640 taataacaat taggagggaa agttatgtca aaagtaaata aagctgtatt taacgctgcc 11700 gtattaaatg gtggaggtaa attaaatcca gaacaagcta acgcttttat taaaatgata 11760 attgatgagc caacaatatt aaaagaggct agaactgtac caatgaaagg agactctaaa 11820 aagattgagt caataggatt tggtcaaaga gtattaaatc ctgcagctga aggaacagat 11880 ccaggacaat ctaaatattc agctccaact actggaacag ttactttaca agctaaagag 11940 tttatagctg tggttaagtt aactgatgat actatagaaa caaatataga aggaaaatca 12000 atagaagaca ctgtaatgca attattagct gagagagttg ctcttgacat ggaagaaata 12060 attgtaaatg gtgatactgg acatagtagc gatactttct tacaacaatt agatggggtt 12120 agaaaacaat taactgctca tgtagtagat gctaacaaag ctgctttaag taaagaccta 12180 ttaaaacaag ctaagaaagc tgtaccagct aagtatttaa gaaacccagg tgaatggaga 12240 ttctactgct caaatggttt agaaactgat tgggtagact tagtaggaga aagattaact 12300 gctaatgggg atgactcatt tataaatggt actgttaaat ctgcttatgg agtacctgtt 12360 aaaggtatag ctatgctaca agattatgac gactcttcta ctcaagtatc agacataatc 12420 ttaactcacc ctaagaatat agttataggt atctcaagag atatcagaat agagttggaa 12480 agagctccta gagaaagatc tacttatgta gttttaactc ttaagatgga tgttaagttt 12540 gaagatccta aagctgctgc taagatagtt aaagtaaaag cttaatcatg aaggcccact 12600 gtaaatgtgg gctctttttc attagaaagg agagatatta tggctaaggt caaattagca 12660 aagggtaaat tacaaactct ggttattaaa ggtcatagaa ttacccaaga ccatgtagaa 12720 gaagttaaag atagtctagt aaattattta aaacagaact atgatgtgat tgtcatagat 12780 gaacctgttg aggatgaacc tctagaagat tttactgaag aagaagtacc ttctacagat 12840 gacaataaag aagaacctgc tagaagattt acaaacaaga ataaaagaaa cagatagggg 12900 tggctcatat gaatattact ccggatgaaa taattgactt ctcaagtaat cctgagatta 12960 ggaaaagaga ccccaaaaat attgagatgg atatattaag agctaaagcc aagatctcag 13020 gaatttttaa aaattcaagt aaagactatg atactattgt ggagaaaaat cccaatagta 13080 aagaattaag gttagcccat atactgtatg ctgagtatta tggactacaa tccttggtca 13140 ttagcaaggc tgatattcaa tccgagacct atgatgacta ctcatataca agaagaactg 13200 ataaggctgt tatagagcct gatattatgc ctctaattaa acctttctta gaagatgata 13260 gtccatctat cagaacaact actatgagat taagggtact gtaatatgat tagaaattca 13320 tataacaagc tcttaaatca tagatgtaat atataccacc tattagaaga tcctgaagat 13380 cccggatatg ggctagccca tacccctagc tttagttatc ctgatgagcc agacatgatt 13440 gaggtaccct gtcattttaa atcaggaact ctagaaaaac ttgagtacaa agaacctagg 13500 tttaagatta cccaggagta ctcagtagat tttgctattg gaactgatgt tagacaaaat 13560 gataaaattg tggaccttga gacgggtatc acatatatag ctaaagctcc tagggatata 13620 agaggtctaa aaattaaggt tggtatagag gctaaggatt ttagatagtt atggcttcaa 13680 attctggggg catagaaata gatgctaaac aaatagaaag gtttataaag aagctaagag 13740 atgccgctaa tagtgatttc aaaaagcaat tagcagtatg gtttgaagct gcagggatag 13800 acttccttga tgtggttcaa gatgaaataa ttagactaca agtagtagac actagattat 13860 tacttaactc ctttcaaaaa gggggtaaga atggtatatg ggtaagtagt gatggaggac 13920 taactttaga agttggtact aacctaaaat atgctaagtg ggtaaacgat ggtcactata 13980 ctaatccaca aggacaatca tttagattcg ttcctggcat atggtcagga gataaattta 14040 tttatcaacc aggagctaag actggtatgg tactaagaca aaagaaagta cctggtaaac 14100 cctattggga caatgcagta aaaatatatg aaaaaatatt tgaacaggct ttccaaagaa 14160 aactagaaga ttggttagcc gaagttttag ggggtagtta ttaatgacag cagatatagg 14220 gagtattgct aaattcatat ctgatctaac tggaattact accctctatt ttgagcgaat 14280 acctgaaaag ttcaagtatc cctcattata tttcaagaag gtatttaatc ctcagaatga 14340 tactctttgt agttatagaa ctgagaatac tttaagagtt aaggtatttg aacctaagga 14400 ctctcttgaa attgctcaga atatagctaa tgctatatac ttgagaaata gaaaaattcc 14460 tatacttaat agggatggta gtaagaccaa caaaaatctc aaaataaaga atgtggaagt 14520 atttgaaggt gaaactgaag aagattctca ggtagagatc aactatgata ttgtggttag 14580 tctttataga gatccaactt acttcgaaaa ctttgagata acaaaggggg taaaatagtg 14640 aaagtacaaa aattttccat acctcaatta agagcccatt gtcaacaact tttccaggtt 14700 gaggagtttg tcttcaacgc tgcagttcaa tcccttccac ctgatgaaga gttaacaata 14760 aaacaaacta gtcagattat aaaagactgg ctaaagaagg aggtaaaata gatggcagga 14820 ggaactttta ttccaggaca aaataaagta agacccggta tgtatactag atttgtaagt 14880 ggtggactta atgagcctaa gcttaataat ctaggagtta tgatcttacc attacaactt 14940 aagtggggta atgctggaga tgttatcaag gtaaccaatg aaggtggcct agaagcatta 15000 tatcaacttg ggtatgacat aactcaccct actgtattac tggttaaaga ggctttaaaa 15060 gaagctgcaa ctgttttagt atataaccta acaggtggaa ctaaagccaa agctgtagta 15120 gggtctctaa catttacagc tactaaagct ggagaaaggg gtaataaatt atctctaact 15180 attaagacca atcctcttgg aggaaaggat atattattat atcttgaagg atctaaaatc 15240 tttgaggcta caggagtaac tgaaatatca acagttaagt ctcaatggtt tacagtaagt 15300 ggaagtggtt ctatagatga gactgctaag actgagttta gtactggtac ttctactcag 15360 atgaataact ctgatgttat taacttctta gataactgtg agttatacaa ctttaaaact 15420 atggccttcc catttactga gagtgaatta cacactgctc tattatcaaa acttaaatac 15480 ttcagagatg aggttggtaa gaatattgtg gcagttactt ctgattttgc tacttgtgat 15540 tacccatata tcatcaatgt taagaatggg gttgtattag atggaggcac taagctaaca 15600 gcttctcaag caactgcatg ggtagctggt gctagagctg cagctggata tactgaagac 15660 cttacttata aagcatatcc tggagcagta gatgctaatc caagactaaa acactctcaa 15720 atagttgatg gattaaaaac aggtcaattt atattctcaa ctcaaactga ctcaaattct 15780 gaaagagtaa aggtagtagt tgaggaagat ataaatacac accatacttt tacagaagaa 15840 tgggatgaaa gctttacaga taacaaagtt gtggctgtat tagatgctgt tatagatcta 15900 ggatctcaga tcttagttcc taatgttttc cctaaccaca aagatggtat aaagctagct 15960 caggatagac taataagttt attaactaca ctacaagctc aaggagcatt aaagaatgtg 16020 gaccctgact ctgatgtagt ggtagatgaa gaagctagta aaggaaagac tatgtatgct 16080 actatactta cacaacctgt aagaactttc aagaaatact ttgtaactgt taagaataat 16140 taaggaggta tagcccatga gtgaagttaa tagaatgtct gctagggagg gtagagcttt 16200 tgtatcaggt aagcaagtaa ctagcttaat aaaactagaa gctctattta ctcctgaggt 16260 aactaagaaa agggttatgg gtcagaaagg attaacctca aaagttatag gttatgatat 16320 aactggaact ataactcaat tcaaagctac tccaatgata agagaagcta ttaaagaata 16380 cttaaaaact ggtatattcc ctaagatgga tatacaagct gtatgtgatg acccagattc 16440 tgactatgtt aagaattatg gtgaagacag agtacaatta ctaggagtac aattagaagg 16500 agatttaccc ctaattggac tagatgctga gggagaagaa attcaagaag aaatatcatt 16560 ctcagcttca gaagtaagat atttataaaa gattaagaat attgtggagg tagattatta 16620 tgtcaaaatt tagtatggaa tttttcatgg aaaatgataa gggtaaacaa tcacccgaaa 16680 ttgaagttg aggggtagct agatttagtt tccctgaggg tcatgaatta gcgggccaac 16740 ctataccttg gatattaaag ccaatcacta ctaagcaaat ggaggatatc caaaagagaa 16800 atacagtaat tactagaaac aaaggtataa ctaactcaat aacagaccaa agaagagcta 16860 ctagggaaat gatactagaa acaatagtat tccctgactt taaagatcca aaatggttag 16920 agtctactaa gtttgtggat ccagtagatt tacttatgga tgtactaagc ttaccagggg 16980 atttcactag aataactagt gaggttatgg atataaatgg attaggcgat aataaggatg 17040 atctaatcaa agaagcaaaa aactaataaa ggccgacact agtgcatttt gggctcatgt 17100 gttattccag aagcataata tactaccctg ggaatttgat aaactatctc ccaataggaa 17160 agcttttctt atagcttcat gtgaactggc tttagaagaa gataaaaaag ctagggataa 17220 attaaagaaa ggctcttaat cataaaggag ggatataatg gcaagtcaag ctgcatttac 17280 tgccatattc tctgcccaag ataagatctc tggtgcattt aaccagatgg ctgatagtgg 17340 taattccttc atgggtactc ttgccaaaat tggtggagta gctgctggag tattctcagc 17400 taaggctgtc tgggatttta ctaaggacag tgttaaagcc tttggggatt ttgagcaagg 17460 tttaaatgag gttatgactc tattacccaa ctctactcaa gagacctttg ataagatgtc 17520 cggccaggta aaaaagttaa gttctgatat agggatattg cctgataaag tagtacctgc 17580 tctctatcaa gccatatcct catcagttcc agaggataac gtttttagtt tcttagaaac 17640 agctggtaaa gcttctatag gtggtatcac atcaatagaa actgcggttg atggactaac 17700 tactgttatt aactcatatg gagctaagaa tatggatgtt caaagggcct cagaccttat 17760 gtttactaca gtaagacttg gtaagactaa ctttgagcaa ctatcacaat ccctattcaa 17820 tgtgctacct tcagcctcag ctgctggggt atcttttgaa gaagtatctg cctctatagc 17880 tgcattaaca gctcaggggg ttccaacatc tgtagctact actaagataa gagctgctat 17940 agatgagcta tctaaatcgg gaactaagac tgataaggtc ttccgggaaa tagctggtaa 18000 gggtttcaaa gacttcatta aaggtggagg taatatccaa gaggcccttc agatgttaga 18060 aaaacacgcc cagaaaagta atctgggtat caatgacctc tttggttctg tagaagctgg 18120 ttcagctgca ctagccttga ctggtgaagg tacggaggcc tttactaatg ctcttaatga 18180 gatgaaggat agtgctgggg ccactgaagc tgcctatgaa aaaatggata agggctttaa 18240 taggaccatg gagagaataa aaaccaaagc tgctgtagtt atgttagatg tgggagatgc 18300 cctagcccct gccgttgagg ttatagctaa tgctgtccta aatggtatca catttataca 18360 agaaggcttt gaaaaactag gacctatatt ctcaatggtt gtttctgctt tccaaggttt 18420 aggagagact atatattaca tcatgaatgg ttgggagttt ggggatgcta tgcaaccctt 18480 tatagatcta accaacgata tattatataa gttcttacct gccgatgtgg cagatcaaat 18540 taatgagaag atatggagtc ttggggaaga catatataat aattttggag agatcttagt 18600 aggtctacca gattatttat cagaggctaa caataatata caagaagcct ttagtggttt 18660 agctgagcct atcatgggag tattagctcc tataggtcaa gtacttcttg atatgtttgg 18720 aactttagct tctatattta tgaataacct aattccagcc atagacttag tagttcttta 18780 tgttcaagga tttactcaag tatggcaaac tgtattggaa gtgttaggtc ctatattaga 18840 agtattaggt tctatgttct ctgctacttt tacagctctt ggagaattga taaccaatgt 18900 tttcatgcca attgctactt ctttagctga ggcctttgct acttatatag ctcctgcaat 18960 ttctcaagtt gtggaagcta taggtaactt cctaatgcca atattacaag gactagcaga 19020 ttggtggaat aatacactac tacctatgtt cttaagcttt gctgagaatg tagttccata 19080 tattcaatca gctattgaga gttttggtat gatattccaa gggttggtgg atatgctaac 19140 tccactactt aaccttttag ccgcatcttt ccaaacagta tggcctgtta tatcctcagt 19200 tgttactggg gccgttcaaa ctatatcagg agtactacag ggattaatgc agatgttgag 19260 tggtttaata gcttttgtgg ctggagtatt cacaggtaat tggtctaaag cttgggaagg 19320 tgttaagaac atctttggag gtattttcca aggtatgtta tccctagcaa aagctcctat 19380 taatagtatt attggtatga ttaataactt aattggtgct cttaatacta ttaagatacc 19440 tgactgggta cctattattg ggggtaaggg tattaatatt ccccagatac caatgcttgc 19500 tgaaggtact gacttcttta cagataaagc tgccatggtt ggtgaagctg gtcctgaatt 19560 agttatgggt cctacattct caaaaatgcc tagaggctct caagtattac ctgcagataa 19620 aactcaggat ttactaagac aatctggagc aggaagtgat gaaggatcta aagttatgaa 19680 cattaacaac tcatttaatt tcacagtaca ggctgaagat ggaacaatct cagaaagtga 19740 attaatgaga gtatacagat ttattgtgga aatgttaaat caggaaatgt tcgagggagg 19800 tgaactggac cttgcatagt aaatatcaat tctggttaat acagggagat actaggttta 19860 ggttaccagt aaatcccact gatataaata tctctgatag tagagataat agtacagttt 19920 ccattgaagg cttaggggaa ctctctttca taggagaccc taagctaaaa tctttttctt 19980 ttaattttga gttaccaagc tcctactacg atggatgtga gtatactgat attgcagacc 20040 cccgagtaag ttatgctaaa tttattgaaa tgggtaaaga tccagtaaga tttactatta 20100 ctagtactga tattaactta ttagttacta ttgataatat ctcatatact gagaaaggtg 20160 gagatattgg tactcttcaa gtaaaaataa aactaactga gtataagaag cctgaggtta 20220 aatcagttag tttcacaatt ccaaaaccta aactagccta tagtggtaat acaggaggta 20280 gttcttctgg aggtactcaa ccttgggcag ctactactac taatgtttac tcatggctat 20340 gggtaagaac tggaccaggt atcaataatc ccactaaggg taagatatgg cctaatgaac 20400 actttacagt attaaggtct caaaatggat ggcattatgt aaagtataaa ttagataatg 20460 gttcaggata taaagaaggt tggagttctg gtgattatat aaggaggtta taatatgcaa 20520 ttaattgtgg tacaaggaga taaatccttt gatattacaa atacctccag tggtaagagt 20580 tgggcagggt cttcttcatc tgcccctcgt accctatcat tccaatataa tgctcaagat 20640 atacatccaa tttatgatgg agatcaggta gtactagtcc atgagggtaa gaatatcttt 20700 aatggtatta ttatgaatat ctcagataat gagaagaagc atttacttaa ggttaaagct 20760 tatgacccta tgatttattg ggctaataat aaagatagtt ttgtatttga aaaccaaact 20820 ggcgaagcta tatttgagtc attatgtaaa gcctttgggg ttgcccctgg agtagtagct 20880 aaaactggat atactttctc tagtttagta tgtgagaaac aaactcccta tgatattatg 20940 aagaaggtaa taaaagaggt ttatgaacaa actggagaac agttttatat ttactataat 21000 tctattacag gtaagtttga ttttataaac cgaagaacca acaatgaatt gtggaaattc 21060 gaatatggta aaaatattgg aactttttca agaagtagaa actttgatag tattgtcaac 21120 caggtaaagc tagtatgtaa atataaacag ggtaagggta aagatgatcc cgagattact 21180 ttatttgcct ctgctaaaga tgatagcagc caaaagaaat atggggtatt tcaattatac 21240 gatactatca gtgatgaact aaaccaagca caattaaatg agaaggctag aataaagcta 21300 gaaacaggta agagacctaa aactgatatc tctataactg ttataggtgt acctgactgt 21360 attagtggta aagccattgt ggttagtata ccacatttaa aaattaataa ggctttctac 21420 attgagtctg atacccataa atttgatact gactacacca tgaatttaaa acttaaatgt 21480 actgaggagg gtgatattta atggctaata atttattaga ttgtgtaaaa caagtatata 21540 gtaatatgtc caccccccta aatatttata gagctaaggt acttagtact agccctttaa 21600 aatgtgagtg tcttcacaat agaaaattag tactatctgg aaatattata gatgttccaa 21660 aacatattga gtctattagt gagggcgata ctgtatatct tatgccctat gatgatggac 21720 aaagatattt tatgttaggg atcttaggag gttgatataa tgctacctaa tggctttaaa 21780 gataccctta tagttgatga gaaaacaatg gagaatacca agacctataa actagatcta 21840 aatcttaata gaatttatga taatgtggat ggtcttgagg ccttaaaaca atctttatat 21900 aaaagattaa taactcctaa aaatgcctat atgatttatc aagatacttc ttatggttgt 21960 gatattagag ctttgattaa tgactctcta gtaactcctg aacttcttga aatggagatt 22020 aatagggcta tccctgaatg tctcttagaa gacgatagag tgactaaagt aactgatcta 22080 aagtataatg tgcaaggcaa gaatgtatat attgaagtat ggataaatac catatatgga 22140 gatacaaaac tggaaggggt gatataatgg gattttctga agatttattc tcccgaacac 22200 ataaagaatt attagaagag atgataggcc tatcccctga tgatattgac actagaactg 22260 gaagtttcac ctatgatatg gctatggttt ttgctgttaa gctagctgag ttttatgctg 22320 ttaatgttaa tcagatatta gactcagtat ttctaagtac tgctacaggt aagttcttag 22380 atgagagag tgccaatgta gggatatcca gaataattcc aactaagtgt agaaggtcag 22440 ctaagtttgt tggctatgaa cccgatgttg gttcaagatt ccaaggagga gattatttct 22500 ggcaagtgat agagcctaat attattgaat gtgagacctt aggttcagga cccaataagg 22560 tatcaacagg tacacatcta atccctatca atgctatagc agaacttaag agtgctacta 22620 tgggagatat acttgttatt ggtagagatc aagagacaga tgatgagtta agagaaagag 22680 ctaccaaagc cactatacaa ccatcgggag atggaaatgt tcaacagtta actgattggt 22740 gtaaaagtgt agctggagta ggtaaagctg tggttcttcc ctgttggaat ggtataaata 22800 ctgctaaagg tattataacc acagttgaga ataagcctgc caatcctgaa ttagttaata 22860 aagttcaaga atacatggat ccaggcaaga aaggtttagg tgaaggtaaa gctcctatag 22920 gatgtgtttt tactgttgag tcagctcaaa cctttaatat cactgtaaga gctcaagtta 22980 agaaattgag ttcagttaca gaagaacaaa taaaattaaa cttcaagaaa gaactagaag 23040 aatttctaaa tagtcaagta tttaaaatta ctagcctatc tataaataga ataggggcta 23100 tgcttttaaa tattgaggga gtagaagatt atacaaccat tacacttaat gggagtagtc 23160 aatctttaac tattcctgct aatcaaatac caaccattac tgaggtggtg gtaacttatg 23220 tatagtctag aacaggttca aaaagaatta ctattgtacg ttcctgaatt ttatgataat 23280 ataaaagagg ctgataactt agtaaacact attatggcta ctgaacttca gaagctatat 23340 actaagggac aggaattaca agactatgtg gatcccaaga ccactcctaa ggaaggtcta 23400 gaaagatttt tcactgacct tgaattatat aaatactttg attacccacc aactgaccaa 23460 gagaaaagag ctattattac agtagttcta tctgcttgga aaaagtttaa tagtacatct 23520 atcaaaaatc ttactaaggc ctatacgggt ggaaaggtaa aggtagagtt catagctgaa 23580 gagagtaaga tcaaaattag ctttactgac ctaccagggg taccacctaa cctaaaggct 23640 ctagaaagag tattcagaga aagaatgcct gctcatttaa tatatcaaat tatattactt 23700 tatattactt gggagatgtt cgataggtat gattatatgt ggagcaagtg ggacagtcta 23760 gaccttacct gggaaaaatg ggaagaatat gctataactg aataaaggag gtatgcaaaa 23820 tgccaagtac tggtaaaact cctactctag gtttaaataa gtgggagtta acagataagc 23880 caaaaatgaa tgattttaac aaggacaatg agaacattga gaacttcgct gccgaacata 23940 aacagaaaat gactgaggta gaaaagaagt ctaaatctaa tgagaatagg ctagatgctt 24000 taagggttgg tactattaat aggcttgagt ggtataattc tgagtcttat actggtcctg 24060 gtaatatcac atctattaca gatgctccaa ctaagagacg aggttttaaa tggtctggta 24120 tgtcaacagg tccacaaact cttgaaatta aaactctgcc tatagaacca aatagtgatt 24180 atgtattagc ctttctagaa gatattgaag gtagttttac tagctatgaa attaaactgg 24240 cttacactaa tgggggtcaa caatataaga gtattgtaaa acaaggtagt acaccacaga 24300 gtaatgggta caacttaaaa actcataaat ttagtattcc ttctggagct actgatacta 24360 aaattcaatt tgtggtacag ggtgcaacct caagcacatc attaaaggta ttaaatatct 24420 tattagctag gggtaatgta gtacccgact ataacctagc tcctgaggat gtgctaaaaa 24480 gattagacct attatttgag aaagacacta gtttacaaaa tcaagtaaac caaaaagctg 24540 ctaaaactga tgtactttta aaggttagtg ctcccctgga tagagattgt aattcattta 24600 aggatattaa tactttttgt acctttgata caggagatgg taattttaag aatacccctg 24660 atggtactct atcccaagga agtgctcagg tatttatggt agttaataga ggtcatcata 24720 gtactagatt acaacaggag tttatatacc tatatcccaa ggatagtgta gctcgctata 24780 agagaaactt tgacggaact cgctggggta actggtataa ggaatatgat agtgctaata 24840 agcccactgc agaagaacta ggagtagctg gggtgaatat gcccatggct gatacagatc 24900 taaataatca taagaatcct ggtttctatt ggggttatca aaatatggtc aatgcccctg 24960 tacccaatgg tgtagcaatt atggaggtag ttaagtataa taatgattgt gtattacaaa 25020 gatttacctc agttagagca gatagtaacc atactactta tattagaacc tgctattcaa 25080 ataaatggtc tagctggaag aagatttatg atgagggtaa taagcctacc ccagcagata 25140 taggggccat accccagata cccattagaa taactcagga tcttaacaca ttcactacac 25200 ctgggttata tattgtggat agagcaactc atgctccaca atcatatgga agattattag 25260 ttctaggttg ggacaacgac cagaagtggg taactcaggt attctatgct gatattacta 25320 ataaagtata tgttagatgt agcactaaca acactgctac tgaatggact tcttgggctg 25380 aacagtactc aactagtttt aaaccaactc cccaagacat tggggcttct cctagtaacc 25440 ataatcataa taaggtatat taccctatac ctaaacaggc tattactgat tttaattccg 25500 gtatagctat aactgagggt caacatctag tatcttctag tacagctata cctaattcac 25560 ctcctttata tgctcaagat ggtagtagta tatatggtat acttcatgta tatgttagtg 25620 ttggtgattc ttataatggt gtgaataact ggatatggca agactttcat gatactaggg 25680 gtaatcacta ctggaggtat aaaattaata atgctgactg gagtccttgg agagctgtat 25740 ataattctgt taataaacca accccccagg ctataggggc tgtagctact gaaggtgatg 25800 gagtaataaa tggaactcta gaacttagaa aagccgataa gctaataagg tttgctactg 25860 agagaggctg gattttaaaa cagggaggaa ctgcagcaga aacctccttg gatctagttt 25920 ctgaaactga tggaaaggcc tttagggtat tagaaccttc gggaaaaaag ggcatctata 25980 tgtgggtatc atcaggggga acacaattta atgtagatgg caattctgtg taccatcaag 26040 gatttaaacc tactccatct gatattggag cagcctcttc tagccataac catgatagtg 26100 cttacctggg taaaactgct acagctgcta acagtaataa acttaagaac tgggacttag 26160 ctgaagggag taataacttt gtgggtatac catttatacc ccatgatgga gtcatggagg 26220 taggtagaat ggttgacttc catgaacctg ggacaaataa agactttaat actagattag 26280 agtctattaa tggagctcta aaatgctggc aagatttctc agctgaaaac ttatgggcca 26340 gaaaatatat gagagtaaat gactggtatg gtggatctga agatggccga ttctactata 26400 aacaagatgg aaaaggacta tatactgaga atgtggaaaa cctctatgta aaaggtagtc 26460 ctgtggctaa aggagactat tgggatgtag gggataatac cagagtagtt agtctggggg 26520 gaggactaag attagttcgt caaatagtta agacaggcta tgctaactca ggtggaggtg 26580 taaattatac tatacatttc cctaaggctt ggaattgggt attacctata tctctagtat 26640 cacataacta taatgacgct tctaataata cctcaggtta ttgtactatt gactattgga 26700 ataactcata tttgaatggt ggatgttatc aaatggtagc tggtaagcca actcaaataa 26760 tactaaccta tttagctact gagtaaggga gttgataaaa catgagaata gtaggattta 26820 attattctgt ggaaactaat aaggttatag accttatatt ctatgatgat gaaaaagaaa 26880 tgctggtaaa tcaggcagaa gactcagaag atgtaagatc atttatagat aaagaccatg 26940 aagaagttac taaaatatta gacctaatta atagacctga acctctatac tatagagagg 27000 atagtgaaat attaaataat atagaagata ttgtatgtct ccatatacct cataattcac 27060 aagggcattt agcctttgtg gataatactg aggttattga tctagacaat cttcctgtta 27120 tacagatgac tataagtcta tttgaccaca taaagggtgg cccaactaaa tttgatatca 27180 gtaaaaagag atcagattta tacagtatca aagataagga cctatttact tcatatgagg 27240 ctatagatcc taatcctcta gatattaata aattaattgg ttatcaagcc caggctcttg 27300 ctaatactaa attagagcta atgcaaacta aagccataat gaaagtatta acacaagaac 27360 tagctgatat aaaaattaaa ttgatgaata aataaggagg tagagattat gaatttctgg 27420 gaaatggctt atagaattgg agcaataggt aaggacaaat tacatcaggc agtaaaatgc 27480 tcttctaatc cagaaggaga aattacacca gaggactata aaagaatatg tggtgaagac 27540 tttaatgaaa gggtgtaatc tatggaaaaa gaactactaa ctcaagttct tagccaaggt 27600 atcttttgg gtctatttgt atggctatta tatgacacta gaaaagaggc tagagaaaga 27660 gagaataaat taaatcaagt gatagataag ctagttgata aattcaacgt agttgaggat 27720 attaaagagg atgtagatac tataaaagat cacctattta gtaacaaagg ttaaattata 27780 agcctagcag gtcatttaag gcctgttagg ttttatataa gaaaatatat taaggaggtg 27840 gttaaaatgc tgtagcgagc ctctcaggcc aaataaaaca cttataatat ttaaaggaga 27900 tgtttattat gataatcaat aaaagattaa gtactactaa tgttacctta aacgctaata 27960 atccagccta tataattatg cacgaaactg ataacactga tagaggagga ggggctgaaa 28020 gacactgtag ggctcaagct aatggaaatt tagaagatgc tagtgttcac tattacgttg 28080 atgacaccgg agtataccaa gctgctgagc ataaacacgc tacttggaat tgtggagatg 28140 gccataatag atatggtata aataatagaa atacaatatc tatagaaata tgtgttaatc 28200 ctgactctga ttataataag gcagttgata atgctgtaga gctagttaga tacctaaaaa 28260 atggctacta ttctaattgt aaagtagtaa gacactatga tgctagtaga aaaaattgtc 28320 ctagaagaat gatagctaat ggttactgga atacattcct agaaagagta aattcaggag 28380 agagctcaag ctcaccagta attaataatc ctaagggatt ctatgagtct aatgagacaa 28440 gaactaatgc tactttagtg ggggaaggtt caatagaagt tctagatgag gattgtaagc 28500 cagtccctgg tagatttatt gatagcctag ataatttatt tgttcttgga atatatcctt 28560 ctagaaattt catagaagtt gtatatcctg gaaaggataa gaaataccat gcttacattg 28620 acataaaaca ttatagcaga ttaagttttg actaccacat gaaatatcaa aatgataatg 28680 gaataactta tgtgtggtgg aaccctgaag acattaatgt caaagatcat aatgaagaat 28740 tacagcctgg tcaaaaggct agcccaatgt atagaactaa gggctggcta agaataacat 28800 tctatagaga ggatggaact ccatctgatg gatacgttcg ctatgagggt gagcaaagcc 28860 agaagttcta tgaggatatt aaacaaggaa tagttaaggt taacacttct cttaatgtaa 28920 gagatgatgt gaatggtaat ataataggct cagtatttaa taatgagaga gttactatat 28980 taggaagtaa gaatggttgg tatcatatag aatataatac tagccatggt aaaaagcaag 29040 gttatgtaag ttcaaaatat gtagagatag tttagggggt atttatcatg gaagaaaatt 29100 tagtaaactt aatattcaat tacattgatc cacagctttt aataatcatt gtatcatgtt 29160 actgtttagg tctgtttatc aaggctaagc cttatatacc agattggtct ataccattaa 29220 tacttttggt attcagtata tcattgtcaa tcttatatat ggcaattcaa ctagaattag 29280 gttttacagc taagaccttt ttgaatggct ttatacaagg tttaatatgt gcagctgtag 29340 cttcctttgg taatcaagta tggaagcagt taatggataa gagaaaatca gatcctcctg 29400 taaatcaata ggggatagag gttgggccta ctttatttag aaagtaggtc cttaaccttt 29460 ttaaattctt cttcattatc ccattcccat tgctttcctg gttttacaac ttctttgttt 29520 cttaagattc ttcttgcctt tttaccattc atatttaatt cttcaagtat catctttaag 29580 gtgaacttac cttccttagt ttttggttca tccttttcct ctggttcctt aacatctgtt 29640 agatcaagag taattttagt ttgttcagcc ttttcttcgg gttcagctag ttgttgtacc 29700 tcttcaactt tttcttcttg tggttgttgg atttgttgga tagcagctat tagatcagcc 29760 tttttcatag tccaccagtt tttaactttt aatttctttg ctaattcttt aatttcaaca 29820 gctttcatag tatttaaatc tttcatatca tttacctctt ctttcttatt tgttttatct 29880 acattttcat tataagagat ttcctctggt ttgtcaggga tttcttttaa taattttgga 29940 cctccccagt tacattcttc tccatcagca ttatcaaaga cctttcttcc ttcttctgtg 30000 taacctatta cctctccatc agcaaatact aaaccctttt taacaaggga tgagatcaca 30060 cctctagcct tagttccttc tatacctgag ttttctatta ctgagaatac ccatacacag 30120 tcaacatcta gtccatcatc aaattcattc tttctcatag catttaatat ctttatttcc 30180 attgcagtta agttcatatt taacatctcc tctttgtttt tgtttacaag tttattataa 30240 gagttattgc cagagatgtc aaaggttttc tccacagatt tataaaattt ctttgagatc 30300 ttagttggta tcacatattt aaagccgtta catttaaagc aagtagtgtt accatctgag 30360 gttcttgagt aatggccagt gcctccacat ctagggcaag ttttaagtaa acctttctta 30420 gcaacctttg acatagtgct agcccatatc ttaccaaagc catatttgat tttagcttta 30480 tctgaaagat cagggtacac catattaatt atatcaccct ttaatggatc atccttgata 30540 taactattat accaatccaa tttaacttgt gtatgtgatc tcctacttat taccttagta 30600 acctcatact cctttaacca tccatctatg gcctcattgg ataatttatc agcctcttca 30660 aaagttatgc tattaacagg tatactaaat gctcctgttg ctacccagct tccatcttct 30720 ttttggcaac caaagaagtt ttcagcttct acttgaatac cataatatat atctatcatg 30780 tgatttacct cctaagtttt ttgttaatat aattatacca gcactctcga gatatgtcaa 30840 cgaaaaaaga gaagaagttt taccttcttc ccctaacctt taagttgatt aatagatcat 30900 atatatcttt aggcatacct tgctcccact cccactgttt acctggtttc tcaatgttat 30960 tctttcttaa tatacttctt gcctttcttg gatctagttc tatcaattca cataaatctt 31020 ttaaggtgta tatatcactt ctagtttgtg ctacagctga gttttcaccc tttctagcct 31080 gttgcacttt gttgtaataa agactagtta attttttatg taaaggacta tgtgaccaat 31140 aagttaagtg agctaattca tggcatatag tttcaagaac acagtcctta atatcccagc 31200 caaatttttt gattttatca taagcttttt ctccttcctg tgcaaacctg aaagggaaaa 31260 ggttaattgt aatcccagta aatttattac ctgatctatg gcatagacca tagtagttag 31320 catccacatc agattctata taaagatcaa cagcacttac accaaatgct tttcctaaga 31380 tcattgtggc ctcattgtat aactccttta cttcatattc agctaattga ttttctatca 31440 tatctctcat tttgtttacc tccaatagtt aatttatgat ttaattataa gagagaactc 31500 tggatctgtc aacgaaaaac ttcgaagaaa aaaaggatgg ggttttacca tcctagatca 31560 tctcttcttc ctatatcata tattgcacag atgcaagcct caagatgttt ttcaatctca 31620 gcttttttat gatgtttaca tacataatat ttagttactc ccatttgttg tagtagttct 31680 atcaccttat ccaagaattg tttttggaag ttcttttcaa aaaccacctt atttattttt 31740 tctatctcat ttaacatttc taatttagtc atttttcatt cctcctaaga attaattaat 31800 attgattaca atattattat accagagggc ccagactatg tcaacgaggt tctggagatt 31860 tttttcgaaa aaaaatagat ggccttttgg ccacccatat taattggatt ttttcatttt 31920 taggataacc tcctcatagg caaccatgca gcaagaaagg tagtctccta attcactttc 31980 cctaactttg ttgcttagtc caaaggattt tacctgttta tctagttctt ttctaaattg 32040 tggatagatt ttattatcca tagccttttg agcaattcct tttattacat taagtttttc 32100 tgatcttgtt agttccatag ttaacatctc cttcaattaa ttttgattac aatattatta 32160 tactagatag tcccgatcct gtcaacgatt aaatccagat tttttcgaaa aaataaaaaa 32220 ggaggttttt acacctcctt agttttatag atccttgaac atttcagggt agttacataa 32280 agtatcttcc acatattgtt ctgggggtaa tcatattcta tgatgtattc aacagcagct 32340 attaagtcat tctcatcatt aaattctacc ccttgtattt ctagttcctc ctttaattgt 32400 tctaaagttt cttgaaattt tttgttattc ataattaaca actcctttga attaattttg 32460 tttacaagat tattatacca gactctggag tctatgtcaa agaaaatccc gaatttcttt 32520 ttggcatgaa agttgctagg tatcggatcg ctttaaaaag aagcaactat ttagttgctc 32580 ccactggggt taatgaaaac cctgatagtt ttttattctt tccccaccta gcacaagatt 32640 ccattgtggg ctcaataccc aagatattgc atatatccaa atattctttt aaccataata 32700 acctcataat aactaatcct cctttgagta tcttaacttc ttagattttt tattcttata 32760 catcattgag tcggcctgta ctatagcttc ttcaatagtt ttacctttac catagccaca 32820 tgatagctca gggtatatat tcaaggtatt gcacagatta taagctatat cactgataac 32880 tacaaactca tcaccacccc atctaatcaa tatatccttc tcaggatgta aatgggcctt 32940 aataagattt acagtgtatt gtatatacac atctccttct agatgaccta atttatcatt 33000 gaactctttt agatcattaa gatcaatata tacaatctca tactcatcaa ggtcaatctt 33060 atagtttcta ttataggctc ctgtccaatc aatagataac tcctttacaa gttcaatcaa 33120 ttcttccctt gtaagttttt caagatcttt catattcagt tctctccttt ttgtttgatt 33180 acaatattta tattaccata tactgtggat attgtcaacg aaaaagaaga ggtttttaaa 33240 cctccccctt attttaacat tttcttaata tcctcaagtt cagatcctga ccatttatag 33300 ggtttgttta ccccttgctt tcttagcttg gctctggctt tacttggatc taatcctaat 33360 gactcacata aagattttaa atcaactgtt cccataggtg cattacttcc aggagtagta 33420 gggtaatctc tcttggcctc aactttattc agatcaacct tctttatagg tttgtccttt 33480 ttctttcttg gcttcttggg ctcattagga ttaaattctt tcattaattc tctctccttc 33540 ttcttttcct catttaggag tcttaacctt tcaagtccct tttgtttttt cttttcctcc 33600 ttgactttaa tcttggttag ggctttaata gttttaccag cctcaattaa agctttagct 33660 atttcagtat gattacttat tctaggttgt tcttttctca ttacttatca cctcaattct 33720 atattaccac attacttatc aatagtcaac agtttttctg ctctatccct tttaagctgt 33780 attgatttta atattttctc atcaatagta ttctcagtta ttaagtgtat tatttgccaa 33840 ggctcctttt gaccattcct tttgattctg gcaatagctt gcacataatc atcatattta 33900 tagttccaac tatagaatat taaccttgag gccctgaaga ggtttagacc aacactacct 33960 gaacttattt gtagtattaa tatgtcaatc ttgccctcct gaaaatcctt attgatctca 34020 gctctattct tagtagatcc tgttaagcat tctactctgt gggtcttttt aaaagtatca 34080 tgtataagtt ttatttcctc aagaaaatta catattacaa caacaggctt tttatatcct 34140 tctagatagt cctgtaataa gcctaatttg cctttatcaa tattaatact atttcctaca 34200 ttgtttgtaa taaatcctcc tgagatctgc tgtagcctca aatattgggt aatggtaaga 34260 tcagctgtta attggttgat aaacatatta gcccttaaga ataaatcctc atatggctca 34320 ttactctcat agggtatacc attagacctt aataaggcct ttaattgctt cctaggaatg 34380 ttttcctgta ctatatctag ttgtgctatc atttcctttc taaggtcatt ataggccttt 34440 ctggctttat tatttaattc acaggttaaa tacagatcag gaagttgttc tggtaaatca 34500 atacaatctt ctattcttac tatatagcaa ttctggtgca taaggttctt taatatttcc 34560 tcattcttat accccacaat ttcatagttc atataacctc ctttgattat aaagtcatct 34620 tcaaactgtt tccaattaac tcctagaata tttgagttca taattttata ttgcataaat 34680 aaatcttcat aaccattagg agtaggagtc cctgtcatta tcaatctata tctacattta 34740 gtagcaattt tatatagagc cttagatatc tgggcatttc tattctttac cttatgggat 34800 tcgtcaacaa caataaactg tggctttaag ttcataaggt atttatccat caacctagcc 34860 ttctcaaaat tggcaataat aatctttaat cctgagtagg attttatagc tctataggtt 34920 tccataacct tggttttatt accctgtagt ttaaatattt tatactcctt aagtttagta 34980 aatttatgga tctcagactc ccagttaaat acaatagatt taggtgagaa tattagtaca 35040 gtatctagac cttcaagttt ctcaagggcc acaataggaa ctaaggtctt acccgtaccc 35100 atggccaa atagggcaaa cttttcttta tttaaaaata gttgtaatgc ttctttctga 35160 tgtttaaata attttctctt catactaaaa aagaagaggc attaaagcct cctccttata 35220 cctccttagc cagttttaca agttcatctc ttgaggcctt actatggaat ttaaccccat 35280 tatcttttag gtattgcttt aaggattctt tattcatact atcaaaatct aattcttctt 35340 cgtcgtaaac attgtcatca tcatctaagt cctcatcgtc attatcatct atgtcttctt 35400 catcatattc ttctttacta agatcatgta tttctgagtt atcatcacta gttaattcca 35460 tagcagccat aatagcatag ttagctaagt ctagtaaagt atcaaccaca gattcttttt 35520 gagcagcata tgaagggaag tcatcttgat ctccgagtaa taaactttta gctcttccta 35580 gcttatcttc aagtcttata catagaacag ggtttccata ttcaagataa gtcttgtgga 35640 aattatcccc atatagatta ttctttaatt catacagatc tcctagttct cctaggaatt 35700 gttggtgtct ttctaaattt ttcatgtaat ctcctccttg tatattcagc tgaggtagcc 35760 ataaaagcta ttcctcctgc tagatttatt ttatttataa tgtctttttg tttttgagtt 35820 aggttctttt cttttccagg aagtttaact tcaataccaa taaacttacc tttgtgacaa 35880 cctagtatat caggtaagcc agctatttga aagggtcccc catgagtttt aaaccaaaaa 35940 ccagggtatt cttttttaag catattcatt atggatttaa ccacatctct ttcaagcata 36000 atagtagcct ccaagggtaa ataaaggagg accttaaagg ccctcatatt ttaataatta 36060 aattttattc ttcatcatct tcttctagca tttctatgat gtcatcctta gacattcctt 36120 ttttaacctt taagcctctg tccttagctt cagctttaag ttcttttaaa gtcattgact 36180 catagtcatt ttcttcttca tcttcctctt cttcatcctc atcttcttcg tcatcttcat 36240 cagagtctcc gaataggtca ttatattgtt cagctatttc ttcttcgtca gcttcttctt 36300 caattagatc aattagatct tcttcagttt tagccttctt gattttctta gctgggattt 36360 ttaaagcctt agctaattct tttagctcat ccttgtctaa ttcttcaaga tcaacttcag 36420 actcatcgtc ttcatcatct tcttcgtcat catcagagtc agcttcgtca aggtttataa 36480 aatcagttat tcttgatttc ttcttacctt catatatttc atgtgatact tctaccccac 36540 attgtaaacc aactagagct tctgtatcta attcgaactc ctcatcaggg atatcaaatc 36600 ctaaagcaac taacacacct tttaagttaa ataaagcttg tggttgaagt gagcaaatgt 36660 ggtataactt ttgtcctttc atctttcctt ctgctattac aaaagtaaat gataaataat 36720 tgcttccact attttcagaa accttaacct ctacatcttc tacttccaca atgtaatttc 36780 cttctgggat agtaactgag gtttcaacgt ttgaaagatc aatctttaat cctcctttgt 36840 ttttgttacc tctcttagta gtctttttag ccatgttatt gtcctcctta aattttaaaa 36900 tatcattatg agataaatat ttgtcaagat attgtacctc tttgtttaca agtttatatt 36960 atcagatttt taattggttg tcaaaggtat tttttagaaa atttttattt tttcttttta 37020 ccagatttct tactggtttt tttaccagat ttagctggct tattattcca ttcaccagcc 37080 agaatagttt caatatcctc ccagatagtt gttggtgatt ttaaatcatg taccagataa 37140 gctggacatt cagatccttg aggtctagta aatttagtaa tataataagg gttaggtcct 37200 agtcttagtc tatactggat ttctttccta accttcatac cctctttttc tgagtgttca 37260 tataaataag tatgacctat tactcttgat acagcacata aatatttaga tacagatggc 37320 attaatcctg gtcctacctc aggcattaat tggtcttctc cttcaccatc accagattct 37380 aacctatctt gtactataaa gcatggtagt ataccttcct catttaattc tttataaagg 37440 tttataacct ctttcatata attagccaca tttccaaaca ttctttggct catctggtcc 37500 ttaccttcct cagctttgac cttttcatta cctaattctt gtaaagcagt taaatggtct 37560 ataactactg ttttaaattt atcagtgttt tctgataggt aatcataagc ctcgtatatg 37620 tcatcaaaac tctttagact aaatacttgt atatctcctc ttttaactct aagcttgttt 37680 ctagctgata gagtaccttt gtcctttaca tcaataaaga atattggttt gggtagtgtt 37740 cctgagatag tagttttacc tgaaccagat cttccataaa gtgttaatat tgtgggctca 37800 tctatatcat atagatccac taatctgtcc tcaaaactat ttgattttga tttttgcttt 37860 tttgccattg tcactttcct ccttttgttt atagtcagcc ttaataata attctttatc 37920 aagatttaaa agttcagctt gacataaaga cttatattca cagaaagagc aaccttttcc 37980 aaggttcttt tcctttcttt tgtggccctc atctctaata agtttagcta tctcaatagt 38040 atcatccatt acactgttaa gtatattctt tgataatatt attgggtgtc ttacaaagaa 38100 gtcttcccat ctagcagaat taatataaga ctcataatct ttaggatcta atcctaattc 38160 tcttattcct ctttctaact ctaaaggtgt ggatttaatc ctcttttgag ataatacccc 38220 tttttgagtt aattctggtc tacctggttg ctgagctaat attatatccc atatagtacc 38280 acttactttt gggtattcat gtagaagagc ccaggtatat atagcagatt gtctattgaa 38340 gatcaggaaa tttctatcag gcatcttttt gtaggtttta gtttccttag cccatatctt 38400 tccatccaca tctaagacac tatcaatata gccttctaga gttacaccct tacatagttt 38460 taattcgaag tgatgttcat tttccacata ggttacatta tcttcttttt catcataata 38520 gtggaaatag ttatcacata aagaataaac catttttgga atatctccaa attcaaggat 38580 ctcttcttta aatgtatttt tgtaaaactc tttggaaaat ttatcaacag attttttcca 38640 actcttccca ttatggtagt cttctatagc ctggtggata gcagagcccc tttgtaaggc 38700 agctccactt cttttaggta ccagaccttc atagtattta acataatggg cataaggaca 38760 agacaagaat ttgttaattc tggaattact tatttttaac atccttatac ctccttttca 38820 ttacaatatt tatattatca gataattatt catctgtcaa cacttttctt atctggcttt 38880 tccaatcacc tttaaaatcc agttcaacac atcctcccca agaataacca actgatatat 38940 ccatgattat tgggactgtt ggggtaaatt taaatacttt ctttaatact ttattatttt 39000 caactattga ttttactttc caagctaatt cttccacata gtctttatct acttcaaata 39060 atattgagtc atgtactgag cctaatacat caaacttaga cttatctaat acatattcag 39120 gattaacata tttagcatag cctgtaatct cacataggcc taataaagtg atatctgaac 39180 caaaaccttg aactggtgag ttaatacatt gtctttcggc ttcagcagct ttagatctat 39240 cagatgaata tatatctggt aatcttctaa ttcttcctat tggagatctt acttcaccat 39300 tggctttaac aatctttctt tgtttatcat gccatgttaa tagtttagga tatttattaa 39360 agaagttcct tctatatgtt ttggcttcct catctgttaa tttaagacca tagttgtctc 39420 ttccataatc cttgaatttt ttccattgca taccatataa gaaaccaaag tttacagcct 39480 tagccttttt ccttttctcc ttcttaactg ctggatcttt ctcatcattg atatcttccc 39540 cagatataat attatatgtg gagtcatgta tatctccacc agtctggtaa ataaatttca 39600 tattatcctc atcggccatc atagcagcta ttctaagttc ggcctgggat agatcggctt 39660 caatgaatac tcttccagga gatggtccta ataaactcct gattttctta tctctaggaa 39720 cttgctgtag attaggatta ttacttgagg ttcttcctgt tacagtccca tgaagtttaa 39780 agtttggata taatctatga ttgtgcatcc tatttagcca accatctata aagtgagaaa 39840 tttgtatatg aactccccta tgctgtaata gtagttctac tgctggatgt ttatctctta 39900 gtcttaataa tacactttct ccagtagctg gtgctccaga gtcagtaacc tcaatcacag 39960 gaagaccaag tgtatcatat aaaaattctc caacctgttt aggtgagctc cagttaatat 40020 cctggccatt agtatacttc ttaagtccag tctcaatctt atctagttca gatcttaagt 40080 atttttcaac ctctttgaat ttctcctgat ttacatatat acccttgaac tgaacatcct 40140 cataagcttt actagctggg atgtataagt ggtggaatag tttatataag ctctcatccc 40200 ttttaagtct tttattaaat attttatata gagcaaatgt atagtatata tcatacccca 40260 gatacctaac ataatcctca aagtctgatt tagtctccac atttccagtc tttaatttct 40320 tatttatatc ccagtcttta gcattacaat gtaataaagc attctcctta agaccattag 40380 gagtattctc atctaaacaa tgtgaggcta gtaaagtatc aaaggttatg attggcttaa 40440 taccatattt ctcctttaag aaattatcat caaacttacc attctgggct attctttcct 40500 tcatctcaga attaagtctt ttaattaagg atttagctaa ccttctttgt gctatgggct 40560 taccccttaa tgggctatat ttgacttcta aaggcagtat atattgtact tgagtattac 40620 caaaaccaaa tagagtaatg tcattattaa acctatctaa gcctctagtt tctatatcat 40680 atgatactct attataacct tcctctctta aatatttgaa agcctggttt aactctttca 40740 tactggtaat aagttttatg ttaagctctg gtaattcatg ctctttacct tttaccatgg 40800 ctttaaagtt attaaaggcc tgatctacaa attgtgcttt agttggatca tatgcaacta 40860 cactgggtga atagctggca ataaacttaa agccatattt ctcatgcttc ttgggtactc 40920 cacatattga ggttatacct tcattactta gcatctttaa agcttgagat cctaagatca 40980 taacatattc aggtttatat ttttcaattt ccttaagtaa atgttcttgg catttcttaa 41040 tctctgatac tttgatttta gtacccttag gagttctaca cttaatagca ttggtataat 41100 aaaagtctag accttccatt ttatcaagta atgatttatc aggagtagct attctattgg 41160 cctcatccct atcattagct aaactattta tgaccattat tcttggcttc tttttctttc 41220 ctacacaagt tccttctata catgagggag ctgagtgagt atataggtta cagttgttac 41280 atctcattta aaatattacc tcctaaatag aaaggacccc taaaggccct tatctttatt 41340 ttatgattga gttacatact ggacagacct tatgtttttt gaagggtatg gttactaccc 41400 ataggcctcc agttaagaaa atcatgatcc aaaacatagc tccatatttt ctttttaatg 41460 ttactgagtt tttacacata ggacaaatta tatttttcat caggatagcc ccctttaaat 41520 tagaatatca tatgatatat tatattggca atatcaaaaa ttattaccca taatcctata 41580 gctgctaaga ttataattaa acaaccaaga cagccattgt tattgttctt aggtctgtta 41640 tcatcactat aattatacat attacttact ccacagctca ttcatataat ttatatagtt 41700 attaagttca ttcctagtca tgtccccaag gttagctggc tccaaatcaa attccttttc 41760 agtattccta ataaattgct ttagggtttc cttcttgtta accttattat taagatcaga 41820 atgtttatat ctttcttttc tcatataact catttattag aataccccct taatatcttt 41880 gttgtaaaca tgaagagaga aaattgtatg agagaaactt cctggttcat agccagtttg 41940 ttcagctacc caatgtaata gtttcatagc aagataaaca tcattagcaa agtgagtagc 42000 aaaatcagaa cttctcataa cataatggat atttagctta ccttctctta cttggaagtt 42060 atatcccaat gaacatggta ctctagttac tcctcctata ttgaatggat caatagctgg 42120 attccacaga gataaccata attgtcttga gcctggatct tctttaattc tattgatgat 42180 atcctcaatt tgtctgtatt gccaaattct ttcattgtaa gaatatccaa attttccatc 42240 tcttagatat tcattccata cctcaggtct taattgataa gctttaccag gatttatgaa 42300 agcctcccta gtatgatgtg gatgtatacc aaataattct tctctttctt caagagagta 42360 ttgattgatg acaccttgtg gatcgtagat cctttctcta aattcagcat ctgcccaagg 42420 ttgagataca cctggaactt cttcaggttt actttctaga attgtgtaac tatagttttg 42480 taactccttt gtgaaataat ctggattacc ttctataact ttatcttgca tagttttggg 42540 tcttacctca atccccattt cacataaatc tctttttact tcttcaaatg cttcttgagc 42600 attaatataa attctcatag tttaattcct ccatttattt aattttgtta agttaattat 42660 actacttatt aaagtacttg tcaatagata atttattgat atctatcttt ggaaatttct 42720 ccagaccaaa atgtaataac tgcatcttct ggagagcttt ataagagtgt atctggtcct 42780 cagatttaaa gaacctgtta tagttactat tcaaggattt atgccaagga tgtttactat 42840 tagcaatact ctttcttggt accttaaaat aattgaagta tccatttata aacattcctg 42900 acaaataagc ttgagacata tgaaaagtaa tattatctat ctcacatact tcttggggta 42960 gttcttcaat gaacctgttt attagtacta gatcagctgc gaatttcttt tgtaattcac 43020 atactctata cataattttt aagtgtttcc aaggtccttt tctttttggt ctagttacta 43080 ctgcagctat caagcaagaa ccattagtag cctttttcct attgaaataa taagtatagc 43140 tcataccact tatcttggta agcttttctc taaattcaat taattgtgga taatcaatgt 43200 atgatcttaa taggtgtccc cacttacctt ctttatatcc aaagtcccct agatttaaag 43260 tacatttatg ggacttacag gagagaacta agtcctcctg aaagccctgt atgttctgag 43320 tatacatcat agtttcttct ggattggtaa tcatttccct attaagattt aagaataagt 43380 catcaaaatc gttatagtct cttattagca tattaccttc tcctaagtat ttattttagt 43440 aattagatct ttggcggaat ttatttacct gtgatttctt aaggtatata tttactatgt 43500 catctacctc taagccagat acagctaata atttaatata acatacccat acattaaata 43560 gtctttggtt aaaagcattt acatctgtta gcattgaagt ttgtttccaa ggtttattct 43620 taaggcaatt acagcacata cccatttctc ttaccaggtc agttacagct tcttctacta 43680 acatttcagg aaattctggt gatgtagtag ccttatgata taagttctct aggctatcaa 43740 ctagtatagc attatcatct atagtattat aatccttacc agctaggatt gtaaattcag 43800 ttaagaagtg taatccatca attaattctt cttggtaatg gtcttcatga ttataagcat 43860 ctaatgcttc tcccacttct tcagtcattc tccaagcaaa atctttaagt ctagcttgac 43920 ctcttttatc atttaagtta actggacagt cttcagtttg gcataatcct gatctagctt 43980 ctatatgatg atatttctcc atcaattcct tttgtctatt aaagatagct tgtaatctat 44040 ctccttctat tttttcttct actgcatggt ttatattcat aacaatctct cctcttatct 44100 tgatatattt ataaattggt ttatagcttc tttagcactt gagtcatcta ttaactggga 44160 tagcttctcc acattggaag tagtataatc atatactagt acattccatc catcacagaa 44220 taacttccat attagttcat catatctagc cagtagatgt tcagctcttt caattactcc 44280 tggcatctgt tctctatcac caaaattgaa gatcacttcc ctaggtggtc tagcatatac 44340 aatagttggc ttacatagat gttttaagat ctttagctcc ttactgtcaa ggttgaaatt 44400 actactatct ctaatgatag gtccatacac catttcttca aagcaagtaa atctatcatg 44460 tatagcttcc ctttcacctt ttctcacaat ttgagttaat acccattctc tttgttcttc 44520 ataagaacca ggtcccattg attggatata ttctcctcct ctgtgtttag ttaggttctt 44580 agctgttgtg gtcttacctg agttatccat tcctaaaatg ataatcatta aagctctcct 44640 ccattcttta gatcttctac tattaagttt agtatatatt gtactccttt atcattgtat 44700 tcaaataaag gatttataag ggacttactt ggagttacat aataaccaag ttctttagca 44760 aggattgggt ctattatttc aagtgtatgt tcattatacc ttctggcaaa tccctttctt 44820 actaaatacc tatttagttt tatcacacta actcctaatg ctcctgctat ctgagtagca 44880 gtggttaaac ttcccttcca tagttggggt ttgtcatcat taaggtcttt caagatgtaa 44940 tactttttat tgaccattaa attctcctcc ttctacattt ttatattacc ataattaatt 45000 tattatgtca aaggttattt tagaattttt aggtatatct ataacagcca gaagattttc 45060 tggtctatta ctatcataaa caagtatagt ataattctga gctcctaagt tattgaccat 45120 aacctcatta gctaacttag ataaattgtt aggggtaagg tccttaggac tattaagatt 45180 tagtttgatg ataagatcct ccccatctag tccagatgga gaagtctcaa taacattata 45240 gtctaaagca ttatgaagct tttctactaa cttttcttga ctttttcttt ggtgatgact 45300 ttctattagt tttgccccct gagttagcat tatccccatt gacaatgata tccctagaat 45360 tagtactact tccttcaagt ttcttcttcg cattagccat agcctcccta tactttatat 45420 tcatacattt tacataaaag tcttggtcac ttataccaca gatagggcaa ttcttgtaac 45480 ctccttggct tctcttaaat tcttgaatag tttgacaata tggacaccac atcttctttt 45540 taggtatttc ttcgccttca gggatcttat gagcttcatg gacatccacc caataaaatt 45600 cagcccaggg acattcttcc ataactttgt ctataaataa tttagctgat ttataggtat 45660 agaaatattt caatcctgaa tgataggtga ttttattctt cttaccccta gcaataatta 45720 ctactacaaa ggcctttaca tgattatgac ctgctggcct cttacatttt attatcatgc 45780 ctttttcctc ccaatattta gtttcttact tatattataa cctttatccg gagaatatac 45840 aatagtatct tttaaagtat ctagaactag attaaccttc tgtgaattta atcctgatat 45900 tgacactatc tcctccttag tatgagctcc tgattttaca gcctctataa ctttggcttt 45960 gtcctcaggt ttcacaatta cttcttttcc aggtattgct acttctacat gataatatgg 46020 atctccaaat tctcccattg ataaattcaa ttcaatttga gggaattgtc ctgcaagacg 46080 gaactctcta tccataatga ttttacttgg taaatggctt tgtttatcaa gattatctat 46140 atctacttct atttcctcat catcatcttc taagtcatca ggtcttttta gatatagagc 46200 agattctacc caaccatgta atatgaatga accagccatc ttttgaccac ctctagtttg 46260 agtagcattt ccacctttat tatagtggtg tattagcata actcctgtat gcttctcatt 46320 cttaagttta agacaccatt gtagtacagg attaagatca gctgcattat taagatctcc 46380 actaaacata agatataatg ggtcgaatat tactagtaca ggttttatct catcaattaa 46440 ttcttctatc tgttttctat gggattcttc atctaacata aatccctgtt gattaataaa 46500 agtgatagga agatctggag caaacctaac tttaagtctt ctcttacctt ttatatccac 46560 attaccaact actcctctgt ggctaattat cttttcagtt ctatctctca taatccagtc 46620 agcattctca ttttgaacta caattactgg gccaggttct agaacgggat attgacctag 46680 gaatggtctt ccactagcaa ctgatatagc taagtcctgt gtaaatgtgg atttaaatac 46740 ctttggttgt ccagccacaa ttccatgaga tcttctaccc cagaaacctt gtactaacca 46800 accagggaag gctccattat tacccataac atcctgataa ctatctattc tcagtttagt 46860 actttcagcc ttttcaatat cagcctctat ttctcctcct atgattttat ccaattcttt 46920 tcttaatctt gtttcttcat cttttcttcc cttatattta ttaaaagctg agcccttaac 46980 taataatata atctcattag gctctaatcc tatttcatgt aatttattct caatatacca 47040 tatagtagag cttctatcta aagaagtaat atcatctaaa gccagtaagt ctctaacctt 47100 ctttggtata ttatatttag catatatctt tctttcagat tgtgaggcct cagatcctcc 47160 tatattgacc ttaacactat cattagattt acttacagcc ttaacagcct tagcgatcac 47220 cttaggctta tatatctcct tagtgtgtat aggtcttttt atttttggtt tattcttgta 47280 tttgtagtta atagttcctg gtattctata tacatgagta acatcaaagc aggagtcaca 47340 gcctatatgt ttagctaatg ctggatttat ttcgtcatac tgattagcct ctatatacct 47400 atccatttcc catagcccct gatattttcc aggagaactt tcccacagat aacttggttt 47460 aggtttaatc cccaaggggt cagtatgctc atctatatct tgtattaagt atttagtctc 47520 tatgaagtta actatccttc tttgaggatt actatagggc attgggctcc aatataaatc 47580 ataaccctta tactgttgtt caaagtcctt tagttttttg tcaatgttat tcttattata 47640 tttgattggt acatctttcc attctttacc ttttctagct gctaggataa tgtaatctcc 47700 attttcacat tgtttagaga atatatcctt caagaacttt atgtccatat attatacccc 47760 ctattataag tctatattat cccaacctaa ttctttcaag tatggaacta cttctcctct 47820 tggctttaat tttctagtgc cttcctctat ctcagcgaat actaatcccc ctatatgtgg 47880 tttagtcccg ttaactactc tcattatact aacagccgat actcctaatt gtttagctgc 47940 atatctgaag gatctatact ccctattatt gttgtaacaa tatactgctt tagtataagt 48000 tgaatttcca cctcttgcca tttatacatc cccctctaac atctgtttct aaaataagta 48060 aaaaatttga ggagcaagtt tgctcccctt attatttgat gagaattatt cttcatcatc 48120 agcttctaat aattcaatta tctcagctgg tttcattttc ttattaactt ttattcctct 48180 ttctttagca agagcaatta agtctttctt agtcatttca gagtagtcag gagtttcatc 48240 ctcttcttct tcctcttcgt cttcttcttc accttgagct tctttgatta aagcgattaa 48300 ctcatcttta tcttttttct tagctccttt gatacctaag tctttagcca tagcagctaa 48360 ttccttttta gataatgatt ctaaatcatt ttcatcttct tcttctgctt cgtcttcatc 48420 aacttcttcg aagtcagcat ctactacatc ttcatcttcc acagcatcta tttctgcttg 48480 gtatgcagcc tttacagttc ctacagccat ttctaatgat tctaatgctt cctctacagt 48540 tatttgtcca tctgtgtcag gagttctttc atctaattcc acaatagctt ttaaagcctt 48600 tttaacttct ttctcatctc cagaatttaa agcagattgt actattacaa ttaactcctt 48660 attatttgct tcctctaacc attttgcttt tccttttgcc ataataatca ttcctccata 48720 tttttaattt ttaaataaca aatttagtat agtttgaaat taactaatta taaagttttg 48780 gttggtgttt gtaatttgat tacaatacta taatatcata tcctatatag cctgtcaacg 48840 agttatttag aaaaaattaa tttattataa cattcattac aaacggcttc atattcaaca 48900 ctttcgggct ggttatcaat ttgtatctgg ttaccctggt cagtatattt accttgatat 48960 ttcaatattg agaatgtggc ttttttacca cattgacaaa tgttctttaa ttcttcaatg 49020 ttatgggcca cttgtaataa tcgggtactt ccttcaaatc cttccctttt aaagtcagtt 49080 ctcaatccat aacatatgat aggataatgt aatgataaat ttaacagatc atcaacctgt 49140 tctctggata agaattgtac ctcatcaatt aatataacat ctactgccca ataattagac 49200 tcaaagtcag aaagtatttt tgataattta acgttaggag gtagaagaac atctaccttt 49260 cttgctaccc ctattctact gattatacat ggtccgcctt tagtatcaat tccgggctta 49320 actactaagg gcttcattcc tctttcttca tagttatgtg ctacctggat taactgagta 49380 gtctttccag agttcatagc actataccta aaatataact tattgctcac aactgatcgc 49440 ctccattata caccatacta ttacaagtat gcctaatatt ctaaatgccc gtaataaata 49500 tcccaagatt atcactcctt tgactcctcc tctaaatcaa atatacctgg tggtactaaa 49560 tccttaatct ctcccattat ggtttcatag ttaagtgttc ccttctcaat ttcctctgtg 49620 tgtttactca tctcttccac aaacctagtc attctagtct taccataacc ttccttcttt 49680 aaggtatagc atgataaggc taacatgata tccacagctt taagaacatt aagatttttg 49740 ttctctcttt cagctcttct tcttgcagct cttgctcttg acataataat ctactccttt 49800 atatatttag tttctaaaaa tattgatact attatagcta gtatcgcaaa acctagttcc 49860 atttcttcac ctcaaaacct ataaacactc ctagtaaaaa tattgatact agggttatta 49920 cccctagcac catttacttc tcctcccata acaggtcaga ataatcagtt ccttggtcat 49980 atgagaatat taatcctttc tgtgaactga tataatcagc caagtgtact attctctttt 50040 gataagtatt gattgggtac tctccccatt gaccatgatg agactcaatt agttctatta 50100 ccttttcata ggttaatgga aactcttcct ttagtggtaa tagtgtctct ctaaccatat 50160 atggatgctc cttagcaggt ttattaatat catgtagtat cagagcagcc tgtatcagat 50220 ccttatcttc ctgacatatc atctcagctt cgcataatgc tttacctata taacaagcag 50280 ctttagtatg tcttaacaaa cctcctaccc ctgcactata agttggatga tatttcccag 50340 tactacttgc aggtattata aagaacttct catcaactat ctcatttagg gcctttctta 50400 caaaattttg tatatcccta tctcttatat catgtatttc gggtattagc attctcatgc 50460 ccatttctct tatcataact attcctccta tcccatgtag tagcctctat attttatctt 50520 ctcccactca tcaacagtaa atactaattt agttctttgt ggtatgttag agtaacaaag 50580 ataatcaagg tctacttcta catattgttt acctttagcc caaggtttat tatcaaattc 50640 atggtactct cttacactgt gtaccttttt tatataatgc tcaagtaggt tcatttcttt 50700 ctcctccttt tctttttctt actctttcca tcaagtcctc gtttaacatg aaagtcaagt 50760 aagttttgag cccaccttag catcttaata tcttgcatac atgaaagatt aaagccagtg 50820 gatccatcaa aaactttaag gtcattccta aatggcatac tgtttactaa acctggattc 50880 tcattcttaa gtttaactac ctctctggca gtctcatccc attgttggtc tgtccagata 50940 ttgttgtcat acctataata tattattgaa tgaagacaaa cctgtaacca cttctttcgt 51000 aattcaattc tttctggagt ttcttcgcag tcatacttga gaattaacat agcacttctt 51060 ctccccaata ttatcacctc cttttctatt ataccatatt cttttacctt ctacaataga 51120 aaaagtttga ccaatatgtt ctggttcaat ttcctcaatg gtaagtgttc ccccatgctt 51180 acatagtatc ttagctaata cttgtcccat gtctgtatct gctaaagata ctccactatt 51240 ttttctggtt attacatacg acttcataac ttagccccac tctcaaattt attaactgat 51300 tatttatctc ttgtatttct tttaacttct tctcagtcaa tcctaagaac tgtcctaatt 51360 ttatcttctg ttcttttgtt aaatgacaat actgtttctc aatcaaatat ctggcatagt 51420 ttaggttctt tcttatctca tcagtatttt gcttatgctt ttcctctatc ccttccagtc 51480 tctttaataa ggatgtgttt atcatgtgat acctccctaa taagatattt tgttatcttt 51540 ggctttctgt cactaaagtc cattactgtg gctacatccc catcgatgaa tactactgat 51600 ttaccaccaa acttatcttt taagtcattt ttattcttga ctattttacc attacataaa 51660 tacatgatat acctccttta aaggcctcta gcagctcctg taaggtgtct ctaccacatt 51720 ataagtaaaa tcacattacc acatatctaa aaggccttag aattgattgt gggctcttat 51780 atactgtatc ttatgatagc tctattgcca ataacctcat ataaatgagt attatcctta 51840 tataagtatc ttggtatgga tttatcctca tatataagaa tacagtactt aggtacttga 51900 gatactaatt ctccgcaatt cttatctatt gctctagata gaataaatct gtcctcttcc 51960 attattatga agaagtcctt gttttcaatg ataatttctt tttcattaga taatataact 52020 ttcatcatgg ttatccctcc tttactgtca ttataagaga agtaagta 52068

Claims (9)

클로스트리디움 퍼프린젠스(Clostridium perfringens)에 특이적인 사멸능을 갖고, 서열번호 1로 표시되는 염기서열로 이루어지며, 수탁번호 KFCC11699P인, 박테리오파지 CP3.
Bacteriophage CP3, which has a killing activity specific to Clostridium perfringens and is composed of the nucleotide sequence shown in SEQ ID NO: 1 and has the accession number KFCC11699P.
삭제delete 제1항의 박테리오파지 CP3을 포함하는, 항생용 조성물.
An antibiotic composition comprising the bacteriophage CP3 of claim 1.
제1항의 박테리오파지 CP3을 포함하는, 사료 첨가용 조성물.
A composition for feed addition comprising the bacteriophage CP3 of claim 1.
제4항의 사료 첨가용 조성물을 포함하는, 사료.
A feed comprising the feed additive composition of claim 4.
제1항의 박테리오파지 CP3을 포함하는, 소독제.
A disinfectant comprising bacteriophage CP3 of claim 1.
제1항의 박테리오파지 CP3을 포함하는, 세척제.
A cleansing agent comprising the bacteriophage CP3 of claim 1.
제1항의 박테리오파지 CP3을 인간을 제외한 개체에 투여하는 단계를 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법.
A method for preventing or treating an infectious disease caused by Clostridium perfringens, comprising the step of administering the bacteriophage CP3 of claim 1 to a subject other than a human.
제8항에 있어서, 상기 클로스트리디움 퍼프린젠스에 의한 감염성 질병은 식중독, 괴사성 감염, 가스괴저병, 연조직 감염 및 화농성 근육염으로 이루어진 군에서 선택된 것인, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법.
9. The method of claim 8, wherein the infectious disease caused by Clostridium perfringens is selected from the group consisting of food poisoning, necrotizing infections, gas gaegeosis, soft tissue infections and pyogenic myositis. Infectious diseases caused by Clostridium perfringens, &Lt; / RTI &gt;
KR1020170172207A 2016-12-23 2017-12-14 Novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the same KR101993121B1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020160178172 2016-12-23
KR20160178172 2016-12-23

Publications (2)

Publication Number Publication Date
KR20180074583A KR20180074583A (en) 2018-07-03
KR101993121B1 true KR101993121B1 (en) 2019-06-26

Family

ID=62918544

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1020170172207A KR101993121B1 (en) 2016-12-23 2017-12-14 Novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the same

Country Status (1)

Country Link
KR (1) KR101993121B1 (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102586837B1 (en) * 2021-02-23 2023-10-10 씨제이제일제당 주식회사 Novel bacteriophage having a specific bactericidal activity against Clostridium perfringens and antibacterial composition comprising the same
KR102586836B1 (en) * 2021-02-23 2023-10-10 씨제이제일제당 주식회사 Novel bacteriophage having a specific bactericidal activity against Clostridium perfringens and antibacterial composition comprising the same
KR102586839B1 (en) * 2021-02-25 2023-10-10 씨제이제일제당 주식회사 Novel bacteriophage having a specific bactericidal activity against Clostridium perfringens and antibacterial composition comprising the same
KR102586838B1 (en) * 2021-02-25 2023-10-10 씨제이제일제당 주식회사 Novel bacteriophage having a specific bactericidal activity against Clostridium perfringens and antibacterial composition comprising the same
CN118086222B (en) * 2024-02-01 2024-08-06 青岛润达生物科技有限公司 Clostridium perfringens bacteriophage suitable for oral administration and application thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150018171A (en) * 2013-08-09 2015-02-23 주식회사 인트론바이오테크놀로지 Method for prevention and treatment of Clostridium perfringens
KR101649849B1 (en) * 2014-12-29 2016-08-22 주식회사 인트론바이오테크놀로지 Novel Clostridium perfringens bacteriophage Clo-PEP-1 and its use for preventing proliferation of Clostridium perfringens

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Clostridium phage CP3, partial genome, GenBank: MF001357.1. 2017.09.16.*

Also Published As

Publication number Publication date
KR20180074583A (en) 2018-07-03

Similar Documents

Publication Publication Date Title
KR101993121B1 (en) Novel Clostridium perfringens specific bacteriophage CP3 and antibacterial composition comprising the same
CN107109372B (en) Novel Clostridium perfringens bacteriophage Clo-PEP-1 and use thereof for inhibiting proliferation of Clostridium perfringens
EP2961834B1 (en) Novel bacteriophage and antibacterial composition comprising the same
KR102341100B1 (en) Novel Salmonella specific bacteriophage SS1 and antibacterial composition comprising the same
KR102193499B1 (en) Novel Bacillus cereus bacteria specific bacteriophage BC13 and antibacterial composition comprising the same
KR102016920B1 (en) Novel Clostridium perfringens specific bacteriophage CP5 and antibacterial composition comprising the same
KR20180042748A (en) Novel Staphylococcus specific bacteriophage SA7 and antibacterial composition comprising the same
KR102016922B1 (en) Novel pathogenic Escherichia coli specific bacteriophage EC10 and antibacterial composition comprising the same
KR101830900B1 (en) Novel avian pathogenic Escherichia coli specific bacteriophage EC13 and antibacterial composition comprising the same
KR101871342B1 (en) Novel Salmonella specific bacteriophage ST1 and antibacterial composition comprising the same
KR102066920B1 (en) Novel Cronobacter sakazakii specific bacteriophage CS3 and antibacterial composition comprising the same
KR101863593B1 (en) Novel Salmonella specific bacteriophage SP1 and antibacterial composition comprising the same
KR102193477B1 (en) Novel Salmonella specific bacteriophage ST20 and antibacterial composition comprising the same
KR20180073489A (en) Novel Pseudomonas aeruginosa specific bacteriophage PA4 and antibacterial composition comprising the same
KR102193495B1 (en) Novel Acinetobacter baumannii specific bacteriophage AB63 and antibacterial composition comprising the same
KR102037383B1 (en) Novel Vibrio specific bacteriophage VP2 and antibacterial composition comprising the same
KR102037398B1 (en) Novel Salmonella specific bacteriophage SG102 and antibacterial composition comprising the same
KR102016919B1 (en) Novel Salmonella specific bacteriophage SC1 and antibacterial composition comprising the same
KR101842673B1 (en) Novel Salmonella specific bacteriophage SE1 and antibacterial composition comprising the same
KR101871338B1 (en) Novel pathogenic Escherichia coli specific bacteriophage EC14 and antibacterial composition comprising the same
KR101842668B1 (en) Novel Salmonella specific becteriophage SG2 and antibacterial composition comprising the same
KR101842667B1 (en) Novel Salmonella specific bacteriophage ST3 and antibacterial composition comprising the same
KR102203670B1 (en) Novel Enterococcus specific bacteriophage EH11 and antibacterial composition comprising the same
KR102003786B1 (en) Novel shiga toxin producing Escherichia coli specific bacteriophage ECOH7 and antibacterial composition comprising the same
KR102359655B1 (en) Novel Listeria specific bacteriophage OPT-LM1 and antibacterial composition comprising the same

Legal Events

Date Code Title Description
A201 Request for examination
E902 Notification of reason for refusal
E701 Decision to grant or registration of patent right
GRNT Written decision to grant