KR102016920B1 - Novel Clostridium perfringens specific bacteriophage CP5 and antibacterial composition comprising the same - Google Patents

Novel Clostridium perfringens specific bacteriophage CP5 and antibacterial composition comprising the same Download PDF

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KR102016920B1
KR102016920B1 KR1020170178596A KR20170178596A KR102016920B1 KR 102016920 B1 KR102016920 B1 KR 102016920B1 KR 1020170178596 A KR1020170178596 A KR 1020170178596A KR 20170178596 A KR20170178596 A KR 20170178596A KR 102016920 B1 KR102016920 B1 KR 102016920B1
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김영주
연영은
한범구
김현일
박철세
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Abstract

본 발명은 클로스트리디움 퍼프린젠스 (Clostridium perfringens)에 특이적인 사멸능을 갖는 신규 박테리오파지 CP5, 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 CP5는 기존의 항생제 등의 화학물질에 비해 클로스트리디움 퍼프린젠스에 대한 특이성이 매우 높고, 상기 균에 감염하여 균 내에서 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있다. 또한, 본 발명의 박테리오파지 CP5는 사람, 동물, 식물 등 세균외의 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 CP5는 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.The present invention provides a novel bacteriophage CP5 having a specific killing ability to Clostridium perfringens , an antibiotic composition comprising the bacteriophage, a feed addition composition, a feed, a disinfectant or a cleaning agent, and the bacteriophage to a subject. It relates to a method for preventing or treating an infectious disease caused by Clostridium perfringens comprising the step of. Bacteriophage CP5 of the present invention has a very high specificity for Clostridium perfringens compared to conventional chemicals such as antibiotics, and is excellent in lytic activity and resistant to physicochemical stimulation by proliferating in bacteria. Have In addition, the bacteriophage CP5 of the present invention is not infected with other hosts other than bacteria such as humans, animals, plants, and the like, which can solve problems of antibiotic resistant bacteria due to misuse of antibiotics, residues of antibiotics in food, and a wide range of hosts. There is this. Therefore, the bacteriophage CP5 of the present invention can be used in various fields in the field of antibiotic composition, feed composition, feed, disinfectant, or detergent.

Description

신규한 클로스트리디움 퍼프린젠스 특이 박테리오파지 CP5 및 이를 포함하는 항균 조성물{Novel Clostridium perfringens specific bacteriophage CP5 and antibacterial composition comprising the same}Novel Clostridium perfringens specific bacteriophage CP5 and antibacterial composition comprising the same} Novel Clostridium perfringens specific bacteriophage CP5

본 발명은 클로스트리디움 퍼프린젠스 (Clostridium perfringens)에 특이적인 사멸능을 갖는 신규 박테리오파지 CP5, 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 provides a novel bacteriophage CP5 having a specific killing ability to Clostridium perfringens , an antibiotic composition comprising the bacteriophage, a feed addition composition, a feed, a disinfectant or a cleaning agent, and the bacteriophage to a subject. It relates to a method for preventing or treating an infectious disease caused by Clostridium perfringens comprising the step of.

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. 이중 A형은 다양한 곳에서 분리 및 보고되며, 특히 사람에게는 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 found individually in England, Germany, and France, and was formerly called Clostridium welchii . Clostridium perfringens is the most frequently isolated bacterium in the genus Clostridium bacteria that is isolated from clinical materials. The characteristics of Clostridium perfringens are gram-positive, non-motile anaerobic follicle-forming bacilli, which are found in various environments in the natural world, such as in the intestines of healthy people and animals, soil, sewage and dust. The bacteria have toxin types such as A, B, C, D, and E depending on the ratio of the types and amounts of the toxins. Among them, type A is isolated and reported in various places, and is a representative cause of food poisoning, especially in humans. The optimum growth temperature of these bacteria grows well at 43 ~ 45 ℃ or 37 ℃, which is higher than normal bacteria. In particular, the generation time of Clostridium perfringens is very short (10-12 minutes), so even if a small amount of bacteria is infected, it is likely to increase to a number that can cause disease in a short time. In addition, the apo is produced by the bacterium is high heat resistance and does not die easily even when heated at 100 ℃ for 1 to 4 hours, it can withstand various disinfectants and chemicals.

클로스트리디움 퍼프린젠스에 오염된 식품이나 물 등을 통하여 사람이 섭취할 경우, 식중독이 흔히 발생하며, 급작스런 복통을 일으키는 괴저성 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 계 군에 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. Food poisoning often occurs when a person consumes food or water contaminated with Clostridium perfringens, causing necrotic enteritis that causes abdominal pain, and necrosis, cellulitis, or cut surface when infected with skin or soft tissue. It can cause infections, abscesses and the like. In addition, Clostridium perfringens is infected with a variety of warm-blooded animals in addition to humans, causing diseases, especially in poultry such as broilers, laying hens and breeders, resulting in decreased productivity, increased mortality, and reduced animal welfare in chickens. It is the cause. Necrotic enteritis (NE) is a chronic disease bacterium that continues to affect the poultry and causes growth inhibition around 3 weeks of age despite the development of the poultry industry. Recent studies show that necrotizing enteritis is more than 80% worldwide, with 94% in Europe, the highest incidence compared to 89% in the United States, 75% in South America and 65% in Asia. In addition, Clostridium perfringens type A, which produces alpha toxin, is the most relevant bacterium and is associated with most asymptomatic (latent) infections. Infection with this bacterium damages the intestinal mucosa, impairing digestive absorption of nutrients, leading to poor feed efficiency and lower 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 used as feed additives to remove causative bacteria, but antibiotics increase the frequency of antibiotic-resistant bacteria on farms, and antibiotics are delivered unintentionally to humans due to antibiotics remaining through livestock products. Possible side effects are appearing. In particular, the ban on growth-promoting antibiotics (AGPs) in the EU and the ban on the use of antibiotics in feeds in Korea have made it difficult to control intestinal diseases in poultry. Therefore, the poultry industry needs a new strategy for the prevention and control of necrotic enteritis, and methods such as vaccines, prebiotics and microbial agents against Clostridium perfringens are emerging as solutions.

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.Meanwhile, bacteriophage refers to a bacterial specific virus that infects certain bacteria to inhibit and inhibit the growth of infected bacteria. Bacteriophages have the ability to infect bacteria by propagating inside the bacterial cells and then killing the bacteria by destroying the cell wall of the host bacteria when the progeny bacteriophages come out of the bacteria. Bacteriophage bacterial infections are highly specific, so there are only a few types of bacteriophages that can infect certain bacteria. That is, certain bacteriophages can infect only a specific category of bacteria, so that certain bacteriophages kill only certain bacteria and do not affect other bacteria. Therefore, recently, the use of bacteriophage (bacteriophage) as a countermeasure against bacterial diseases has attracted much attention. In particular, bacteriophages have attracted attention as anti-bacterial drugs since 2000, due to the increase of antibiotic-resistant bacteria, the limitations of existing antibiotics appeared, and the possibility of developing them as substitutes for conventional antibiotics emerged.

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, to date, there is still a lack of research on novel bacteriophages for preventing or treating infection caused by Clostridium perfringens, and the technology of bacteriophages having lytic activity specific to Clostridium perfringens and its application Need development

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 can isolate bacteriophages having lytic activity specific to Clostridium perfringens and analyze the morphological and genetic characteristics of the isolated bacteriophages to selectively kill Clostridium perfringens. By confirming that the present invention was completed.

본 발명의 목적은 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 CP5를 제공하는 것이다.An object of the present invention is to provide a bacteriophage CP5 consisting of the nucleotide sequence represented by SEQ ID NO: 1 having a specific killing ability to Clostridium perfringens.

본 발명의 다른 목적은 상기 박테리오파지 CP5를 포함하는, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제를 제공하는 것이다.Another object of the present invention is to provide an antibiotic composition, feed composition, feed, disinfectant or cleaning agent containing the bacteriophage CP5.

본 발명의 또 다른 목적은 상기 박테리오파지 CP5를 개체에 투여하는 단계를 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법을 제공하는 것이다.Still another object of the present invention is to provide a method for preventing or treating an infectious disease caused by Clostridium perfringens, comprising administering the bacteriophage CP5 to a subject.

상기 목적을 달성하기 위하여, 본 발명은 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 CP5를 제공한다.In order to achieve the above object, the present invention provides a bacteriophage CP5 consisting of the nucleotide sequence represented by SEQ ID NO: 1 having a specific killing ability to Clostridium perfringens.

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 항생용 조성물을 제공한다.The present invention also provides an antibiotic composition comprising the bacteriophage CP5.

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 사료 첨가용 조성물을 제공한다.In addition, the present invention provides a composition for feed addition, comprising the bacteriophage CP5.

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 소독제를 제공한다.The present invention also provides a disinfectant comprising the bacteriophage CP5.

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 세척제를 제공한다.The present invention also provides a cleaning agent comprising the bacteriophage CP5.

또한, 본 발명은 상기 박테리오파지 CP5를 개체에 투여하는 단계를 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.The present invention also provides a method for preventing or treating an infectious disease caused by Clostridium perfringens, comprising administering the bacteriophage CP5 to a subject.

본 발명의 박테리오파지 CP5는 기존의 항생제 등의 화학물질에 비해 클로스트리디움 퍼프린젠스에 대한 특이성이 매우 높고, 상기 균에 감염하여 균 내에서 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있다. 또한, 본 발명의 박테리오파지 CP5는 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 CP5는 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.Bacteriophage CP5 of the present invention has a very high specificity for Clostridium perfringens compared to conventional chemicals such as antibiotics, and is excellent in lytic activity and resistant to physicochemical stimulation by proliferating in bacteria. Have In addition, the bacteriophage CP5 of the present invention is not infected with other hosts other than bacteria such as humans, animals, plants, and the like, which can solve problems of antibiotic resistant bacteria due to misuse of antibiotics, residues of antibiotics in food, and a wide range of host problems. There is this. Therefore, the bacteriophage CP5 of the present invention can be used in various fields in the field of antibiotic composition, feed composition, feed, disinfectant, or detergent.

도 1은 박테리오파지 CP5의 클로스트리디움 퍼프린젠스에 대한 용균반 형성 활성을 확인한 결과를 나타낸 도이다.
도 2는 전자 현미경을 통해 박테리오파지 CP5의 형태학적 특성을 확인한 결과를 나타낸 도이다.
도 3은 박테리오파지 CP5의 단백질 패턴을 SDS-PAGE를 통해 확인한 결과를 나타낸 도이다.
도 4는 박테리오파지 CP5의 pH 안정성을 확인한 결과를 나타낸 도이다.
도 5는 박테리오파지 CP5의 열 안정성을 확인한 결과를 나타낸 도이다.1 is a diagram showing the results of confirming the lytic plaque formation activity against Clostridium perfringens of the bacteriophage CP5.
Figure 2 shows the results of confirming the morphological characteristics of the bacteriophage CP5 through an electron microscope.
3 is a diagram showing the results of confirming the protein pattern of the bacteriophage CP5 through SDS-PAGE.
4 is a diagram showing the results of confirming the pH stability of the bacteriophage CP5.
5 is a view showing the results of confirming the thermal stability of the bacteriophage CP5.

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 CP5를 제공한다.The present invention provides bacteriophage CP5 consisting of the nucleotide sequence represented by SEQ ID NO: 1 having a killing ability 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” is a bacterial specific virus that infects a particular bacterium and inhibits and inhibits the growth of the bacterium, and refers to a virus including a single or double chain DNA or RNA as a genetic material. .

본 발명에서, 상기 클로스트리디움 퍼프린젠스는 생산하는 외독소와 효소에 의하여 A, B, C, D, E, F 형 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 제한없이 포함된다. In the present invention, the Clostridium perfringens can be divided into A, B, C, D, E, F type and the like by the exotoxin and enzyme to produce, more specifically, type A bacteria are alpha toxin, type B The bacterium produces beta and epsilon toxin, the type C bacteria are alpha and beta toxin, the type D bacteria are alpha and epsilon toxin, and the type E bacteria produce alpha and iota toxin, respectively, without being limited to the scope of the present invention. do.

본 발명의 박테리오파지 CP5는 약 100-120 nm의 크기로 정이십면체의 머리와 수축성 꼬리를 가지는 마이오비리대(Myoviridae)에 속한다. 상기 박테리오파지 CP5의 단백질 패턴 분석 결과 110 kDa, 54 kDa, 48 kDa, 42 kDa, 33 kDa, 그리고 30 kDa 크기의 단백질을 포함함을 확인하였으며, 박테리오파지 CP5는 열 및 pH에 대한 안정성이 우수한 특성을 가지고 있다.The bacteriophage CP5 of the present invention belongs to Myoviridae having a head and contractile tail of icosahedron with a size of about 100-120 nm. Protein pattern analysis of the bacteriophage CP5 was confirmed to include 110 kDa, 54 kDa, 48 kDa, 42 kDa, 33 kDa, and 30 kDa protein, bacteriophage CP5 has excellent stability against heat and pH have.

본 발명의 박테리오파지 CP5는 서열번호 1로 표시되는 염기서열을 전체 유전자의 전체 또는 일부로서 포함할 수 있다. 또한, 본 발명의 박테리오파지 CP5는 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.Bacteriophage CP5 of the present invention may include the nucleotide sequence represented by SEQ ID NO: 1 as a whole or part of the entire gene. In addition, the bacteriophage CP5 of the present invention may be composed of a nucleotide sequence represented by SEQ ID NO: 1, and a functional equivalent of the nucleotide sequence. The functional equivalent is at least 70%, preferably at least 80%, more preferably at least 90%, even more preferably at least 95% of the nucleotide sequence represented by SEQ ID NO: 1 as a result of modification or substitution of the nucleotide sequence. The above sequence homology means a sequence that exhibits substantially homogeneous physiological activity with the nucleotide sequence represented by SEQ ID NO: 1.

상기와 같은 클로스트리디움 퍼프린젠스 특이적 용균활성, 내산성, 내염기성 및 내열성은 본 발명의 박테리오파지 CP5를 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 및 치료용 조성물, 및 상기 박테리오파지 CP5를 유효성분으로 포함하는 다양한 제품에 적용함에 있어, 다양한 온도 및 pH 범위의 적용이 가능하게 한다.Clostridium perfringens specific lytic activity, acid resistance, base resistance and heat resistance, the bacteriophage CP5 of the present invention, the composition for the prevention and treatment of infectious diseases caused by Clostridium perfringens, and the bacteriophage CP5 effective In application to a variety of products comprising as components, it is possible to apply a variety of temperature and pH ranges.

본 발명의 박테리오파지 CP5는 도축장 및 양돈 농장 부근의 하수로부터 시료를 채취하여 분리한 박테리오파지로서, 본 발명자는 이를 박테리오파지 CP5로 명명하고 2016년 07월 22일에 한국미생물보존센터에 수탁번호 KFCC11676P로 기탁하였다. The bacteriophage CP5 of the present invention is a bacteriophage obtained by separating samples from the sewage near the slaughterhouses and pig farms. .

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 항생용 조성물을 제공한다. The present invention also provides an antibiotic composition comprising the bacteriophage CP5.

본 발명에서, 용어 “항생용 조성물”은 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 an agent that can be provided to a subject in the form of a drug to kill bacteria, and generically refers to preservatives, fungicides, antibiotics and antibacterial agents.

본 발명의 박테리오파지 CP5는 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 bacteriophage CP5 of the present invention has a very high specificity for Clostridium perfringens as compared to conventional antibiotics, beneficial bacteria are not killed, only certain pathogens can be killed, and drug resistance or resistance is not induced. In comparison, life cycle can be used as a new novel antibiotic.

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 사료 첨가용 조성물을 제공한다.In addition, the present invention provides a composition for feed addition, comprising the bacteriophage CP5.

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 CP5를 이용할 수 있다.Feeding antibiotics used in livestock and fisheries are used for the prevention of diseases, and the administration of antibiotics for prevention is a problem because it increases the likelihood of developing resistant bacteria and the antibiotics remaining in livestock can be delivered to humans. When antibiotics are absorbed into the body through meat, they can cause antibiotic resistance, which can lead to the spread of disease. In addition, there are many kinds of antibiotics to be mixed in the feed, which has a problem of increasing the probability of occurrence of multi-drug-resistant bacteria is more natural and yet to solve the problems arising from the use of conventional antibiotics, the bacteriophage CP5 of the present invention as a new feed additive antibiotics 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 건조상태일 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 composition for feed addition, the feed of the present invention can be prepared by separately mixing the feed in the form of feed additives, or in the preparation of the feed to produce the bacteriophage. . Bacteriophage in the feed of the present invention may be a liquid or dried state, preferably in the form of a dry powder. Drying methods may be, but not limited to, ventilation drying, natural drying, spray drying and freeze drying. Bacteriophage of the present invention may be mixed in the form of a powder of 0.05 to 10% by weight, preferably 0.1 to 2% by weight of the feed in the form of a powder. In addition, the feed may further include conventional additives to increase the shelf life of the feed in addition to the bacteriophage of the present invention.

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 feed composition of the present invention, other non-pathogenic microorganisms may be further added. Microorganisms that may be added include Lactobacillus, which has physiological activity and organic degradability under anaerobic conditions such as Bacillus subtilis , Bacillus subtilis , which can produce proteolytic enzymes, lipolytic enzymes and sugar converting enzymes, and bovine stomach. Strain ( Lactobacillus sp.), Filamentous fungi and Saccharomyces cerevisiae , such as Aspergillus oryzae , which increase the weight of livestock, increase milk yield and increase digestibility of feed Can be selected from the group consisting of yeast.

본 발명의 박테리오파지 CP5를 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.In the feed containing the bacteriophage CP5 of the present invention, the plants include grains, root fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils, starches, gourds, grain by-products, etc. Proteins, minerals, fats, oils, minerals, single-cell proteins, zooplankton, leftovers, and the like are not limited thereto.

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 feed composition of the present invention may include a binder, an emulsifier, a preservative, and the like, added to prevent quality deterioration, and include amino acids, vitamins, enzymes, probiotics, flavors, and non-proteins, which are added to the feed to increase its efficacy. Nitrogen compounds, silicates, buffers, colorants, extractants, oligosaccharides, and the like may be included, in addition to the feed mixture may be further included.

또한, 본 발명은 박테리오파지 CP5를 포함하는, 음용수 첨가제를 제공한다.The present invention also provides a drinking water additive, including bacteriophage CP5.

본 발명의 음용수 첨가제는 상기 박테리오파지 CP5 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.Drinking water additives of the present invention may be used in the form of mixing the bacteriophage CP5 or a composition comprising the same separately in the form of drinking water additives to feed or drinking water, or may be used in the manner of adding directly in the preparation of drinking water. By supplying the mixture to the drinking water as described above there is an effect that can continuously reduce the number of Clostridium perfringens.

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 conventionally used in the art may be used.

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 소독제를 제공한다.The present invention also provides a disinfectant comprising the bacteriophage CP5.

클로스트리디움 퍼프린젠스에 특이적 사멸능을 갖는 본 발명의 박테리오파지 CP5를 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 bacteriophage CP5 of the present invention having specific killing ability in Clostridium perfringens can be usefully used as a disinfectant for hospitals and health care to prevent hospital infections, general household disinfectants, food and cooking places and equipment It can be used for disinfection of various livestock products such as disinfectant, poultry farm, barn, building, shaft, drinking water, litter, stool, transport vehicle, tableware.

또한, 본 발명은 상기 박테리오파지 CP5를 포함하는, 세척제를 제공한다.The present invention also provides a cleaning agent comprising the bacteriophage CP5.

본 발명의 박테리오파지 CP5는 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 bacteriophage CP5 of the present invention has specific killing ability to Clostridium perfringens, it is also used for cleaning the skin surface or parts of the body of individuals exposed to or possibly exposed to Clostridium perfringens. Can be used.

또한, 본 발명은 상기 박테리오파지 CP5를 개체에 투여하는 단계를 포함하는, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.The present invention also provides a method for preventing or treating an infectious disease caused by Clostridium perfringens, comprising administering the bacteriophage CP5 to a subject.

본 발명의 박테리오파지 CP5는 클로스트리디움 퍼프린젠스에 특이적인 사멸능을 가지므로, 클로스트리디움 퍼프린젠스에 의한 감염성 질병의 예방 또는 치료에 이용될 수 있다. Bacteriophage CP5 of the present invention has a specific killing ability to Clostridium perfringens, it 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, necrotic infection, gas necrosis disease, soft tissue infection, purulent myositis, and the like, secondary to Clostridium perfringens Infections are included.

본 발명의 용어 “예방”이란 조성물의 투여로 질병을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.The term "prevention" of the present invention means any action that inhibits or delays the onset of the disease by administration of the composition.

본 발명의 용어 “치료”란 조성물의 투여로 상기 질병의 증세가 호전되거나 상기 질병의 억제 또는 경감 및 이롭게 변경되는 모든 행위를 의미한다.The term “treatment” of the present invention means any action in which administration of the composition improves or suppresses or alleviates and beneficially alters the symptoms of the disease.

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" means any individual capable of developing an infectious disease caused by Clostridium perfringens, and includes, but is not limited to, mammals, livestock or poultry, including humans.

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" refers to a useful animal that is purified and improved by humans to share a common life with humans, including, but not limited to, pigs, cattle, chickens, horses, ducks, or dogs. It doesn't happen.

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” is a concept that collectively refers to animals belonging to birds in livestock, and includes, but is not limited to, for example, chicken, duck, pheasant, quail, ostrich, goose or turkey.

본 발명의 방법에서 상기 박테리오파지 CP5 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 CP5 or the composition may be administered to an animal in the form of a pharmaceutical preparation, or may be administered through a method of feeding on a feed or drinking water of a livestock.

본 발명의 약학적 제제(약학적 조성물과 혼용됨)는 유효성분 외에 약학적으로 허용 가능한 담체를 추가로 포함할 수 있다.The pharmaceutical formulation of the present invention (mixed with the pharmaceutical composition) may further include a pharmaceutically acceptable carrier in addition to the active ingredient.

본 발명에서 용어, “약학적으로 허용 가능한 담체”란 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 1 성분 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.As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not irritate an organism and does not inhibit the biological activity and properties of the administered compound. Acceptable pharmaceutical carriers in compositions formulated in liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, 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 to a diseased site, and can also be administered through nasal spray, oral administration or parenteral administration, and in the case of parenteral administration, intravenous administration, intraperitoneal administration Administration may be by administration, intramuscular administration, subcutaneous administration or topical administration.

상기 비강 분무의 경우 일정량의 박테리오파지 CP5를 물에 희석하여 분무기나 분무 시스템을 통해 비강으로 흡수되도록 할 수 있다. 상기 비강 분무를 위한 비강 분무제 또는 호흡기용 제형으로는 에어로졸제 등을 들 수 있다.In the case of nasal spray, a certain amount of bacteriophage CP5 may be diluted in water to be absorbed into the nasal cavity through a nebulizer or a spray system. Examples of the nasal spray or respiratory formulation for nasal spray include aerosols and the like.

상기 비강 분무제 또는 호흡기용 제형은, 수분산된 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 to disperse the dispersed dispersion or the 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.Suitable applications, sprays, and dosages of the pharmaceutical compositions of the present invention may be formulated by the method of formulation, mode of administration, age, weight, sex, degree of disease symptom, food, time of administration, route of administration, rate of excretion and Depending on factors such as response responsiveness, usually an experienced physician or veterinarian can readily determine and prescribe a dosage 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 마스네슘, 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.Formulations for oral administration comprising the pharmaceutical composition of the present invention as an active ingredient include, for example, tablets, troches, lozenges, water-soluble or oily suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or elixirs It can be formulated to zero. For formulation into tablets and capsules, lactose, saccharose, sorbitol, mannitol, starch, amylopectin, binders such as cellulose or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, stearic acid masne It may include lubricating oils such as calcium, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax, and in the case of capsule formulations, it may further contain a liquid carrier such as fatty oil in addition to the above-mentioned materials.

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 제제화할 수 있다. 에어로졸제 등의 스프레이용으로 제형화하는 경우, 수분산된 농축물 또는 습윤 분말이 분산되도록 추진제 등이 첨가제와 함께 배합될 수 있다.Formulations for parenteral administration comprising the pharmaceutical composition of the present invention as an active ingredient include, for example, injectable forms such as subcutaneous injection, intravenous injection or intramuscular injection, suppository injection method, or aerosol for inhalation through the respiratory system. It can be formulated for spraying. To formulate injectable formulations, the compositions of the present invention may be mixed in water with stabilizers or buffers to prepare solutions or suspensions, which may be formulated for unit administration of ampoules or vials. When formulated for spraying such as aerosols, a propellant or the like may be combined with the additives to disperse the dispersed dispersion or wet powder.

본 발명의 방법에서 상기 박테리오파지 CP5 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 CP5 or the composition may be administered through various routes orally or parenterally as long as it can reach the target tissue, and specifically, oral, rectal, topical, intravenous, intraperitoneal, Intramuscular, intraarterial, transdermal, intranasal, inhalation and the like can be administered in conventional manner.

본 발명의 방법에 투여되는 상기 박테리오파지 CP5의 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 usage of the bacteriophage CP5 administered in the methods of the invention can be determined by the practitioner within the correct medical judgment. The specific therapeutically effective amount for a particular individual may be determined in conjunction with the type and severity of the response to be achieved, the age, body weight, general health, sex and diet, time of administration, route of administration and rate of composition, duration of treatment, and specific composition of the patient. It is desirable 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 defined otherwise in this specification are intended to have a meaning commonly used in the art to which the present invention pertains.

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 by way of examples and preparation examples. However, the following Examples and Formulation Examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following Examples and Formulation Examples.

실시예Example 1:  One: 신규한New 박테리오파지의 분리 및 배양 Isolation and Culture of Bacteriophage

1.1 박테리오파지의 스크리닝 및 분리1.1 Screening and Isolation of Bacteriophage

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.A lytic plaque assay was performed to screen and isolate bacteriophages specific for Clostridium perfringens. Samples were collected from the sewage near the slaughterhouses and pig farms where bacteriophage was expected to be present, and Clostridium perfringens used for the separation of bacteriophages was identified and isolated from outdoor samples collected from Optifam Co., Ltd.

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 um인 필터를 이용하여 여과하여 시료 여과액을 얻었다. Specifically, the sample was homogenized and then shaken overnight at 35 ° C with Clostridium perfringens. At this time, Clostridium perfringens is anaerobic bacteria that inhibit the growth if oxygen is present, all cultured in 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 um to obtain a sample filtrate.

전배양된 클로스트리디움 퍼프린젠스 150 uL를 취해서 멸균된 튜브에 넣고 3-5 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 uL씩 점적하고, 다시 실온에서 30분간 방치한 후 35℃에서 24시간 동안 배양하여 박테리오파지의 유무를 확인하였다. Take 150 uL of pre-cultured Clostridium perfringens and place in a sterile tube in 3-5 mL of BHI (Brain Heart Infusion) 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) was added and mixed well, and then layered onto a 2% BHI agar plate prepared in advance. After standing at room temperature for 30 minutes, each sample filtrate was added dropwise by 10 uL, and then left at room temperature for 30 minutes, and then cultured at 35 ° C. for 24 hours to confirm the presence of bacteriophage.

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 uL를 취하고, 대상 세균(클로스트리디움 퍼프린젠스) 배양액 150 uL에 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 uL의 SM 완충액 (NaCl 5.8 g/L; MgSO47H2O 2 g/L; 1M Tris-HCl (pH 7.5) 50 mL)에 현탁하고, 실온에서 4시간동안 방치하였다. 상기 박테리오파지액 100 uL를 0.6% BHI 탑 아가와 세균 배양액 500 uL와 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 0.2 um 필터로 여과하여 최종 박테리오파지 용액을 회수하였다. 이렇게 얻어진 최종 박테리오파지의 용균반을 확인한 결과를 도 1에 나타내었다.A soft agar overlay method was used to purely separate the bacteriophages from the samples confirming the presence of the bacteriophages. Dilute the sample identified as bacteriophage with saline, take 100 uL of sample dilution from each dilution stage, mix it with 150 uL of the target bacterial (Clostridium perfringens) culture, and add 3 mL of 0.6% BHI top agar. After the addition, the mixture was mixed well, and the mixture was left to stand on a previously prepared 2% BHI agar plate and incubated at 35 ° C for 24 hours. After identifying the lysate formed on each plate incubated, it was selected and suspended in 400 uL of SM buffer (NaCl 5.8 g / L; MgSO 4 7H 2 O 2 g / L; 50 mL of 1M Tris-HCl (pH 7.5)). And left at room temperature for 4 hours. 100 uL of the bacteriophage solution was mixed with 0.6% BHI top agar and 500 uL of bacterial culture solution, and then aliquoted into a 150 mm diameter BHI plate and incubated at 35 ° C until complete lysis occurred. After the incubation was completed, 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. After centrifugation at 4,000 rpm for 20 minutes, only the supernatant was collected and filtered through a 0.2 um filter to obtain the final bacteriophage. The solution was recovered. The results of confirming the lysate plaque of the final bacteriophage thus obtained are shown in FIG. 1.

도 1에 나타낸 바와 같이, 약 2-3 mm의 크기를 가지는 용균반을 형성하는 박테리오파지를 확인하였고, 상기 박테리오파지는 클로스트리디움 퍼프린젠스에 특이적인 용균능을 갖는 박테리오파지임을 확인하였다.As shown in Figure 1, it was confirmed that the bacteriophage forming a lytic plaque having a size of about 2-3 mm, the bacteriophage was confirmed to be a bacteriophage having a lytic ability specific to Clostridium perfringens.

1.2 박테리오파지의 대량 배양 및 정제1.2 Mass Cultivation and Purification of Bacteriophage

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 um 필터로 여과하여 최종 박테리오파지 배양액을 회수하였다. 분리된 박테리오파지를 “박테리오파지 CP5(Bacteriophage CP5)”로 명명하고, 2016년 07월 22일 한국미생물보존센터 (Korean Culture Center of Microorganisms, 서울, 한국)에 기탁하고 수탁번호 KFCC11676P를 부여받았다.In order to mass-produce the Clostridium perfringens specific bacteriophage identified in Example 1.1, the final bacteriophage culture solution recovered in 1.1 was 1.0 × 10 8. After diluting with PFU / mL, mixed with pre-cultured Clostridium perfringens 1.0 × 10 9 CFU / mL, the MOI (Multiplicity of infection) became 0.1, and incubated at 35 ° C. at 120 rpm for 9 hours. After the incubation was completed, the entire culture was centrifuged at 6,000 rpm for 30 minutes to obtain a supernatant, which was filtered through a 0.2 um filter to recover the final bacteriophage culture. The separated bacteriophage was named “Bacteriophage CP5” and was deposited on July 22, 2016 at the Korean Culture Center of Microorganisms (Seoul, Korea) and was assigned accession number KFCC11676P.

실시예Example 2: 박테리오파지 CP5의 전체 유전자 분석 2: Whole Gene Analysis of Bacteriophage CP5

박테리오파지 CP5의 전체 유전자를 분석하기 위해, 박테리오파지 CP5 배양액에 20 mM EDTA, 50 ug/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 uL가 되도록 TE 완충액 (Tris-EDTA, pH8.0)에 용해시킨 후 DNA 농도를 측정하였다. 전체 유전자 분석을 위해 HiSeq 2000(Illumina, USA)를 이용하여 유전자 시퀀싱을 시행하였다 (마크로젠, 한국). To analyze the entire gene of bacteriophage CP5, 20 mM EDTA, 50 ug / mL proteinase K, 0.5% (w / v) SDS was added to the bacteriophage CP5 culture, and then allowed to stand at 50 ° C for 1 hour. The supernatant was recovered by mixing with an equal amount of phenol-chloroform-isopropyl alcohol (25: 24: 1) and centrifuging at 12,000 rpm for 10 minutes at room temperature. The recovered supernatant was mixed with an equal volume of phenol-chloroform-isopropyl alcohol (25: 24: 1), centrifuged at 12,000 rpm for 10 minutes at room temperature, and then the supernatant was recovered, and 3M sodium acetate was added to the supernatant. 10% (v / v) was added and mixed. Two volumes of cold 95% ethanol were added thereto and mixed, followed by standing at -20 ° C for 1 hour. After the reaction, the precipitate was obtained by centrifugation at 12,000 rpm for 10 minutes at 0 ° C. After washing twice with cold 70% ethanol, the ethanol was completely removed to recover the DNA pellet. The recovered pellet was dissolved in TE buffer (Tris-EDTA, pH8.0) to a final volume of 50 uL, and then the DNA concentration was measured. Gene sequencing was performed using HiSeq 2000 (Illumina, USA) for total gene analysis (Macrogen, Korea).

그 결과, 박테리오파지 CP5의 전체 유전자의 크기는 51,600 bp 이며, GC 함량이 34%, 전체 ORFs의 수는 79개임을 확인하였다. 박테리오파지 CP5의 전체 유전자 서열을 서열번호 1로 나타내었다.As a result, the total gene size of the bacteriophage CP5 was 51,600 bp, GC content was 34%, the total number of ORFs was confirmed to be 79. The entire gene sequence of bacteriophage CP5 is shown in SEQ ID NO: 1.

실시예Example 3: 박테리오파지 CP5의 형태학적 특성 3: Morphological characteristics of bacteriophage CP5

박테리오파지 CP5의 형태학적 특성을 알아보기 위해, 다음과 같은 실험을 수행하였다. 먼저, 박테리오파지 CP5 배양액을 2.5% 파라포름알데히드-글루타르알데히드(4℃, phosphate buffer, pH 7.2) 고정액에서 2시간 전고정하고, 완충액(0.1M phosphate buffer, pH 7.2)으로 10분씩 3회 세척한 후, 1% OsO4(25℃, 0.1M phosphate buffer, pH 7.2)에서 2시간동안 후고정하였다. 고정이 끝난 시료를 동일 완충용액으로 수회 세척한 후, 에탄올 농도 상승 순으로 탈수하였으며, 프로필렌 옥사이드로 치환하여 Epon-812로 포매한 다음, 60℃ 오븐에서 36시간 중합반응시켰다. 포매된 조직은 울트라마이크로톰(ULTRACUT E, Leica)을 이용하여 초박절편으로 제작하였으며, 아세트산 우라닐(uranyl acetate)과 납 시트레이트(lead citrate)로 이중염색한 후, 투과전자현미경 (CM 20, Philips)으로 100kV에서 120,000~250,000배 사이로 관찰하였다. 그 결과를 도 2에 나타내었다.In order to determine the morphological characteristics of the bacteriophage CP5, the following experiment was performed. First, the bacteriophage CP5 culture was pre-fixed in 2.5% paraformaldehyde-glutaraldehyde (4 ° C., phosphate buffer, pH 7.2) fixed solution for 2 hours, and washed three times for 10 minutes with buffer (0.1 M phosphate buffer, pH 7.2). Then, it was post-fixed in 1% OsO 4 (25 ° C., 0.1 M phosphate buffer, pH 7.2) for 2 hours. The fixed sample was washed several times with the same buffer solution, dehydrated in the order of increasing ethanol concentration, substituted with propylene oxide, embedded with Epon-812, and then polymerized in an oven at 60 ° C. for 36 hours. Embedded tissues were prepared in ultra-thin sections using ultramicrotoms (ULTRACUT E, Leica), and double-stained with uranyl acetate and lead citrate, followed by transmission electron microscopy (CM 20, Philips). ) Was observed between 120,000 and 250,000 times at 100kV. The results are shown in FIG.

도 2에 나타낸 바와 같이, 박테리오파지 CP5는 약 100-120 nm의 크기로 정이십면체의 머리와 수축성 꼬리를 가지는 마이오비리대(Myoviridae)에 속함을 확인하였다. As shown in FIG. 2, the bacteriophage CP5 was found to belong to Myoviridae having a head and contractile tail of icosahedron with a size of about 100-120 nm.

실시예Example 4: 박테리오파지 CP5의 단백질 패턴 분석 4: Protein pattern analysis of bacteriophage CP5

박테리오파지 CP5의 단백질 패턴을 분석하기 위해, 1.0×1010 PFU/mL로 준비된 박테리오파지 CP5 40 uL를 10 uL의 5X 로딩 완충액과 혼합하고, 10분간 끓인 다음 12.5% SDS-PAGE를 수행하였다. SDS-PAGE 수행 후 단백질 전기영동 결과를 도 3에 나타내었다.To analyze the protein pattern of bacteriophage CP5, 40 uL of bacteriophage CP5 prepared at 1.0 × 10 10 PFU / mL was mixed with 10 uL of 5 × loading buffer, boiled for 10 minutes and then subjected to 12.5% SDS-PAGE. Protein electrophoresis results after SDS-PAGE are shown in FIG. 3.

도 3에 나타낸 바와 같이, 박테리오파지 CP5는 110 kDa, 54 kDa, 48 kDa, 42 kDa, 33 kDa, 그리고 30 kDa 크기의 주요 단백질을 가지는 것을 확인하였다.As shown in Figure 3, bacteriophage CP5 was confirmed to have a major protein of 110 kDa, 54 kDa, 48 kDa, 42 kDa, 33 kDa, and 30 kDa.

실시예Example 5: 박테리오파지 CP5의 다양한 세균에 대한 감염성 조사 5: Investigation of various bacteria of bacteriophage CP5

박테리오파지 CP5의 다양한 세균에 대한 감염성을 조사하였다. The bacterial nature of the bacteriophage CP5 was investigated.

우선, 박테리오파지 CP5의 클로스트리디움 퍼프린젠스에 대한 용균 활성을 확인하기 위하여, ㈜옵티팜에서 분리하여 보유하고 있는 클로스트리디움 퍼프린젠스 균주를 대상으로 소프트 아가 오버레이법을 수행하였다. 그 결과를 표 1에 나타내었다.First, in order to confirm the lytic activity of Clostridium perfringens of the bacteriophage CP5, the soft agar overlay method was performed on the Clostridium perfringens strains isolated and retained in Optipal. The results are shown in Table 1.

대상 세균명Target bacteria name 대상 세균수Target bacteria 용균 활성이
확인된 수Lytic activity
Verified number 용균 활성이
확인되지 않은 수Lytic activity
Unverified number 감수성 비율Sensitivity ratio Clostridium Clostridium perfringensperfringens 32 균주32 strains 30 균주30 strains 2 균주2 strains 93.8%93.8%

상기 표 1에 나타낸 바와 같이, 본 발명의 박테리오파지 CP5는 클로스트리디움 퍼프린젠스에 대해 높은 감수성을 보이며, 이를 통해 클로스트리디움 퍼프린젠스에 대한 용균활성이 탁월함을 확인하였다.As shown in Table 1, the bacteriophage CP5 of the present invention showed high sensitivity to Clostridium perfringens, through which it was confirmed that the lytic activity against Clostridium perfringens is excellent.

다음으로, 박테리오파지 CP5가 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 2에 나타내었다.Next, in order to confirm whether bacteriophage CP5 showed lytic activity against other bacteria, the infectivity was confirmed through the soft agar overlay method for the strains isolated from Optifam Inc. and the strains distributed in the microbial resource center. The results are shown in Table 2.

Figure 112017128490983-pat00001
Figure 112017128490983-pat00001

표 2에 나타낸 바와 같이, 본 발명의 박테리오파지 CP5는 클로스트리디움 퍼프린젠스 외에 대장균, 살모넬라 등의 다른 속 균주에 대해서는 용균활성이 없는 것을 확인하였다.As shown in Table 2, it was confirmed that bacteriophage CP5 of the present invention had no lytic activity against other genus strains such as Escherichia coli and Salmonella, in addition to Clostridium perfringens.

따라서, 본 발명의 박테리오파지 CP5는 클로스트리디움 퍼프린젠스에 특이적으로 용균활성을 나타내는 박테리오파지임을 확인하였다.Therefore, the bacteriophage CP5 of the present invention was confirmed to be a bacteriophage showing lytic activity specifically to Clostridium perfringens.

실시예Example 6: 박테리오파지 CP5의 pH 안정성 조사 6: pH stability investigation of bacteriophage CP5

박테리오파지 CP5의 pH 안정성을 확인하기 위하여, 다양한 pH 범위 (pH 2, 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 uL로 분주하여 준비하였다. 박테리오파지 CP5 배양액을 2.0×108 PFU/mL로 준비한 다음, 상기 배양액을 10 uL씩 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 4에 나타내었다.In order to confirm the pH stability of the bacteriophage CP5, the number of bacteriophages was measured in various pH ranges (pH 2, 3, 4, 5, 6, 7, 8, 9, 10, 11). Sodium acetate buffer, sodium phosphate buffer, Tris-HCl was prepared for each pH and then prepared by dispensing at 990 uL. Bacteriophage CP5 culture solution was prepared at 2.0 × 10 8 PFU / mL, and the culture solution was added to each of the buffers prepared according to pH by taking 10 uL and left at room temperature for 2 hours. 4 shows the result of performing titer measurement by soft agar overlay method by diluting each reaction solution after standing.

도 4에 나타낸 바와 같이, 본 발명의 박테리오파지 CP5는 pH 3-11의 범위에서 2시간 동안 안정한 것을 확인하였다. 이를 통해 박테리오파지 CP5는 pH 안정성이 우수한 박테리오파지임을 알 수 있다.As shown in Figure 4, the bacteriophage CP5 of the present invention was confirmed to be stable for 2 hours in the range of pH 3-11. This shows that bacteriophage CP5 is a bacteriophage with excellent pH stability.

실시예Example 7: 박테리오파지 CP5의 열 안정성 조사 7: Investigation of thermal stability of bacteriophage CP5

박테리오파지 CP5의 열 안정성을 확인하기 위하여, 40℃, 50℃, 60℃, 및 70℃ 등 다양한 온도 범위에서 일정 시간 방치 후 박테리오파지의 수를 측정하였다. 보다 구체적으로, 박테리오파지 CP5 배양액을 2.0×106 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 5에 나타내었다.In order to confirm the thermal stability of the bacteriophage CP5, the number of bacteriophages was measured after standing for a certain time in various temperature ranges such as 40 ℃, 50 ℃, 60 ℃, and 70 ℃. More specifically, after diluting the Bacteriophage CP5 culture to 2.0 × 10 6 PFU / mL, 5 mL of the culture was dispensed into sterile tubes and left at each temperature for 30 minutes, 1 hour, and 2 hours. After standing, each reaction solution was diluted with a step to perform titer measurement by a soft agar overlay method.

도 5에 나타낸 바와 같이, 본 발명의 박테리오파지 CP5는 40-70℃의 범위에서 약 60분 이상 안정한 것을 확인하였다. 이를 통해 박테리오파지 CP5는 내열성이 우수한 박테리오파지임을 알 수 있다.As shown in Figure 5, the bacteriophage CP5 of the present invention was confirmed to be stable for about 60 minutes or more in the range of 40-70 ℃. It can be seen that the bacteriophage CP5 is a bacteriophage having excellent heat resistance.

상기 실험으로부터 박테리오파지 CP5는 클로스트리디움 퍼프린젠스에 대한 특이성이 매우 높고, 균 내에서 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있음을 확인하였다. 또한 본 발명의 박테리오파지 CP5는 사람, 동물, 식물 등 세균외의 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 CP5는 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.From the above experiments, it was confirmed that bacteriophage CP5 had a very high specificity for Clostridium perfringens, proliferated in bacteria, and excellent in lytic activity and strong in physicochemical stimulation. In addition, since the bacteriophage CP5 of the present invention is not infected with other hosts other than bacteria such as humans, animals, plants, and the like, the problems of antibiotic resistance bacteria due to the misuse of antibiotics, the residue of antibiotics in food, and the problems of a wide range of host ranges are solved. have. Therefore, the bacteriophage CP5 of the present invention can be used in various fields in the field of antibiotic composition, feed composition, feed, disinfectant, or detergent.

이하, 제제예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 제제예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 제제예에 의해 제한되는 것으로 해석되지 않는다.Hereinafter, the present invention will be described in more detail with reference to the formulation examples. The formulation examples are only for illustrating the present invention, and the scope of the present invention is not to be construed as being limited by the formulation examples.

제제예 1. 사료 첨가용 조성물Formulation Example 1 Composition for Feed Addition

박테리오파지 CP5 1x107 PFU/mLBacteriophage CP5 1x10 7 PFU / mL

비타민 믹스 100 mg100 mg of vitamin mix

미네랄 믹스 100 mg100 mg of mineral mix

스테아린산 마그네슘 2 mg2 mg magnesium stearate

통상의 사료 첨가제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.It is prepared by mixing the above components according to a conventional feed additive manufacturing method.

제제예 2. 소독제 조성물Formulation Example 2 Disinfectant Composition

박테리오파지 CP5 1x107 PFU/mLBacteriophage CP5 1x10 7 PFU / mL

에탄올 적량Ethanol

멸균 증류수 적량Sterile distilled water

pH 조절제 적량pH adjuster

통상의 소독제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.It is prepared by mixing the above components according to a conventional disinfectant production method.

제제예Formulation example 3. 세척제 조성물 3. Cleanser Composition

박테리오파지 CP5 1x107 PFU/mLBacteriophage CP5 1x10 7 PFU / mL

에탄올 적량Ethanol

효소 복합물 적량Enzyme Complex Proper

멸균 증류수 적량Sterile distilled water

통상의 세척제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.It is prepared by mixing the above components in accordance with a conventional cleaning method.

이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다. As mentioned above, specific parts of the present invention have been described in detail, and it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

한국미생물보존센터(국내)Korea Microbial Conservation Center KFCC11676PKFCC11676P 2016072220160722

<110> Optipharm.CO.,LTD <120> Novel Clostridium perfringens specific bacteriophage CP5 and antibacterial composition comprising the same <130> OTP1-38p <150> KR 10-2016-0178220 <151> 2016-12-23 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 51600 <212> DNA <213> Artificial Sequence <220> <223> bacteriophage CP5 <400> 1 aaaaaaacca gtaagaaatc tggtaaaaag aaaaaataaa attttctaaa aatacctttg 60 acaaccaatt aaaaatctga taatataaac ttgtaaacaa agaggtacaa tatcttgaca 120 aatatttatc tcataatgat attttaaaat ttaaggagga caataacatg gctaaaaaga 180 ctactaagag aggtaacaaa aacaaaggag gattaaagat tgatctttca aacgttgaaa 240 cttcagttac tatcccagaa ggaaattaca ttgtggaagt agaagatgta gaggttaagg 300 tttctgaaaa tagtggaagc aattatttat catttacttt tgtaatagca gaaggaaaga 360 tgaaaggaca aaagttatac cacatttgct cacttcaacc acaagcttta tttaacttaa 420 aaggtgtgtt agttgcttta ggatttgata tccctgatga ggagttcgaa ttagatacag 480 aagctctagt tggtttacaa tgtggggtag aagtatcaca tgaaatatat gagggtaaga 540 agaaatcaag aataactgat tttataaacc ttgacgaagc tgactctgat gatgacgaag 600 atgaggatga tgactcagat gatgatgaag aagatgatga agacgatgag tctgaagttg 660 atcttgaaga attagacaag gatgagctaa aagaattagc taaggcttta aaaatcccag 720 ctaagaaaat caagaaggct aaaactgaag aagatctaat tgatctaatt gaagaagaag 780 ctgacgaaga agaaatagct gaacaatata atgacctatt cggagactct gatgaagatg 840 acgaagaaga tgaggatgaa gaagaggaag atgaagaaga aaatgactat gagtcaatga 900 ctttaaaaga acttaaagct gaagctaagg acagaggctt aaaggttaaa aaaggaatgt 960 ctaaggatga catcatagaa atgctagaag aagatgatga agaataaaat ttaattatta 1020 aaatatgagg gcctttaagg tcctccttta tttacccttg gaggctacta ttatgcttga 1080 aagagatgtg gttaaatcca taatgaatat gcttaaaaaa gaatacccag gtttttggtt 1140 taaaactcat gggggaccat ttcaaatagc tggcttacct gatatactag gttgccacaa 1200 aggtaagttt attggtattg aagttaaact tcctggaaaa gaaaagaacc taactcaaaa 1260 acaaaaagac attataaata aaataaatct agcaggagga atagctttta tggctacctc 1320 agctgaatat acaaggagga gattacatga aaaatttaga aagacaccaa caattcctag 1380 gagaactagg agatctgtat gaattaaaga ataatctata tggggataat ttccacaaga 1440 cttatcttga atatggaaac cctgttctat gtataagact tgaagataag ctaggaagag 1500 ctaaaagttt attactagga gatcaagatg acttcccttc atatgctgct caaaaagaat 1560 ctgtggttga tactttacta gacctagcta actatgccat tatggctgct atggaactaa 1620 ctagtgatga taactcagaa atacatgatc ttagtaaaga agaatatgat gaagaagacc 1680 tagatgataa tgacgatgat gacttagatg atgatgacaa tgtttacgac gaagaagaat 1740 tagattttga tagtatgaat aaagaatcct taaaacaata tctaaaagat aatggggtta 1800 aattccatag taaggcctca agagatgaac ttgtaaaact ggctaaggag gtataaggag 1860 gaggctttaa tgcctcttct tttttagtat gaagagaaaa ttatttaaac atcagaaaga 1920 agcattacaa ctatttttaa ataaagaaaa gtttgcccta tttatggaca tgggtacggg 1980 taagacctta gtccctattg tggcccttga gaaacttgaa ggtctagata ctgtactaat 2040 attctcacct aaatctattg tatttaactg ggagtctgag atccataaat ttactaaact 2100 taaggaatat aggatattta aattacaggg taataagacc aaggttatgg aaacctatag 2160 agctataaaa tcctactcag gattaaagat tattattgcc aattttgaga aggctaggtt 2220 gatggataaa taccttatga acttaaagcc acagtttatt gttgttgacg aatcccataa 2280 ggtaaagaat agaaatgccc agatatctaa ggctctatat aaaattgcta ctaaatgtaa 2340 atataggttg ataatgacag ggactcccac tcctaatggt tatgaagatt tatttatgca 2400 atataaaatt atgaactcaa atattctagg agttaattgg aaacagtttg aagatgactt 2460 tataatcaaa ggaggttata tgaactatga aattgtgggg tataagaatg aggaaatatt 2520 aaagaacctt atgcaccaga attgctatat agtaagaata gaagattgta ttgatttacc 2580 agaacaactt cctgatctgt atttaacctg tgaattaaat agtaaagcca gaaaggccta 2640 taatgacctt agaaaggaaa tgatagcaca actagatata gtacaggaaa acattcctag 2700 gaagcaatta aaggccttat taaggtctaa tggtataccc tatgagagta atgagcccta 2760 tgaggattta ttcttaaggg ctaatatgtt tatcaaccaa ttaacagctg atcttactat 2820 tacccaatat ttgaggctac agcagatctc aggaggattt atcacaaaca atgtaggaaa 2880 taatattaat attgataaag gcaaattagg cttgttacag gactatctag aaggatataa 2940 aaagcctgtt gttgtaatat gtaattttct tgaggaaata aaacttatac atgatacttt 3000 taaaaagacc cacagagtag aatgcttaac aggatctact aagaatagag ctgagatcaa 3060 taaggatttt caggagggca agattgatat attaatacta caaataagct caggtagtgt 3120 tggtctaaac ctcttcaggg cctcaaggtt aatattctat agttggaact ataaatatga 3180 tgattatgtg caagctattg ccagaatcaa aaggaatggt caaaaggagc cttggcaaat 3240 aatacactta ataacggaga atactattga tgagaaaata ttaaaatcaa tacagcttaa 3300 aagggataga gcagaaaaac tgttgactat tgataagtaa tgtggtaata tagaattgag 3360 gtgataagta atgagaaaag aacaacctag aataagtaat catactgaaa tagctaaggc 3420 tttaattgag gctggtaaaa ctattaaagc tctaaccaag attaaagtca aggaggaaaa 3480 gaaaaaacaa aaaagacttg aaaggttaag actcctaaat gaggaaaaga agaaggagag 3540 agaattaatg aaagaattta atcccgatga gcctaagaag ccaagaaaga aaaaggacaa 3600 acctataaag aaggttgatc tgaataaagt tgaggctaag ggaacttacc ctacaccttc 3660 tggaggtaat gcacctacgg gaacagttga cttaaaaatt ttatgtgagt cattaggatt 3720 agatccaagt aaagccaggg ccaagctaag aaagcaaggt gtaaacaaac cctataaatg 3780 gtcaggatct gaacttgagg atattaagaa aatgttaaaa taagggggag gtttaaaaac 3840 ctcttctttt tcgttgacaa tatccacagt atatggtaat ataaatattg taatcaaaca 3900 aaaaaggaga gaactgaata tgaaagatct tgaaaaactt acaagggaag aattgattga 3960 acttgtaaag gagttatcta ttgattggac aggagcctat aatagaaact ataagattga 4020 ccttgatgag tatgagattg tatatattga tcttaatgat ctaaaagagt tcaatgataa 4080 attaggtcat ctagaaggag atgtgtatat acaatacacg gtaaatctta ttaaggccca 4140 tttacatcct gagaaggata tattgattag agggggtggt gatgagtttg tagttatcag 4200 tgatatagct tataatctgt gcaatacctt gaatatatac cctgagctat catgtggcta 4260 gggtaaaggt aaaactattg aagaagctat agtacaggcc gactcaatga tgtataagaa 4320 taaaaaatct aagaagttaa gatactcaaa ggaggattag ttattatgag gttattatgg 4380 ttaaaagaat atttggatat atgcaatatc ttgggtattg agcccacaat ggaatcttgt 4440 gctaggtggg gaaagaataa aaaactatca gggttttcat taaccccagt gggagcaact 4500 aaatagttgc ttctttttaa agcgatccga tacctagcaa ctttcatgcc aaaaagaaat 4560 tcgggatttt ctttgacata gactccagag tctggtataa taatcttgta aacaaaatta 4620 attcaaagga gttgttaatt atgaataaca aaaaatttca agaaacttta gaacaattaa 4680 aggaggaact agaaatacaa ggggtagaat ttaatgatga gaatgactta atagctgctg 4740 ttgaatacat catagaatat gattaccacc cagaacaata tgtggaagat actttatgta 4800 actaccctga aatgttcaag gatctataaa actaaggagg tgtaaaaacc tcctttttta 4860 tttttctaaa aatctggatt taatcgttga caggatcggg actatctagt ataataatat 4920 tgtaatcaaa attaattgaa ggagatgtta actatggaac taacaagatc agaaaaactt 4980 aatgtaataa aaggaattgc tcaaaaggct atggataata aaatctatcc acaatttaga 5040 aaagaactag ataaacaggt aaaatccttt ggactaagca acaaagttag ggaaagtgaa 5100 ttaggagact acctttcttg ctgtatggtt gcctatgaag aggttatact aaaaatgaaa 5160 aaagccaatt aatatgggtg gccaaaaggc catctatttt tttcgaaaaa atctccagaa 5220 tctcgttgac atagtctggg ccctctggta taataatatt gtaatcaata ttaattaatt 5280 cttaggagga atgaaaaatg actaaattag aaatgttaaa tgagatagaa aaaataaata 5340 aggtggtttt tgaaaagaac ttccaaaaac aattcttgga taaggtgata gaactactac 5400 aacaaatggg agtaactaaa tattatgtat gtaaacatca taaaaaagct gagattgaaa 5460 aacatcttga ggcttgcatc tgtgcaatat atgatatagg aagaagggat gatctaggat 5520 ggtaaacccc atcctttttc ttcgaagttt ttcgttgaca gatccagagt tctctcttat 5580 aattaaatca taaattaact attggaggta aacaaaatga gagatatgat agaaaatcaa 5640 ttagctgaat atgaggtaaa ggagttatac aatgaggcca caatgatctt aggaaaagca 5700 tttggtgtaa gtgctgttga tctttatata gaatctgatg tggatgctaa ctactatggt 5760 ctatgccata gatcaggtaa taaatttact gggattacaa ttaacctttt ccctttcagg 5820 tttgcacagg aaggagaaaa agcttatgat aaaatcaaaa aatttggctg ggatattaag 5880 gactgtgttc ttgaaactat atgccatgaa ttggctcact taacttattg gtcacatagt 5940 cctttacata aaaaattaac tagtctttat tacaacaaag tgcaacaggc tagaaaaggt 6000 gaaaactcag ctgtagcaca aactagaagt gatatataca ctttaaaaga tttatgtgaa 6060 ttaatagaac tagatccaag aaaggctaga agtatattaa gaaagaataa tattgagaaa 6120 cctggtaaac agtgggagtg ggagcaaggt atgcctaaag atatatatga tctattaatc 6180 aacttaaagg ttaggggaag aaggtaaact tcttctcttt tttcgttgac atatctcgag 6240 agtgctggta taattatatt aacaaataac ttagggggta aatcacatga tagatatata 6300 ttatagtatt caagtagaag ctgaaaactt ctttggttgt caaaaagaag atggaagctg 6360 ggtagcaaca ggagcattta gtatacctgt taatagcata acttttgaag aggctgatca 6420 attatccaat gaggccatag atgggtggtt aaaggagtat gaggttacta aggtaataag 6480 taggagatca catacacagg ttaaattaga ttggtataat agttatatca aggatgatcc 6540 attaaagggt gatataatta atatggtgta ccctgatctt tcagataaag ctaaaatcaa 6600 atatggcttt ggtaagatat gggctagcac tatgtcaaag gttgctaaga agggtttact 6660 taaaacttgc cctagatgtg gaggcactgg ccattactca agaacctcag atggtaacac 6720 tacttgcttt aaatgtaagg gctttaaata tgtgatacca actaagatct caaagaaatt 6780 ttataaatct gtggagaaaa cctttgacat ctctggcaat aactcttata ataaacttgt 6840 aaacaaaaac aaagaggaga tgttaaatat gaacttaact gcaatggaaa caaagatatt 6900 aaatgctatg agaaagaatg aatttgatga tggactagat gttgactgtg tatgggtatt 6960 ctcagtaata gaaaactcag gtatagaagg aactaaggct agaggtgtga tctcatcact 7020 tgttaaaaaa ggtttagtat ttgctgatgg agaggtaata ggttacacag aagaaggaag 7080 aaaggtcttt gataatgctg atggagaaga atgtaactgg ggaggtccaa aattattaaa 7140 agaaatccct gacaaaccag aggaaatctc ttataatgaa aatgtagata aaacaaataa 7200 gaaagaagag gtaaatgata tgaaagattt aaatactatg aaagctgttg aaattaaaga 7260 attagcaaag gaattaaaag ttaaaaactg gtggactatg aaaaaggctg atctaatagc 7320 tgctatccaa caaatccaac aaccacaaga agaaaaagtt gaagaggtac aacaaccagc 7380 tgaacctgaa gaaaaggctg aacaaactaa aattactctt gatctaacag atgttaagga 7440 acctgaagaa gaaaaggttg aaactaagaa aaaaccaaaa actaaagaag gtgtatttac 7500 actaaagatg atacttgaag aattaaatat gaatggtaaa aaggcaagaa ggatcttaag 7560 aaacaaagag gttgtaaaac caggaaagca atgggaatgg gataatgaag aagaatttaa 7620 aaaggttaag gacctacttt ctaaataaag taggcccaac ctctatcccc tattgattta 7680 caggaggatc tgattttctc ttatccatta actgcttcca tacttgatta ccaaaggaag 7740 ctacagctgc acatattaaa ccctgtataa agccattcaa aaaggtcttg gctgtaaaac 7800 ctaattctaa ttgaattgcc atatataaga ttgataatga tatactgaat atcaaaagta 7860 ttaatggtat agaccaatct ggtatataag gcttagcctt gataaacaga cctaaacagt 7920 aacatgatac aatgattatt aaaagctgtg gatcaatgta attgaatatt aagtttacta 7980 aattttcttc catgataaat accccctaaa ttatctctac atattttgaa cttacataac 8040 cttgcttctt accgtggcta gtattatatt ctatatgata ccaaccattc ttacttccca 8100 atatagtaac tctctcatta ctaaatactg agcctattat attaccattc acatcatctc 8160 ttacattaag agaagtgtta actttaacta ttccttgttt aacatcctca tagaatttct 8220 ggctttgctc accctcgtag cgaacgtatc catcagatgg agtaccatcc tttctataaa 8280 atgttattct taaccagcca ttagttctat acattgggct agccttttga ccaggctgta 8340 gttcttcatt atgatctttg atgttaacgt cttcagggtt ccaccacaca taagttactc 8400 cattgtcatt ttgatatttc atgtggtagt caaaacttaa tctgctgtaa tgttttatgt 8460 caatataagc atggtacttc ttattctttc caggatatac aacttctatg aaatttctag 8520 aaggatatat tccgagaaca aataaattat ctaggctatc aataaatcta ccagggactg 8580 gcttacaatc ctcatctaga acttctattg aaccttcccc cactaaagta gcattagttc 8640 ttgtctcatt agactcatag aatcctttgg ggttattgat tactggtgag cttgagctct 8700 ctcctgaatt tactctttct aggaatgtat tccagtaacc attggctatc attcttctag 8760 gacaattttt tctactggca tcatagtgtc ttactacttt acaattagaa tagtagccat 8820 tctttaggta tctaactagc tctacagcat tatcaactgc cttattgtag tcagagtcag 8880 gattaacaca tatttctata gatattgtat ttctattatt gataccgtat ctattatgac 8940 catctccaca attccaagtg gcgtgtttgt gttctacagc ttgatatacc cctgtatcat 9000 cgacataata atggacactg gctttaccta tatttccatt tgcttgagcc ttacagtggg 9060 tttcggctcc agcaccccaa gaggtattat cagtttcgtg gataattata tacccaggat 9120 tatttgcatt tagagtaaca ttcgtagtac ttaatctttt attgattttc ataataaaca 9180 tctcctttat atattatagt gttttattgg gcctgagagg ctcgctacag cattttaacc 9240 acctccttaa tatatttcct tatataaaac ctaacaggcc ttaaatggcc tgctaggctt 9300 ataagttaac ctttgttact aaataggtga tcttttatag tatctacatc ctctttgata 9360 tcctcaacta cattgaattt atcaactagc ttatctatca cttgatttaa tttattctcc 9420 ctttctctag cttcttgtct agtgtcatat aatagccata caaatagact acaaaagata 9480 ccttggctaa gaacttgagt tagtatttct ttttccatag attacaccct ttccttaaag 9540 tcttcaccac atattctttt atagtcctct ggtgtaattt ctccttccgg attagaagag 9600 cattttaccg cctgatgtaa tttgtcctta cctattgctc caattctata agccatttcc 9660 cagaaattca taatccctac ctccttattt attcatcatt ttaatcttta tatcagctag 9720 ttcttgtgtt aatactttca ttatggcctt agtttgcatt agctctaatt tagtatcagc 9780 aagagcctgg gcttgataac caattaattt attaatatcc agaggattag gatctatagc 9840 ctcataggaa gtaaataggt ccttatcttt gatactgtat agatctgatc tctttttact 9900 aatatcaaat ttggttgggc caccctttat gtggtcaaat aaacttacag tcatttgtat 9960 ggctgaaaga ttgtctagat caattacctc agtgttatcc acaaaggcta aatgcccttg 10020 tgaattatga ggtatatgga gacatataat atcttctata ttatttaata tttcactatc 10080 ctctctatag tatagaggtt caggcctatt aattaggtct aatatcttag taacttcttc 10140 atggtcttta tctataaatg attttacatc ttctgagtct tctgcctgat ttatcagcat 10200 ttctttttca tcatcataga atataaggtc tatgatttta ttagtttcca cagaataatt 10260 aaatcctact attctcatgt tttatcaact cctttactca gtggctaaat aggttagtat 10320 tatttgagtt ggcttaccag ctaccatttg ataacatcca ccattcaaat atgagttatt 10380 ccaatagtca atagtacaat aacctgaggt attattagaa gcgtcattat agttatgtga 10440 tactagagat ataggtaata cccaattcca agccttagga aaatgtatag tataacttac 10500 acctccacct gagttagcat aacccgtctt aactatttga cggactaatc ttagtcctcc 10560 cccaagatga catactctaa cgttaccatt gtcattatcc caatagtctc ctttagcaat 10620 agatctatta tttacaacga agtcagtaac attctcagta tatagctttt tgttctcttg 10680 tttgaaatac aatcttccat cttcagctcc accataccag tcatttattc ttaggtattt 10740 tctagcccat aggttttcag ctgatagagc cttatcaacc tctagagatc cgtctctagt 10800 agttatcctc atatcaaaat ctttcttact accctttaag tggaaatcta atataggacc 10860 tagttccata actccatcat gtgatataaa agggatacct gcatagtcat tactagctcc 10920 agctaagtcc cattctttta gtttattact gttagcagct gtagcggttt tacctaaata 10980 agcactatca tggttatggt ctgaggctgc agctcctata tcggatggct taggtttatt 11040 gaggttacta taagctatac accattcgga ccatatgcct ttaaaactac aaaacctagt 11100 atatgttttg ccataactat ctgtaaatag ttgagttacc tcacccccct tgggtaatac 11160 caatagagtt ccataaatac catatgtgta aactacaggg gagtttggta aggtatctac 11220 actagatact gagtatatcc cctcttcctt acaaagatta aaatctacta tatcatcttt 11280 tctaatagtt aaataaactc cctcatgggt atggttagtt ggggatgccc ccacatctgc 11340 tgcagttggt ttattatctt catgatagac cttcttattg ttcttattaa caatatccag 11400 agagtctatc tttacaggcc ttccagcata ccctatatgg actttattat ctgcatctat 11460 atagatcaca tagtcatcac tgccttcttt agtcctacag ctaaatccat aattgtgggc 11520 cccaataaat ttctgacccg gtctcatgat tagatggcct gataattctc cacccgtagt 11580 aggtaaggca ccaatatctg ctggagtagg cttattttgt ttactatata tttcagccca 11640 attcgtccat ccggtaccct caatgtttgt agctgttcta gtaaaaactc ttccaccatc 11700 tgcaaagaac atttgagaag cccagtgtcc tgtattccag gataaaacaa tcaatctccc 11760 atctgctagg attggggcat ttctatcgac agaatttcta actgtagaat aaatacctcc 11820 ctctgtaatg gtgtttagat ctacaccctc tagtcttttg gctgttttat cccaggcccc 11880 aatatctgcg ggtgttggct tagctccgga gttataatct tttctccaag ggaaccaatc 11940 cgtattattt actttatacc tccaatactt attaccatta gtatcataat actcttgcca 12000 tatccagtta ttagtattat tatgcttagt ctctgtacta acatatacta tcagtatacc 12060 atattttgta ccagttagtg gggcattagg taactcacct gagatcaggt attgcccctc 12120 agttaaagct gtattaaaat cagttatggc gtccttagga atgggataag cctttaggtc 12180 agcaggggta ggcttattag cctcatcata aaccttatac cacttactcc aagaattatt 12240 tcctgatgtg tagtttctaa catatctagt tattttattg gttgggtata ggttaataaa 12300 ctcctgttgg aatcccctct cacctgtatt tctaaccata aaaaccctat tactcatatg 12360 ggctaaggtt ccctcggggg tatttttaaa gtcaccagtt cctgtatcaa aatagaagaa 12420 ggcattcgct tgtttaaagg aattacaatc tctatctaag ggagcactaa ccttaataag 12480 tgcttgatct tgattagcct ttcggtctac ttgattttgt aaactggtgt ctttctcaaa 12540 taataggtct aatcttttta gcacatcctc aggggctaga ttataatcag gtactacatt 12600 acccctagct aatatgaggt ttagtacctt taatgaggta cttgaggttg caccctgtac 12660 cacaaattgt attttggtat cagtagctcc agatggaata ctaaatttat gagtttttaa 12720 cttgtaccca ttactctgtg gtgtactacc ttgttttaca atactcctat attgttgacc 12780 tccattagtg taagccagtt taatttcata gctagtaaaa ctaccttcaa tatcttctag 12840 aaaggctagt atataatcac tatttggttc tataggcaga gttttaatct caagagtttg 12900 tggacctgtt gacataccag accatttaaa acctcgtctc tgggttggag tatctgtaat 12960 gggtgtgata ttaccaggac cagtataaga ctcagagttg taccactcaa gcctattaat 13020 agtaccaacc cttaaagcat ctagcctatc ctcattagac ttagacttct tttctacctc 13080 agtcattttc tgtttatgtt cggtagcaaa gttctcaatg ttctcattgt ccttgttaaa 13140 atcattcatt tttggcttat ctgttaactc ccacttattt aaacctaggg taggagtttt 13200 actagtactt ggcattttgc atacctcctt tattcagtta tagcatattc ttcccatttt 13260 tcccaggtaa gatctagact gtcccacttg ctccacatat aatcatatct atcgaacatc 13320 tcccaagtaa tataaagtaa tataatttga tatattaggt gagcaggcat tctttccctg 13380 aatactcttt ctagagcctt taggttaggt ggtacccctg gtaggtcagt aaagctaatc 13440 ttgatcttac tctcttcagc tatgaactct acctttacct ttccacctgt ataggcctta 13500 gtaagatttt tgatagatgt actattaaac tttttccaag cagatagaac tactgtaata 13560 atagctcttt tctcttggtc agttggtggg taatcaaagt atttatataa ctcaagatca 13620 gtaaaaaatc tttctagacc ttccttagga gtggtcttgg gatccacata gtcttgtaat 13680 tcctgtccct tagtatataa cttctgaagt tcagtagcca taatggtgtt tactaagtta 13740 tcagcctctt ttatattatc ataaaattca ggaacataca atagtaattc tttttgaacc 13800 tgttctagac tatacataag ttaccaccac ctcagtaatg gttggtattt gattagcagg 13860 aatagttaaa gattgactac ttccattaag tgtaatggtt gtataatctt ctactccctc 13920 aatatttaaa agcatagccc ctattctatt tatagatagg ctagtaattt taaatacttg 13980 actatttaga aattcttcta gttctttctt gaagtttaat tttatttgtt cttctgtaac 14040 tgaactcaat ttcttaactt gagctcttac agtgatatta aaggtttggg ctgactcaac 14100 agtaaaaaca catcctatag gagctttacc ctcacctaaa cctttcttgc ctggatccat 14160 gtattcttga actttattaa ctagctcagg attagcaggc ttattttcaa ctgtggttat 14220 aataccttta gcagtattta ttccatccca acagggaaga accacagctt tacccactcc 14280 agctacactt ttacaccaat cagttaactg ttgaacattc ccatctccag atggttgtat 14340 agtggcttta gtagctcttt ctcttaactc atcatctgtc tcctgatctc taccaataac 14400 aagtatatct cccatagtag cacttctaag ttctgctata gcattgatag gaattaaatg 14460 tgtacctgtt gatactttat tgggtcctga acctaaggtc tcacattcaa taatattagg 14520 ctctatcact tgccagaaat aatctccccc ttggaatctt gaaccaacat cgggttcata 14580 gccaacaaat atagctgacc ttctacactt agttggaatt attctggata tccctacatt 14640 ggcagctctc tcatctaaga acttacctgt agcagtactt agaaatactg agtctaatat 14700 ttgattaaca ttaacagcat aaaactcagc tagcttaaca gcaaaaacca tagccatatc 14760 ataggtgaaa cttccagttc tagtgtcaat atcatcaggg gataggccta tcatttcttc 14820 taataattct ttatgtgttc gggagaataa atcttcagaa aatcccatta tatcacccct 14880 tccagttttg tatctccata tatggtattt atccatactt caatatatac attcttacct 14940 tgcacattat actttagatc agttacttta gtcactctat catcttctaa gagacattca 15000 gggatagccc tattaatctc catttcaaga agttcaggag ttactagaga gtcattaatc 15060 aaagctctaa tatcacaacc ataagaagta tcttgataaa tcatataggc atttttagga 15120 gttattaatc ttttatataa agattgtttt aaggcctcaa gaccatccac attatcataa 15180 atcctattaa ggtttagatc cagtttatag gtcttggtat tctccattgt tttctcatca 15240 actataagag tatctttaaa gccattaggt agcattatat caacctccta agatccctaa 15300 cataaaatat ctttgtccat catcataggg cataagataa acagtatcgc cctcactaat 15360 agactcaata tgttttggaa catctataat atttccagat agtactaatt ttctattgtg 15420 aagacactca cattttagag ggctagtact aagtacctta gctctataaa tatttagagg 15480 ggtggacata ttactatata cttgttttac acagtctaat aaattattag ccattaaata 15540 tcaccctcct cagtacattt aagttttaaa ttcatggtgt agtcagtatc aaatttatgg 15600 gtatcagact caatgtagaa agccttgtta atttttaaat gtggtatact aaccacaatg 15660 gctttgccac taatacaatc aggtacacct ataacagtta tagagatatc agttttaggt 15720 ctcttacctg tttctagctt tattctagcc ttctcattta attgggcttg gtttagttca 15780 tcactgatag tatcgtataa ttgaaataca ccatatttct tttggctgct atcatcttta 15840 gcagaggcaa ataaagtaat ctcgggatca tctttaccct taccctgttt atatttacat 15900 actagcttta cctggttgac aatactatca aagtttctac ttcttgaaaa ggttccaata 15960 tttttaccat attcgaattt ccacaattca ttattggttc ttcggtttat aaaatcaaac 16020 ttacctgtaa tagaattata gtaaatataa aactgttctc cagtttgttc ataaacctct 16080 tttattacct tcttcataat atcataggga gtttgtttct cacatactaa actagagaaa 16140 gtatatccag ttttagctac tactccaggg gcaaccccaa aggctttaca taatgactca 16200 aatatagctt cgccagtttg gttttcgaat acaaagctat ctttattatt agcccaataa 16260 atcatagggt cataagcttt aaccttaagt aaatgctttt tctcattatc tgagatattc 16320 ataataatac cattaaagat attcttaccc tcatggacta gtactacctg atctccatca 16380 taaattggat gtatatcttg agcattatat tggaatgata gggtacgagg ggcagatgaa 16440 gaagaccctg cccaactctt accactggag gtatttgtaa tatcaaagga tttatctcct 16500 tgtaccacaa ttaattgcat attataacct ccttatataa tcaccagaac tccaaccttc 16560 tttatatcct gaaccattat ctaatttata ctttacataa tgccatccat tttgagatct 16620 taatactgta aagtgttcat taggccatat cttaccctta gtgggattat tgatacctgg 16680 tccagttctt acccatagcc atgagtaaac attagtagta gtagctgccc aaggttgagt 16740 acctccagaa gaactacctc ctgtattacc actataggct agtttaggtt ttggaattgt 16800 gaaactaact gatttaacct caggcttctt atactcagtt agttttattt ttacttgaag 16860 agtaccaata tctccacctt tctcagtata tgagatatta tcaatagtaa ctaataagtt 16920 aatatcagta ctagtaatag taaatcttac tggatcttta cccatttcaa taagtttagc 16980 ataacttact ctagggtctg caatatcagt atactcacat ccatcgtagt atgagcttgg 17040 taactcaaaa ttaaaagaaa aagattttag cttagggtct cctatgaaag agagttcccc 17100 taagccttca atggaaactg tactattatc tctactatca gagatattta tatcagtggg 17160 atttactggt aacctaaacc tagtatctcc ctgtattaac cagaattgat atttactatg 17220 caaggtccag ttcacctccc tcgaacattt cctgatttaa catttccaca ataaatctgt 17280 atactctcat taattcactt tctgagattg ttccatcttc agcctgtact gtgaaattaa 17340 atgagttgtt aatgttcata actttagatc catcatcact tcctgctcca gattgtctta 17400 gtaaatcctg agttttatct gcaggtaata cttgagagcc tctaggcatt tttgagaatg 17460 taggacccat aactaattca ggaccagctt caccaaccat ggcagcttta tctgtaaaga 17520 agtcagtacc ttcagcaagc attggtatct ggggaatatt aataccctta cccccaataa 17580 taggtaccca gtcaggtatc ttaatagtat taagagcacc aattaagtta ttaatcatac 17640 caataatact attaatagga gcttttgcta gggataacat accttggaaa atacccccaa 17700 agatgttctt aacaccttcc caagctttag accaattacc tgtgaatact ccagccacaa 17760 aagctattaa accactcaac atctgcatta atccctgtag tattcctgat atagtttgaa 17820 cggccccagt aacaactgag gagataacag gccatactgt ttggaaagat gcggctaaaa 17880 ggttaagtag tggagttagc atatccacca acccttggaa tatcatacca aaactctcaa 17940 tagctgattg aatatatgga actacattct cagcaaagct taagaacata ggtagtagtg 18000 tattattcca ccaatctgct agtccttgta atattggcat taggaagtta cctatagctt 18060 ccacaacttg tgatatagct ggggctatgt aggtgttaaa ggcttcagct aatgatacag 18120 ctattggcat gaaaacatta gttatcaatt ctccaagagc tgtaaaagta gcagagaaca 18180 tagaacctaa tacttctaat atagggccta acacttccaa tacagtttgc catacttgag 18240 taaagccttg aacataaaga actactaagt ctatggctgg aattaggtta ttcataaata 18300 tagaagctaa agttccaaac atatcaagaa gtacttgacc tataggagct aatactccca 18360 tgataggctc agctaaacca ttaaaggctt cttgtatatt attattagcc tctgataaat 18420 aatctggtag acctattaag atctctccaa aattattata gatgtcttct ccaagactcc 18480 atatcttctc attaatttga tctgccacat cagcaggtaa gaacttatat aatatatcat 18540 tggttagatc tataaagggt tgcatagcat ccccaaactc ccaaccattc atgatgtaat 18600 atatagtctc tcctaaacct tggaaagcag aaacaaccat tgagaatata ggtcctagtt 18660 tttcaaagcc ttcttgtata aatgtgatac catttagaac agcattagct ataacctcaa 18720 cggcaggggc tagggcatct cccacatcta acataactac agcagctttg gtttttattc 18780 tctccatggt tctattaaag cccttatcca ttttttcata ggcagcttca gtggccccag 18840 cactatcctt catctcatta agagcattag taaaggcctc agtaccttca ccagtcaagg 18900 ctagtgcagc tgaaccagct tctacagaac caaagaggtc attgataccc agattacttt 18960 tctgggcatg tttttctaac atctgaaggg cctcttggat attacctcca cctttaatga 19020 agtctttgaa acccttacca gctatttccc ggaagacttt atcagtcttg gttcccgatt 19080 tagatagctc atctatagca gctcttatct tagtagtagc tacagatgtt ggaaccccct 19140 gagctgttaa tgcagctata gaggcagata cttcttcaaa agatacccca gcagctgagg 19200 ctgaaggtag cacattgaat agggattgtg atagttgctc aaagttagtc ttaccaagtc 19260 ttactgtagt aaacataagg tctgaggccc tttgaacatc catattttta gctccatatg 19320 agttaataac agtagttagt ccatcaaccg cagtttctat tgatgtgata ccacctatag 19380 aagctttacc agctgtttct aagaaactaa aaacgttatc ctctggaact gatgaggata 19440 tagcttgata aagagcaggt actactttat caggcaatat ccctatatca gaacttaact 19500 tttttacctg gccggacatc ttatcaaagg tctcttgagt agagttgggt aatagagtca 19560 taacctcatt taaaccttgc tcaaaatccc caaaggcttt aacactgtcc ttagtaaaat 19620 cccagacagc cttagctgag aatactccag cagctactcc accaattttg gcaagagtac 19680 ccatgaagga attaccacta tcagccatct ggttaaatgc accagagatc ttatcttggg 19740 cagagaatat ggcagtaaat gcagcttgac ttgccattat atccctcctt tatgattaag 19800 agcctttctt taacttatcc ctagcttttt tatcttcttc taaagccagt tcacatgagg 19860 ctataagaaa ggctttccta ttgggagata gtttatcaaa ttcccagggt agtatattat 19920 gcttctggaa taacacatga gcccaaaatg cactagtgtc ggcctttatt agttttttgc 19980 ttctttgatt agatcatcct tattatcgcc taatccattt atgtccataa cctcactagt 20040 tattctagtg aaatctcctg gtaagcttag tacatccata agtaaatcta ctggatccac 20100 aaacttagta gactctaacc attttggatc tttaaagtca gggaatacta ttgtttctag 20160 tatcatttcc ctagtagctc ttctttggtc tgttattgag ttagttatac ctttgtttct 20220 agtaattact gtatttctct tttggatatc ttccatttgc ttagtagtga ttggctttaa 20280 tatccaaggt ataggttggc ccgctaattc atgaccctca gggaaactaa atctagctac 20340 cccttcaact tcaatttcgg gtgattgttt acccttatca ttttccatga aaaattccat 20400 actaaatttt gacataataa tctacctcca caatattctt aatcttttat aaatatctta 20460 cttctgaagc tgagaatgat atttcttctt gaatttcttc tccctcagca tctagtccaa 20520 ttagtggtaa atctccttct aattgtactc ctagtaattg tactctgtcc tcaccataat 20580 tcttaacata gtcagaatct ggatcatcac atacagcttg tatatccatc ttagggaata 20640 taccagtttt taagtattcc ttaatagctt ctcttatcat tggagtagct ttgaattgag 20700 ttatagttcc agttatatca taacctataa cttttgaggt taatcctttc tgacccataa 20760 ctcttttctt agtcacctca ggagtaaata gagcttctag ttttattaag ctagttactt 20820 gcttacctga tacaaaagct ctaccctctc tagcagacat tctattaact tcactcatgg 20880 gctatacctc cttaattatt cttaacagtt acaaagtatt tcttgaaagt tcttacaggt 20940 tgtgtaagta tagtagcata catagtcttt cctttactag cttcttcatc taccactaca 21000 tcagagtcag ggtccacatt ctttaatgct ccttgagctt gtagtgtagt taatagactt 21060 attagtctat cctgagctag ctttatacca tctttgtggt tagggaaaac attaggaact 21120 aagatctgag atcctagatc tataacagca tccaatacag ccacaacttt gttatctgta 21180 aagctttcat cccattcttc tgtaaaagta tggtgtgtat ttatatcttc ctcaactact 21240 acctttactc tttcagaatt tgagtcagtt tgagttgaga atataaattg acctgttttt 21300 aatccatcaa ctatttgaga gtgttttagt cttggattgg catctactgc tccaggatat 21360 gctttataag taaggtcttc agtatatcca gctgcagctc tagcaccagc tacccatgca 21420 gttgcttgag aagctgttaa cttagtgccc ccttctaata caacaccatt cttaacattg 21480 ataatgtatg ggtaatcaca agtagcaaag tcagaagtaa ctgccacaat attcttacca 21540 acttcatctc tgaagtattt aagttttgat aatagagcag tgtgtaattc actatcagta 21600 aatgggaagg ccatagtttt aaagttgtat aactcacagt tatctaagaa gttgataaca 21660 tcagagttat tcatctgagt agaagtacca gtactaaact cagtcttagc agtctcatct 21720 atagaaccac ttccacttac tgtaaaccat tgagacttaa ctgttgatat ttcagttact 21780 cctgtagcct caaagatttt agatccctca agatataata atatatcctt tcctccaaga 21840 ggattggtct taatagttag agataactta ttacctcttt ctccagcttt agtagctgta 21900 aatgttagag accctactac agctttggct ttagttccac ctgttaggtt atatactaaa 21960 acagttgcag cttcttttaa agcctcttta actagtaata cagtagggtg agttatgtca 22020 tacccaagtt gatataatgc ttctaggcca ccttcattgg ttaccttgat aacatctcca 22080 gcattacccc atttaagttg taatggtaag atcataactc ctagattatt aagcttaggc 22140 tcattaagtc caccacttac gaatctagta tacataccag gtcttacttt attttgtcct 22200 ggaataaaag ttcctcctgc catctatttt acctccttct ttagccagtc ttttataatc 22260 tgactagttt gttttattgt taactcttca tcaggtggaa gggattgaac tgcagcgttg 22320 aagacaaact cctcaacctg gaaaagttgt tgacaatggg ctcttaattg aggtatggaa 22380 aatttttgta ctttcactat tttaccccct ttgttatctc aaagttttcg aagtaagttg 22440 gatctttata aagactaacc acaatatcat agtttatctc tacctgagaa tcttcttcag 22500 tttcaccttc aaatacttcc acattcttta ttttgagatt tttgttggtc ttactaccat 22560 ccctattaag tataggaatt tttctatttc tcaagtatat agcattagct atattctgag 22620 caatttcaag agagtcttta ggttcaaata ccttaactct taaagtattc tcagttctat 22680 aactacaaag agtatcattc tgaggattaa ataccttctt gaaatataat gagggatact 22740 tgaacttttc aggtattcgc tcaaaataga ggatagtaat tccagttaga tcagatatga 22800 atttagcaat actccctata tctgctgtca ttaataacta ccccctaaaa cttcggctaa 22860 ccaatcttct aattttcttt ggaaagcctg ttcaaatatt ttttcatata tttttactgc 22920 attgtcccaa taaggtttac caggtacttt cttttgtctt agtaccatac cagtcttagc 22980 tcctggttga taaataaatt tatctcctga ccatatgcca ggaacgaatc taaatgattg 23040 tccttgtgga ttagtgtagt gaccatcgtt tacccactta gcatatttta ggttagtacc 23100 aacttctaaa gttagtcctc catcactact tacccatata ccattcttac ctcccttttg 23160 aaaggagtta agtaataatc tagtgtctac tacttgtagt ctaattattt catcttgaac 23220 cacatcaagg aagtctatcc ctgcagcttc aaaccatact gctaattgct ttttgaaatc 23280 actattagct gcatctctta gcttctttat aaacctttct atttgtttag catctatttc 23340 tatgcctcca gaatttgaag ccataactat ctaaaatcct tagcctctat accaacctta 23400 atttttagac ctcttatatc cctaggagct ttagctatat atgtgatacc agtctcaagg 23460 tccacaattt tatcattttg tctaacatca gttccaatag caaaatctac tgagtactcc 23520 tgggtaatct taaacctagg ttctttgtac tcaagttttt ctagagttcc tgacttaaaa 23580 tgacagggta cctcagtcat gtctggctca tcaggataac taaagctagg ggtatgggct 23640 agcccatatc cgggatcttc agggtcttct aataggtggt atatattaca tctatgattt 23700 aagagcttgt tatatgaatt tccaatcata ttacagtacc cttaatctca tagtagttgt 23760 tctgatagat ggactatcat cttctaagaa aggtttaatc agaggcataa tatcaggctc 23820 tatgacagcc ttatcagttc ttcttgtata tgagtaatca tcataggtct cggattgaat 23880 atcagccttg ctaatgacca aggattgtag tccataatac tcagcataca atatatgggc 23940 taaccttaat tctttactat taggattttt ctccacaata gtatcatagt ctttatttga 24000 atttttaaaa attcctgaga tcttggcttt agctcttagt atatccatct caatattttt 24060 ggggtctctt ttcttaatct caggattact tgaaaagtca attatttcat ccggagtaat 24120 attcatatga gccaccccta tctgtttctt ttattcttgt ttgtaaatct tctagcaggt 24180 tcttctttac tgtcatctgt agaaggtatt tcttctccag taaaatcttc tagaggttca 24240 tcctcaacag gttcatctat gacaatcaca tcatagttct gttttaaata atttactagg 24300 ttatctttaa cttcttctac atggtcttga gtaattctat gacccttaat aaccagagtt 24360 tgtaatttac cctttgctaa tttgacctta gccataatat ctctcctttc taatgaaaaa 24420 gagcccacat ttatagtggg ccttcatgat taagctttta ccttgattat cttagcagca 24480 gctttaggat cttcaaactt aacatccatc ttaagagtta aaactacata agtagctctt 24540 tctctaggag ctctttctaa ttcaattctg atatctcttg atatacctat aactatattc 24600 ttagggtgag ttaagattat atctgatact tgagtagaag tatcttcata atcttgtagc 24660 atagctatac ctttaacagg tacaccataa gcagatttaa cagagccatt tataaatgag 24720 tcatcaccat tagcagttaa tctctctcct actaagtcta cccaatcagt ttctaaacca 24780 tttgagcagt agaatctcca ctcacttggg tttcttaaat acttagctgg tacagctttc 24840 ttagcttgtt ttaataggtc tttacttaaa gcaccccttt gagcatctac tacatgagca 24900 gttaactgtt ttctaacccc atctaattgt tgtaagaact catcactacc ctcactagta 24960 tcaccattaa caattatttc ttccatatca agagcaactc tttcagctaa taattgcatt 25020 acagtgtctt ctattgattt tccttctata tttgtttcta tagtatcatc agttaactta 25080 accacagcta taaattcttt agcttgtaaa gtaactgttc cagtagttgg agctgaatat 25140 ttagactccc ctggatcttt tccttcagtt gcaggattta atactctttg accaaatcct 25200 attgactcaa tctttttaga gtctcctttc attggtacag ttctagcctc ttttaatatt 25260 gttggctcat caattatcat tttaataaaa gcgttagctt gttctgtgtt taatttacct 25320 ccaccactta atactgtaga gttaaatact gctttattta ttttagccat aatcttttcc 25380 tcctatcaat tattcttggt ttccaccaaa taatccatta aatactgatt tttgaacttc 25440 ttgtccttct cctccatttt taacaatttg tttacttcct acatatggag ccatggcctt 25500 agctactgcc tcagctattt tttcttcctc agttttttct tcttgtttct ggattaagcc 25560 agcctctaac atagcttttt ccacagcctt agcaactata tcttcagtag tagcttgttg 25620 accttcactc tttttaactt caccctcaac aggtttagtt ccatctaatg cctttttgat 25680 agcttcgtct atcatttttt gtacttgagt ttcattcatc tcttcttccc cctccactga 25740 gtctaataat tcattaatag cactttgtgc tgtttttaac ttttccacat tttttgaaga 25800 tattttctta cctgccttat tgactggtaa ctcaaaggat ttagctactt cctcgggagt 25860 tcctaataat attccctcca tagtttcaga aaattcttga agagcttcct ttatcttatc 25920 tacatcactt tcaaaagatc tagtgtaatc ccaactagaa tatggatata gaatatcttc 25980 taaggtacta actgcattcc agaaattatc tctttttgca gtctctttat atttttcaga 26040 gtatgcacct ttctgtatat gctttggagt taaccctaag gcctttccaa tcttatcaag 26100 tatagattga cctatagact tctcaatctc atcaatatct acatcttgtg tagaatattt 26160 accagtacca cccatagacc atccagtaat ttcgcctttc tcaatcttgc tccatagatc 26220 atcatcgaca atctcagctt tagctagcca agttccagct ttaactactg taccttctat 26280 tgtagtatct tcctcagtta cccatgattt tacaatcttg acaccctcaa gagtgttctc 26340 attgtgttgt aaatcactac caaggccatt ttcattaaac cactcacaag ctttcttgat 26400 ctcttcctca gtcataaaat tatcatggta gtcagcagtc attggctcat atactacacc 26460 gataacaaaa tgattatctc cttcagactc agactttaag attggcttat catatctaaa 26520 tctattagga gccccaagtt ctttagccac aatccattgc catttattag ctgctttatc 26580 aactaatgaa aggtaatcaa ttttagcatc tgagatctca atagctttct taactgattt 26640 tactggcatt tactctccct cctttacaaa ttatattctg ctatcacttg atctctgata 26700 gcttcttttt cttctttaga taatcctagt atatccttat caattacagg cccatgggta 26760 caatgacaat tcacagtctc agaagctggt aagttgatat ccctaggaaa tctagcttca 26820 taaattccta cagtgaagta ctcaccttta ttaatgacag taccatccag atcgacatgg 26880 tgttgccttg ggtgaatacc ccttccccct gagtgtttcc actgtattcc tgttactgcg 26940 gggcattgat taaaagcctc atccttagca tatgaatgag ctcttaacat ttcagtaata 27000 gcagtagctc tagctctagt tctactaaag ccataagtct cagataattt accaactaca 27060 tcttctacag actctccatc ttcaagagcc ttatccagaa tattctgtag tttttcatgg 27120 gaagttaatt tcatcatttc ccctagttta gaagaccact catctataaa ggtagtagta 27180 tatggagaaa agactttggt tagggataag tccttatcaa taccttgtaa ataagtatct 27240 cctaaatttt ctatagcttc ttgataacaa ctcttaagag tttctgctaa agaagtatcg 27300 atattatctg tacctatttg agactccaca tattccataa attcttcaat ggtttcaaag 27360 tctccatctt ttataagatg ttttaattct tctgtaagga gatcagaaat agtgtcttct 27420 aattcagaga tcttttctat tgtataatca atctcctcat agccagcttc tttcaattct 27480 ttcttaaggc tatcatcttc attctttaag atcttatcaa ttttagcgat caactcatca 27540 gtacccacta ttacttacct ccttttaatt gatagtttaa taactgtttc ttagcctctt 27600 taagtacagc tatcacctca tcactcccat tagagctttg agccttctca attatacttg 27660 ttaagttatt actcattgaa gaggcaactg agtaagtagg gatatttgcc cattcttcat 27720 caataggagt atagtcctta ttgattgatg aatagtactc ctcattaatc tgattaggag 27780 tcatagctct agaggctata gcaaacattc ctttctgata atcagagttc tcaatattag 27840 gactcttaaa gtaagcttcc acatatttaa aaccatagcc agatagaata acattattta 27900 gtatccatgt taggttttct ctttcaggaa caaatacttg ctcttcagtt tttcttgtag 27960 cctctctggc tgtagccaca ttataatctt gggagtaccc tacatatata tctggtagta 28020 agaaagaaga ttgtatcttt ttcctagtat tatccatgta gtcttggaat aactcatcct 28080 tctgtaatat ttcagatagg ggcttgacct caacagtaac tggcttagaa tcatcaaggg 28140 cagtgtctct ctctgcagct tcagtttcta atactaagaa accatgctgg gaattttcac 28200 cttttacatt attcatgtac tcagttaatt cattccaaga tttatcatta agagtaccac 28260 cttgaattag taacattaat ggagtatgtt taccattagt aaaatatctc cagtttaata 28320 attcggcctt tcttgaacct tgagcggcta acataggacc ttcccatctt ggtgtaccat 28380 agacaccata acctgtttta aaatgaagta tactgttagc tcgatcttta agaggtatct 28440 ctcccttttc ataatacttt ccagtattca tatctagatc tctaggatct ccccactctt 28500 taaagaagat tgattgatta tttacagtct gcttaaatct tctgaacctt tttcttctag 28560 tgtactgttt accatatcgg gttacagtgt actctgtact ctcatcatct agcttaagaa 28620 ttttaattgt cttagggtct tcaatttctt ttatctcttt tacgagagct gagtctttag 28680 gatcttctac gatctctaaa aaagaatacc caagtccttc tctccaatca actatctctt 28740 caaagacatt cttggttggt ttatcaaaag acagggtatc aattaattct tccagctgtt 28800 tatattcagc ttccatttct ggagtaggtt catctacctt ctcaatatac ctaataccta 28860 ttccgaatcc agcaatatta tttcggtagg cctttttgca ttggggtatt attgtagact 28920 cccttataat cacttctaat agttgggggt catattcagg ggatatctca tcaacttcaa 28980 ttccaagtac actggtttgt tgattagatt tatccactgg atccaaagac ttttggatag 29040 ggtctccaca tatacttctg gcggatatct ttttcaaagt tactttacct cctttcttac 29100 cttatatatc aaattataaa atccagatgg cctcatattt cagggaggat tacatcaact 29160 ccacatatag tgtcaaatac ggccattcac atgatttaaa aataactata atttgatacg 29220 tttattataa cataattcac aaaaatttgt caacgtttta tttttacatt attttccgga 29280 agattttccg catatattcc ccatataaaa ttatataatt attaatgtat actatattac 29340 tatattaatt tatgaaataa attatatagt atatagtata cttatattac atatatataa 29400 aggtggtaac tctgatatgc cttctatatc acattattac taatatagag cctagtcaat 29460 ttcaaaactg atatctcaca tatcagaata aaaattattg gctttaatag gcatatcagt 29520 tacttagatt tttcttaatt tagatccaac aggaaggcaa gctaagaata gagagtctgc 29580 taagtctgga gaatgtagtc ctcttttctt catttcatcc ttactctcca gctttatttg 29640 acctttggag ttgaatactt taaactttct ggtagtcaat tctcccacaa gatctgtatt 29700 atctggtagt cttaactcat catcctccag taattcctta actacagagg ctatatatgt 29760 ggttgtatca taataaaagg cataagcctg ggaccccctc tttaatggca taccaaattt 29820 acatggtact accttcaacc agtccagttt ttcttcaaat tttatttcct ttaatctatc 29880 agttactccc ccacctacac cagtgtcgtc tattttcaca actactgcta gtttagggaa 29940 ctcaccatgt agggatttag cctttcttag cacttctcct actgttttca tagtgtcctt 30000 accttggtaa taaataggtt cttccacata tccatcaatt ttggtcataa tagaggtctt 30060 atctgaacca aatctagcta cgtctacacc catatctata gtaactggtt ctgtaggtct 30120 ggttatttca ctattaatac ctctatatac tgtggataat tgtataaata tgtcatcttc 30180 ttctagtggg aagtcccctt ttaccagaca tctgtagaca ttggactctt ctccatactt 30240 tttccttaac attttatggt tttctttatc agttctatca acttttgtag cgtcaatatg 30300 aatacattta aaattttcct taagactgtg gtgagaatca aagaagatac ctgtattttt 30360 aacagggtta ccaatcaaaa ttatcttatt ttcctcatcg gatatagtac caagtaatgc 30420 ttctagtatc tcatcatcta ttccaggagc ctcatcacat ataattagca taaactcact 30480 atgaagacca agcatattct cttttctatt actagtttta cacatagcaa accaagtcat 30540 aggactacaa ttcctagtca tagtagtttt atttactgta aagtaatctt ccacaatagt 30600 tccttgtagc cattttgata tttcaggcca taatacagat gttaactgtg aaaatgtagg 30660 agctgtagca attaccttac attcaggcct aaaggtcata taccaaatta taagacaagc 30720 acaggccatg gtcttaccta ccccctgtcc tgaccttaca gaggttctct tattgtctct 30780 tacactcaat aaaatatctt tctgatgtgg atctaagtgt actcctagta tatcttccga 30840 gaatccacaa ggatcgtcat agtatagatc tactaattct tcaaaggata tttcttccag 30900 agcttccttg ttaatgtgga acttatcaac taaagcatct agctctaatt tctcaattat 30960 gctatctaaa tcaaacaatt atatacacct ccttagtaaa gtgctttatt tagtcctgta 31020 ataccttttt acttagtggt attagatttt cctttacctg cattcttttt ctttctatca 31080 gccattttgg ctctaagaac ctcaagatct tttcccttac cagcattatt ctgattttcc 31140 tgtatttgta aaagtaattt aaccttttca agttcaactt tcatcacatc agtacttaag 31200 ccgaagtata cctcaagtct ttttagtgcc tgaagtctat cctccatttt aactgatacg 31260 ccatctttac tttgtttaat ctcattgatt aaagtaccat ctacagcctt actagacctt 31320 aatagaacta gatcatcctt gaattggcac acatcagtta tatctgcaaa ggctattttc 31380 atgtactgcc ttattacatc cataccatcg catagtaatt cttcatctct atcggctctg 31440 atagccttta gagcttcctg tattttcttc ttctttaata gtttagaccc atgtgcatga 31500 gaattactca caggataacc tgctctagca gctgctgtag ttgcattaaa gcacttcatg 31560 taatgaaata caaacaactc ttctttctca tcaagatcat acttatcagc cagggtctta 31620 atagtctttt ttgttttgct actcaatgcc acaatatcac cttcttcttc taaaaatgtt 31680 atgccctgct ctttaaactc tcttagacac tgcctaacaa aggtctgtga taccttagca 31740 aggtttctta gttgtttagg atgtgtaaag ggatcagcct tcagggcatt atatatcaat 31800 tttttggctt tcttcaaatg ttcaggagta cagtactctg gttcagtgta tgacttatag 31860 ctcataatac aatatcaccc ccttaaaatc cagactttac tacgtgaaaa atggttggca 31920 cactaacctt aatgctagta ataccaacca aatggcaaaa ttaactatgg atttatacca 31980 ctattatacc acatctggat taattatgtc aacgttttat aattttattt cacctgtcaa 32040 tattttttca tacatatcca catctattct tccttcctgt agcaggttat cagctgcaat 32100 ttgtatctct tctggggaat atactggatt acctttagta tcaagttcta taccagtata 32160 tgagtctacc ttacctctgt tcataccata tccaactgat ttatgaactt ctagtttttc 32220 attatggtct tcttgccagt gttttaattc ttcctgatat tccttattct cctctttagt 32280 ttgtttacat tcaataggtc tattatcttt catcttttgc tttagagctg ctgtattagg 32340 tttacagttc ttagatcctc cacctgaacc tctattctta tcttcctcat aagctttatt 32400 aatctcaatc ccaattatta gatccatctt ctctaccaag taaagacatt cttttactat 32460 tggattgtta ggttgatttt tagaaatgat gtcttgacac tctttcttta attgatgtgg 32520 attattacat tggctagggc atttttctcc tggcccatat ggaattgtat atgctaagca 32580 gttacctgct ggatcccttc tataacatct tcttttacat actattttac tcatggttaa 32640 tacctcctaa tatattttct ttctcttcta ataatttttc aagttcctca agattttctt 32700 ctttatcact attaccagct ttagctatat ttagaatagc ttctagttgt tcctcactca 32760 aaaactcctt taatttctca tcctctaata tattagatac ctctttactc atattgttct 32820 acctccacat cattacaaat taatttagct attccctcac ttaatctgta tttctcaaga 32880 tataccactt tcttaattcc tgcattgatt attaaattag cacagctctc acatggggat 32940 aaggttacat acatagtagt tccttcagtt gagatcccat gtttagcaca gtaacttatt 33000 aagccagctt ctgcatggat cacaatatca catccaggac ctaaacaagt tttacaatgt 33060 tctactcctg aaggtactcc attatatgaa gtagcaacta tcctgttatc tctaactaat 33120 acacaaccaa ctgcttttct taggcaagta gatcttcctg tcataagttt agcaacttcc 33180 atgtaaagtg tttctctgct tattctgttg ttattatttt ccatagttta tcacctcaat 33240 tttattatat actattccct atcatatgtc aagagaaact ttagaaaggt atatcaccat 33300 aatccagatc aatacattgg gctggctttg gttttacttt tttaggctct ggagttttct 33360 tgactttatg atactttgga gctctggcta aatccttttt taactttctg actctcttca 33420 tatcccactc atgtatttca tggtctctaa aatattcatt caatactctt actgcggatc 33480 tctttgaagg tgtctttaca gagcagttat taggagcaag acagcatcct gtctccttat 33540 ctgatatagt ataacatact gtattatgag gggctcctac aggcctatga atgaataagg 33600 ttctgcccat ataatcaaac tcacataata cttccacata tcttttatac tgttgtcctt 33660 ggtcattctt acataataca taagtatatt ctttcatgtt aattctcctt tctatgttaa 33720 aagggagatc atctctcccc ttaatatctt tttattcttc tgttgttctc atcaacctca 33780 atcacttcat accttctaac cttatcagat ataccttcct cctgtaatac tttaggaata 33840 tatcctatgg cctgtctttt gttatcaact tcaaaatatc tggtcaagat cctttttgat 33900 gttacaatca ctaatcttac tttcatatta tctacctcca tgtggcctga actacagctc 33960 caggtctatg cttagcttca atcttagctt cttcctcagt atcaaaatag attatatctt 34020 ccttgtgttc atccttaaca gatacccagt gtactcctaa atatttgtta tgtgctaata 34080 cactaacatc agggttccaa tccacatcaa ctgtaacttg taaaacgata tatctccttt 34140 tcatatcagt tctccttttt gtatttattt gttgatttta ttataagaga gcctcggaag 34200 tttgtcaaag attttctcga aggaattcgt ataatatctt ccgagctgag cataatatag 34260 actataaaac aaaatcaagg agttgtaata aatgaaagat aataatttaa taccattaag 34320 agatattgcc aaagaggttg gaataaatga aaggaaagcc agaaagctat taagatcaca 34380 tgagaataaa gcaaatcaca attattattg gatgtttaca ccaagagaga aaagacatat 34440 tgtatcagtt ctaaagagac tgcaataggt tttctaaaaa taagggaggg attaaccctc 34500 ctttttataa ttttttgagt tctggaaaaa tccttaatta gttcaagttt cgtaaagttc 34560 tcttgatctt ttccattctc ttcttaactt ggatagcttc ttcttcgcaa ggagcatggc 34620 cctctaggaa ttggtgccat cttctttcaa gttctctgta ttcagattga agttccatga 34680 actcaagtga atgattgtcc atactatcac ctcctcaaca ctattataag tgatactgtc 34740 atattctgtc aaataaaaaa gagagaatgt tattctcccc atcttaatat ctttatagtt 34800 gctccatcaa ctcccttgcc atccacatca tggacatact tatcaagtct cattgttgag 34860 tcctgtataa ttcttttgtc ttctctgtca aaaaccttaa tgacatcagg gttacccctg 34920 tcaatgatgt aagcaatctc ttgagttcca tctgcatatg ttacttgaag taatgaatgt 34980 agaggaataa agtctggagc tgcccaatca ccaatcttaa gatcccctcc tatactattc 35040 ttggtctcat atcctgccac atctgtaggc tcatctgatg agtaataagt aacttcaact 35100 ttgaactcca ttcctttaaa tgaatttatc atctcaatta gattttgttt atcagttctc 35160 agtttatcat tctgatattt taatccttct acattgtcca tcagtatttc agtctgctca 35220 gttaattgtt tattcgactc aatatattct tggacagcat ttccccaatg attgtttagg 35280 ctctgtatgg ttaacagtag ccctcctatc acaattattg ttagtaatga cataattctt 35340 ttaatcattc tcttttaact cctttatttt taatttgaat ttctcaattc cagatttata 35400 cttacccaca atatcaaagg tatcggttag tattgcatag gtttcctcat cagaattaaa 35460 tctgaagctt cctctcttga tgtctcctag cttattaaag gtcttaccct ccttgtccag 35520 atatccttta tagtccatat agcacagaca agcagtaaca aactcctgct tattcctgaa 35580 attatgcttg atacccatac catctaccca tttacttaat acaaatagac tttcattagt 35640 cttctttact ttactattct ggattagatc cttccacaca taatcattag cctctttaaa 35700 tgacttaaat ctcttgcctt tcagtatctt actactctgg atcttggctt tataatcctc 35760 aaatatactt ggccagaagt cttttagata taaatacaag gaaatatcat ctaatttagc 35820 attcggatcg ttaatctgga ttacaggatg accatccatt gctctgatat tattccttag 35880 ttgggtattc tctctcttca ttttagatac ttcaatatat aatttaacga atgcgtcaga 35940 caattcatga ttggttaatt ccctttttag gggcttacca tctttacttt ctttagtagt 36000 tacaatatta aagtagtctt gtaacctatc ccagttattc ttcattagat ccacctccta 36060 tgcttacatt atacatatat tgtactgata tgtcaaagtc caggcattaa ctgatatctg 36120 atatgcctat atttttaccc catatcagtc tgatatccca catatcagtt gaaactacaa 36180 aatcacaaag gcattggtat cactggcgta gagggatatc agttatgccc cctttatata 36240 tacatagtat tatatatact atataataat atattattat atatagtata tatatattac 36300 atggtgatta catcaatggc aggctattgc cttgcctcca ttatcattac ggcaggcata 36360 tccatgcttc cgagcttttg ccctatatag aaaaacccag acggtcagcg atcgagctct 36420 tgattttgtt ttgtcgaggg agagaacagt aagagagaac ccccgaacgt tgacagttgt 36480 acttacttct cttataatga cagtaaagga gggataacca tgaaaacatt atttactgat 36540 acaaaactag tattaattga aactactaac ttcttcatta tgttacaaga agaaggtaag 36600 tatgtactat ctaaaacaat cgataaggag atgggagaac aaatacacag agtatctaag 36660 ttagctgccc ttgtatatac taatgaggag tatactaact ctaatgaacc aagatctata 36720 tacttagatg acaatttctt atacgaagtg cttggtaatg ggaagtatat tataagatac 36780 agtatataag agcccacaat caattctaag gccttttaga tatgtggtaa tgtgatttta 36840 cttataatgt ggtagagaca ccttacagga gctgctagag gcctttaaag gaggtatatc 36900 atgtatttat gtaatggtaa aatagtcaag aataaaaatg acttaaaaga taagtttggt 36960 ggtaaatcag tagtattcat cgatggggat gtagctacag taatggactt tagtgacaga 37020 aagccaaaga taacaaaata tcttattagg gaggtatcac atgataaaca catcattatt 37080 gaagagactg gaagggatag aggaaaagca taagcaaaat actgatgaga taagaaagaa 37140 cctaaactat gccagatatt tgattgagaa acagtattgt catttaacaa aagaacagaa 37200 gataaaattg ggacagttct taggattgac tgagaagaag ttaaaagaaa tacaagagat 37260 aaataatcag ttaataaatt tgagagtggg gctaagttat gaagtcgtat gtaataacca 37320 gaaaaaatag tggggtatct ttagcagata cagacatggg acaagtatta gctaagatac 37380 tatgtaagca tgggggaaca cttaccattg aggaaattga accagaacat attggtcaaa 37440 ctttttctat tgtagaaggt aaaagaatat ggtataatag aaaaggaggt gataatattg 37500 gggagaagaa gtgctatgtt aattctcaag tatgactgcg aagaaactcc agaaaggatt 37560 gaattacgaa agaagtggtt acagatttgt cttcattcaa taatatatta tagatatgac 37620 aacaatatct ggacagacca acaatgggat gagactgcca gagaggtagt taaacttaag 37680 aatgagaatc caggtttagt aaacagtatg ccatttagga atgaccttaa agtctttgat 37740 ggatccactg gctttaacct ttcatgtatg caagatatta agatgttaag gtgggctcaa 37800 aacttacttg actttcatgt taaacgagga cttgatggaa agagtaagaa aaagaagaag 37860 aagaggagga aaaagaaatg aacctacttg agcactatat aaaaaaggta cacagtgtaa 37920 gagagtacca tgaatttgat aatgaacctt gggctaaagg taaacaatat gtagaagtag 37980 acatggacta tatttgttat tctaatacac cacaaagagt gaagatggta tttactgttg 38040 atgagtggga gaagataaaa tatagaggct actacatggg ataggaggaa tagttatgat 38100 aagagaaatg ggcatgagaa tgctaatacc agaaatacat gatataaaag atagggatat 38160 acaaaatttt gtaagaaagg ccctaaatga gatagttgat gaaaagttct ttataatacc 38220 tgcaagtagt actgggaaat atcatccaac ttatagtgca ggggtaggag gtttgttaag 38280 acatactaaa gctgcttgtt atataggtaa agcattatgc gaagctgaga tgatatgttc 38340 aagggacaga gatatgataa tagctgcttt aatattacat gacattaata agccagataa 38400 agaacatcca tatatggtta gagagacact atcaccccta gaggaagagt ttccaataat 38460 atacaaagtg gtagttgaac taattgagtc tcatcatggt caatggggta gtgttcctat 38520 taatagttat tcccagagaa tagtacactt ggctgattat atcagttcac agaaaggttt 38580 aatattctca tatgaccaag gaactgatta ttctgacctg ttatgggagg agaagtaaat 38640 ggtgctaggg gtaataaccc tagtatcaat atttttacta ggagtgttta taggttttga 38700 ggtgaagaaa tggaactagg ttttgcgata ctagctataa tagtatcaat atttttagaa 38760 actaaatata taaaggagta gattattatg tcaagagcaa gagctgcaag gagaagagcc 38820 gaaagagaga acaaaaatct taatgttctt aaagctgtgg acatcatgtt agccttatca 38880 tgctatacat taaagaagga aggttatggt aagactagaa tgactaggtt tgtggaaggg 38940 atgagtaaac acacagagga gattgaaaag ggaacactta actatgaaac cataatggga 39000 gagattaagg atttagtacc accaggtata tttgatttag gggaggagtc aaaggagtga 39060 tagtattggg atatttatta tgggcattta gaatattagg catacttgta atagtatggt 39120 gtataatgga ggcgatcagt tgtgagcaat aagttatatt ttaggtatag tgctatgaac 39180 tctggaaaga ctactcagtt aatccaggta gcacataact atgaagaaag gggaatgaag 39240 ccattagtag ttaagcccgg aattgatact aaaggtggac catgtataat cagtagaata 39300 ggggtagcaa gaaaggtgga tgttcttcta cctcctagag ttaaattatc aaaaatactt 39360 tctgactttg agtctaatta ttgggcagta gatgttatat taattgatga ggtacaattc 39420 ttatccagag aacaggttga tgatctgtta aatttatcat tacattatcc tattatatgt 39480 tatggattga gaactgactt taaaagggaa ggatttgaag gaagtaccag attattacaa 39540 gtggcccata atattgaaga attaaagaac atttgtcaat gtggtaaaaa agccacattc 39600 tcaatattaa aataccaggg taaatatact gaccagggta accagataca aattgataac 39660 cagcccgaaa gtgttgaata tgaagccgtt tgtaatgaat gttataataa attaattttt 39720 tctaaataac tcgttgacag gctatatagg atatgatatt atagtattgt aatcaaatta 39780 caaacaccaa ccaaaacttt ataattagtt aatttcaaac tatactaaat ttgttattta 39840 aaaattaaaa atatggagga atgattatta tggcaaaagg aaaagcaaaa tggttagagg 39900 aagcaaacaa taaggagcta attgtaatcg tacaatctgc tttaaattct ggagatgaga 39960 aagaagttaa aaaggcttta aaagctattg tggaattaga tgaaagaact cctgacacag 40020 atggacaaat aactgtagag gaagcattag aatcattaga aatggctgta ggaactgtaa 40080 aggctgcata ccaagcagaa atagatgctg tggaagatga aggtgtagta gatgctgact 40140 tcgaagaagt tgatgaagac gaagcagaag aagaagatga aaatgattta gaatcattat 40200 ctaaaaggga attagctgct atggctaaag acttaggtat caaaggagct aagaaaaaag 40260 ataaagatga gttaatcgct ttaatcaaag aagctcaagg tgaagaagaa gacgaagagg 40320 aagaagaaga ggatgaaact cctgactact ctgaaatgac taagaaagac ttaattgctc 40380 ttgctaaaga aagaggaata aaagttaata agaaaatgaa accagctgag ataattgaat 40440 tattagaagc tgatgatgaa gaataattct catcaaataa taaggggagc aaacttgctc 40500 ctcaaatttt ttacttattt tagaaacaga tgttagaggg ggatgtataa atggcaagag 40560 gtggaaattc aacttatact aaagcagtat attgttacaa caataatagg gagtatagat 40620 ccttcagata tgcagctaaa cagttaggag tatcggctgt tagtataatg agagtagtta 40680 acgggactaa accacatata gggggattag tattcgctga gatagaggaa ggcactagaa 40740 aattaaagcc aagaggagaa gtagttccat acttgaaaga attaggttgg gataatatag 40800 acttataata gggggtataa tatatggaca taaagttctt gaaggatata ttctctaaac 40860 aatgtgaaaa tggagattac attatcctag cagctagaaa aggtaaagaa tggaaagatg 40920 taccaatcaa atataataag aataacattg acaaaaaact aaaggacttt gaacaacagt 40980 ataagggtta tgatttatat tggagcccaa tgccctatag taatcctcaa agaaggatag 41040 ttaacttcat agagactaaa tacttaatac aagatataga tgagcatact gacccgttgg 41100 ggattaaacc taaaccaagt tatctgtggg aaagttctcc tggaaaatat caggggctat 41160 gggaaatgga taggtatata gaggctaatc aatatgacga aataaatcca gcattagcta 41220 aacatatagg ctgtgactcc tgctttgatg ttactcatgt atatagaata ccaggaacta 41280 ttaactataa atacaagaat aaaccaaaag taaaaagacc tatacacact aaggagatat 41340 ataagcctaa ggtgatcgct aaggctgtta aggctgtaag taaatctaat gatagtgtta 41400 aggtcaatat aggaggatct gaggcctcac aatctgaaag aaagatatat gctaaatata 41460 atataccaaa gaaggttaga gacttactgg ctttagatga cattacttct ttagatagaa 41520 gttctactat atggtatatt gagaataaat tacatgaaat aggactagag cctaatgaga 41580 ttatattatt agttaagggc tcagctttta ataaatataa gggaagaaaa gatgaagaaa 41640 caaggttaag aaaagaattg gataaaatca taggaggaga aatagaggct gatattgaaa 41700 aggctgaaag tactaaactg agaatagata gttatcagga tgttatgggt aataatggtg 41760 ccttcccagg ttggttagta caaggtttct ggggtagaag atctcatgga attgtggctg 41820 gacaaccaaa ggtatttaaa tccacattta cacaggactt agctatatca gttgctagtg 41880 gaagaccctt cctaggtcaa tatcctgttc tagaaccagg cccagtaatt gtagttcaaa 41940 atgagaatgc tgactggatt atgagggata gaactgaaaa gataattagc cacagaggag 42000 tagttggtaa tgtggatata aagggtaaga gaagacttaa agttaggttt gctccagatc 42060 ttcctatcac ttttattaat caacagggat ttatgttaga tgaagaatcc catagaaaac 42120 agatagaaga attaattgat gagataaaac ctgtactagt aatattcgac ccattatatc 42180 ttatgtttag tggagatctt aataatgcag ctgatcttaa tcctgtacta caatggtgtc 42240 ttaaacttaa gaatgagaag catacaggag ttatgttaat acaccactat aataaaggtg 42300 gaaatgccac tcaaaccaga ggtggtcaaa agatggctgg ttcattcata ttacatggtt 42360 gggtagaatc agcattatat ttaaaaagac ctgatgactt agaaggtgac gatgaggaga 42420 ttgaagtaga tatagataac cttgataaac aaagccattt accaagtaaa atcattatgg 42480 atagagagtt ccgtcttgca ggacaattcc ctcaaattga attgaattta tcaatgggag 42540 aatttggaga tccatattat catgtagaag tagcaatacc cggaaaagaa gtaattgtga 42600 aacctgagga caaagccaaa gttatagagg ctgtaaaatc aggagctcat accaaggagg 42660 agatagtgtc aatatcagga ttaaattcac agaaggttaa tctagttcta gatactttaa 42720 aagatactat tgtatattct ccggataaag gttataatat aagtaagaaa ctaaatattg 42780 ggaggaaaaa ggcatgataa taaagtgtaa gaggccagca ggtcataatc atataaaggc 42840 ctttgtagta gtaattattg ctagaggtaa gaagaataaa atcacttatc attcaggatt 42900 gaaatatttc tatacctata aatcagctaa attatttata gacaaagtta tggaagaatg 42960 tccctgggct gaattttatt gggtggatgt ccatgaagcc cataagatcc ctgaaggcga 43020 agagatacct aagaagaaaa tgtggtgtcc atattgtcaa actattcaag aatttaagag 43080 aagccaagga ggttacaaga attgccctat ctgtggtata agtgaccaag acttttatgt 43140 aaaatgtatg aatataaaat atagggaggc taaggctaat gcgaagaaga aacttgaagg 43200 aagtaatact aatcctaggg atatcattgt caatggggat aatgctaact cagggggcaa 43260 aactaataga aagtcatcac caaagaaaaa gtcaagaaaa gttagtagaa aagcttcata 43320 atgctttaga ctataatgtt attgagactt ctccatctgg actagatggg gaggatctta 43380 ttatcaaact aaaccttaat agtcctaaag accttacccc taacaactta tctaagttag 43440 ctaatgaggt tatggtcaat aacttaggag ctcagaatta tactatactt gtttatgata 43500 gtaatagacc agaaaatctt ctggctgtta tagatatacc taaaaattct aaaataacct 43560 ttgacataat aaattaatta tggtaatata aaaatgtaga aggaggagaa tttaatggtc 43620 aataaaaagt attacatctt gaaagacctt aatgatgaca aaccccaact atggaaggga 43680 agtttaacca ctgctactaa gatagcagga gcattaggag ttagtgtgat aaaactaaat 43740 aggtatttag taagaaaggg atttgccaga aggtacaatg agcatacact tgagataata 43800 gacccaatcc ttgctaaaga acttggttat tatgtaactc caagtaagtc ccttataaat 43860 cctttatttg aatataatga taaaggagta caatatatac taaacttaat agtagaagat 43920 ctaaagaatg gaggagagct ttaatgatta tcattttagg aatggataac tcaggtaaga 43980 ccacaacagc taagaaccta actaaacaca gaggaggaga atatatccaa tcaatgggac 44040 ctggttctta tgaagaacaa agagaatggg tattaactca aattgtgaga aaaggtgaaa 44100 gggaagctat acatgataga tttacttgct ttgaagaaat ggtgtatgga cctatcatca 44160 gagatagtag taatttcaac cttgacagta aggagctaaa gatcttaaaa catctatgta 44220 agccaactat tgtatatgct agaccaccta gggaagtgat cttcaatttt ggtgatagag 44280 aacagatgcc aggagtaatt gaaagagctg aacatctact ggctagatat gatgaactga 44340 tatggaagtt attctgtgat ggatggaatg tactggtata tgattatact acttccaatg 44400 tggagaagct atcccagtta atagatgact caagtgctaa agaagctata aaccaattta 44460 taaatatatc aagataagag gagagattgt tatgaatata aaccatgcag tagaagaaaa 44520 aatagaagga gatagattac aagctatctt taatagacaa aaggagttaa tggagaaata 44580 tcatcatata gaagctagat caggattatg ccaaactgaa gactgtccag ttaacttaaa 44640 tgataaaaga ggtcaagcta gacttaaaga ttttgcttgg agaatgactg aagaagtagg 44700 agaagcattg gatgcttata atcatgaaga ccattaccaa gaagaattaa ttgatggatt 44760 acacttctta actgaattta caattctagc tggtaaggat tataatacta tagataataa 44820 tgctatatta gttgatagcc tagagaactt atatcataag gctactacat caccagaatt 44880 tcctgaaatg ttagtagaag aagctgtaac tgacctggta agagaaatgg gtatgtgctg 44940 taactgcctt aagaataaac cttggaaaca aacttcaatg ctaacagatg taaatgcttt 45000 taatcaaaga ttattcaatg tatgggtatg ttacattaaa ttattagctg tatctggctt 45060 agaggtagat gacatagtaa atatatacct taagaaatca caggtaaata aattccgcca 45120 aagatctaat tactaaaata aatacttagg agaaggtaat atgctaataa gagactataa 45180 tgattttgat gacttattct taaaccttaa tagggaaatg attaccaatc cagaagaaac 45240 tatgatgtat actcagaaca tacagggctt tcaggaggac ttagttctct cctgtaagtc 45300 ccataaatgt actttaaatc taggggactt tggatataaa gaaggtaagt ggggacacct 45360 attaagatca tacattgatt atccacaatt aattgaattt agagaaaagc ttaccaagat 45420 aagtggtatg agctatactt attatttcaa caggaaaaaa gctactaatg gttcttgctt 45480 gatagctgca gtagtaacta gaccaaaaag aaaaggacct tggaaacact taaaaattat 45540 gtatagagta tgtgaattac aaaagaaatt cgcagctgat ctagtattaa taaacaggtt 45600 cattgaagaa ctacctcaag aagtatgtga gatagataat attacttttc atatgtctca 45660 agcttatttg tcaggaatgt ttataaatgg atatttcaat tattttaagg taccaagaaa 45720 gagtattgct aatagcaaac atccttggca taaatccttg aatagtaact ataacaggtt 45780 ctttaaatca gaggaccaga tacactctta taaagctctc cagaagatgc agttattaca 45840 ttttggtctg gagaaatttc caaagataga tatcaataaa ttatctattg acaagtactt 45900 taataagtag tataattaac ttaacaaaat taaataaatg gaggaattaa actatgagaa 45960 tttatattaa tgctcaagaa gcatttgaag aagtaaaaag agatttatgt gaaatgggga 46020 ttgaggtaag acccaaaact atgcaagata aagttataga aggtaatcca gattatttca 46080 caaaggagtt acaaaactat agttacacaa ttctagaaag taaacctgaa gaagttccag 46140 gtgtatctca accttgggca gatgctgaat ttagagaaag gatctacgat ccacaaggtg 46200 tcatcaatca atactctctt gaagaaagag aagaattatt tggtatacat ccacatcata 46260 ctagggaggc tttcataaat cccggtaaag cttatcaatt aagacctgag gtatggaatg 46320 aatatctaag agatggaaaa tttgggtatt cttacaatga aagaatttgg caatacagac 46380 aaattgagga tatcatcaat agaattaaag aagatccagg ctcaagacaa ttatggttat 46440 ctctgtggaa tccagctatt gatccattca atataggagg agtaactaga gtaccatgtt 46500 cattaggata taacttccaa gtaagagaag gtaagctaaa tatccattat gttatgagaa 46560 gttctgattt tgctactcac tttgctaatg atgtttatct tgctatgaaa ctattacatt 46620 gggtagctga acaaactggc tatgaaccag gaagtttctc tcatacaatt ttctctcttc 46680 atgtttacaa caaagatatt aagggggtat tctaataaat gagttacatg agaaaagaaa 46740 gatataaaca ttctgatctt aataataagg ttaacaagaa ggaaacccta aagcaattta 46800 ttaggaatac tgaaaaggaa tttgatttag aaccagctaa ccttggggac atgactagga 46860 atgaacttaa taactatata aattatatgg atgaactgtg gagtaagtaa tatgtataat 46920 tatagtgatg ataacagacc taagaacaat aacaatggct gtcttggttg tttaattata 46980 atcttagcag ctataggatt atgggtaata atttttgata ttgccaatat aatatatcat 47040 atgatattct aaatttaaag ggggctatcc taatgaaaaa tataatttgt cctatgtgta 47100 aaaactcagt aacattaaaa agaaaatatg gagctatgtt ttggatcatg attttcttaa 47160 ctggaggcct atgggtagta accataccct tcaaaaaaca taaggtctgt ccagtatgta 47220 actcaatcat aaaataaaga taagggcctt taggggtcct ttctatttag gaggtaatat 47280 tttaaatgag atgtaacaac tgtaacctat atactcactc agctccctca tgtatagaag 47340 gaacttgtgt aggaaagaaa aagaagccaa gaataatggt cataaatagt ttagctaatg 47400 atagggatga ggccaataga atagctactc ctgataaatc attacttgat aaaatggaag 47460 gtctagactt ttattatacc aatgctatta agtgtagaac tcctaagggt actaaaatca 47520 aagtatcaga gattaagaaa tgccaagaac atttacttaa ggaaattgaa aaatataaac 47580 ccgaatatgt tatgatctta ggatctcaag cattaaagat gctaagtaat gaaggtataa 47640 cttcaatatg tggagtacct aagaagcatg agaaatacgg ctttaagttt attgccagct 47700 attcacccgg tgtagttgca tatgacccaa ctaaagcaca atttgtagat caggccttta 47760 ataactttaa agccatggta aaaggtaaag agcatgaatt acctgatctt aacataaaac 47820 ttattactag tatgaaagaa ctaaaccagg ctttcaaata tttaagggat gagggttata 47880 atagagtatc atatgatata gaaactagag gcttagatag gtttaataat aacattactc 47940 tatttggttt tggtaatacc caagtacagt atatactgcc tttagaagtc aaatatagcc 48000 cattaagggg taagcccata gcacaaagaa gattagctaa atccttgatt aaaagactta 48060 attctgagat gaaggaaaga atagcccaga atggtaagtt tgatgataat ttcttaaagg 48120 agaaatatgg tattaagcca atcataacct ttgatacttt actagcctca cattgtttag 48180 atgagaatac tcctaatggt cttaaggaga atgctttatt acattgtaat gctaaagact 48240 gggatataaa taagaaatta aagactggaa atgtggagac taaatcagac tttgaggatt 48300 atgttaggta tctggggtat gatatatact atacatttgc tctatataaa atatttaata 48360 aaagacttaa aagggatgag agcttatata aactattcca ccacttatac atcccagcta 48420 gtaaagctta tgaggatgtt cagttcaagg gtatatatgt aaatcaggag aaattcaaag 48480 aggttgaaaa atacttaaga tctgaactag ataagattga gactggactt aagaagtata 48540 ctaatggcca ggatattaac tggagctcac ctaaacaggt tggagaattt ttatatgata 48600 cacttggtct tcctgtgatt gaggttactg actctggagc accagctact ggagaaagtg 48660 tattattaag actaagagat aaacatccag cagtagaact actattacag cataggggag 48720 ttcatataca aatttcccac tttatagatg gttggctaaa taggatgcac aatcatagat 48780 tatatccaaa ctttaaactt catggaactg taacaggaag aacctcaagt aataatccta 48840 atctacagca agttcctaga gataagaaaa tcaggagttt attaggacca tctcctggaa 48900 gagtattcat tgaagccgat ctatcccagg ccgaacttag aatagctgct atgatggccg 48960 atgaggataa tatgaagttt atttaccaga ctggtggaga tatacatgac tccacatata 49020 atattatatc tggggaagat atcaatgatg agaaagatcc agcagttaag aaggagaaaa 49080 ggaaaaaggc taaggctgta aactttggtt tcttatatgg tatgcaatgg aaaaaattca 49140 aggattatgg aagagacaac tatggtctta aattaacaga tgaggaagcc aaaacatata 49200 gaaggaactt ctttaataaa taccctaaac tattaacatg gcatgataaa caaagaaaaa 49260 ttgttaaagc caatggtgaa gtaagatctc taataggaag aattagaaga ttacccgata 49320 tatattcatc tgatagatct aaagctgctg aagccgaaag acaatgtatt aactcaccag 49380 ttcaaggttt tggttcagat atcactttat taggcctatg tgagattaca ggctatgcta 49440 aatatgttaa tcctgaatat gtattagata agtctaagtt tgatgtatta ggctcagtac 49500 atgactcaat attatttgaa gtagataaag actatgtgga agaattagct tggaaagtaa 49560 aatcaatagt tgaaaataat aaagtattaa agaaagtatt taaatttacc cccacagtcc 49620 caataatcat ggatatatca gttggttatt cttggggagg atgtgttgaa ctggatttta 49680 aaggtgattg gaaaagccag ataagaaaag tgttgacaga tgaataatta tctgataata 49740 taaatattgt aatgaaaagg aggtataagg atgttaaaaa taagtaattc cagaattaac 49800 aaattcttgt cttgtcctta tgcccattat gttaaatact atgaaggtct ggtacctaaa 49860 agaagtggag ctgccttaca aaggggctct gctatccacc aggctataga agactaccat 49920 aatgggaaga gttggaaaaa atctgttgat aaattttcca aagagtttta caaaaataca 49980 tttaaagaag agatccttga atttggagat attccaaaaa tggtttattc tttatgtgat 50040 aactatttcc actattatga tgaaaaagaa gataatgtaa cctatgtgga aaatgaacat 50100 cacttcgaat taaaactatg taagggtgta actctagaag gctatattga tagtgtctta 50160 gatgtggatg gaaagatatg ggctaaggaa actaaaacct acaaaaagat gcctgataga 50220 aatttcctga tcttcaatag acaatctgct atatatacct gggctcttct acatgaatac 50280 ccaaaagtaa gtggtactat atgggatata atattagctc agcaaccagg tagaccagaa 50340 ttaactcaaa aaggggtatt atctcaaaag aggattaaat ccacacccct agagttagaa 50400 agaggaataa gagaattagg attagatcct aaagattatg agtcttatat taattctgct 50460 agatgggaag acttctttgt aagacaccca ataatattat caaagaatat acttaacagt 50520 gtaatgattg atactattga gatagctaaa cttattagag atgagggcca caaaagaaag 50580 gaaaagaacc ttggaaaagg ttgctctttc tgtgaatata aatctttatg ccaagctgaa 50640 cttttaaatc ttgataaaga atttattatt aaggctgact acaaacaaag ggaggaaagt 50700 gacaatggca aaaaagcaaa aatcaaaatc aaatagtttt gaggacagat tagtggatct 50760 atatgatata gatgagccca caatactaac actttatgga agatctggtt caggtaaaac 50820 tactatctca ggaacactac ccaaaccaat attctttatt gatgtaaagg acaaaggtac 50880 tctatcagct agaaacaagc ttagagttaa aagaggagat atacaagtat ttagtctaaa 50940 gagttttgat gacatatacg aggcttatga ttacctatca gaaaacactg ataaatttaa 51000 aacagtagtt atagaccatt taactgcttt acaagaatta ggtaatgaaa aggtcaaagc 51060 tgaagaaggt aaggaccaga tgagccaaag aatgtttgga aatgtggcta attatatgaa 51120 agaggttata aacctttata aagaattaaa tgaggaaggt atactaccat gctttatagt 51180 acaagatagg ttagaatctg gtgatggtga aggagaagac caattaatgc ctgaggtagg 51240 accaggatta atgccatctg tatctaaata tttatgtgct gtatcaagag taataggtca 51300 tacttattta tatgaacact cagaaaaaga gggtatgaag gttaagaaag aaatccagta 51360 tagactaaga ctaggaccta acccttatta tattactaaa tttactagac ctcaaggatc 51420 tgaatgtcca gcttatctgg tacatgattt aaaatcacca acaactatct gggaggatat 51480 tgaaactatt ctggctggtg aatggaataa taagccagct aaatctggta aaaaaaccag 51540 taagaaatct ggtaaaaaga aaaaataaaa ttttctaaaa atacctttga caaccaatta 51600 51600 <110> Optipharm.CO., LTD <120> Novel Clostridium perfringens specific bacteriophage CP5 and          antibacterial composition comprising the same <130> OTP1-38p <150> KR 10-2016-0178220 <151> 2016-12-23 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 51600 <212> DNA <213> Artificial Sequence <220> <223> bacteriophage CP5 <400> 1 aaaaaaacca gtaagaaatc tggtaaaaag aaaaaataaa attttctaaa aatacctttg 60 acaaccaatt aaaaatctga taatataaac ttgtaaacaa agaggtacaa tatcttgaca 120 aatatttatc tcataatgat attttaaaat ttaaggagga caataacatg gctaaaaaga 180 ctactaagag aggtaacaaa aacaaaggag gattaaagat tgatctttca aacgttgaaa 240 cttcagttac tatcccagaa ggaaattaca ttgtggaagt agaagatgta gaggttaagg 300 tttctgaaaa tagtggaagc aattatttat catttacttt tgtaatagca gaaggaaaga 360 tgaaaggaca aaagttatac cacatttgct cacttcaacc acaagcttta tttaacttaa 420 aaggtgtgtt agttgcttta ggatttgata tccctgatga ggagttcgaa ttagatacag 480 aagctctagt tggtttacaa tgtggggtag aagtatcaca tgaaatatat gagggtaaga 540 agaaatcaag aataactgat tttataaacc ttgacgaagc tgactctgat gatgacgaag 600 atgaggatga tgactcagat gatgatgaag aagatgatga agacgatgag tctgaagttg 660 atcttgaaga attagacaag gatgagctaa aagaattagc taaggcttta aaaatcccag 720 ctaagaaaat caagaaggct aaaactgaag aagatctaat tgatctaatt gaagaagaag 780 ctgacgaaga agaaatagct gaacaatata atgacctatt cggagactct gatgaagatg 840 acgaagaaga tgaggatgaa gaagaggaag atgaagaaga aaatgactat gagtcaatga 900 ctttaaaaga acttaaagct gaagctaagg acagaggctt aaaggttaaa aaaggaatgt 960 ctaaggatga catcatagaa atgctagaag aagatgatga agaataaaat ttaattatta 1020 aaatatgagg gcctttaagg tcctccttta tttacccttg gaggctacta ttatgcttga 1080 aagagatgtg gttaaatcca taatgaatat gcttaaaaaa gaatacccag gtttttggtt 1140 taaaactcat gggggaccat ttcaaatagc tggcttacct gatatactag gttgccacaa 1200 aggtaagttt attggtattg aagttaaact tcctggaaaa gaaaagaacc taactcaaaa 1260 acaaaaagac attataaata aaataaatct agcaggagga atagctttta tggctacctc 1320 agctgaatat acaaggagga gattacatga aaaatttaga aagacaccaa caattcctag 1380 gagaactagg agatctgtat gaattaaaga ataatctata tggggataat ttccacaaga 1440 cttatcttga atatggaaac cctgttctat gtataagact tgaagataag ctaggaagag 1500 ctaaaagttt attactagga gatcaagatg acttcccttc atatgctgct caaaaagaat 1560 ctgtggttga tactttacta gacctagcta actatgccat tatggctgct atggaactaa 1620 ctagtgatga taactcagaa atacatgatc ttagtaaaga agaatatgat gaagaagacc 1680 tagatgataa tgacgatgat gacttagatg atgatgacaa tgtttacgac gaagaagaat 1740 tagattttga tagtatgaat aaagaatcct taaaacaata tctaaaagat aatggggtta 1800 aattccatag taaggcctca agagatgaac ttgtaaaact ggctaaggag gtataaggag 1860 gaggctttaa tgcctcttct tttttagtat gaagagaaaa ttatttaaac atcagaaaga 1920 agcattacaa ctatttttaa ataaagaaaa gtttgcccta tttatggaca tgggtacggg 1980 taagacctta gtccctattg tggcccttga gaaacttgaa ggtctagata ctgtactaat 2040 attctcacct aaatctattg tatttaactg ggagtctgag atccataaat ttactaaact 2100 taaggaatat aggatattta aattacaggg taataagacc aaggttatgg aaacctatag 2160 agctataaaa tcctactcag gattaaagat tattattgcc aattttgaga aggctaggtt 2220 gatggataaa taccttatga acttaaagcc acagtttatt gttgttgacg aatcccataa 2280 ggtaaagaat agaaatgccc agatatctaa ggctctatat aaaattgcta ctaaatgtaa 2340 atataggttg ataatgacag ggactcccac tcctaatggt tatgaagatt tatttatgca 2400 atataaaatt atgaactcaa atattctagg agttaattgg aaacagtttg aagatgactt 2460 tataatcaaa ggaggttata tgaactatga aattgtgggg tataagaatg aggaaatatt 2520 aaagaacctt atgcaccaga attgctatat agtaagaata gaagattgta ttgatttacc 2580 agaacaactt cctgatctgt atttaacctg tgaattaaat agtaaagcca gaaaggccta 2640 taatgacctt agaaaggaaa tgatagcaca actagatata gtacaggaaa acattcctag 2700 gaagcaatta aaggccttat taaggtctaa tggtataccc tatgagagta atgagcccta 2760 tgaggattta ttcttaaggg ctaatatgtt tatcaaccaa ttaacagctg atcttactat 2820 tacccaatat ttgaggctac agcagatctc aggaggattt atcacaaaca atgtaggaaa 2880 taatattaat attgataaag gcaaattagg cttgttacag gactatctag aaggatataa 2940 aaagcctgtt gttgtaatat gtaattttct tgaggaaata aaacttatac atgatacttt 3000 taaaaagacc cacagagtag aatgcttaac aggatctact aagaatagag ctgagatcaa 3060 taaggatttt caggagggca agattgatat attaatacta caaataagct caggtagtgt 3120 tggtctaaac ctcttcaggg cctcaaggtt aatattctat agttggaact ataaatatga 3180 tgattatgtg caagctattg ccagaatcaa aaggaatggt caaaaggagc cttggcaaat 3240 aatacactta ataacggaga atactattga tgagaaaata ttaaaatcaa tacagcttaa 3300 aagggataga gcagaaaaac tgttgactat tgataagtaa tgtggtaata tagaattgag 3360 gtgataagta atgagaaaag aacaacctag aataagtaat catactgaaa tagctaaggc 3420 tttaattgag gctggtaaaa ctattaaagc tctaaccaag attaaagtca aggaggaaaa 3480 gaaaaaacaa aaaagacttg aaaggttaag actcctaaat gaggaaaaga agaaggagag 3540 agaattaatg aaagaattta atcccgatga gcctaagaag ccaagaaaga aaaaggacaa 3600 acctataaag aaggttgatc tgaataaagt tgaggctaag ggaacttacc ctacaccttc 3660 tggaggtaat gcacctacgg gaacagttga cttaaaaatt ttatgtgagt cattaggatt 3720 agatccaagt aaagccaggg ccaagctaag aaagcaaggt gtaaacaaac cctataaatg 3780 gtcaggatct gaacttgagg atattaagaa aatgttaaaa taagggggag gtttaaaaac 3840 ctcttctttt tcgttgacaa tatccacagt atatggtaat ataaatattg taatcaaaca 3900 aaaaaggaga gaactgaata tgaaagatct tgaaaaactt acaagggaag aattgattga 3960 acttgtaaag gagttatcta ttgattggac aggagcctat aatagaaact ataagattga 4020 ccttgatgag tatgagattg tatatattga tcttaatgat ctaaaagagt tcaatgataa 4080 attaggtcat ctagaaggag atgtgtatat acaatacacg gtaaatctta ttaaggccca 4140 tttacatcct gagaaggata tattgattag agggggtggt gatgagtttg tagttatcag 4200 tgatatagct tataatctgt gcaatacctt gaatatatac cctgagctat catgtggcta 4260 gggtaaaggt aaaactattg aagaagctat agtacaggcc gactcaatga tgtataagaa 4320 taaaaaatct aagaagttaa gatactcaaa ggaggattag ttattatgag gttattatgg 4380 ttaaaagaat atttggatat atgcaatatc ttgggtattg agcccacaat ggaatcttgt 4440 gctaggtggg gaaagaataa aaaactatca gggttttcat taaccccagt gggagcaact 4500 aaatagttgc ttctttttaa agcgatccga tacctagcaa ctttcatgcc aaaaagaaat 4560 tcgggatttt ctttgacata gactccagag tctggtataa taatcttgta aacaaaatta 4620 attcaaagga gttgttaatt atgaataaca aaaaatttca agaaacttta gaacaattaa 4680 aggaggaact agaaatacaa ggggtagaat ttaatgatga gaatgactta atagctgctg 4740 ttgaatacat catagaatat gattaccacc cagaacaata tgtggaagat actttatgta 4800 actaccctga aatgttcaag gatctataaa actaaggagg tgtaaaaacc tcctttttta 4860 tttttctaaa aatctggatt taatcgttga caggatcggg actatctagt ataataatat 4920 tgtaatcaaa attaattgaa ggagatgtta actatggaac taacaagatc agaaaaactt 4980 aatgtaataa aaggaattgc tcaaaaggct atggataata aaatctatcc acaatttaga 5040 aaagaactag ataaacaggt aaaatccttt ggactaagca acaaagttag ggaaagtgaa 5100 ttaggagact acctttcttg ctgtatggtt gcctatgaag aggttatact aaaaatgaaa 5160 aaagccaatt aatatgggtg gccaaaaggc catctatttt tttcgaaaaa atctccagaa 5220 tctcgttgac atagtctggg ccctctggta taataatatt gtaatcaata ttaattaatt 5280 cttaggagga atgaaaaatg actaaattag aaatgttaaa tgagatagaa aaaataaata 5340 aggtggtttt tgaaaagaac ttccaaaaac aattcttgga taaggtgata gaactactac 5400 aacaaatggg agtaactaaa tattatgtat gtaaacatca taaaaaagct gagattgaaa 5460 aacatcttga ggcttgcatc tgtgcaatat atgatatagg aagaagggat gatctaggat 5520 ggtaaacccc atcctttttc ttcgaagttt ttcgttgaca gatccagagt tctctcttat 5580 aattaaatca taaattaact attggaggta aacaaaatga gagatatgat agaaaatcaa 5640 ttagctgaat atgaggtaaa ggagttatac aatgaggcca caatgatctt aggaaaagca 5700 tttggtgtaa gtgctgttga tctttatata gaatctgatg tggatgctaa ctactatggt 5760 ctatgccata gatcaggtaa taaatttact gggattacaa ttaacctttt ccctttcagg 5820 tttgcacagg aaggagaaaa agcttatgat aaaatcaaaa aatttggctg ggatattaag 5880 gactgtgttc ttgaaactat atgccatgaa ttggctcact taacttattg gtcacatagt 5940 cctttacata aaaaattaac tagtctttat tacaacaaag tgcaacaggc tagaaaaggt 6000 gaaaactcag ctgtagcaca aactagaagt gatatataca ctttaaaaga tttatgtgaa 6060 ttaatagaac tagatccaag aaaggctaga agtatattaa gaaagaataa tattgagaaa 6120 cctggtaaac agtgggagtg ggagcaaggt atgcctaaag atatatatga tctattaatc 6180 aacttaaagg ttaggggaag aaggtaaact tcttctcttt tttcgttgac atatctcgag 6240 agtgctggta taattatatt aacaaataac ttagggggta aatcacatga tagatatata 6300 ttatagtatt caagtagaag ctgaaaactt ctttggttgt caaaaagaag atggaagctg 6360 ggtagcaaca ggagcattta gtatacctgt taatagcata acttttgaag aggctgatca 6420 attatccaat gaggccatag atgggtggtt aaaggagtat gaggttacta aggtaataag 6480 taggagatca catacacagg ttaaattaga ttggtataat agttatatca aggatgatcc 6540 attaaagggt gatataatta atatggtgta ccctgatctt tcagataaag ctaaaatcaa 6600 atatggcttt ggtaagatat gggctagcac tatgtcaaag gttgctaaga agggtttact 6660 taaaacttgc cctagatgtg gaggcactgg ccattactca agaacctcag atggtaacac 6720 tacttgcttt aaatgtaagg gctttaaata tgtgatacca actaagatct caaagaaatt 6780 ttataaatct gtggagaaaa cctttgacat ctctggcaat aactcttata ataaacttgt 6840 aaacaaaaac aaagaggaga tgttaaatat gaacttaact gcaatggaaa caaagatatt 6900 aaatgctatg agaaagaatg aatttgatga tggactagat gttgactgtg tatgggtatt 6960 ctcagtaata gaaaactcag gtatagaagg aactaaggct agaggtgtga tctcatcact 7020 tgttaaaaaa ggtttagtat ttgctgatgg agaggtaata ggttacacag aagaaggaag 7080 aaaggtcttt gataatgctg atggagaaga atgtaactgg ggaggtccaa aattattaaa 7140 agaaatccct gacaaaccag aggaaatctc ttataatgaa aatgtagata aaacaaataa 7200 gaaagaagag gtaaatgata tgaaagattt aaatactatg aaagctgttg aaattaaaga 7260 attagcaaag gaattaaaag ttaaaaactg gtggactatg aaaaaggctg atctaatagc 7320 tgctatccaa caaatccaac aaccacaaga agaaaaagtt gaagaggtac aacaaccagc 7380 tgaacctgaa gaaaaggctg aacaaactaa aattactctt gatctaacag atgttaagga 7440 acctgaagaa gaaaaggttg aaactaagaa aaaaccaaaa actaaagaag gtgtatttac 7500 actaaagatg atacttgaag aattaaatat gaatggtaaa aaggcaagaa ggatcttaag 7560 aaacaaagag gttgtaaaac caggaaagca atgggaatgg gataatgaag aagaatttaa 7620 aaaggttaag gacctacttt ctaaataaag taggcccaac ctctatcccc tattgattta 7680 caggaggatc tgattttctc ttatccatta actgcttcca tacttgatta ccaaaggaag 7740 ctacagctgc acatattaaa ccctgtataa agccattcaa aaaggtcttg gctgtaaaac 7800 ctaattctaa ttgaattgcc atatataaga ttgataatga tatactgaat atcaaaagta 7860 ttaatggtat agaccaatct ggtatataag gcttagcctt gataaacaga cctaaacagt 7920 aacatgatac aatgattatt aaaagctgtg gatcaatgta attgaatatt aagtttacta 7980 aattttcttc catgataaat accccctaaa ttatctctac atattttgaa cttacataac 8040 cttgcttctt accgtggcta gtattatatt ctatatgata ccaaccattc ttacttccca 8100 atatagtaac tctctcatta ctaaatactg agcctattat attaccattc acatcatctc 8160 ttacattaag agaagtgtta actttaacta ttccttgttt aacatcctca tagaatttct 8220 ggctttgctc accctcgtag cgaacgtatc catcagatgg agtaccatcc tttctataaa 8280 atgttattct taaccagcca ttagttctat acattgggct agccttttga ccaggctgta 8340 gttcttcatt atgatctttg atgttaacgt cttcagggtt ccaccacaca taagttactc 8400 cattgtcatt ttgatatttc atgtggtagt caaaacttaa tctgctgtaa tgttttatgt 8460 caatataagc atggtacttc ttattctttc caggatatac aacttctatg aaatttctag 8520 aaggatatat tccgagaaca aataaattat ctaggctatc aataaatcta ccagggactg 8580 gcttacaatc ctcatctaga acttctattg aaccttcccc cactaaagta gcattagttc 8640 ttgtctcatt agactcatag aatcctttgg ggttattgat tactggtgag cttgagctct 8700 ctcctgaatt tactctttct aggaatgtat tccagtaacc attggctatc attcttctag 8760 gacaattttt tctactggca tcatagtgtc ttactacttt acaattagaa tagtagccat 8820 tctttaggta tctaactagc tctacagcat tatcaactgc cttattgtag tcagagtcag 8880 gattaacaca tatttctata gatattgtat ttctattatt gataccgtat ctattatgac 8940 catctccaca attccaagtg gcgtgtttgt gttctacagc ttgatatacc cctgtatcat 9000 cgacataata atggacactg gctttaccta tatttccatt tgcttgagcc ttacagtggg 9060 tttcggctcc agcaccccaa gaggtattat cagtttcgtg gataattata tacccaggat 9120 tatttgcatt tagagtaaca ttcgtagtac ttaatctttt attgattttc ataataaaca 9180 tctcctttat atattatagt gttttattgg gcctgagagg ctcgctacag cattttaacc 9240 acctccttaa tatatttcct tatataaaac ctaacaggcc ttaaatggcc tgctaggctt 9300 ataagttaac ctttgttact aaataggtga tcttttatag tatctacatc ctctttgata 9360 tcctcaacta cattgaattt atcaactagc ttatctatca cttgatttaa tttattctcc 9420 ctttctctag cttcttgtct agtgtcatat aatagccata caaatagact acaaaagata 9480 ccttggctaa gaacttgagt tagtatttct ttttccatag attacaccct ttccttaaag 9540 tcttcaccac atattctttt atagtcctct ggtgtaattt ctccttccgg attagaagag 9600 cattttaccg cctgatgtaa tttgtcctta cctattgctc caattctata agccatttcc 9660 cagaaattca taatccctac ctccttattt attcatcatt ttaatcttta tatcagctag 9720 ttcttgtgtt aatactttca ttatggcctt agtttgcatt agctctaatt tagtatcagc 9780 aagagcctgg gcttgataac caattaattt attaatatcc agaggattag gatctatagc 9840 ctcataggaa gtaaataggt ccttatcttt gatactgtat agatctgatc tctttttact 9900 aatatcaaat ttggttgggc caccctttat gtggtcaaat aaacttacag tcatttgtat 9960 ggctgaaaga ttgtctagat caattacctc agtgttatcc acaaaggcta aatgcccttg 10020 tgaattatga ggtatatgga gacatataat atcttctata ttatttaata tttcactatc 10080 ctctctatag tatagaggtt caggcctatt aattaggtct aatatcttag taacttcttc 10140 atggtcttta tctataaatg attttacatc ttctgagtct tctgcctgat ttatcagcat 10200 ttctttttca tcatcataga atataaggtc tatgatttta ttagtttcca cagaataatt 10260 aaatcctact attctcatgt tttatcaact cctttactca gtggctaaat aggttagtat 10320 tatttgagtt ggcttaccag ctaccatttg ataacatcca ccattcaaat atgagttatt 10380 ccaatagtca atagtacaat aacctgaggt attattagaa gcgtcattat agttatgtga 10440 tactagagat ataggtaata cccaattcca agccttagga aaatgtatag tataacttac 10500 acctccacct gagttagcat aacccgtctt aactatttga cggactaatc ttagtcctcc 10560 cccaagatga catactctaa cgttaccatt gtcattatcc caatagtctc ctttagcaat 10620 agatctatta tttacaacga agtcagtaac attctcagta tatagctttt tgttctcttg 10680 tttgaaatac aatcttccat cttcagctcc accataccag tcatttattc ttaggtattt 10740 tctagcccat aggttttcag ctgatagagc cttatcaacc tctagagatc cgtctctagt 10800 agttatcctc atatcaaaat ctttcttact accctttaag tggaaatcta atataggacc 10860 tagttccata actccatcat gtgatataaa agggatacct gcatagtcat tactagctcc 10920 agctaagtcc cattctttta gtttattact gttagcagct gtagcggttt tacctaaata 10980 agcactatca tggttatggt ctgaggctgc agctcctata tcggatggct taggtttatt 11040 gaggttacta taagctatac accattcgga ccatatgcct ttaaaactac aaaacctagt 11100 atatgttttg ccataactat ctgtaaatag ttgagttacc tcacccccct tgggtaatac 11160 caatagagtt ccataaatac catatgtgta aactacaggg gagtttggta aggtatctac 11220 actagatact gagtatatcc cctcttcctt acaaagatta aaatctacta tatcatcttt 11280 tctaatagtt aaataaactc cctcatgggt atggttagtt ggggatgccc ccacatctgc 11340 tgcagttggt ttattatctt catgatagac cttcttattg ttcttattaa caatatccag 11400 agagtctatc tttacaggcc ttccagcata ccctatatgg actttattat ctgcatctat 11460 atagatcaca tagtcatcac tgccttcttt agtcctacag ctaaatccat aattgtgggc 11520 cccaataaat ttctgacccg gtctcatgat tagatggcct gataattctc cacccgtagt 11580 aggtaaggca ccaatatctg ctggagtagg cttattttgt ttactatata tttcagccca 11640 attcgtccat ccggtaccct caatgtttgt agctgttcta gtaaaaactc ttccaccatc 11700 tgcaaagaac atttgagaag cccagtgtcc tgtattccag gataaaacaa tcaatctccc 11760 atctgctagg attggggcat ttctatcgac agaatttcta actgtagaat aaatacctcc 11820 ctctgtaatg gtgtttagat ctacaccctc tagtcttttg gctgttttat cccaggcccc 11880 aatatctgcg ggtgttggct tagctccgga gttataatct tttctccaag ggaaccaatc 11940 cgtattattt actttatacc tccaatactt attaccatta gtatcataat actcttgcca 12000 tatccagtta ttagtattat tatgcttagt ctctgtacta acatatacta tcagtatacc 12060 atattttgta ccagttagtg gggcattagg taactcacct gagatcaggt attgcccctc 12120 agttaaagct gtattaaaat cagttatggc gtccttagga atgggataag cctttaggtc 12180 agcaggggta ggcttattag cctcatcata aaccttatac cacttactcc aagaattatt 12240 tcctgatgtg tagtttctaa catatctagt tattttattg gttgggtata ggttaataaa 12300 ctcctgttgg aatcccctct cacctgtatt tctaaccata aaaaccctat tactcatatg 12360 ggctaaggtt ccctcggggg tatttttaaa gtcaccagtt cctgtatcaa aatagaagaa 12420 ggcattcgct tgtttaaagg aattacaatc tctatctaag ggagcactaa ccttaataag 12480 tgcttgatct tgattagcct ttcggtctac ttgattttgt aaactggtgt ctttctcaaa 12540 taataggtct aatcttttta gcacatcctc aggggctaga ttataatcag gtactacatt 12600 acccctagct aatatgaggt ttagtacctt taatgaggta cttgaggttg caccctgtac 12660 cacaaattgt attttggtat cagtagctcc agatggaata ctaaatttat gagtttttaa 12720 cttgtaccca ttactctgtg gtgtactacc ttgttttaca atactcctat attgttgacc 12780 tccattagtg taagccagtt taatttcata gctagtaaaa ctaccttcaa tatcttctag 12840 aaaggctagt atataatcac tatttggttc tataggcaga gttttaatct caagagtttg 12900 tggacctgtt gacataccag accatttaaa acctcgtctc tgggttggag tatctgtaat 12960 gggtgtgata ttaccaggac cagtataaga ctcagagttg taccactcaa gcctattaat 13020 agtaccaacc cttaaagcat ctagcctatc ctcattagac ttagacttct tttctacctc 13080 agtcattttc tgtttatgtt cggtagcaaa gttctcaatg ttctcattgt ccttgttaaa 13140 atcattcatt tttggcttat ctgttaactc ccacttattt aaacctaggg taggagtttt 13200 actagtactt ggcattttgc atacctcctt tattcagtta tagcatattc ttcccatttt 13260 tcccaggtaa gatctagact gtcccacttg ctccacatat aatcatatct atcgaacatc 13320 tcccaagtaa tataaagtaa tataatttga tatattaggt gagcaggcat tctttccctg 13380 aatactcttt ctagagcctt taggttaggt ggtacccctg gtaggtcagt aaagctaatc 13440 ttgatcttac tctcttcagc tatgaactct acctttacct ttccacctgt ataggcctta 13500 gtaagatttt tgatagatgt actattaaac tttttccaag cagatagaac tactgtaata 13560 atagctcttt tctcttggtc agttggtggg taatcaaagt atttatataa ctcaagatca 13620 gtaaaaaatc tttctagacc ttccttagga gtggtcttgg gatccacata gtcttgtaat 13680 tcctgtccct tagtatataa cttctgaagt tcagtagcca taatggtgtt tactaagtta 13740 tcagcctctt ttatattatc ataaaattca ggaacataca atagtaattc tttttgaacc 13800 tgttctagac tatacataag ttaccaccac ctcagtaatg gttggtattt gattagcagg 13860 aatagttaaa gattgactac ttccattaag tgtaatggtt gtataatctt ctactccctc 13920 aatatttaaa agcatagccc ctattctatt tatagatagg ctagtaattt taaatacttg 13980 actatttaga aattcttcta gttctttctt gaagtttaat tttatttgtt cttctgtaac 14040 tgaactcaat ttcttaactt gagctcttac agtgatatta aaggtttggg ctgactcaac 14100 agtaaaaaca catcctatag gagctttacc ctcacctaaa cctttcttgc ctggatccat 14160 gtattcttga actttattaa ctagctcagg attagcaggc ttattttcaa ctgtggttat 14220 aataccttta gcagtattta ttccatccca acagggaaga accacagctt tacccactcc 14280 agctacactt ttacaccaat cagttaactg ttgaacattc ccatctccag atggttgtat 14340 agtggcttta gtagctcttt ctcttaactc atcatctgtc tcctgatctc taccaataac 14400 aagtatatct cccatagtag cacttctaag ttctgctata gcattgatag gaattaaatg 14460 tgtacctgtt gatactttat tgggtcctga acctaaggtc tcacattcaa taatattagg 14520 ctctatcact tgccagaaat aatctccccc ttggaatctt gaaccaacat cgggttcata 14580 gccaacaaat atagctgacc ttctacactt agttggaatt attctggata tccctacatt 14640 ggcagctctc tcatctaaga acttacctgt agcagtactt agaaatactg agtctaatat 14700 ttgattaaca ttaacagcat aaaactcagc tagcttaaca gcaaaaacca tagccatatc 14760 ataggtgaaa cttccagttc tagtgtcaat atcatcaggg gataggccta tcatttcttc 14820 taataattct ttatgtgttc gggagaataa atcttcagaa aatcccatta tatcacccct 14880 tccagttttg tatctccata tatggtattt atccatactt caatatatac attcttacct 14940 tgcacattat actttagatc agttacttta gtcactctat catcttctaa gagacattca 15000 gggatagccc tattaatctc catttcaaga agttcaggag ttactagaga gtcattaatc 15060 aaagctctaa tatcacaacc ataagaagta tcttgataaa tcatataggc atttttagga 15120 gttattaatc ttttatataa agattgtttt aaggcctcaa gaccatccac attatcataa 15180 atcctattaa ggtttagatc cagtttatag gtcttggtat tctccattgt tttctcatca 15240 actataagag tatctttaaa gccattaggt agcattatat caacctccta agatccctaa 15300 cataaaatat ctttgtccat catcataggg cataagataa acagtatcgc cctcactaat 15360 agactcaata tgttttggaa catctataat atttccagat agtactaatt ttctattgtg 15420 aagacactca cattttagag ggctagtact aagtacctta gctctataaa tatttagagg 15480 ggtggacata ttactatata cttgttttac acagtctaat aaattattag ccattaaata 15540 tcaccctcct cagtacattt aagttttaaa ttcatggtgt agtcagtatc aaatttatgg 15600 gtatcagact caatgtagaa agccttgtta atttttaaat gtggtatact aaccacaatg 15660 gctttgccac taatacaatc aggtacacct ataacagtta tagagatatc agttttaggt 15720 ctcttacctg tttctagctt tattctagcc ttctcattta attgggcttg gtttagttca 15780 tcactgatag tatcgtataa ttgaaataca ccatatttct tttggctgct atcatcttta 15840 gcagaggcaa ataaagtaat ctcgggatca tctttaccct taccctgttt atatttacat 15900 actagcttta cctggttgac aatactatca aagtttctac ttcttgaaaa ggttccaata 15960 tttttaccat attcgaattt ccacaattca ttattggttc ttcggtttat aaaatcaaac 16020 ttacctgtaa tagaattata gtaaatataa aactgttctc cagtttgttc ataaacctct 16080 tttattacct tcttcataat atcataggga gtttgtttct cacatactaa actagagaaa 16140 gtatatccag ttttagctac tactccaggg gcaaccccaa aggctttaca taatgactca 16200 aatatagctt cgccagtttg gttttcgaat acaaagctat ctttattatt agcccaataa 16260 atcatagggt cataagcttt aaccttaagt aaatgctttt tctcattatc tgagatattc 16320 ataataatac cattaaagat attcttaccc tcatggacta gtactacctg atctccatca 16380 taaattggat gtatatcttg agcattatat tggaatgata gggtacgagg ggcagatgaa 16440 gaagaccctg cccaactctt accactggag gtatttgtaa tatcaaagga tttatctcct 16500 tgtaccacaa ttaattgcat attataacct ccttatataa tcaccagaac tccaaccttc 16560 tttatatcct gaaccattat ctaatttata ctttacataa tgccatccat tttgagatct 16620 taatactgta aagtgttcat taggccatat cttaccctta gtgggattat tgatacctgg 16680 tccagttctt acccatagcc atgagtaaac attagtagta gtagctgccc aaggttgagt 16740 acctccagaa gaactacctc ctgtattacc actataggct agtttaggtt ttggaattgt 16800 gaaactaact gatttaacct caggcttctt atactcagtt agttttattt ttacttgaag 16860 agtaccaata tctccacctt tctcagtata tgagatatta tcaatagtaa ctaataagtt 16920 aatatcagta ctagtaatag taaatcttac tggatcttta cccatttcaa taagtttagc 16980 ataacttact ctagggtctg caatatcagt atactcacat ccatcgtagt atgagcttgg 17040 taactcaaaa ttaaaagaaa aagattttag cttagggtct cctatgaaag agagttcccc 17100 taagccttca atggaaactg tactattatc tctactatca gagatattta tatcagtggg 17160 atttactggt aacctaaacc tagtatctcc ctgtattaac cagaattgat atttactatg 17220 caaggtccag ttcacctccc tcgaacattt cctgatttaa catttccaca ataaatctgt 17280 atactctcat taattcactt tctgagattg ttccatcttc agcctgtact gtgaaattaa 17340 atgagttgtt aatgttcata actttagatc catcatcact tcctgctcca gattgtctta 17400 gtaaatcctg agttttatct gcaggtaata cttgagagcc tctaggcatt tttgagaatg 17460 taggacccat aactaattca ggaccagctt caccaaccat ggcagcttta tctgtaaaga 17520 agtcagtacc ttcagcaagc attggtatct ggggaatatt aataccctta cccccaataa 17580 taggtaccca gtcaggtatc ttaatagtat taagagcacc aattaagtta ttaatcatac 17640 caataatact attaatagga gcttttgcta gggataacat accttggaaa atacccccaa 17700 agatgttctt aacaccttcc caagctttag accaattacc tgtgaatact ccagccacaa 17760 aagctattaa accactcaac atctgcatta atccctgtag tattcctgat atagtttgaa 17820 cggccccagt aacaactgag gagataacag gccatactgt ttggaaagat gcggctaaaa 17880 ggttaagtag tggagttagc atatccacca acccttggaa tatcatacca aaactctcaa 17940 tagctgattg aatatatgga actacattct cagcaaagct taagaacata ggtagtagtg 18000 tattattcca ccaatctgct agtccttgta atattggcat taggaagtta cctatagctt 18060 ccacaacttg tgatatagct ggggctatgt aggtgttaaa ggcttcagct aatgatacag 18120 ctattggcat gaaaacatta gttatcaatt ctccaagagc tgtaaaagta gcagagaaca 18180 tagaacctaa tacttctaat atagggccta acacttccaa tacagtttgc catacttgag 18240 taaagccttg aacataaaga actactaagt ctatggctgg aattaggtta ttcataaata 18300 tagaagctaa agttccaaac atatcaagaa gtacttgacc tataggagct aatactccca 18360 tgataggctc agctaaacca ttaaaggctt cttgtatatt attattagcc tctgataaat 18420 aatctggtag acctattaag atctctccaa aattattata gatgtcttct ccaagactcc 18 480 atatcttctc attaatttga tctgccacat cagcaggtaa gaacttatat aatatatcat 18540 tggttagatc tataaagggt tgcatagcat ccccaaactc ccaaccattc atgatgtaat 18600 atatagtctc tcctaaacct tggaaagcag aaacaaccat tgagaatata ggtcctagtt 18660 tttcaaagcc ttcttgtata aatgtgatac catttagaac agcattagct ataacctcaa 18720 cggcaggggc tagggcatct cccacatcta acataactac agcagctttg gtttttattc 18780 tctccatggt tctattaaag cccttatcca ttttttcata ggcagcttca gtggccccag 18840 cactatcctt catctcatta agagcattag taaaggcctc agtaccttca ccagtcaagg 18900 ctagtgcagc tgaaccagct tctacagaac caaagaggtc attgataccc agattacttt 18960 tctgggcatg tttttctaac atctgaaggg cctcttggat attacctcca cctttaatga 19020 agtctttgaa acccttacca gctatttccc ggaagacttt atcagtcttg gttcccgatt 19080 tagatagctc atctatagca gctcttatct tagtagtagc tacagatgtt ggaaccccct 19140 gagctgttaa tgcagctata gaggcagata cttcttcaaa agatacccca gcagctgagg 19200 ctgaaggtag cacattgaat agggattgtg atagttgctc aaagttagtc ttaccaagtc 19260 ttactgtagt aaacataagg tctgaggccc tttgaacatc catattttta gctccatatg 19320 agttaataac agtagttagt ccatcaaccg cagtttctat tgatgtgata ccacctatag 19380 aagctttacc agctgtttct aagaaactaa aaacgttatc ctctggaact gatgaggata 19440 tagcttgata aagagcaggt actactttat caggcaatat ccctatatca gaacttaact 19500 tttttacctg gccggacatc ttatcaaagg tctcttgagt agagttgggt aatagagtca 19560 taacctcatt taaaccttgc tcaaaatccc caaaggcttt aacactgtcc ttagtaaaat 19620 cccagacagc cttagctgag aatactccag cagctactcc accaattttg gcaagagtac 19680 ccatgaagga attaccacta tcagccatct ggttaaatgc accagagatc ttatcttggg 19740 cagagaatat ggcagtaaat gcagcttgac ttgccattat atccctcctt tatgattaag 19800 agcctttctt taacttatcc ctagcttttt tatcttcttc taaagccagt tcacatgagg 19860 ctataagaaa ggctttccta ttgggagata gtttatcaaa ttcccagggt agtatattat 19920 gcttctggaa taacacatga gcccaaaatg cactagtgtc ggcctttatt agttttttgc 19980 ttctttgatt agatcatcct tattatcgcc taatccattt atgtccataa cctcactagt 20040 tattctagtg aaatctcctg gtaagcttag tacatccata agtaaatcta ctggatccac 20100 aaacttagta gactctaacc attttggatc tttaaagtca gggaatacta ttgtttctag 20160 tatcatttcc ctagtagctc ttctttggtc tgttattgag ttagttatac ctttgtttct 20220 agtaattact gtatttctct tttggatatc ttccatttgc ttagtagtga ttggctttaa 20280 tatccaaggt ataggttggc ccgctaattc atgaccctca gggaaactaa atctagctac 20340 cccttcaact tcaatttcgg gtgattgttt acccttatca ttttccatga aaaattccat 20400 actaaatttt gacataataa tctacctcca caatattctt aatcttttat aaatatctta 20460 cttctgaagc tgagaatgat atttcttctt gaatttcttc tccctcagca tctagtccaa 20520 ttagtggtaa atctccttct aattgtactc ctagtaattg tactctgtcc tcaccataat 20580 tcttaacata gtcagaatct ggatcatcac atacagcttg tatatccatc ttagggaata 20640 taccagtttt taagtattcc ttaatagctt ctcttatcat tggagtagct ttgaattgag 20700 ttatagttcc agttatatca taacctataa cttttgaggt taatcctttc tgacccataa 20760 ctcttttctt agtcacctca ggagtaaata gagcttctag ttttattaag ctagttactt 20820 gcttacctga tacaaaagct ctaccctctc tagcagacat tctattaact tcactcatgg 20880 gctatacctc cttaattatt cttaacagtt acaaagtatt tcttgaaagt tcttacaggt 20940 tgtgtaagta tagtagcata catagtcttt cctttactag cttcttcatc taccactaca 21000 tcagagtcag ggtccacatt ctttaatgct ccttgagctt gtagtgtagt taatagactt 21060 attagtctat cctgagctag ctttatacca tctttgtggt tagggaaaac attaggaact 21120 aagatctgag atcctagatc tataacagca tccaatacag ccacaacttt gttatctgta 21180 aagctttcat cccattcttc tgtaaaagta tggtgtgtat ttatatcttc ctcaactact 21240 acctttactc tttcagaatt tgagtcagtt tgagttgaga atataaattg acctgttttt 21300 aatccatcaa ctatttgaga gtgttttagt cttggattgg catctactgc tccaggatat 21360 gctttataag taaggtcttc agtatatcca gctgcagctc tagcaccagc tacccatgca 21420 gttgcttgag aagctgttaa cttagtgccc ccttctaata caacaccatt cttaacattg 21480 ataatgtatg ggtaatcaca agtagcaaag tcagaagtaa ctgccacaat attcttacca 21540 acttcatctc tgaagtattt aagttttgat aatagagcag tgtgtaattc actatcagta 21600 aatgggaagg ccatagtttt aaagttgtat aactcacagt tatctaagaa gttgataaca 21660 tcagagttat tcatctgagt agaagtacca gtactaaact cagtcttagc agtctcatct 21720 atagaaccac ttccacttac tgtaaaccat tgagacttaa ctgttgatat ttcagttact 21780 cctgtagcct caaagatttt agatccctca agatataata atatatcctt tcctccaaga 21840 ggattggtct taatagttag agataactta ttacctcttt ctccagcttt agtagctgta 21900 aatgttagag accctactac agctttggct ttagttccac ctgttaggtt atatactaaa 21960 acagttgcag cttcttttaa agcctcttta actagtaata cagtagggtg agttatgtca 22020 tacccaagtt gatataatgc ttctaggcca ccttcattgg ttaccttgat aacatctcca 22080 gcattacccc atttaagttg taatggtaag atcataactc ctagattatt aagcttaggc 22140 tcattaagtc caccacttac gaatctagta tacataccag gtcttacttt attttgtcct 22200 ggaataaaag ttcctcctgc catctatttt acctccttct ttagccagtc ttttataatc 22260 tgactagttt gttttattgt taactcttca tcaggtggaa gggattgaac tgcagcgttg 22320 aagacaaact cctcaacctg gaaaagttgt tgacaatggg ctcttaattg aggtatggaa 22380 aatttttgta ctttcactat tttaccccct ttgttatctc aaagttttcg aagtaagttg 22440 gatctttata aagactaacc acaatatcat agtttatctc tacctgagaa tcttcttcag 22500 tttcaccttc aaatacttcc acattcttta ttttgagatt tttgttggtc ttactaccat 22560 ccctattaag tataggaatt tttctatttc tcaagtatat agcattagct atattctgag 22620 caatttcaag agagtcttta ggttcaaata ccttaactct taaagtattc tcagttctat 22680 aactacaaag agtatcattc tgaggattaa ataccttctt gaaatataat gagggatact 22740 tgaacttttc aggtattcgc tcaaaataga ggatagtaat tccagttaga tcagatatga 22800 atttagcaat actccctata tctgctgtca ttaataacta ccccctaaaa cttcggctaa 22860 ccaatcttct aattttcttt ggaaagcctg ttcaaatatt ttttcatata tttttactgc 22920 attgtcccaa taaggtttac caggtacttt cttttgtctt agtaccatac cagtcttagc 22980 tcctggttga taaataaatt tatctcctga ccatatgcca ggaacgaatc taaatgattg 23040 tccttgtgga ttagtgtagt gaccatcgtt tacccactta gcatatttta ggttagtacc 23100 aacttctaaa gttagtcctc catcactact tacccatata ccattcttac ctcccttttg 23160 aaaggagtta agtaataatc tagtgtctac tacttgtagt ctaattattt catcttgaac 23220 cacatcaagg aagtctatcc ctgcagcttc aaaccatact gctaattgct ttttgaaatc 23280 actattagct gcatctctta gcttctttat aaacctttct atttgtttag catctatttc 23340 tatgcctcca gaatttgaag ccataactat ctaaaatcct tagcctctat accaacctta 23400 atttttagac ctcttatatc cctaggagct ttagctatat atgtgatacc agtctcaagg 23460 tccacaattt tatcattttg tctaacatca gttccaatag caaaatctac tgagtactcc 23520 tgggtaatct taaacctagg ttctttgtac tcaagttttt ctagagttcc tgacttaaaa 23580 tgacagggta cctcagtcat gtctggctca tcaggataac taaagctagg ggtatgggct 23640 agcccatatc cgggatcttc agggtcttct aataggtggt atatattaca tctatgattt 23700 aagagcttgt tatatgaatt tccaatcata ttacagtacc cttaatctca tagtagttgt 23760 tctgatagat ggactatcat cttctaagaa aggtttaatc agaggcataa tatcaggctc 23820 tatgacagcc ttatcagttc ttcttgtata tgagtaatca tcataggtct cggattgaat 23880 atcagccttg ctaatgacca aggattgtag tccataatac tcagcataca atatatgggc 23940 taaccttaat tctttactat taggattttt ctccacaata gtatcatagt ctttatttga 24000 atttttaaaa attcctgaga tcttggcttt agctcttagt atatccatct caatattttt 24060 ggggtctctt ttcttaatct caggattact tgaaaagtca attatttcat ccggagtaat 24120 attcatatga gccaccccta tctgtttctt ttattcttgt ttgtaaatct tctagcaggt 24180 tcttctttac tgtcatctgt agaaggtatt tcttctccag taaaatcttc tagaggttca 24240 tcctcaacag gttcatctat gacaatcaca tcatagttct gttttaaata atttactagg 24300 ttatctttaa cttcttctac atggtcttga gtaattctat gacccttaat aaccagagtt 24360 tgtaatttac cctttgctaa tttgacctta gccataatat ctctcctttc taatgaaaaa 24420 gagcccacat ttatagtggg ccttcatgat taagctttta ccttgattat cttagcagca 24480 gctttaggat cttcaaactt aacatccatc ttaagagtta aaactacata agtagctctt 24540 tctctaggag ctctttctaa ttcaattctg atatctcttg atatacctat aactatattc 24600 ttagggtgag ttaagattat atctgatact tgagtagaag tatcttcata atcttgtagc 24660 atagctatac ctttaacagg tacaccataa gcagatttaa cagagccatt tataaatgag 24720 tcatcaccat tagcagttaa tctctctcct actaagtcta cccaatcagt ttctaaacca 24780 tttgagcagt agaatctcca ctcacttggg tttcttaaat acttagctgg tacagctttc 24840 ttagcttgtt ttaataggtc tttacttaaa gcaccccttt gagcatctac tacatgagca 24900 gttaactgtt ttctaacccc atctaattgt tgtaagaact catcactacc ctcactagta 24960 tcaccattaa caattatttc ttccatatca agagcaactc tttcagctaa taattgcatt 25020 acagtgtctt ctattgattt tccttctata tttgtttcta tagtatcatc agttaactta 25080 accacagcta taaattcttt agcttgtaaa gtaactgttc cagtagttgg agctgaatat 25140 ttagactccc ctggatcttt tccttcagtt gcaggattta atactctttg accaaatcct 25200 attgactcaa tctttttaga gtctcctttc attggtacag ttctagcctc ttttaatatt 25260 gttggctcat caattatcat tttaataaaa gcgttagctt gttctgtgtt taatttacct 25320 ccaccactta atactgtaga gttaaatact gctttattta ttttagccat aatcttttcc 25380 tcctatcaat tattcttggt ttccaccaaa taatccatta aatactgatt tttgaacttc 25440 ttgtccttct cctccatttt taacaatttg tttacttcct acatatggag ccatggcctt 25500 agctactgcc tcagctattt tttcttcctc agttttttct tcttgtttct ggattaagcc 25560 agcctctaac atagcttttt ccacagcctt agcaactata tcttcagtag tagcttgttg 25620 accttcactc tttttaactt caccctcaac aggtttagtt ccatctaatg cctttttgat 25680 agcttcgtct atcatttttt gtacttgagt ttcattcatc tcttcttccc cctccactga 25740 gtctaataat tcattaatag cactttgtgc tgtttttaac ttttccacat tttttgaaga 25800 tattttctta cctgccttat tgactggtaa ctcaaaggat ttagctactt cctcgggagt 25860 tcctaataat attccctcca tagtttcaga aaattcttga agagcttcct ttatcttatc 25920 tacatcactt tcaaaagatc tagtgtaatc ccaactagaa tatggatata gaatatcttc 25980 taaggtacta actgcattcc agaaattatc tctttttgca gtctctttat atttttcaga 26040 gtatgcacct ttctgtatat gctttggagt taaccctaag gcctttccaa tcttatcaag 26100 tatagattga cctatagact tctcaatctc atcaatatct acatcttgtg tagaatattt 26160 accagtacca cccatagacc atccagtaat ttcgcctttc tcaatcttgc tccatagatc 26220 atcatcgaca atctcagctt tagctagcca agttccagct ttaactactg taccttctat 26280 tgtagtatct tcctcagtta cccatgattt tacaatcttg acaccctcaa gagtgttctc 26340 attgtgttgt aaatcactac caaggccatt ttcattaaac cactcacaag ctttcttgat 26400 ctcttcctca gtcataaaat tatcatggta gtcagcagtc attggctcat atactacacc 26460 gataacaaaa tgattatctc cttcagactc agactttaag attggcttat catatctaaa 26520 tctattagga gccccaagtt ctttagccac aatccattgc catttattag ctgctttatc 26580 aactaatgaa aggtaatcaa ttttagcatc tgagatctca atagctttct taactgattt 26640 tactggcatt tactctccct cctttacaaa ttatattctg ctatcacttg atctctgata 26700 gcttcttttt cttctttaga taatcctagt atatccttat caattacagg cccatgggta 26760 caatgacaat tcacagtctc agaagctggt aagttgatat ccctaggaaa tctagcttca 26820 taaattccta cagtgaagta ctcaccttta ttaatgacag taccatccag atcgacatgg 26880 tgttgccttg ggtgaatacc ccttccccct gagtgtttcc actgtattcc tgttactgcg 26940 gggcattgat taaaagcctc atccttagca tatgaatgag ctcttaacat ttcagtaata 27000 gcagtagctc tagctctagt tctactaaag ccataagtct cagataattt accaactaca 27060 tcttctacag actctccatc ttcaagagcc ttatccagaa tattctgtag tttttcatgg 27120 gaagttaatt tcatcatttc ccctagttta gaagaccact catctataaa ggtagtagta 27180 tatggagaaa agactttggt tagggataag tccttatcaa taccttgtaa ataagtatct 27240 cctaaatttt ctatagcttc ttgataacaa ctcttaagag tttctgctaa agaagtatcg 27300 atattatctg tacctatttg agactccaca tattccataa attcttcaat ggtttcaaag 27360 tctccatctt ttataagatg ttttaattct tctgtaagga gatcagaaat agtgtcttct 27420 aattcagaga tcttttctat tgtataatca atctcctcat agccagcttc tttcaattct 27480 ttcttaaggc tatcatcttc attctttaag atcttatcaa ttttagcgat caactcatca 27540 gtacccacta ttacttacct ccttttaatt gatagtttaa taactgtttc ttagcctctt 27600 taagtacagc tatcacctca tcactcccat tagagctttg agccttctca attatacttg 27660 ttaagttatt actcattgaa gaggcaactg agtaagtagg gatatttgcc cattcttcat 27720 caataggagt atagtcctta ttgattgatg aatagtactc ctcattaatc tgattaggag 27780 tcatagctct agaggctata gcaaacattc ctttctgata atcagagttc tcaatattag 27840 gactcttaaa gtaagcttcc acatatttaa aaccatagcc agatagaata acattattta 27900 gtatccatgt taggttttct ctttcaggaa caaatacttg ctcttcagtt tttcttgtag 27960 cctctctggc tgtagccaca ttataatctt gggagtaccc tacatatata tctggtagta 28020 agaaagaaga ttgtatcttt ttcctagtat tatccatgta gtcttggaat aactcatcct 28080 tctgtaatat ttcagatagg ggcttgacct caacagtaac tggcttagaa tcatcaaggg 28140 cagtgtctct ctctgcagct tcagtttcta atactaagaa accatgctgg gaattttcac 28200 cttttacatt attcatgtac tcagttaatt cattccaaga tttatcatta agagtaccac 28260 cttgaattag taacattaat ggagtatgtt taccattagt aaaatatctc cagtttaata 28320 attcggcctt tcttgaacct tgagcggcta acataggacc ttcccatctt ggtgtaccat 28380 agacaccata acctgtttta aaatgaagta tactgttagc tcgatcttta agaggtatct 28440 ctcccttttc ataatacttt ccagtattca tatctagatc tctaggatct ccccactctt 28500 taaagaagat tgattgatta tttacagtct gcttaaatct tctgaacctt tttcttctag 28560 tgtactgttt accatatcgg gttacagtgt actctgtact ctcatcatct agcttaagaa 28620 ttttaattgt cttagggtct tcaatttctt ttatctcttt tacgagagct gagtctttag 28680 gatcttctac gatctctaaa aaagaatacc caagtccttc tctccaatca actatctctt 28740 caaagacatt cttggttggt ttatcaaaag acagggtatc aattaattct tccagctgtt 28800 tatattcagc ttccatttct ggagtaggtt catctacctt ctcaatatac ctaataccta 28860 ttccgaatcc agcaatatta tttcggtagg cctttttgca ttggggtatt attgtagact 28920 cccttataat cacttctaat agttgggggt catattcagg ggatatctca tcaacttcaa 28980 ttccaagtac actggtttgt tgattagatt tatccactgg atccaaagac ttttggatag 29040 ggtctccaca tatacttctg gcggatatct ttttcaaagt tactttacct cctttcttac 29100 cttatatatc aaattataaa atccagatgg cctcatattt cagggaggat tacatcaact 29160 ccacatatag tgtcaaatac ggccattcac atgatttaaa aataactata atttgatacg 29220 tttattataa cataattcac aaaaatttgt caacgtttta tttttacatt attttccgga 29280 agattttccg catatattcc ccatataaaa ttatataatt attaatgtat actatattac 29340 tatattaatt tatgaaataa attatatagt atatagtata cttatattac atatatataa 29400 aggtggtaac tctgatatgc cttctatatc acattattac taatatagag cctagtcaat 29460 ttcaaaactg atatctcaca tatcagaata aaaattattg gctttaatag gcatatcagt 29520 tacttagatt tttcttaatt tagatccaac aggaaggcaa gctaagaata gagagtctgc 29580 taagtctgga gaatgtagtc ctcttttctt catttcatcc ttactctcca gctttatttg 29640 acctttggag ttgaatactt taaactttct ggtagtcaat tctcccacaa gatctgtatt 29700 atctggtagt cttaactcat catcctccag taattcctta actacagagg ctatatatgt 29760 ggttgtatca taataaaagg cataagcctg ggaccccctc tttaatggca taccaaattt 29820 acatggtact accttcaacc agtccagttt ttcttcaaat tttatttcct ttaatctatc 29880 agttactccc ccacctacac cagtgtcgtc tattttcaca actactgcta gtttagggaa 29940 ctcaccatgt agggatttag cctttcttag cacttctcct actgttttca tagtgtcctt 30000 accttggtaa taaataggtt cttccacata tccatcaatt ttggtcataa tagaggtctt 30060 atctgaacca aatctagcta cgtctacacc catatctata gtaactggtt ctgtaggtct 30 120 ggttatttca ctattaatac ctctatatac tgtggataat tgtataaata tgtcatcttc 30180 ttctagtggg aagtcccctt ttaccagaca tctgtagaca ttggactctt ctccatactt 30240 tttccttaac attttatggt tttctttatc agttctatca acttttgtag cgtcaatatg 30300 aatacattta aaattttcct taagactgtg gtgagaatca aagaagatac ctgtattttt 30360 aacagggtta ccaatcaaaa ttatcttatt ttcctcatcg gatatagtac caagtaatgc 30420 ttctagtatc tcatcatcta ttccaggagc ctcatcacat ataattagca taaactcact 30480 atgaagacca agcatattct cttttctatt actagtttta cacatagcaa accaagtcat 30540 aggactacaa ttcctagtca tagtagtttt atttactgta aagtaatctt ccacaatagt 30600 tccttgtagc cattttgata tttcaggcca taatacagat gttaactgtg aaaatgtagg 30660 agctgtagca attaccttac attcaggcct aaaggtcata taccaaatta taagacaagc 30720 acaggccatg gtcttaccta ccccctgtcc tgaccttaca gaggttctct tattgtctct 30780 tacactcaat aaaatatctt tctgatgtgg atctaagtgt actcctagta tatcttccga 30840 gaatccacaa ggatcgtcat agtatagatc tactaattct tcaaaggata tttcttccag 30900 agcttccttg ttaatgtgga acttatcaac taaagcatct agctctaatt tctcaattat 30960 gctatctaaa tcaaacaatt atatacacct ccttagtaaa gtgctttatt tagtcctgta 31020 ataccttttt acttagtggt attagatttt cctttacctg cattcttttt ctttctatca 31080 gccattttgg ctctaagaac ctcaagatct tttcccttac cagcattatt ctgattttcc 31140 tgtatttgta aaagtaattt aaccttttca agttcaactt tcatcacatc agtacttaag 31200 ccgaagtata cctcaagtct ttttagtgcc tgaagtctat cctccatttt aactgatacg 31260 ccatctttac tttgtttaat ctcattgatt aaagtaccat ctacagcctt actagacctt 31320 aatagaacta gatcatcctt gaattggcac acatcagtta tatctgcaaa ggctattttc 31380 atgtactgcc ttattacatc cataccatcg catagtaatt cttcatctct atcggctctg 31440 atagccttta gagcttcctg tattttcttc ttctttaata gtttagaccc atgtgcatga 31500 gaattactca caggataacc tgctctagca gctgctgtag ttgcattaaa gcacttcatg 31560 taatgaaata caaacaactc ttctttctca tcaagatcat acttatcagc cagggtctta 31620 atagtctttt ttgttttgct actcaatgcc acaatatcac cttcttcttc taaaaatgtt 31680 atgccctgct ctttaaactc tcttagacac tgcctaacaa aggtctgtga taccttagca 31740 aggtttctta gttgtttagg atgtgtaaag ggatcagcct tcagggcatt atatatcaat 31800 tttttggctt tcttcaaatg ttcaggagta cagtactctg gttcagtgta tgacttatag 31860 ctcataatac aatatcaccc ccttaaaatc cagactttac tacgtgaaaa atggttggca 31920 cactaacctt aatgctagta ataccaacca aatggcaaaa ttaactatgg atttatacca 31980 ctattatacc acatctggat taattatgtc aacgttttat aattttattt cacctgtcaa 32040 tattttttca tacatatcca catctattct tccttcctgt agcaggttat cagctgcaat 32100 ttgtatctct tctggggaat atactggatt acctttagta tcaagttcta taccagtata 32160 tgagtctacc ttacctctgt tcataccata tccaactgat ttatgaactt ctagtttttc 32220 attatggtct tcttgccagt gttttaattc ttcctgatat tccttattct cctctttagt 32280 ttgtttacat tcaataggtc tattatcttt catcttttgc tttagagctg ctgtattagg 32340 tttacagttc ttagatcctc cacctgaacc tctattctta tcttcctcat aagctttatt 32400 aatctcaatc ccaattatta gatccatctt ctctaccaag taaagacatt cttttactat 32460 tggattgtta ggttgatttt tagaaatgat gtcttgacac tctttcttta attgatgtgg 32520 attattacat tggctagggc atttttctcc tggcccatat ggaattgtat atgctaagca 32580 gttacctgct ggatcccttc tataacatct tcttttacat actattttac tcatggttaa 32640 tacctcctaa tatattttct ttctcttcta ataatttttc aagttcctca agattttctt 32700 ctttatcact attaccagct ttagctatat ttagaatagc ttctagttgt tcctcactca 32760 aaaactcctt taatttctca tcctctaata tattagatac ctctttactc atattgttct 32820 acctccacat cattacaaat taatttagct attccctcac ttaatctgta tttctcaaga 32880 tataccactt tcttaattcc tgcattgatt attaaattag cacagctctc acatggggat 32940 aaggttacat acatagtagt tccttcagtt gagatcccat gtttagcaca gtaacttatt 33000 aagccagctt ctgcatggat cacaatatca catccaggac ctaaacaagt tttacaatgt 33060 tctactcctg aaggtactcc attatatgaa gtagcaacta tcctgttatc tctaactaat 33120 acacaaccaa ctgcttttct taggcaagta gatcttcctg tcataagttt agcaacttcc 33180 atgtaaagtg tttctctgct tattctgttg ttattatttt ccatagttta tcacctcaat 33240 tttattatat actattccct atcatatgtc aagagaaact ttagaaaggt atatcaccat 33300 aatccagatc aatacattgg gctggctttg gttttacttt tttaggctct ggagttttct 33360 tgactttatg atactttgga gctctggcta aatccttttt taactttctg actctcttca 33420 tatcccactc atgtatttca tggtctctaa aatattcatt caatactctt actgcggatc 33480 tctttgaagg tgtctttaca gagcagttat taggagcaag acagcatcct gtctccttat 33540 ctgatatagt ataacatact gtattatgag gggctcctac aggcctatga atgaataagg 33600 ttctgcccat ataatcaaac tcacataata cttccacata tcttttatac tgttgtcctt 33660 ggtcattctt acataataca taagtatatt ctttcatgtt aattctcctt tctatgttaa 33720 aagggagatc atctctcccc ttaatatctt tttattcttc tgttgttctc atcaacctca 33780 atcacttcat accttctaac cttatcagat ataccttcct cctgtaatac tttaggaata 33840 tatcctatgg cctgtctttt gttatcaact tcaaaatatc tggtcaagat cctttttgat 33900 gttacaatca ctaatcttac tttcatatta tctacctcca tgtggcctga actacagctc 33960 caggtctatg cttagcttca atcttagctt cttcctcagt atcaaaatag attatatctt 34020 ccttgtgttc atccttaaca gatacccagt gtactcctaa atatttgtta tgtgctaata 34080 cactaacatc agggttccaa tccacatcaa ctgtaacttg taaaacgata tatctccttt 34140 tcatatcagt tctccttttt gtatttattt gttgatttta ttataagaga gcctcggaag 34200 tttgtcaaag attttctcga aggaattcgt ataatatctt ccgagctgag cataatatag 34260 actataaaac aaaatcaagg agttgtaata aatgaaagat aataatttaa taccattaag 34320 agatattgcc aaagaggttg gaataaatga aaggaaagcc agaaagctat taagatcaca 34380 tgagaataaa gcaaatcaca attattattg gatgtttaca ccaagagaga aaagacatat 34440 tgtatcagtt ctaaagagac tgcaataggt tttctaaaaa taagggaggg attaaccctc 34500 ctttttataa ttttttgagt tctggaaaaa tccttaatta gttcaagttt cgtaaagttc 34560 tcttgatctt ttccattctc ttcttaactt ggatagcttc ttcttcgcaa ggagcatggc 34620 cctctaggaa ttggtgccat cttctttcaa gttctctgta ttcagattga agttccatga 34680 actcaagtga atgattgtcc atactatcac ctcctcaaca ctattataag tgatactgtc 34740 atattctgtc aaataaaaaa gagagaatgt tattctcccc atcttaatat ctttatagtt 34800 gctccatcaa ctcccttgcc atccacatca tggacatact tatcaagtct cattgttgag 34860 tcctgtataa ttcttttgtc ttctctgtca aaaaccttaa tgacatcagg gttacccctg 34920 tcaatgatgt aagcaatctc ttgagttcca tctgcatatg ttacttgaag taatgaatgt 34980 agaggaataa agtctggagc tgcccaatca ccaatcttaa gatcccctcc tatactattc 35040 ttggtctcat atcctgccac atctgtaggc tcatctgatg agtaataagt aacttcaact 35100 ttgaactcca ttcctttaaa tgaatttatc atctcaatta gattttgttt atcagttctc 35160 agtttatcat tctgatattt taatccttct acattgtcca tcagtatttc agtctgctca 35220 gttaattgtt tattcgactc aatatattct tggacagcat ttccccaatg attgtttagg 35280 ctctgtatgg ttaacagtag ccctcctatc acaattattg ttagtaatga cataattctt 35340 ttaatcattc tcttttaact cctttatttt taatttgaat ttctcaattc cagatttata 35400 cttacccaca atatcaaagg tatcggttag tattgcatag gtttcctcat cagaattaaa 35460 tctgaagctt cctctcttga tgtctcctag cttattaaag gtcttaccct ccttgtccag 35520 atatccttta tagtccatat agcacagaca agcagtaaca aactcctgct tattcctgaa 35580 attatgcttg atacccatac catctaccca tttacttaat acaaatagac tttcattagt 35640 cttctttact ttactattct ggattagatc cttccacaca taatcattag cctctttaaa 35700 tgacttaaat ctcttgcctt tcagtatctt actactctgg atcttggctt tataatcctc 35760 aaatatactt ggccagaagt cttttagata taaatacaag gaaatatcat ctaatttagc 35820 attcggatcg ttaatctgga ttacaggatg accatccatt gctctgatat tattccttag 35880 ttgggtattc tctctcttca ttttagatac ttcaatatat aatttaacga atgcgtcaga 35940 caattcatga ttggttaatt ccctttttag gggcttacca tctttacttt ctttagtagt 36000 tacaatatta aagtagtctt gtaacctatc ccagttattc ttcattagat ccacctccta 36060 tgcttacatt atacatatat tgtactgata tgtcaaagtc caggcattaa ctgatatctg 36 120 atatgcctat atttttaccc catatcagtc tgatatccca catatcagtt gaaactacaa 36180 aatcacaaag gcattggtat cactggcgta gagggatatc agttatgccc cctttatata 36240 tacatagtat tatatatact atataataat atattattat atatagtata tatatattac 36300 atggtgatta catcaatggc aggctattgc cttgcctcca ttatcattac ggcaggcata 36360 tccatgcttc cgagcttttg ccctatatag aaaaacccag acggtcagcg atcgagctct 36420 tgattttgtt ttgtcgaggg agagaacagt aagagagaac ccccgaacgt tgacagttgt 36 480 acttacttct cttataatga cagtaaagga gggataacca tgaaaacatt atttactgat 36540 acaaaactag tattaattga aactactaac ttcttcatta tgttacaaga agaaggtaag 36600 tatgtactat ctaaaacaat cgataaggag atgggagaac aaatacacag agtatctaag 36660 ttagctgccc ttgtatatac taatgaggag tatactaact ctaatgaacc aagatctata 36720 tacttagatg acaatttctt atacgaagtg cttggtaatg ggaagtatat tataagatac 36780 agtatataag agcccacaat caattctaag gccttttaga tatgtggtaa tgtgatttta 36840 cttataatgt ggtagagaca ccttacagga gctgctagag gcctttaaag gaggtatatc 36900 atgtatttat gtaatggtaa aatagtcaag aataaaaatg acttaaaaga taagtttggt 36960 ggtaaatcag tagtattcat cgatggggat gtagctacag taatggactt tagtgacaga 37020 aagccaaaga taacaaaata tcttattagg gaggtatcac atgataaaca catcattatt 37080 gaagagactg gaagggatag aggaaaagca taagcaaaat actgatgaga taagaaagaa 37140 cctaaactat gccagatatt tgattgagaa acagtattgt catttaacaa aagaacagaa 37200 gataaaattg ggacagttct taggattgac tgagaagaag ttaaaagaaa tacaagagat 37260 aaataatcag ttaataaatt tgagagtggg gctaagttat gaagtcgtat gtaataacca 37320 gaaaaaatag tggggtatct ttagcagata cagacatggg acaagtatta gctaagatac 37380 tatgtaagca tgggggaaca cttaccattg aggaaattga accagaacat attggtcaaa 37440 ctttttctat tgtagaaggt aaaagaatat ggtataatag aaaaggaggt gataatattg 37500 gggagaagaa gtgctatgtt aattctcaag tatgactgcg aagaaactcc agaaaggatt 37560 gaattacgaa agaagtggtt acagatttgt cttcattcaa taatatatta tagatatgac 37620 aacaatatct ggacagacca acaatgggat gagactgcca gagaggtagt taaacttaag 37680 aatgagaatc caggtttagt aaacagtatg ccatttagga atgaccttaa agtctttgat 37740 ggatccactg gctttaacct ttcatgtatg caagatatta agatgttaag gtgggctcaa 37800 aacttacttg actttcatgt taaacgagga cttgatggaa agagtaagaa aaagaagaag 37860 aagaggagga aaaagaaatg aacctacttg agcactatat aaaaaaggta cacagtgtaa 37920 gagagtacca tgaatttgat aatgaacctt gggctaaagg taaacaatat gtagaagtag 37980 acatggacta tatttgttat tctaatacac cacaaagagt gaagatggta tttactgttg 38040 atgagtggga gaagataaaa tatagaggct actacatggg ataggaggaa tagttatgat 38100 aagagaaatg ggcatgagaa tgctaatacc agaaatacat gatataaaag atagggatat 38160 acaaaatttt gtaagaaagg ccctaaatga gatagttgat gaaaagttct ttataatacc 38220 tgcaagtagt actgggaaat atcatccaac ttatagtgca ggggtaggag gtttgttaag 38280 acatactaaa gctgcttgtt atataggtaa agcattatgc gaagctgaga tgatatgttc 38340 aagggacaga gatatgataa tagctgcttt aatattacat gacattaata agccagataa 38400 agaacatcca tatatggtta gagagacact atcaccccta gaggaagagt ttccaataat 38460 atacaaagtg gtagttgaac taattgagtc tcatcatggt caatggggta gtgttcctat 38520 taatagttat tcccagagaa tagtacactt ggctgattat atcagttcac agaaaggttt 38580 aatattctca tatgaccaag gaactgatta ttctgacctg ttatgggagg agaagtaaat 38640 ggtgctaggg gtaataaccc tagtatcaat atttttacta ggagtgttta taggttttga 38700 ggtgaagaaa tggaactagg ttttgcgata ctagctataa tagtatcaat atttttagaa 38760 actaaatata taaaggagta gattattatg tcaagagcaa gagctgcaag gagaagagcc 38820 gaaagagaga acaaaaatct taatgttctt aaagctgtgg acatcatgtt agccttatca 38880 tgctatacat taaagaagga aggttatggt aagactagaa tgactaggtt tgtggaaggg 38940 atgagtaaac acacagagga gattgaaaag ggaacactta actatgaaac cataatggga 39000 gagattaagg atttagtacc accaggtata tttgatttag gggaggagtc aaaggagtga 39060 tagtattggg atatttatta tgggcattta gaatattagg catacttgta atagtatggt 39120 gtataatgga ggcgatcagt tgtgagcaat aagttatatt ttaggtatag tgctatgaac 39180 tctggaaaga ctactcagtt aatccaggta gcacataact atgaagaaag gggaatgaag 39240 ccattagtag ttaagcccgg aattgatact aaaggtggac catgtataat cagtagaata 39300 ggggtagcaa gaaaggtgga tgttcttcta cctcctagag ttaaattatc aaaaatactt 39360 tctgactttg agtctaatta ttgggcagta gatgttatat taattgatga ggtacaattc 39420 ttatccagag aacaggttga tgatctgtta aatttatcat tacattatcc tattatatgt 39 480 tatggattga gaactgactt taaaagggaa ggatttgaag gaagtaccag attattacaa 39540 gtggcccata atattgaaga attaaagaac atttgtcaat gtggtaaaaa agccacattc 39600 tcaatattaa aataccaggg taaatatact gaccagggta accagataca aattgataac 39660 cagcccgaaa gtgttgaata tgaagccgtt tgtaatgaat gttataataa attaattttt 39720 tctaaataac tcgttgacag gctatatagg atatgatatt atagtattgt aatcaaatta 39780 caaacaccaa ccaaaacttt ataattagtt aatttcaaac tatactaaat ttgttattta 39840 aaaattaaaa atatggagga atgattatta tggcaaaagg aaaagcaaaa tggttagagg 39900 aagcaaacaa taaggagcta attgtaatcg tacaatctgc tttaaattct ggagatgaga 39960 aagaagttaa aaaggcttta aaagctattg tggaattaga tgaaagaact cctgacacag 40020 atggacaaat aactgtagag gaagcattag aatcattaga aatggctgta ggaactgtaa 40080 aggctgcata ccaagcagaa atagatgctg tggaagatga aggtgtagta gatgctgact 40140 tcgaagaagt tgatgaagac gaagcagaag aagaagatga aaatgattta gaatcattat 40200 ctaaaaggga attagctgct atggctaaag acttaggtat caaaggagct aagaaaaaag 40260 ataaagatga gttaatcgct ttaatcaaag aagctcaagg tgaagaagaa gacgaagagg 40320 aagaagaaga ggatgaaact cctgactact ctgaaatgac taagaaagac ttaattgctc 40380 ttgctaaaga aagaggaata aaagttaata agaaaatgaa accagctgag ataattgaat 40440 tattagaagc tgatgatgaa gaataattct catcaaataa taaggggagc aaacttgctc 40500 ctcaaatttt ttacttattt tagaaacaga tgttagaggg ggatgtataa atggcaagag 40560 gtggaaattc aacttatact aaagcagtat attgttacaa caataatagg gagtatagat 40620 ccttcagata tgcagctaaa cagttaggag tatcggctgt tagtataatg agagtagtta 40680 acgggactaa accacatata gggggattag tattcgctga gatagaggaa ggcactagaa 40740 aattaaagcc aagaggagaa gtagttccat acttgaaaga attaggttgg gataatatag 40800 acttataata gggggtataa tatatggaca taaagttctt gaaggatata ttctctaaac 40860 aatgtgaaaa tggagattac attatcctag cagctagaaa aggtaaagaa tggaaagatg 40920 taccaatcaa atataataag aataacattg acaaaaaact aaaggacttt gaacaacagt 40980 ataagggtta tgatttatat tggagcccaa tgccctatag taatcctcaa agaaggatag 41040 ttaacttcat agagactaaa tacttaatac aagatataga tgagcatact gacccgttgg 41100 ggattaaacc taaaccaagt tatctgtggg aaagttctcc tggaaaatat caggggctat 41160 gggaaatgga taggtatata gaggctaatc aatatgacga aataaatcca gcattagcta 41220 aacatatagg ctgtgactcc tgctttgatg ttactcatgt atatagaata ccaggaacta 41280 ttaactataa atacaagaat aaaccaaaag taaaaagacc tatacacact aaggagatat 41340 ataagcctaa ggtgatcgct aaggctgtta aggctgtaag taaatctaat gatagtgtta 41400 aggtcaatat aggaggatct gaggcctcac aatctgaaag aaagatatat gctaaatata 41460 atataccaaa gaaggttaga gacttactgg ctttagatga cattacttct ttagatagaa 41520 gttctactat atggtatatt gagaataaat tacatgaaat aggactagag cctaatgaga 41580 ttatattatt agttaagggc tcagctttta ataaatataa gggaagaaaa gatgaagaaa 41640 caaggttaag aaaagaattg gataaaatca taggaggaga aatagaggct gatattgaaa 41700 aggctgaaag tactaaactg agaatagata gttatcagga tgttatgggt aataatggtg 41760 ccttcccagg ttggttagta caaggtttct ggggtagaag atctcatgga attgtggctg 41820 gacaaccaaa ggtatttaaa tccacattta cacaggactt agctatatca gttgctagtg 41880 gaagaccctt cctaggtcaa tatcctgttc tagaaccagg cccagtaatt gtagttcaaa 41940 atgagaatgc tgactggatt atgagggata gaactgaaaa gataattagc cacagaggag 42000 tagttggtaa tgtggatata aagggtaaga gaagacttaa agttaggttt gctccagatc 42060 ttcctatcac ttttattaat caacagggat ttatgttaga tgaagaatcc catagaaaac 42120 agatagaaga attaattgat gagataaaac ctgtactagt aatattcgac ccattatatc 42180 ttatgtttag tggagatctt aataatgcag ctgatcttaa tcctgtacta caatggtgtc 42240 ttaaacttaa gaatgagaag catacaggag ttatgttaat acaccactat aataaaggtg 42300 gaaatgccac tcaaaccaga ggtggtcaaa agatggctgg ttcattcata ttacatggtt 42360 gggtagaatc agcattatat ttaaaaagac ctgatgactt agaaggtgac gatgaggaga 42420 ttgaagtaga tatagataac cttgataaac aaagccattt accaagtaaa atcattatgg 42480 atagagagtt ccgtcttgca ggacaattcc ctcaaattga attgaattta tcaatgggag 42540 aatttggaga tccatattat catgtagaag tagcaatacc cggaaaagaa gtaattgtga 42600 aacctgagga caaagccaaa gttatagagg ctgtaaaatc aggagctcat accaaggagg 42660 agatagtgtc aatatcagga ttaaattcac agaaggttaa tctagttcta gatactttaa 42720 aagatactat tgtatattct ccggataaag gttataatat aagtaagaaa ctaaatattg 42780 ggaggaaaaa ggcatgataa taaagtgtaa gaggccagca ggtcataatc atataaaggc 42840 ctttgtagta gtaattattg ctagaggtaa gaagaataaa atcacttatc attcaggatt 42900 gaaatatttc tatacctata aatcagctaa attatttata gacaaagtta tggaagaatg 42960 tccctgggct gaattttatt gggtggatgt ccatgaagcc cataagatcc ctgaaggcga 43020 agagatacct aagaagaaaa tgtggtgtcc atattgtcaa actattcaag aatttaagag 43080 aagccaagga ggttacaaga attgccctat ctgtggtata agtgaccaag acttttatgt 43140 aaaatgtatg aatataaaat atagggaggc taaggctaat gcgaagaaga aacttgaagg 43200 aagtaatact aatcctaggg atatcattgt caatggggat aatgctaact cagggggcaa 43260 aactaataga aagtcatcac caaagaaaaa gtcaagaaaa gttagtagaa aagcttcata 43320 atgctttaga ctataatgtt attgagactt ctccatctgg actagatggg gaggatctta 43380 ttatcaaact aaaccttaat agtcctaaag accttacccc taacaactta tctaagttag 43440 ctaatgaggt tatggtcaat aacttaggag ctcagaatta tactatactt gtttatgata 43500 gtaatagacc agaaaatctt ctggctgtta tagatatacc taaaaattct aaaataacct 43560 ttgacataat aaattaatta tggtaatata aaaatgtaga aggaggagaa tttaatggtc 43620 aataaaaagt attacatctt gaaagacctt aatgatgaca aaccccaact atggaaggga 43680 agtttaacca ctgctactaa gatagcagga gcattaggag ttagtgtgat aaaactaaat 43740 aggtatttag taagaaaggg atttgccaga aggtacaatg agcatacact tgagataata 43800 gacccaatcc ttgctaaaga acttggttat tatgtaactc caagtaagtc ccttataaat 43860 cctttatttg aatataatga taaaggagta caatatatac taaacttaat agtagaagat 43920 ctaaagaatg gaggagagct ttaatgatta tcattttagg aatggataac tcaggtaaga 43980 ccacaacagc taagaaccta actaaacaca gaggaggaga atatatccaa tcaatgggac 44040 ctggttctta tgaagaacaa agagaatggg tattaactca aattgtgaga aaaggtgaaa 44100 gggaagctat acatgataga tttacttgct ttgaagaaat ggtgtatgga cctatcatca 44160 gagatagtag taatttcaac cttgacagta aggagctaaa gatcttaaaa catctatgta 44220 agccaactat tgtatatgct agaccaccta gggaagtgat cttcaatttt ggtgatagag 44280 aacagatgcc aggagtaatt gaaagagctg aacatctact ggctagatat gatgaactga 44340 tatggaagtt attctgtgat ggatggaatg tactggtata tgattatact acttccaatg 44400 tggagaagct atcccagtta atagatgact caagtgctaa agaagctata aaccaattta 44460 taaatatatc aagataagag gagagattgt tatgaatata aaccatgcag tagaagaaaa 44520 aatagaagga gatagattac aagctatctt taatagacaa aaggagttaa tggagaaata 44580 tcatcatata gaagctagat caggattatg ccaaactgaa gactgtccag ttaacttaaa 44640 tgataaaaga ggtcaagcta gacttaaaga ttttgcttgg agaatgactg aagaagtagg 44700 agaagcattg gatgcttata atcatgaaga ccattaccaa gaagaattaa ttgatggatt 44760 acacttctta actgaattta caattctagc tggtaaggat tataatacta tagataataa 44820 tgctatatta gttgatagcc tagagaactt atatcataag gctactacat caccagaatt 44880 tcctgaaatg ttagtagaag aagctgtaac tgacctggta agagaaatgg gtatgtgctg 44940 taactgcctt aagaataaac cttggaaaca aacttcaatg ctaacagatg taaatgcttt 45000 taatcaaaga ttattcaatg tatgggtatg ttacattaaa ttattagctg tatctggctt 45060 agaggtagat gacatagtaa atatatacct taagaaatca caggtaaata aattccgcca 45120 aagatctaat tactaaaata aatacttagg agaaggtaat atgctaataa gagactataa 45180 tgattttgat gacttattct taaaccttaa tagggaaatg attaccaatc cagaagaaac 45240 tatgatgtat actcagaaca tacagggctt tcaggaggac ttagttctct cctgtaagtc 45300 ccataaatgt actttaaatc taggggactt tggatataaa gaaggtaagt ggggacacct 45360 attaagatca tacattgatt atccacaatt aattgaattt agagaaaagc ttaccaagat 45420 aagtggtatg agctatactt attatttcaa caggaaaaaa gctactaatg gttcttgctt 45 480 gatagctgca gtagtaacta gaccaaaaag aaaaggacct tggaaacact taaaaattat 45540 gtatagagta tgtgaattac aaaagaaatt cgcagctgat ctagtattaa taaacaggtt 45600 cattgaagaa ctacctcaag aagtatgtga gatagataat attacttttc atatgtctca 45660 agcttatttg tcaggaatgt ttataaatgg atatttcaat tattttaagg taccaagaaa 45720 gagtattgct aatagcaaac atccttggca taaatccttg aatagtaact ataacaggtt 45780 ctttaaatca gaggaccaga tacactctta taaagctctc cagaagatgc agttattaca 45840 ttttggtctg gagaaatttc caaagataga tatcaataaa ttatctattg acaagtactt 45900 taataagtag tataattaac ttaacaaaat taaataaatg gaggaattaa actatgagaa 45960 tttatattaa tgctcaagaa gcatttgaag aagtaaaaag agatttatgt gaaatgggga 46020 ttgaggtaag acccaaaact atgcaagata aagttataga aggtaatcca gattatttca 46080 caaaggagtt acaaaactat agttacacaa ttctagaaag taaacctgaa gaagttccag 46140 gtgtatctca accttgggca gatgctgaat ttagagaaag gatctacgat ccacaaggtg 46200 tcatcaatca atactctctt gaagaaagag aagaattatt tggtatacat ccacatcata 46260 ctagggaggc tttcataaat cccggtaaag cttatcaatt aagacctgag gtatggaatg 46320 aatatctaag agatggaaaa tttgggtatt cttacaatga aagaatttgg caatacagac 46380 aaattgagga tatcatcaat agaattaaag aagatccagg ctcaagacaa ttatggttat 46440 ctctgtggaa tccagctatt gatccattca atataggagg agtaactaga gtaccatgtt 46500 cattaggata taacttccaa gtaagagaag gtaagctaaa tatccattat gttatgagaa 46560 gttctgattt tgctactcac tttgctaatg atgtttatct tgctatgaaa ctattacatt 46620 gggtagctga acaaactggc tatgaaccag gaagtttctc tcatacaatt ttctctcttc 46680 atgtttacaa caaagatatt aagggggtat tctaataaat gagttacatg agaaaagaaa 46740 gatataaaca ttctgatctt aataataagg ttaacaagaa ggaaacccta aagcaattta 46800 ttaggaatac tgaaaaggaa tttgatttag aaccagctaa ccttggggac atgactagga 46860 atgaacttaa taactatata aattatatgg atgaactgtg gagtaagtaa tatgtataat 46920 tatagtgatg ataacagacc taagaacaat aacaatggct gtcttggttg tttaattata 46980 atcttagcag ctataggatt atgggtaata atttttgata ttgccaatat aatatatcat 47040 atgatattct aaatttaaag ggggctatcc taatgaaaaa tataatttgt cctatgtgta 47100 aaaactcagt aacattaaaa agaaaatatg gagctatgtt ttggatcatg attttcttaa 47160 ctggaggcct atgggtagta accataccct tcaaaaaaca taaggtctgt ccagtatgta 47220 actcaatcat aaaataaaga taagggcctt taggggtcct ttctatttag gaggtaatat 47280 tttaaatgag atgtaacaac tgtaacctat atactcactc agctccctca tgtatagaag 47340 gaacttgtgt aggaaagaaa aagaagccaa gaataatggt cataaatagt ttagctaatg 47400 atagggatga ggccaataga atagctactc ctgataaatc attacttgat aaaatggaag 47460 gtctagactt ttattatacc aatgctatta agtgtagaac tcctaagggt actaaaatca 47520 aagtatcaga gattaagaaa tgccaagaac atttacttaa ggaaattgaa aaatataaac 47580 ccgaatatgt tatgatctta ggatctcaag cattaaagat gctaagtaat gaaggtataa 47640 cttcaatatg tggagtacct aagaagcatg agaaatacgg ctttaagttt attgccagct 47700 attcacccgg tgtagttgca tatgacccaa ctaaagcaca atttgtagat caggccttta 47760 ataactttaa agccatggta aaaggtaaag agcatgaatt acctgatctt aacataaaac 47820 ttattactag tatgaaagaa ctaaaccagg ctttcaaata tttaagggat gagggttata 47880 atagagtatc atatgatata gaaactagag gcttagatag gtttaataat aacattactc 47940 tatttggttt tggtaatacc caagtacagt atatactgcc tttagaagtc aaatatagcc 48000 cattaagggg taagcccata gcacaaagaa gattagctaa atccttgatt aaaagactta 48060 attctgagat gaaggaaaga atagcccaga atggtaagtt tgatgataat ttcttaaagg 48120 agaaatatgg tattaagcca atcataacct ttgatacttt actagcctca cattgtttag 48180 atgagaatac tcctaatggt cttaaggaga atgctttatt acattgtaat gctaaagact 48240 gggatataaa taagaaatta aagactggaa atgtggagac taaatcagac tttgaggatt 48300 atgttaggta tctggggtat gatatatact atacatttgc tctatataaa atatttaata 48360 aaagacttaa aagggatgag agcttatata aactattcca ccacttatac atcccagcta 48420 gtaaagctta tgaggatgtt cagttcaagg gtatatatgt aaatcaggag aaattcaaag 48480 aggttgaaaa atacttaaga tctgaactag ataagattga gactggactt aagaagtata 48540 ctaatggcca ggatattaac tggagctcac ctaaacaggt tggagaattt ttatatgata 48600 cacttggtct tcctgtgatt gaggttactg actctggagc accagctact ggagaaagtg 48660 tattattaag actaagagat aaacatccag cagtagaact actattacag cataggggag 48720 ttcatataca aatttcccac tttatagatg gttggctaaa taggatgcac aatcatagat 48780 tatatccaaa ctttaaactt catggaactg taacaggaag aacctcaagt aataatccta 48840 atctacagca agttcctaga gataagaaaa tcaggagttt attaggacca tctcctggaa 48900 gagtattcat tgaagccgat ctatcccagg ccgaacttag aatagctgct atgatggccg 48960 atgaggataa tatgaagttt atttaccaga ctggtggaga tatacatgac tccacatata 49020 atattatatc tggggaagat atcaatgatg agaaagatcc agcagttaag aaggagaaaa 49080 ggaaaaaggc taaggctgta aactttggtt tcttatatgg tatgcaatgg aaaaaattca 49140 aggattatgg aagagacaac tatggtctta aattaacaga tgaggaagcc aaaacatata 49200 gaaggaactt ctttaataaa taccctaaac tattaacatg gcatgataaa caaagaaaaa 49260 ttgttaaagc caatggtgaa gtaagatctc taataggaag aattagaaga ttacccgata 49320 tatattcatc tgatagatct aaagctgctg aagccgaaag acaatgtatt aactcaccag 49380 ttcaaggttt tggttcagat atcactttat taggcctatg tgagattaca ggctatgcta 49440 aatatgttaa tcctgaatat gtattagata agtctaagtt tgatgtatta ggctcagtac 49500 atgactcaat attatttgaa gtagataaag actatgtgga agaattagct tggaaagtaa 49560 aatcaatagt tgaaaataat aaagtattaa agaaagtatt taaatttacc cccacagtcc 49620 caataatcat ggatatatca gttggttatt cttggggagg atgtgttgaa ctggatttta 49680 aaggtgattg gaaaagccag ataagaaaag tgttgacaga tgaataatta tctgataata 49740 taaatattgt aatgaaaagg aggtataagg atgttaaaaa taagtaattc cagaattaac 49800 aaattcttgt cttgtcctta tgcccattat gttaaatact atgaaggtct ggtacctaaa 49860 agaagtggag ctgccttaca aaggggctct gctatccacc aggctataga agactaccat 49920 aatgggaaga gttggaaaaa atctgttgat aaattttcca aagagtttta caaaaataca 49980 tttaaagaag agatccttga atttggagat attccaaaaa tggtttattc tttatgtgat 50040 aactatttcc actattatga tgaaaaagaa gataatgtaa cctatgtgga aaatgaacat 50 100 cacttcgaat taaaactatg taagggtgta actctagaag gctatattga tagtgtctta 50160 gatgtggatg gaaagatatg ggctaaggaa actaaaacct acaaaaagat gcctgataga 50220 aatttcctga tcttcaatag acaatctgct atatatacct gggctcttct acatgaatac 50280 ccaaaagtaa gtggtactat atgggatata atattagctc agcaaccagg tagaccagaa 50340 ttaactcaaa aaggggtatt atctcaaaag aggattaaat ccacacccct agagttagaa 50400 agaggaataa gagaattagg attagatcct aaagattatg agtcttatat taattctgct 50460 agatgggaag acttctttgt aagacaccca ataatattat caaagaatat acttaacagt 50520 gtaatgattg atactattga gatagctaaa cttattagag atgagggcca caaaagaaag 50580 gaaaagaacc ttggaaaagg ttgctctttc tgtgaatata aatctttatg ccaagctgaa 50640 cttttaaatc ttgataaaga atttattatt aaggctgact acaaacaaag ggaggaaagt 50700 gacaatggca aaaaagcaaa aatcaaaatc aaatagtttt gaggacagat tagtggatct 50760 atatgatata gatgagccca caatactaac actttatgga agatctggtt caggtaaaac 50820 tactatctca ggaacactac ccaaaccaat attctttatt gatgtaaagg acaaaggtac 50880 tctatcagct agaaacaagc ttagagttaa aagaggagat atacaagtat ttagtctaaa 50940 gagttttgat gacatatacg aggcttatga ttacctatca gaaaacactg ataaatttaa 51000 aacagtagtt atagaccatt taactgcttt acaagaatta ggtaatgaaa aggtcaaagc 51060 tgaagaaggt aaggaccaga tgagccaaag aatgtttgga aatgtggcta attatatgaa 51120 agaggttata aacctttata aagaattaaa tgaggaaggt atactaccat gctttatagt 51180 acaagatagg ttagaatctg gtgatggtga aggagaagac caattaatgc ctgaggtagg 51240 accaggatta atgccatctg tatctaaata tttatgtgct gtatcaagag taataggtca 51300 tacttattta tatgaacact cagaaaaaga gggtatgaag gttaagaaag aaatccagta 51360 tagactaaga ctaggaccta acccttatta tattactaaa tttactagac ctcaaggatc 51420 tgaatgtcca gcttatctgg tacatgattt aaaatcacca acaactatct gggaggatat 51480 tgaaactatt ctggctggtg aatggaataa taagccagct aaatctggta aaaaaaccag 51540 taagaaatct ggtaaaaaga aaaaataaaa ttttctaaaa atacctttga caaccaatta 51600                                                                        51600

Claims (8)

클로스트리디움 퍼프린젠스(Clostridium perfringens)에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 CP5(수탁번호 KFCC11676P).
Bacteriophage CP5 (Accession No. KFCC11676P) consisting of the nucleotide sequence represented by SEQ ID NO: 1 having a killing ability specific to Clostridium perfringens .
삭제delete 제1항의 박테리오파지 CP5를 포함하는, 항생용 조성물.
Antibiotic composition comprising the bacteriophage CP5 of claim 1.
제1항의 박테리오파지 CP5를 포함하는, 사료 첨가용 조성물.
Claim 1, comprising the bacteriophage CP5, feed composition.
제4항의 사료 첨가용 조성물을 포함하는, 사료.
Feed comprising the composition for feed addition of claim 4.
제1항의 박테리오파지 CP5를 포함하는, 소독제.
A disinfectant comprising the bacteriophage CP5 of claim 1.
제1항의 박테리오파지 CP5를 포함하는, 세척제.
A detergent comprising the bacteriophage CP5 of claim 1.
제1항의 박테리오파지 CP5를 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 administering the bacteriophage CP5 of claim 1 to an individual except a human.
KR1020170178596A 2016-12-23 2017-12-22 Novel Clostridium perfringens specific bacteriophage CP5 and antibacterial composition comprising the same KR102016920B1 (en)

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