KR102193477B1 - Novel Salmonella specific bacteriophage ST20 and antibacterial composition comprising the same - Google Patents
Novel Salmonella specific bacteriophage ST20 and antibacterial composition comprising the same Download PDFInfo
- Publication number
- KR102193477B1 KR102193477B1 KR1020190076337A KR20190076337A KR102193477B1 KR 102193477 B1 KR102193477 B1 KR 102193477B1 KR 1020190076337 A KR1020190076337 A KR 1020190076337A KR 20190076337 A KR20190076337 A KR 20190076337A KR 102193477 B1 KR102193477 B1 KR 102193477B1
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- South Korea
- Prior art keywords
- bacteriophage
- salmonella
- present
- bacteria
- feed
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Abstract
Description
본 발명은 살모넬라 균에 특이적인 사멸능을 갖는 신규 박테리오파지 ST20, 상기 박테리오파지를 포함하는 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제 및 상기 박테리오파지를 개체에 투여하는 단계를 포함하는 살모넬라 균에 의한 감염성 질병의 예방 또는 치료 방법에 관한 것이다.The present invention is a novel bacteriophage ST20 having a specific killing ability for Salmonella bacteria, an antibiotic composition containing the bacteriophage, a composition for feed addition, a feed, a disinfectant or detergent, and a Salmonella bacteria comprising the step of administering the bacteriophage to an individual. It relates to a method of preventing or treating infectious diseases caused by.
살모넬라(Salmonella)는 장내세균과에 속하는 그람음성의 통성혐기성 세균으로 아포를 형성하지 않는 간균이다. 살모넬라균은 사람뿐만 아니라 돼지, 소, 닭 등을 비롯한 다양한 가축에 감염되어 질병을 일으킨다. 사람에게는 식중독을 일으키는 원인균이며, 동물에게 있어서는 다양한 살모넬라증(Salmonellosis)을 일으키는 병원성 미생물이다. 살모넬라균은 다양한 혈청형을 가지며 현재 2,600여종 이상의 혈청형이 보고되고 있다. 장티푸스의 원인균인 살모넬라 티피(Salmonella Typhi)나 식중독의 원인균인 살모넬라 파라티피(Salmonella Paratyphi) 등은 강한 병원성을 가지며, 또한 대표적으로 살모넬라 티피뮤리움(Salmonella Typhimurium)과 살모넬라 엔테라이티디스(Salmonella Enteritidis)는 사람뿐 아니라 동물에도 동시에 감염될 수 있는 혈청형으로 알려져 있다. 사람에게 있어서는 식중독을 일으키지만, 닭, 돼지, 소 등에 있어서는 살모넬라증을 유발하여 급성 또는 만성의 감염증으로 발생하며, 감염 시 발열, 장염, 패혈증 등이 발생하고 폐렴, 관절염, 유산 등도 일으킬 수 있다. 돼지에서 살모넬라 티피뮤리움은 매우 감염력이 높아 급속도로 감염이 퍼지며 주로 이유 시기나 4개월령 시기에 감염되며, 흔한 임상증상으로는 설사와 장염이 나타난다. 가금류에서 주로 분리되는 살모넬라 엔테라이티디스는 전 세계적으로 살모넬라 유래 식중독과 관련하여 빈번하게 분리된다. 살모넬라 식중독은 주로 계란, 가금육 또는 돼지고기와 같이 살모넬라균에 오염된 식품을 통해 감염된다. 또한 살모넬라 갈리나룸(Salmonella Gallinarum)과 살모넬라 풀로럼(Salmonella Pullorum)은 각각 양계에 있어서 가금티푸스와 추백리를 일으키며 발생 시 대량 폐사를 유발 할 수 있다. 이러한 살모넬라는 동물에 감염 시 세포 내에 침투하여 증식하고 감염하는 경우가 많아 항생제나 약제, 기타 생균제가 침투하여 작용하기가 어렵다.Salmonella ( Salmonella ) is a gram-negative, painless anaerobic bacterium belonging to the intestinal bacteria family, and is a bacillus that does not form spores. Salmonella infects not only humans but also various livestock including pigs, cows, and chickens, causing diseases. It is the causative agent of food poisoning in humans, and it is a pathogenic microorganism that causes various salmonellosis in animals. Salmonella has various serotypes and more than 2,600 serotypes are currently reported. Salmonella Typhi, the causative agent of typhoid fever, and Salmonella Paratyphi, the causative agent of food poisoning, have strong pathogenic properties, and representatively, Salmonella Typhimurium and Salmonella Enteritidis It is known as a serotype that can infect not only humans but also animals at the same time. In humans, it causes food poisoning, but in chickens, pigs, cows, etc., it causes salmonellosis, resulting in acute or chronic infections, and fever, enteritis, sepsis, etc., may occur during infection, and pneumonia, arthritis, and miscarriage may also occur. Salmonella typhimurium in pigs is highly infectious, so the infection spreads rapidly. It is mainly infected during weaning or 4 months of age, and common clinical symptoms include diarrhea and enteritis. Salmonella enteratidis, which is mainly isolated from poultry, is frequently isolated worldwide in connection with food poisoning derived from Salmonella. Salmonella food poisoning is mainly transmitted through foods contaminated with Salmonella bacteria, such as eggs, poultry or pork. In addition, Salmonella Galina room (Salmonella Gallinarum) and the salmonella pool column (Salmonella Pullorum) may cause mortality in the event of a large amount causes the fowl typhoid and chubaekri in each poultry. Such Salmonella penetrates into cells, proliferates and infects when infected with animals, making it difficult for antibiotics, drugs, and other probiotics to penetrate and act.
산업동물분야에서 항생제는 세균성 질병의 치료나 예방, 성장 촉진을 위해 사용되고 있었으나, 가축에서의 무분별한 항생제 사용과 항생제 내성세균의 증가가 관련이 되어있다는 사실이 알려지며 여러 국가에서 항생제를 성장 촉진제로 사용하는 것을 규제하고 있다. 유럽연합에서는 2006년부터, 우리나라에서는 2011년부터 사료 내 항생제 사용을 금지하였으며, 미국 식품의약국(FDA)은 항생제 오남용을 줄이기 위해 항생제의 성장촉진 효과 선전을 불법화하며 질병 예방 용도의 항생제는 수의사의 처방을 받도록 규정하고 있다. 또한 닭과 돼지에서 분리되는 Salmonella Typhimurium 중 항생제 내성이 발견되며, 2종 이상의 항생제에 대하여 내성을 나타내는 다제 내성 균주가 발견되고 있다. 항생제 내성률은 증가하고 있으며 이 때문에 살모넬라증을 치료하기 점점 더 어려워지는 실정이라 항생제를 대체하기 위한 효과적인 치료제의 개발이 요구된다.In the field of industrial animals, antibiotics have been used to treat, prevent, and promote growth of bacterial diseases, but it is known that the reckless use of antibiotics in livestock and the increase of antibiotic-resistant bacteria are known, and antibiotics are used as growth promoters in various countries. Are regulated. The European Union has banned the use of antibiotics in feed since 2006 and in Korea since 2011. The Food and Drug Administration (FDA) has made it illegal to promote the growth-promoting effect of antibiotics in order to reduce the misuse of antibiotics. It is prescribed to receive a prescription. In addition, antibiotic resistance was found among Salmonella Typhimurium isolated from chickens and pigs, and multidrug resistant strains showing resistance to two or more antibiotics were found. The antibiotic resistance rate is increasing, and because of this, it is becoming more and more difficult to treat salmonellosis. Therefore, the development of effective therapeutic agents to replace antibiotics is required.
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 박테리오파지의 활용이 크게 주목을 받고 있다. 자연 친화적 방식의 선호로 인하여 박테리오파지에 대한 관심은 어느 때보다 높다고 할 수 있다. 기존 항생제의 대체 물질로의 개발 가능성이 부각되면서 다시 박테리오파지가 항-박테리아제로 주목을 받고 있다.On the other hand, bacteriophage refers to a bacteria-specific virus that inhibits and inhibits the growth of infected bacteria by infecting specific bacteria. Bacteriophages have the ability to kill bacteria by proliferating inside bacterial cells after infection with bacteria, and destroying the cell walls of host bacteria when progeny bacteriophages come out of the bacteria after proliferation. The bacterial infection method of bacteriophage is very specific, so the types of bacteriophage that can infect specific bacteria are limited to some. That is, a specific bacteriophage can infect only a specific category of bacteria, and because of this, a specific bacteriophage kills only specific bacteria and does not affect other bacteria. Therefore, the use of bacteriophage as a countermeasure for recent bacterial diseases is receiving great attention. It can be said that interest in bacteriophage is higher than ever due to the preference of nature-friendly methods. As the possibility of development as a substitute for existing antibiotics has emerged, bacteriophage is again attracting attention as an anti-bacterial agent.
이에 본 발명자들은 살모넬라(Salmonella) 균에 특이적인 용균활성을 갖는 박테리오파지를 분리하고 분리된 박테리오파지의 형태적 및 유전적 특성을 분석하여 상기 박테리오파지가 살모넬라 균, 특히 살모넬라 티피뮤리움(Salmonella Typhimurium), 살모넬라 엔테라이티디스(Salmonella Enteritidis), 살모넬라 갈리나룸(Salmonella Gallinarum) 및 살모넬라 풀로럼(Salmonella Pullorum)을 선택적으로 사멸할 수 있음을 확인하였으며, 상기 박테리오파지의 pH 안정성과 온도 안정성을 검증함으로써 본 발명을 완성하였다.The present inventors have Salmonella (Salmonella) specific lysis to remove the bacteriophage has an activity and to analyze the morphological and genetic characteristics of the isolated bacteriophage The bacteriophage is Salmonella, in particular Salmonella typhimurium (Salmonella Typhimurium) on bacteria, Salmonella It was confirmed that it can selectively kill Salmonella Enteritidis, Salmonella Gallinarum, and Salmonella Pullorum, and the present invention was completed by verifying the pH stability and temperature stability of the bacteriophage. .
본 발명의 목적은 살모넬라 균에 특이적인 사멸능을 갖는 박테리오파지 ST20을 제공하는 것이다.An object of the present invention is to provide a bacteriophage ST20 having a specific killing ability for Salmonella bacteria.
본 발명의 또 다른 목적은 상기 박테리오파지 ST20를 포함하는, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제를 제공하는 것이다.Another object of the present invention is to provide an antibiotic composition, a composition for feed addition, a feed, a disinfectant or a cleaning agent, including the bacteriophage ST20.
본 발명의 또 다른 목적은 상기 박테리오파지 ST20를 개체에 투여하는 단계를 포함하는, 살모넬라 균에 의한 감염성 질병의 예방 또는 치료 방법을 제공하는 것이다.Another object of the present invention is to provide a method of preventing or treating infectious diseases caused by Salmonella, comprising administering the bacteriophage ST20 to an individual.
상기 목적을 달성하기 위하여, 본 발명은 살모넬라 균에 특이적인 사멸능을 갖는 박테리오파지 ST20 (기탁번호 KFCC11828P)을 제공한다.In order to achieve the above object, the present invention provides a bacteriophage ST20 (accession number KFCC11828P) having a killing ability specific to Salmonella bacteria.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 항생용 조성물을 제공한다. In addition, the present invention provides a composition for antibiotics, including the bacteriophage ST20.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 사료 첨가용 조성물을 제공한다.In addition, the present invention provides a composition for feed addition, comprising the bacteriophage ST20.
또한, 본 발명은 상기 사료 첨가용 조성물을 포함하는 사료를 제공한다.In addition, the present invention provides a feed comprising the composition for the feed addition.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 소독제를 제공한다.In addition, the present invention provides a disinfectant comprising the bacteriophage ST20.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 세척제를 제공한다.In addition, the present invention provides a cleaning agent comprising the bacteriophage ST20.
또한, 본 발명은 상기 박테리오파지 ST20을 인간을 제외한 개체에 투여하는 단계를 포함하는, 살모넬라 균에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.In addition, the present invention provides a method for preventing or treating infectious diseases caused by Salmonella, comprising administering the bacteriophage ST20 to an individual other than humans.
본 발명의 박테리오파지 ST20은 기존의 항생제 등의 화학물질에 비해 살모넬라 균, 특히 살모넬라 티피뮤리움, 살모넬라 엔테라이티디스, 살모넬라 갈리나룸 및 살모넬라 풀로럼에 대한 특이성이 매우 높으므로, 항생제 내성균 문제를 해소할 수 있다. 또한 본 발명의 박테리오파지 ST20은 물리화학적 자극에 강하다는 장점을 가지고 있으며, 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 ST20은 살모넬라 균에 의한 감염성 질병의 예방 또는 치료, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.The bacteriophage ST20 of the present invention has a very high specificity for Salmonella bacteria, in particular Salmonella typhimurium, Salmonella enteratidis, Salmonella galinarum, and Salmonella fullorum, compared to chemical substances such as conventional antibiotics, so it can solve the problem of antibiotic-resistant bacteria. I can. In addition, the bacteriophage ST20 of the present invention has the advantage of being strong against physicochemical stimulation, and does not infect other hosts other than bacteria such as humans, animals, plants, etc., so the problem of antibiotic-resistant bacteria due to abuse of antibiotics, the problem of residual antibiotics in food, It has the advantage of solving the problem of a wide host range. Therefore, the bacteriophage ST20 of the present invention can be used in various fields in the field of prevention or treatment of infectious diseases caused by Salmonella bacteria, antibiotic compositions, feed addition compositions, feed, disinfectants, or cleaning agents.
도 1은 박테리오파지 ST20의 살모넬라 균에 대한 용균반 형성 활성을 확인한 결과를 나타낸 도이다.
도 2는 박테리오파지 ST20의 유전자 맵을 나타낸 도이다.
도 3은 전자 현미경을 통해 박테리오파지 ST20의 형태학적 특성을 확인한 결과를 나타낸 도이다.
도 4는 박테리오파지 ST20의 단백질 패턴을 SDS-PAGE를 통해 확인한 결과를 나타낸 도이다.
도 5는 박테리오파지 ST20의 살모넬라 티피뮤리움에 대한 사멸능을 확인한 결과를 나타낸 도이다.
도 6은 박테리오파지 ST20의 pH 안정성 실험 결과를 나타낸 도이다.
도 7은 박테리오파지 ST20의 온도 안정성 실험 결과를 나타낸 도이다.1 is a diagram showing the results of confirming the lysis plaque formation activity against Salmonella bacteria of bacteriophage ST20.
2 is a diagram showing a gene map of the bacteriophage ST20.
3 is a diagram showing the result of confirming the morphological characteristics of the bacteriophage ST20 through an electron microscope.
4 is a diagram showing the result of confirming the protein pattern of bacteriophage ST20 through SDS-PAGE.
Figure 5 is a diagram showing the result of confirming the killing ability of the bacteriophage ST20 Salmonella typhimurium.
6 is a diagram showing the results of a pH stability experiment of bacteriophage ST20.
7 is a diagram showing the temperature stability test results of the bacteriophage ST20.
이하, 본 발명에 대해 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 살모넬라 균에 특이적인 사멸능을 갖는 박테리오파지 ST20 (기탁번호 KFCC11828P)을 제공한다.The present invention provides a bacteriophage ST20 (accession number KFCC11828P) having a killing ability specific to Salmonella bacteria.
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 inhibits and inhibits the growth of the bacterium by infecting a specific bacterium, and refers to a virus containing a single or double-stranded DNA or RNA as a genetic material.
본 발명의 박테리오파지 ST20은 살모넬라 균에 대해 용균활성이 탁월하다. 상기 박테리오파지 ST20은 170 nm 내지 190 nm의 크기를 가지는 정이십면체의 머리와 비수축성의 꼬리가 있는 형태인 사이포비리대(Siphoviridae)에 속하는 것을 확인하였다.The bacteriophage ST20 of the present invention has excellent lytic activity against Salmonella bacteria. It was confirmed that the bacteriophage ST20 belongs to Siphoviridae , which is a form having a head of a regular icosahedron having a size of 170 nm to 190 nm and a non-shrinkable tail.
본 발명의 박테리오파지 ST20은 단백질 패턴 분석 결과, 72 kDa, 60 kDa, 38 kDa 그리고 36 kDa 크기의 주요 단백질을 포함하는 것을 확인하였다.As a result of protein pattern analysis, the bacteriophage ST20 of the present invention was confirmed to contain major proteins of 72 kDa, 60 kDa, 38 kDa and 36 kDa.
박테리오파지 ST20은 서열번호 1로 표시되는 염기서열을 전체 유전자의 전체 또는 일부로서 포함할 수 있다. 또한, 본 발명의 박테리오파지 ST20은 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 ST20 may include the nucleotide sequence represented by SEQ ID NO: 1 as all or part of the entire gene. In addition, the bacteriophage ST20 of the present invention may consist 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% or more, preferably 80% or more, more preferably 90% or more, even more preferably 95% of the base sequence represented by SEQ ID NO: 1 as a result of modification or substitution of the base sequence. It has the above sequence homology, and refers to a sequence that exhibits substantially the same physiological activity as the nucleotide sequence represented by SEQ ID NO: 1.
상기와 같은 살모넬라 균 특이적 용균활성 또는 살모넬라 균에 특이적인 사멸능은 본 발명의 박테리오파지 ST20을 살모넬라 균에 의한 감염성 질병의 예방 및 치료용 조성물, 및 상기 박테리오파지 ST20을 유효 성분으로 포함하는 다양한 제품에 적용을 가능하게 한다.Salmonella specific lytic activity or Salmonella specific killing ability as described above, the bacteriophage ST20 of the present invention for the prevention and treatment of infectious diseases caused by Salmonella bacteria, and various products containing the bacteriophage ST20 as an active ingredient. Enables application.
본 발명의 박테리오파지 ST20은 상기 박테리오파지가 존재할 것으로 예상되는 양계농장에서 시료를 채취하여 분리한 박테리오파지로서, 본 발명자는 이를 박테리오파지 ST20으로 명명하고 2019년 04월 12일에 한국미생물보존센터에 수탁번호 KFCC11828P로 기탁하였다. The bacteriophage ST20 of the present invention is a bacteriophage obtained by collecting a sample from a poultry farm where the bacteriophage is expected to exist, and the present inventors named it bacteriophage ST20, and on April 12, 2019, the accession number KFCC11828P Deposited.
본 발명에서 pH 안정성은 다양한 pH 범위에서 박테리오파지 ST20의 역가가 안정적으로 유지되는 것을 의미하며, 본 발명의 박테리오파지 ST20은 산성 내지 염기성의 넓은 pH 범위에서 장기간 동안 안정한 것을 특징으로 할 수 있으며, 바람직하게는 pH 3 내지 11범위에서, 더욱 바람직하게는 pH 4 내지 11범위에서 안정한 것을 특징으로 할 수 있다.In the present invention, pH stability means that the titer of the bacteriophage ST20 is stably maintained in various pH ranges, and the bacteriophage ST20 of the present invention may be characterized by being stable for a long period in a wide pH range of acidic to basic, and preferably It may be characterized in that it is stable in the range of
본 발명에서 온도 안정성은 다양한 온도 범위에서 박테리오파지 ST20의 역가가 안정적으로 유지되는 것을 의미하며, 본 발명의 박테리오파지 ST20은 넓은 온도 범위에서 역가가 장기간 동안 안정한 것을 특징으로 할 수 있으며, 바람직하게는 40~70℃범위에서, 더욱 바람직하게는 40~60℃범위에서 안정한 것을 특징으로 할 수 있다.Temperature stability in the present invention means that the titer of the bacteriophage ST20 is stably maintained in various temperature ranges, and the bacteriophage ST20 of the present invention may be characterized in that the titer is stable for a long time in a wide temperature range, preferably 40 ~ It can be characterized in that it is stable in the range of 70 ℃, more preferably in the range of 40 ~ 60 ℃.
본 발명에서 살모넬라 균 특이적 사멸능은 박테리오파지 ST20이 살모넬라 균 이외의 다른 균을 사멸시키지 않으면서, 특이적으로 살모넬라 균만 사멸시키는 능력을 의미하며, 특히 살모넬라 티피뮤리움(Salmonella Typhimurium), 살모넬라 엔테라이티디스(Salmonella Enteritidis), 살모넬라 갈리나룸(Salmonella Gallinarum) 또는 살모넬라 풀로럼(Salmonella Pullorum)을 특이적으로 사멸시키는 것을 특징으로 할 수 있다.In the present invention, the specific killing ability of Salmonella refers to the ability of the bacteriophage ST20 to specifically kill only Salmonella bacteria without killing other bacteria other than Salmonella, and in particular, Salmonella Typhimurium, Salmonella Enteraity. Diss ( Salmonella Enteritidis), Salmonella Gallinarum ( Salmonella Gallinarum) or Salmonella Pullorum ( Salmonella Pullorum) may be characterized by specifically killing.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 항생용 조성물을 제공한다. In addition, the present invention provides a composition for antibiotics, including the bacteriophage ST20.
본 발명에서, 용어 항생용 조성물은 약제 형태로 동물에게 제공되어 균을 사멸시킬 수 있는 제제를 의미하며, 방부제, 살균제, 항생제 및 항균제를 총칭하는 것이다. In the present invention, the term antibiotic composition refers to a preparation that is provided to an animal in a pharmaceutical form to kill bacteria, and refers to preservatives, fungicides, antibiotics and antibacterial agents.
본 발명의 박테리오파지 ST20은 기존 항생제에 비하여 살모넬라 균에 대한 특이성이 매우 높고, 유익균은 죽이지 않으면서 살모넬라 균만 사멸시킬 수 있다. 아울러, 약물 내성 내지 저항성을 유도하지 않아, 기존의 항생물질에 비하여 제품수명(life cycling)이 긴 신규 항생제로서 이용될 수 있다는 이점이 있다.The bacteriophage ST20 of the present invention has very high specificity for Salmonella compared to conventional antibiotics, and can kill only Salmonella bacteria without killing beneficial bacteria. In addition, since it does not induce drug resistance or resistance, there is an advantage that it can be used as a novel antibiotic having a longer life cycle compared to existing antibiotics.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 사료 첨가용 조성물을 제공한다.In addition, the present invention provides a composition for feed addition, comprising the bacteriophage ST20.
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 섞어 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 ST20을 이용할 수 있다.Feed-added antibiotics used in livestock and fishery industries are used for the purpose of preventing diseases, and administration of antibiotics for preventive purposes increases the likelihood of developing resistant bacteria and is a problem because 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, since there are many types of antibiotics mixed with feed and this has a problem of increasing the probability of multidrug resistant bacteria, the bacteriophage ST20 of the present invention is a new feed additive that is more nature friendly and solves the problem arising from the use of existing antibiotics. You can use
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 may provide a feed containing the composition for feed addition, and the feed of the present invention may be prepared by separately preparing bacteriophage in the form of a feed additive and mixing it with the feed, or by directly adding it when preparing the feed. . The bacteriophage in the feed of the present invention may be in a liquid or dry state, preferably in a dried powder form. The drying method may be ventilated drying, natural drying, spray drying, and freeze drying, but is not limited thereto. The bacteriophage of the present invention may be mixed in a powder form in a component ratio of 0.05 to 10% by weight, preferably 0.1 to 2% by weight of the feed weight. In addition, the feed may further include conventional additives that can increase the preservability 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.Other non-pathogenic microorganisms may be additionally added to the composition for feed addition of the present invention. Microorganisms that can be added include Bacillus subtilis , which can produce proteases, lipolytic enzymes, and sugar converting enzymes, and Lactobacillus, which has physiological activity and ability to degrade organic matter under anaerobic conditions such as the stomach of cattle. Strain ( Lactobacillus sp.), filamentous fungi such as Aspergillus oryzae and Saccharomyces cerevisiae showing the effect of increasing the weight of livestock, increasing milk production and increasing the digestion and absorption rate of feed. ) May be selected from the group consisting of yeast.
본 발명의 박테리오파지 ST20을 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.The feed containing the bacteriophage ST20 of the present invention includes grains, root fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starch, meals, grain by-products, etc. Proteins, inorganic logistics, oils and fats, minerals, single-cell proteins, zooplanktons, food leftovers, etc., but 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 있으며, 그 외에도 사료 혼합제 등을 추가로 포함할 수 있다.The composition for feed addition of the present invention may contain binders, emulsifiers, preservatives, etc. added to prevent quality deterioration, and amino acids, vitamins, enzymes, probiotics, flavoring agents, non-proteinaceous agents added to the feed to increase utility Nitrogen compounds, silicates, buffers, colorants, extractants, oligosaccharides, etc. may be included. In addition, feed mixing agents may be additionally included.
또한, 본 발명은 박테리오파지 ST20을 포함하는, 음용수 첨가제를 제공한다.In addition, the present invention provides a drinking water additive, including bacteriophage ST20.
본 발명의 음용수 첨가제는 상기 박테리오파지 ST20 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 살모넬라 균의 숫자를 감소시킬 수 있는 효과가 있다.The drinking water additive of the present invention may be used in a manner in which the bacteriophage ST20 or a composition containing the same is separately prepared in the form of a drinking water additive and used in a manner that is mixed with feed or drinking water, or directly added when preparing drinking water. By mixing and supplying the drinking water as described above, there is an effect of continuously reducing the number of Salmonella bacteria.
본 발명에서 음용수는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 음용수를 사용할 수 있다.In the present invention, drinking water is not particularly limited, and drinking water commonly used in the art may be used.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 소독제를 제공한다.In addition, the present invention provides a disinfectant comprising the bacteriophage ST20.
살모넬라 균에 특이적 사멸능을 갖는 본 발명의 박테리오파지 ST20을 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.The disinfectant containing the bacteriophage ST20 of the present invention having a specific killing ability for Salmonella bacteria can be usefully used as a disinfectant for hospitals and health care to prevent hospital infection, and as a disinfectant for general life, disinfectant for food and cooking places and facilities, and poultry farms , It can be used for disinfection of various growth products such as buildings, livestock bodies, drinking water, litter, nanjwa, transportation vehicles, and tableware.
또한, 본 발명은 상기 박테리오파지 ST20을 포함하는, 세척제를 제공한다.In addition, the present invention provides a cleaning agent comprising the bacteriophage ST20.
본 발명의 박테리오파지 ST20은 살모넬라 균에 특이적 사멸능을 가지므로, 살모넬라 균에 노출되었거나 노출될 가능성이 있는 개체의 피부 표면 또는 신체 각 부위 등을 세척하는 용도로도 사용될 수 있다.Since the bacteriophage ST20 of the present invention has a specific killing ability against Salmonella bacteria, it can also be used to clean the skin surface or parts of the body of individuals exposed to or likely to be exposed to Salmonella bacteria.
또한, 본 발명은 상기 박테리오파지 ST20을 인간을 제외한 개체에 투여하는 단계를 포함하는, 살모넬라 균에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.In addition, the present invention provides a method for preventing or treating infectious diseases caused by Salmonella, comprising administering the bacteriophage ST20 to an individual other than humans.
상기 살모넬라균은 살모넬라 갈리나룸(Salmonella Gallinarum), 살모넬라 풀로럼(Salmonella Pullorum), 살모넬라 콜레라수이스(Salmonella Choleraesuis), 살모넬라 엔테라이티디스(Salmonella Enteritidis) 및 살모넬라 티피뮤리움(Salmonella Typhimurium)으로 구성된 군으로부터 선택된 1종 이상일 수 있으며, 바람직하게는 살모넬라 티피뮤리움(Salmonella Typhimurium), 살모넬라 엔테라이티디스(Salmonella Enteritidis), 살모넬라 갈리나룸(Salmonella Gallinarum) 및 살모넬라 풀로럼(Salmonella Pullorum) 으로 구성된 군으로부터 선택된 1종 이상일 수 있다.The Salmonella bacteria are Salmonella Gallinarum, Salmonella Pullorum, Salmonella Choleraesuis, Salmonella Enteritidis, and Salmonella Typhimurium from the group consisting of Salmonella Enteritidis and Salmonella Tymurium. It may be one or more selected, preferably one selected from the group consisting of Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Gallinarum, and Salmonella Pullorum It can be more than that.
본 발명의 방법은 살모넬라 균에 의한 감염성 질병, 예컨대 유행성 또는 급성으로 발병하는 전염성 질환으로, 살모넬라증 외에 식중독, 위염, 장염, 폐렴, 패혈증, 돼지 콜레라, 추백리 또는 가금 티푸스가 발생한 개체 또는 발생 위험이 있는 개체에 박테리오파지 ST20을 투여하는 단계를 포함한다.The method of the present invention is an infectious disease caused by Salmonella, such as an epidemic or acutely infectious disease, in addition to salmonellosis, food poisoning, gastritis, enteritis, pneumonia, sepsis, pig cholera, chubaekri or poultry typhus developed or at risk. It includes the step of administering bacteriophage ST20 to the subject.
본 발명에서, 용어 "개체"는 살모넬라균에 의한 감염성 질병이 발병될 수 있는 모든 개체를 의미하는 것으로 인간을 포함한 포유동물, 가축류 또는 가금류를 포함하나, 이에 제한되는 것은 아니다. In the present invention, the term "individual" refers to any individual capable of developing an infectious disease caused by Salmonella, and includes, but is not limited to, mammals, livestock, and 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" is a concept that refers to a useful animal that has been purified and improved by humans to live in common with humans, and includes, for example, pigs, cows, chickens, horses, ducks or dogs, but is limited thereto. It does not become.
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 collectively referring to animals belonging to birds among livestock, and includes, for example, chickens, ducks, pheasants, quails, ostrichs, geese or turkeys, but is not limited thereto.
본 발명의 방법에서 상기 박테리오파지 ST20 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 ST20 or the composition may be administered to an animal in the form of a pharmaceutical preparation, or may be mixed with feed or drinking water of livestock and administered through a method of feeding it.
본 발명의 방법에서 상기 박테리오파지 ST20 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 route of administration of the bacteriophage ST20 or the composition may be administered through various oral or parenteral routes as long as it can reach the target tissue, and specifically, oral, rectal, topical, intravenous, intraperitoneal, It can be administered in a conventional manner via intramuscular, intraarterial, transdermal, intranasal, inhalation, and the like.
본 발명의 방법에 투여되는 상기 박테리오파지 ST20의 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 is obvious to those skilled in the art that the appropriate total daily use amount of the bacteriophage ST20 administered in the method of the present invention can be determined by the treating physician within the correct medical judgment range. The specific therapeutically effective amount for a specific individual is determined along with the type and extent of the reaction to be achieved, the patient's age, weight, general health status, sex and diet, administration time, route of administration and secretion rate of the composition, treatment period, and specific composition. It is desirable to apply differently depending on various factors including drugs used or used simultaneously and similar factors well known in the medical field.
중복되는 내용은 본 명세서의 복잡성을 고려하여 생락하며, 본 명세서에서 달리 정의되지 않은 용어들은 본 발명이 속하는 기술분야에서 통상적으로 사용되는 의미를 갖는 것이다.Redundant content is omitted in consideration of the complexity of the present specification, and terms not otherwise defined herein have meanings commonly used in the technical field to which the present invention belongs.
이하, 본 발명을 실시예, 제제예에 의해 상세히 설명한다. 단, 하기 실시예, 제제예는 본 발명을 예시하는 것일 뿐 본 발명의 내용이 하기 실시예, 제제예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by examples and formulation 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.
실시예 1. 신규한 박테리오파지의 분리 및 배양Example 1. Isolation and culture of novel bacteriophage
1.1 박테리오파지의 스크리닝 및 분리1.1 Screening and isolation of bacteriophage
대상세균인 살모넬라 티피뮤리움(Salmonella Typhimurium)은 ㈜옵티팜에서 분리 및 동정한 균주를 사용하였다. 살모넬라 티피뮤리움을 특이적으로 사멸시킬 수 있는 박테리오파지를 분리하기 위해 양계농장에서 시료를 채취하여 대상세균과 함께 35℃에서 하룻밤 동안 진탕배양 하였다. 배양액을 4,000rpm으로 20분간 원심분리하여 상등액을 pore size 0.2μm의 필터를 이용하여 여과한 후, 얻어진 여과액을 이용하여 용균반 분석을 통해 살모넬라 티피뮤리움에 특이적인 박테리오파지를 선별하였다. The target bacteria, Salmonella Typhimurium, were isolated and identified by Optipharm Co., Ltd. In order to isolate the bacteriophage that can specifically kill Salmonella typhimurium, samples were collected from poultry farms and cultured with shaking at 35°C overnight with the target bacteria. The culture solution was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was filtered using a filter with a pore size of 0.2 μm, and then the bacteriophage specific to Salmonella typhimurium was selected through lysis spot analysis using the obtained filtrate.
박테리오파지를 스크리닝 하기 위해서 구체적으로 전배양된 대상세균 150μL를 취해서 멸균된 튜브에 넣고 3mL의 LB 탑 아가 (LB배지 (트립톤 10g/L; 효모 추출물 5g/L; NaCl 10g/L) + 0.6% 아가)를 첨가하여 잘 혼합한 후 미리 만들어 둔 2% LB 아가 플레이트에 중층하였다. 실온에서 30분간 방치한 후 준비된 샘플 여과액을 각각 10μL씩 점적한 후 실온에서 30분간 방치 후 35℃에서 24시간 배양하여 박테리오파지의 유무를 확인하였다. 박테리오파지의 존재를 확인한 샘플로부터 파지를 순수 분리하기 위해서 소프트 아가 오버레이법을 이용하였다. 박테리오파지가 있다고 확인된 샘플을 식염수로 적절히 희석하여 각 희석단의 샘플 희석액 100μL를 취하여 대상 세균 배양액 150μL에 혼합한 후 0.6% LB 탑 아가를 3mL 첨가하여 잘 혼합한 뒤, 미리 만들어 둔 2% LB 아가 플레이트에 중층하여 방치한 후 35℃에서 24시간 배양하였다. 배양한 각 플레이트에서 형성된 용균반을 확인한 후 선택하여 400μL의 SM 완충액(NaCl 5.8g/L; MgSO47H2O 2g/L; 1M Tris-Cl (pH7.5) 50mL)에 현탁하고, 실온에서 4시간동안 방치하였다. 이 박테리오파지 액 100μL를 0.6% LB 탑 아가와 세균배양액 500μL와 혼합한 뒤 150mm 지름의 LB 플레이트에 분주하여 중층한 후 완전히 용균이 일어날 때까지 35℃에서 배양하였다. 배양이 종료된 후, LB 플레이트에 15mL의 SM 완충액을 가하고 실온에서 4시간동안 천천히 교반하면서 박테리오파지 용액을 회수한 후 4,000rpm으로 20분간 원심분리하여 상등액만 취하고 이를 0.2μm 필터로 여과하여 최종 박테리오파지 용액을 회수하였다. 이렇게 얻어진 최종 박테리오파지의 용균반을 확인한 결과를 도 1에 나타내었다.To screen bacteriophage specifically, take 150 μL of pre-cultured target bacteria and put it in a sterilized tube, 3 mL of LB top agar (LB medium (trypton 10 g/L; yeast extract 5 g/L; NaCl 10 g/L) + 0.6% agar ) Was added, mixed well, and then layered on a previously prepared 2% LB agar plate. After standing at room temperature for 30 minutes, 10 μL of each prepared sample filtrate was added dropwise, and then allowed to stand at room temperature for 30 minutes, and then cultured at 35° C. for 24 hours to check the presence or absence of bacteriophage. Soft agar overlay method was used to purely separate phage from a sample confirming the presence of bacteriophage. After appropriately diluting the sample confirmed to have bacteriophage with saline, take 100 μL of the sample dilution solution from each dilution stage, mix it with 150 μL of the target bacterial culture solution, add 3 mL of 0.6% LB top agar, mix well, and mix it well with 2% LB agar prepared in advance. After being stacked on a plate and left to stand, it was incubated at 35°C for 24 hours. After confirming the lysate formed in each cultured plate, select it, and suspend it in 400 μL of SM buffer (NaCl 5.8 g/L; MgSO47H2O 2 g/L; 1M Tris-Cl (pH7.5) 50 mL) and leave at room temperature for 4 hours. I did. 100 μL of this bacteriophage solution was mixed with 500 μL of 0.6% LB top agar and bacterial culture solution, then dispensed on an LB plate having a diameter of 150 mm, layered, and incubated at 35° C. until completely lysis occurred. After the cultivation was completed, 15 mL of SM buffer was added to the LB plate, and the bacteriophage solution was collected while slowly stirring at room temperature for 4 hours, and then centrifuged at 4,000 rpm for 20 minutes to take only the supernatant and filtered through a 0.2 μm filter to final bacteriophage solution Was recovered. The results of confirming the lysis plate of the final bacteriophage thus obtained are shown in FIG. 1.
도 1에 나타낸 바와 같이, 박테리오파지 ST20은 3-4 mm 크기의 용균반을 형성하는 박테리오파지로 확인하였다.As shown in Figure 1, the bacteriophage ST20 was confirmed as a bacteriophage forming a lysis plate of 3-4 mm size.
1.2 박테리오파지의 대량 배양 및 정제1.2 Mass culture and purification of bacteriophage
박테리오파지를 대량 배양하기 위하여, 실시예 1.1에서 회수한 최종 박테리오파지 용액을 1.0x107 PFU/mL로 희석한 후 전배양된 세균배양액(1.0x109 CFU/mL)과 혼합하여 MOI(Multiplicity of infection)를 0.01이 되도록 하여 35℃에서 100rpm으로 18시간 배양하였다. 배양이 종료된 후 전량의 배양액을 6,000rpm으로 30분간 원심분리하여 상등액을 수득하였고, 이를 0.2μm 필터로 여과하여 최종 박테리오파지 배양액을 회수하였다.In order to cultivate a large amount of bacteriophage, the final bacteriophage solution recovered in Example 1.1 was diluted to 1.0x10 7 PFU/mL, and then mixed with a pre-cultured bacterial culture solution (1.0x10 9 CFU/mL) to obtain a MOI (Multiplicity of infection). It was incubated for 18 hours at 100 rpm at 35°C to 0.01. After the cultivation was completed, the entire amount of culture was centrifuged at 6,000 rpm for 30 minutes to obtain a supernatant, which was filtered through a 0.2 μm filter to recover the final bacteriophage culture solution.
분리된 파지를 박테리오파지 ST20로 명명하고, 2019년 4월 12일, 한국미생물보존센터(Korean Culture Center of Microorganisms, 서울, 한국)에 기탁하여 기탁번호 KFCC11828P를 부여받았다.The separated phage was named Bacteriophage ST20, and on April 12, 2019, it was deposited with the Korean Culture Center of Microorganisms (Seoul, Korea) and received the deposit number KFCC11828P.
실시예 2. 박테리오파지 ST20의 전체 유전자 분석Example 2. Whole gene analysis of bacteriophage ST20
파지 배양액에 20mM EDTA, 50μg/mL의 프로테이나제 K(Proteinase K), 0.5%(w/v) SDS를 가한 후, 50℃에서 1시간 동안 정치하였다. 동량의 페놀-클로로포름-이소프로필 알코올(Phenol-Chloroform-Isopropyl alcohol) (25:24:1)과 혼합한 후 실온에서 12,000 rpm으로 10분간 원심분리하여 상등액을 회수하였다. 회수된 상등액에 동일 부피의 페놀-클로로포름-이소프로필 알코올 (25:24:1)과 혼합하여 실온에서 12,000rpm으로 10분간 원심분리한 후 상등액을 회수하고, 상등액에 3M 소듐 아세테이트(sodium acetate)를 전체 부피의 10%(v/v)가 되도록 가한 후 혼합하였다. 여기에 2배 부피의 차가운 95% 에탄올을 가하여 혼합한 뒤 -20℃에서 1시간 동안 정치하였고, 반응이 끝난 후 0℃에서 10분간 12,000rpm으로 원심분리한 후 침전물을 획득하였고, 차가운 70% 에탄올로 2회 세척한 뒤 에탄올을 완전히 제거하여 DNA 펠렛을 회수하였고 이를 최종 50μL TE buffer(Tris-EDTA, pH8.0)에 용해시킨 후 DNA 농도를 측정하였다. 전체 유전자 분석을 위해 PacBioRSII를 이용하여 유전자 시퀀싱을 시행하였다.(마크로젠, 한국). 유전자 분석 결과, 박테리오파지 ST20의 전체 유전자 염기 서열을 서열번호 1로 나타내었으며, 박테리오파지 ST20의 유전자 맵을 도 2에 나타내었다.20 mM EDTA, 50 μg/mL Proteinase K, and 0.5% (w/v) SDS were added to the phage culture, and then allowed to stand at 50° C. for 1 hour. After mixing with the same amount of phenol-chloroform-isopropyl alcohol (25:24:1), the supernatant was recovered by centrifugation at 12,000 rpm for 10 minutes at room temperature. The recovered supernatant was mixed with the same 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. It was added so as to be 10% (v/v) of the total volume and then mixed. Two times the volume of cold 95% ethanol was added thereto, mixed, and allowed to stand at -20°C for 1 hour, and after the reaction was completed, centrifuged at 12,000 rpm for 10 minutes at 0°C to obtain a precipitate, and cold 70% ethanol After washing twice with, ethanol was completely removed and the DNA pellet was recovered. After dissolving it in a final 50 μL TE buffer (Tris-EDTA, pH 8.0), the DNA concentration was measured. Gene sequencing was performed using PacBioRSII for full gene analysis (Macrogen, Korea). As a result of genetic analysis, the entire gene nucleotide sequence of the bacteriophage ST20 was shown in SEQ ID NO: 1, and the gene map of the bacteriophage ST20 is shown in FIG. 2.
도 2에 나타난 바와 같이, 박테리오파지 ST20의 전체 유전자의 크기는 42,267 bp를 가지며, GC함량이 49.93%, 전체 ORFs의 수는 60개로 확인하였다. As shown in FIG. 2, the size of the total genes of the bacteriophage ST20 was 42,267 bp, the GC content was 49.93%, and the total number of ORFs was confirmed to be 60.
실시예 3.Example 3. 박테리오파지 ST20의 형태학적 특성Morphological characteristics of bacteriophage ST20
분리된 박테리오파지의 형태학적 특성을 알아보기 위해, 박테리오파지 ST20을 0.01% 젤라틴 용액에 희석하여 2.5% 글루타르알데하이드(glutaraldehyde) 용액으로 고정하였다. 이를 carbon-coated mica plate(ca. 2.5 x 2.5 mm)에 적가한 후, 10분간 적응시켜 멸균증류수로 세척하였다. 이를 구리 격자(copper grid)에 끼워 2% 우라닐 아세테이트(Uranyl acetate)에서 30-60초간 염색하고 건조시킨 후, 투과전자현미경(Tecnai G2 Spirit Twin, Bio-Transmission electron microscope, 120kV, 120,000~400,000)으로 검경하였으며, 박테리오파지 ST20의 전자현미경 사진을 도 3에 나타내었다.In order to investigate the morphological characteristics of the isolated bacteriophage, bacteriophage ST20 was diluted in 0.01% gelatin solution and fixed with a 2.5% glutaraldehyde solution. This was added dropwise to a carbon-coated mica plate (ca. 2.5 x 2.5 mm), acclimated for 10 minutes, and washed with sterile distilled water. Insert it into a copper grid, dye it in 2% uranyl acetate for 30-60 seconds, dry it, and then dry it under a transmission electron microscope (Tecnai G2 Spirit Twin, Bio-Transmission electron microscope, 120kV, 120,000~400,000). It was examined as, and an electron microscope photograph of the bacteriophage ST20 is shown in FIG. 3.
도 3에 나타낸 바와 같이, 박테리오파지 ST20는 170 내지 190 nm의 크기를 가지는 정이십면체의 머리와 비수축성 꼬리가 있는 형태인 사이포비리대(Siphoviridae)에 속함을 확인하였다.As shown in FIG. 3, it was confirmed that the bacteriophage ST20 belongs to the Siphoviridae , a form having a head of a regular icosahedron having a size of 170 to 190 nm and a non-constricting tail.
실시예 4.Example 4. 박테리오파지 ST20의 단백질 패턴 분석Protein pattern analysis of bacteriophage ST20
1.0x1010 PFU/mL로 준비된 박테리오파지 ST20 40μL와 10μL의 5X 로딩 버퍼(loading buffer)에 혼합하고, 10분간 끓인 다음 12.5% SDS-PAGE를 수행하였으며, 박테리오파지 ST20의 단백질 전기영동 결과를 도 4에 나타내었다.Mixing in 40 μL of bacteriophage ST20 prepared at 1.0 × 10 10 PFU/mL and 10 μL of 5× loading buffer, boiled for 10 minutes, and then 12.5% SDS-PAGE was performed, and the protein electrophoresis results of bacteriophage ST20 are shown in FIG. Done.
도 4에 나타낸 바와 같이, 박테리오파지 ST20은 72 kDa, 60 kDa, 38 kDa 그리고 36 kDa 크기의 주요 단백질을 포함하는 것을 확인하였다.As shown in Figure 4, it was confirmed that the bacteriophage ST20 contains major proteins of 72 kDa, 60 kDa, 38 kDa and 36 kDa.
실시예 5. 박테리오파지 ST20의 다양한 세균에 대한 감수성 확인Example 5. Confirmation of susceptibility of bacteriophage ST20 to various bacteria
분리된 박테리오파지 ST20이 다양한 세균에 대해서 용균활성을 나타내는지 확인하기 위하여 ㈜옵티팜에서 분리하여 보유하고 있는 균주와 미생물 자원센터 등에서 분양 받은 균주를 대상으로 소프트 아가 오버레이법을 이용하여 감수성을 확인하였다. 살모넬라 티피뮤리움 20 균주, 살모넬라 엔테라이티디스(Salmonella Enteritidis) 3 균주 및 살모넬라 종(Salmonella spp.) 9 균주를 대상으로 감수성 검사 진행하였으며, 그 결과를 표 1에 나타내었다.In order to confirm whether the isolated bacteriophage ST20 exhibits lytic activity against various bacteria, the susceptibility was confirmed by using a soft agar overlay method for strains isolated and possessed by Optipharm Co., Ltd. and strains sold in microbial resource centers.
[표 1][Table 1]
표 1에 나타낸 바와 같이, 박테리오파지 ST20은 살모넬라 티피뮤리움과 살모넬라 엔테라이티디스에 100%의 용균활성을 보였으며 살모넬라 종 (Salmonella spp.)에는 88.9%의 용균활성을 보였다. As shown in Table 1, bacteriophage ST20 showed a lytic activity of 100% in Salmonella typhimurium and Salmonella enteratidis, and 88.9% in Salmonella spp.
또한 다른 미생물을 대상으로 용균활성을 확인하는 실험을 실시하였으며, 실험 결과를 표 2에 나타내었다.In addition, an experiment was conducted to confirm the lysis activity for other microorganisms, and the experimental results are shown in Table 2.
[표 2][Table 2]
표 2에 나타낸 바와 같이, 박테리오파지 ST20은 살모넬라 갈리나룸(Salmonella Gallinarum) 및 살모넬라 풀로럼(Salmonella Pullorum)에도 용균활성을 보여 살모넬라에 대한 광범위한 활성을 가진 박테리오파지인 것으로 확인하였다. 다만, 대장균이나 스타필로코커스(Staphylococcus) 종, 바실러스(Bacillus) 종 등 다른 세균에 대해서는 용균활성이 없는 것을 확인하였다.As shown in Table 2, bacteriophage ST20 showed lytic activity in Salmonella Gallinarum and Salmonella Pullorum, and was confirmed to be a bacteriophage having a wide range of activity against Salmonella. However, confirming that there is no lytic activity was for other bacteria such as Escherichia coli and Staphylococcus (Staphylococcus) species, Bacillus (Bacillus) species.
실시예 6.Example 6. 박테리오파지 ST20의 살모넬라 티피뮤리움균에 대한 사멸능의 확인Confirmation of killing ability of bacteriophage ST20 against Salmonella typhimurium bacteria
박테리오파지 ST20의 살모넬라 티피뮤리움에 대한 사멸 능력을 분석하기 위하여, 600nm에서 흡광도가 약 0.5 되는 세균 배양액을 10mL 준비하였다. 세균 배양액에 농도가 1.0x1010 PFU/mL 되는 박테리오파지액을 1 mL 넣어 MOI 10으로 접종하고, 1.0x109 PFU/mL 농도의 박테리오파지액을 1 mL 넣어 MOI 1로 접종하여 두 가지 조건으로 실험하였다. 박테리오파지액을 넣지 않은 시료를 대조군으로 하였다. 3시간 동안 배양하며 1시간 단위로 흡광도 600 nm에서 세균의 배양 상태를 관찰하였으며, 실험 결과를 단위를 OD600nm로 하여 표 3 및 도 5에 나타내었다. In order to analyze the killing ability of the bacteriophage ST20 against Salmonella typhimurium, 10 mL of a bacterial culture solution having an absorbance of about 0.5 at 600 nm was prepared. 1 mL of a bacteriophage solution having a concentration of 1.0x10 10 PFU/mL was added to the bacterial culture solution and inoculated with
[표 3][Table 3]
표 3 및 도 5에 나타낸 바와 같이, 박테리오파지 ST20을 첨가하지 않은 대조군의 경우, 2시간 후 600 nm에서 흡광도가 1.32로 나타나 세균이 잘 성장한 것을 확인하였다. 반면, 박테리오파지 ST20을 첨가한 실험군의 경우, 3시간 경과했을 때 600 nm에서 흡광도가 0.34와 0.4로 배양 시작단계보다 감소한 것을 확인하였다. 따라서, 박테리오파지 ST20이 살모넬라 티피뮤리움의 사멸에 탁월한 효과를 나타내는 것을 다시 한번 확인하였다.As shown in Table 3 and Figure 5, in the case of the control group to which the bacteriophage ST20 was not added, the absorbance at 600 nm was 1.32 after 2 hours, confirming that the bacteria grew well. On the other hand, in the case of the experimental group to which the bacteriophage ST20 was added, it was confirmed that the absorbance at 600 nm was 0.34 and 0.4 after 3 hours had elapsed from the incubation start stage. Therefore, it was confirmed once again that bacteriophage ST20 exhibits an excellent effect on killing Salmonella typhimurium.
실시예 7.Example 7. 박테리오파지 ST20의 pH 안정성 조사Investigation of pH stability of bacteriophage ST20
박테리오파지 ST20의 pH 안정성을 확인하기 위하여 다양한 pH 범위(pH2, 3, 4, 5, 6, 7, 8, 9, 10, 11)에서 시험을 진행하였다. 소듐 아세테이트 버퍼 (Sodium acetate buffer), 소듐 포스페이트 버퍼(Sodium phosphate buffer), Tris-HCl을 이용하여 각각의 pH에 맞게 조제한 후 990μL로 분주하여 준비한 후, 1.0x1010 PFU/mL로 준비한 박테리오파지 ST20 배양액 10μL을 첨가하여 2시간동안 상온에서 방치하였다. 방치 후 각 반응액을 단계 희석하여 소프트 아가 오버레이법에 의해 역가를 측정하였으며, 측정 결과를 도 6에 나타내었다.In order to confirm the pH stability of the bacteriophage ST20, tests were conducted in various pH ranges (pH2, 3, 4, 5, 6, 7, 8, 9, 10, 11). Prepared according to each pH using sodium acetate buffer, sodium phosphate buffer, and Tris-HCl, dispensed to 990 μL, and prepared, 10 μL of bacteriophage ST20 culture solution prepared at 1.0x10 10 PFU/mL Was added and left at room temperature for 2 hours. After standing, each reaction solution was diluted stepwise and the titer was measured by the soft agar overlay method, and the measurement results are shown in FIG. 6.
도 6에 나타낸 바와 같이, 박테리오파지 ST20은 pH4 내지 pH11의 범위에서 2시간까지 안정한 것을 확인하였다.As shown in Figure 6, it was confirmed that the bacteriophage ST20 is stable up to 2 hours in the range of pH4 to pH11.
실시예 8.Example 8. 박테리오파지 ST20의 온도 안정성 조사Investigation of temperature stability of bacteriophage ST20
박테리오파지 ST20의 온도 안정성을 확인하기 위하여 다양한 온도 범위(40℃, 50℃, 60℃ 그리고 70℃)에서 시험하였다. 박테리오파지 ST20 배양액(1.0x108 PFU/mL) 5mL를 멸균된 튜브에 분주하고, 10분, 20분 및 30분 동안 각 온도에서 방치하였다. 방치 후 각 반응액을 단계 희석하여 소프트 아가 오버레이법에 의해 역가를 측정하였으며, 측정 결과를 도 7에 나타내었다.In order to confirm the temperature stability of the bacteriophage ST20, it was tested in various temperature ranges (40°C, 50°C, 60°C and 70°C). 5 mL of the bacteriophage ST20 culture solution (1.0x10 8 PFU/mL) was dispensed into a sterilized tube, and left at each temperature for 10 minutes, 20 minutes and 30 minutes. After standing, each reaction solution was diluted stepwise and the titer was measured by a soft agar overlay method, and the measurement results are shown in FIG. 7.
도 7에 나타낸 바와 같이, 박테리오파지 ST20은 40℃ 내지 70℃의 범위에서 30분까지 안정한 것을 확인하였다. As shown in Figure 7, it was confirmed that the bacteriophage ST20 is stable up to 30 minutes in the range of 40 ℃ to 70 ℃.
이하 본 발명의 박테리오파지 ST20을 포함하는 감염성 질병의 예방 또는 치료용 약학적 조성물의 제제예를 설명하나, 본 발명을 한정하고자 함이 아니고 단지 이를 구체적으로 설명하고자 함이다.Hereinafter, an example of the preparation of a pharmaceutical composition for preventing or treating infectious diseases including the bacteriophage ST20 of the present invention is described, but is not intended to limit the present invention, but only to describe it in detail.
제제예 1. 사료 첨가용 조성물Formulation Example 1. Composition for feed addition
박테리오파지 ST20 1x109 PFU/mLBacteriophage ST20 1x10 9 PFU/mL
비타민 믹스 100 mg100 mg of vitamin mix
미네랄 믹스 100 mg100 mg of mineral mix
스테아린산 마그네슘 2 mg2 mg of magnesium stearate
통상의 사료 첨가제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.It is prepared by mixing the above ingredients according to a conventional feed additive manufacturing method.
제제예 2. 소독제 조성물Formulation Example 2. Disinfectant Composition
박테리오파지 ST20 1x109 PFU/mLBacteriophage ST20 1x10 9 PFU/mL
에탄올 적량Appropriate amount of ethanol
멸균 증류수 적량Proper amount of sterile distilled water
pH 조절제 적량proper amount of pH adjuster
통상의 소독제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.It is prepared by mixing the above components according to a conventional disinfectant manufacturing method.
제제예 3. 세척제 조성물Formulation Example 3. Cleaning agent composition
박테리오파지 ST20 1x109 PFU/mLBacteriophage ST20 1x10 9 PFU/mL
에탄올 적량Appropriate amount of ethanol
효소 복합물 적량Appropriate amount of enzyme complex
멸균 증류수 적량Proper amount of sterile distilled water
통상의 세척제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.It is prepared by mixing the above components according to a conventional cleaning agent manufacturing method.
이상, 본 발명내용의 특정한 부분을 상세히 기술하였는 바, 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.Above, a specific part of the present invention has been described in detail, and for those of ordinary skill in the art, it is obvious that this specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Therefore, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.
<110> Optipharm.CO.,LTD <120> Novel Salmonella specific bacteriophage ST20 and antibacterial composition comprising the same <130> optipharm1-84P <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 42267 <212> DNA <213> Unknown <220> <223> Bacteriophage ST20 <400> 1 cgcttcgatt gttgcgtttg gcgaaataga aaaatattaa caaaaatggt acgctgctac 60 tcagcagcgt aagtcactca gtaaaatcct aaactactgt tacaagcttc ctgcggttcc 120 gataacagcg cccaggtgcg gccccgccac cacgataccc caggaactac ccacccacca 180 tcatcggcgt ccggatgcgg ccccggggac cactaaaacc ctagccataa acggcaacct 240 cctttaagac ccgagaatca ccggcgcgga ctctttagaa tcttttgccg tggggtattt 300 acagggaggt ccccgccccc attagagtgc ctttattccc gtccctcgtc aggtaagcgc 360 ttgccgcaag gcaagtgcgt taattcatga gtagtagact aagaatcttt cggcatacaa 420 gagaacagtt acgtttgccg ttgtccagtg ttgtgccttt attcctatta tctggtaact 480 aaagggcagg tctaaaacct gcccgctaat cgttacattt tacggttgcg aatctacggc 540 acatcaagga attcagatac ttaagagata tgctgccgat gagaatatag ctcgcccagg 600 gcagttcaga tactaaggaa gggaagttac tgagttttaa gtttgcgggt ttcagtaagc 660 cttaacgcgt atccgcttgg ggcggacccg ctaagctctc acaaagtgaa acagaaactt 720 aagtaacttg aagttacaag agacaactta cacgaaaaaa atgaaaatgt caataggtgt 780 gtttatagtt gatttcccat tatgccgtgg gtatacttgc tttactaact gagatggagg 840 atttgagaat gacacagaat gaagtagctg agcttattgg ggttacccgc cgcacgctga 900 ataactggct aagagacggc aagttcccgg attgttgtgt ccggattatg ggtcgcagga 960 tgccgggtac gtttgaccgg gagaaagtgg aagcgtggat tagggagaat gtaaagtgac 1020 cgatttccag aaaagggtat tcagaacggt agtcgcgctt acgtgtaaaa aagggtcgtg 1080 tagtgttctt gacttgcgcc gcacttattt tcagtactat agttcctctc tcatcgagaa 1140 cacgttgaag gttctggtta gagacggagc agttaaaagt aaggccggaa aatattcggc 1200 agtagctgaa gttaaaggct cccaggcgac cccagaagac cttgaataaa agaaaagccc 1260 caacgcgcga acgtcagggc cttaaacact acaaaggaaa agcacatgtc aacaggtaat 1320 ttagcatact catggtcgcg tcgcaatcgc cgtgaggaac gcgctgacat taacgtttac 1380 accacaactt ccggcacctg ggatgagttt gtcgaactga tgcagccgct taaacgttcg 1440 cgccggaatc ccaagacgga cccgggttac attaccgccg cttgtactgc cacggtaagc 1500 tctaccggta aagaagccgc cgaaggtatg ttctaccgct gcaatgcatc tgttacgtct 1560 tcatccctgg cttacgccga tgtggatagt gcgactccgg aagagtttgc caccgactgt 1620 gaaatggtgc gcgagtcacg tttcgctatg atgctctaca ccacagcatc gcacactgcg 1680 gaagcgccgc gctatcgcgt cgttatgccc gtgcgtacac cggtcactgg tggcgacgtt 1740 atccgcatcc ggtacggcct gttgacccac ttccttaaag gacgtgacgt agatagcgcc 1800 gggttcaccc tgtctcagcc gatgtaccgc ccgcctgtcg ggagtcaggt catcgtgtcc 1860 gaaagtagcc gcatgattac ggccagtaag cttatggagg gagttcctga aattaacgtt 1920 acaggcgctt ccgattacaa agtgcccgag ggtgagcaat ccgaattaac tgacctgttt 1980 gaagagttcg ccttcgagtt cggcggccgt atgaccgacc gcggcctgca aatgcccgcc 2040 acgccggagc acgccgccca atacactacc ggtgaaccta agcaggatga cttcctgttc 2100 tgttggccgc gcgacggctt cgagcgaccc aacgttacca tgtaccacga taccgacctg 2160 gtagctacgg gcggtatgaa gcctggcgga cgggatatgt gggcttacgc ttgtgcggcc 2220 accggcttac cttttgaccg tgtggaggtg gcgcttgggt gggcggaggg ggtcgcttgt 2280 gacgaagaag acctggccga cgaagaacca ccagcaccac aagcggactt catcgtcgaa 2340 ggatatatgc cgtctaactg catctgggat atcgtcggtg aatccgggac gtacaaatcg 2400 ttctacacac tcggcatgat gtatatcagt gctgccgggt atcggttcgc gggggcagac 2460 acccgtaaag cccatcattt ctatattgat ggtgagggtg gggagttcac gcacacccgt 2520 atcgctgcgt tggccgctaa atacggcgat gaaggtatgc ggtatatgca tgtgcttgac 2580 gctggtgaat tcgcggatac gaagaaactg gtgcgtaaaa tgcgccagat agcgggcagt 2640 gaacctgtcg ggatggtcgc tttcgacact cttaaccaga ccttcgggaa ctggatagac 2700 aagttcaatg agaactcagc cgggcaagac gggatgggcc gtgtcgtggc tatgcttaaa 2760 gaagtgcgcg acggtactaa aggcgctgta ggcgttgtcc accacacccc gaaaggtggg 2820 agtaaggcgc gcggaagtgg ggcgctgtat gctggtgtcg acgtcgaact tacgcttgtc 2880 cgagcgaccg agaaacaaat aaacgttgcg cacaccaaga acaaaaacgg catgcagcag 2940 aaaacgatag gtatggtgct tgaaccggta cagtttcgtg aagctccacc gccgaaagag 3000 ttccaggctg tagaattcgt tggaggtgag ggatacggtg aaatcgttaa cctcgacctg 3060 ccggaaccac ataaagctct tgtactcatg ccgtggggct tccagccgtt cgaaacggat 3120 gaagaaaaag aacgcaacga aggactcgat ggtaaaggta aagattccgt aaagggtact 3180 gttaaacggt ctaaagatgc ttccgcccgc gaatcagtta tgtctgctct cgaagattta 3240 cagcaagccg atgatacagg acgtggcttc acacaacggc aaatagtcgc aagagcgagg 3300 gaccacagca tcacaaacct ggtacttgag aagatgctac gtgagggtga attgatgctt 3360 ggttgcgatg aaaacggtga agttgtaacg aatacttaca ggttgccgac gggtatagat 3420 gaccgtaagc ggccaaagaa ccggtatgaa cctaacgaca acataaagac aacagaagtg 3480 gatttagagt aaaaagaaag gggccatcag gcccctgtat caccaaagct taaccatcga 3540 gggtcaacac cgagagcatc ggcaatgtgg aagattttaa aacagtctac cttacatacc 3600 cgaccagtcg ccaagtgggt aatcaggctt tgggataccc cggctttgaa ggcaagctca 3660 gtttgaccca tgccgatttc tttgcgacgg cgttctacac gagcgcctag ctcagattgc 3720 tgcatatgtt tactccttag ttagatatgt agtaatagta cagtaataaa atacacggtg 3780 caatttacga aaaagtgctt gcataaataa tagagtacga ttatagttct tatcaccaac 3840 taaacaggag aacaaataat gttagagaaa ttcttggtat tacttgaacg tttcgtagtt 3900 gcccatgaac tgattgctgc gaatagcgcg aaacagactg tgtaccaact taaaagtgat 3960 ataaccacgg taaacaaaga gttgacagaa aaagcggtag aaatggcgat gcaggataat 4020 tataaggcta gttctgcggt acgcaaagca tcagaaaaag aactcccggt agaaggcgaa 4080 agcgtcgttg acactaaacc ggcggaagaa gagaagccga aacgtaagac gcgtaaagct 4140 aaagtagagg agccggaaga agagaaagaa ggagttgact accagtctct tcgcgaccag 4200 attcaggcta tcgacgatgc gattagcgaa ggtcctagcg atgacgctta cgatgattcc 4260 aatgaactgc tgggagagtt caccggtaag aagatgaagc ttgccgcgat taaggacgaa 4320 gacctggccg aatacctgga acgcctgaca gcgatcaaga acaagtattt cgaagaagaa 4380 gaataactat cccgcggcct tcgggccgct ttacatggag attgacggat gaaattaaaa 4440 gaacgcggtg gcaacaacga tgtgcattcc ctgctgtcgc cgtctggggc caaaaagtgg 4500 ctggcgtgca gtgcatcact ggcttgtgaa aaagatatac ccaacacgtc cggtaaagcc 4560 gcggtgttag gaacggcgat gcacacactt gccgagattc atcttaacca gtacatcaaa 4620 ggcacttctt tgccgttaga gcgagatgta ggggcatatg tgctggaaga aggtaaaggc 4680 gcggttaagg ccctaattaa gccgatgaaa ggcgcggtgc tggttacaga tgacatggtc 4740 gagcaggtgc gaaagtacac cgactactgt aaagcggtta tcgatgtagc ggcctacgcc 4800 aagcttgaaa tgcgcgttaa tcttagtgag gtgttgcacc ccggatacga aggcgttgag 4860 acgttcggga ccgctgacct tgttgccgta caggaattgg ctaacactga cgatcacatg 4920 ctcgttattg gcgacctgaa aacagggcgc catagagtag aggcgaaaga aaacaagcaa 4980 cttatgctct atgctctcgg tgtttaccgc cgcctaaaga gacgctataa cataacagct 5040 gttcgtctgg ttatctttca accgtacacc ggtggcgcgt cggagtggga tatctcagtt 5100 gaaggtcttg aactattcgc taagttcgcg cagaaacgtg cggtagccgc acttgacgcg 5160 tatcgacgtg gtaagaaaaa ccttaaacca tcggacttca agccgtcggt tgatggttgc 5220 caatggtgcc gtttctcaga acaatgtgcc gcgcgtacaa aaaccgttaa ttctgtactg 5280 gcggaagaac tggaagacga tacggatgta tatcgtacag ccgtagaacg tgtagctaag 5340 ttctgcaacg acgcggacta tatgagcgtt tgggaggatg ttcagccgag cgatattctt 5400 aagttgctgc ctgatggccc agatttacct aagaagccag tgagagggga cgactttgca 5460 ctggagctta cgccggagca actcgtcgct gagtatgaga agttgccgtt gttgcgccag 5520 cacatcgaca aagtagaaaa agctatggca gccgctttgc attccggtaa gaaagttccc 5580 ggttacaaac tggttgaggg ccgtcctggt aatcgcacgt ggaaagatga agatgctgta 5640 aataaattta gggaaaacca tgtaaacggc tatctgctgg ataagactgt tcccgtatcg 5700 ccgaccgaag cggaaaagat catcggcaag gaagaccccg aactttgggc tgagctatct 5760 aaactcgtaa ctcgtaagcc gggtgtgcca tgtgttgcat cggcggatga caggcgacct 5820 gattggaaaa atattacgga ggaggatttg gaatgaagaa attgacgtgg tggctgcttg 5880 ccaccccact aataccggtg gcttttatat actatctaat gagagcgtta gagaaagttg 5940 gtgaaacgtc ggcgatttta gctgacagga tgctagactc aaaaatgagg atgaaatatt 6000 tagcatggcg taatcgaaaa tttggttgac aagttaatag tactctatta tattcttaat 6060 cactgacctg gcagcttcct ggataaactc taaaacgcga gaaaactaaa atgggaatca 6120 aacttaatct tcgtaaagta caaactgcat ggttgaacgt attcgaacgc gctaaagacc 6180 gtgaaaataa tgacggttcc gttactaaag gtacgtataa cggtactttt atcctgactc 6240 cggaacaccc gcaaattgaa gaacttcgcg atacggtatt cgctgtagtg tcggaagcac 6300 tgggagaggc cgccgcccag aaatggatga agcagaacta cggtgaaggg aagcatatgg 6360 ataaatgcgc cgtgcgcgac atcgccgaac gtgataaccc gttcgaagac ttcccggaag 6420 gtttttactt ccaggcaaag aacaagcaac agccattaat cctgacttcg gttaagggcg 6480 aaaagcaggt agaaccagac ttcaatattg atggtgagca gattgaaggt aagcaggtat 6540 atagcggttg tgtagctaat atatcaatcg aaatttggtt ttccgagcaa tataaagttt 6600 taggcgcaaa actaaacggc atcaaatttg caggtgaagg aaaggcattc ggtggttctg 6660 cggtttctgc cagcgttgac gacctggaag atgatgaaga cgaaacaccg cgtcgcgaac 6720 gccgtcgtaa ccgttaatgt attttctttt caattaaggc ggcttcggtc gccttttcta 6780 taagggtcaa ataaatgaat ctgctttatc tcgatactga aacattctca gaagccgatt 6840 tgaaaaaagt cggttcctat gcctatgccg aacatccgac tactgaaatt gttatctgca 6900 cttatgcttt cgatgaaggc cccgtgcaag tatgggacgc caccgacggc agcgatatgc 6960 cgagagattt gcgcagggcg atgctaaagc tgcaaaaacc agacagcaac ctcaaactgg 7020 taggccaaaa cttccttatg ttcgaccgac cagttattaa gcattgctgg ggattcgaac 7080 tcctggtaga aaacattata gacactatga tagtcgcgtt ccgacatgcc ctcccgggtt 7140 cactggccgc gctgtgtgag gttttaaaca ttgacgcaag catggctaag gataaacgcg 7200 gcaaggcgct catacagcgg ttcagtaagc ctacgcctaa gaactataag attcgacgat 7260 ataccgccaa tacccaccca aaagagtggg cagaatttat cgcatacgca aaaagcgaca 7320 ttacgtctat gcgtgaagtg tataagaaaa tgccgaagtg ggggaattct gagttcgaag 7380 accgcgtgct gtggttagac caagtaatta acgaccgcgg atttaaggtt gacgttgccc 7440 tcgctgaggc cgcgattgaa gcagtgacgc gccacaagga agagctacag gaagaagccc 7500 aacgcaaata cggcggttca ctaaccggca aagacttctt gcctatttta caggaactgg 7560 cgccggcgca ccgcattcac aacgcacaga agtcaacact aaacgatttg ctggcggatg 7620 aggatttacc ggacgacgcc cgcactatta tcgagatgcg tctcggggct gcttctactg 7680 catcgacgaa atacgcgccg ttgctgttag gccgttcttc agatgaccgc cgccgtggtt 7740 gcctgcaata cggaggagcg aagcgaacat tgcggtgggc ggggaaaggt tttcagccgc 7800 aaaacctggc gcgcgggtat tatcacgacg atgaactgga taggggtatt gccgcgttac 7860 ttaaaggccg tgcacaccgc cgttttgacg tggcaaaact aacagcgtct accgtccgaa 7920 gctgcattat cccggaagcc gggcgtaagt ttgttgttgc ggattactct aacgtcgaag 7980 gccgtgggct tgcatggctg gcgggcgaag aaaccgcgct tgatacgttc cgcgccgggc 8040 tggatattta ctgcgtaacc gcaggtaaga tgtttggcat ggacccagat tacattaaga 8100 aagaacgtaa agatttacgc cagattggta aagcctgcct tcaccgccat acccaagttt 8160 tgaccaatgg cggatttaag gatattatgg cggtcacatc gacagataaa gtatggagtg 8220 gtgaaaaatg ggtgaataca aaaggcgcgc accttatggg gtggaaaccg gtgataaatg 8280 tggacggggt gctaatgacc gaagaccaca aaatcttgac gcattcctgg aagcaggcaa 8340 agcaactcgt ttcaaacaaa tatatgatgg accgcgccct ggagataggt atggacgcct 8400 ggttatcctg cgcgagttac caaaacgaca aggccaaaga caactactcg tccaatgtga 8460 ttgtggagag atgcctgggc gggtctatta tgacaatgtc agggcgggta aaaccactca 8520 atgcaaccgc tgcgcccttg aagcaacaaa aaaatatcgt aaacagtatt tctgctacga 8580 agacgcaatg ccggatgatg cgcacagaac gcgattactc aaccggctgt ccgcggcaat 8640 cgtgcggaca acaagccccg gaaacaagag ttacaaaaac tatggggcta ggggtatcac 8700 ggtatttgac cagtggaggg ccgataaaag gtcgtttctt agatatgttc aaactttgga 8760 aggctgggat gacccaaacc tcgaaatgga ccgaatcgac acagacggta actacgagcc 8820 tggcaacatt cggtttgtca gccgttctga aaactgccgt aacagacgtc gaatacctga 8880 gcttcaacga aagtatgatg cagccgttgc ccgcattgct gagcttgaac gggaaattaa 8940 cctactgcga gccagtttac gacctgattg atgtggaaga cggcaaccgt tttcttatag 9000 cctcagactc gggctttctt gtggcgcata actgcgaact gggcctcggc tacgagggag 9060 gtgtcggagc attcgttacg tttgctaaaa acctgggtct tgaccttatt gagatggcaa 9120 aaacaatgga cgggactttc cccgaccaca tctgggctgc tactgcacgt gggtatgagt 9180 gggcgcgtat tcaggaagcc aagagaccac cgcgccctgg cgaaaaggat gacaggccat 9240 cgtatattct ggataagaaa gtgtggcgta cctgtgatgc catcaagcgc atgtggcgcg 9300 agtcacaccc tgaaacagta gcgttctggc gcgaccttaa agacggaatt ttagccgctg 9360 ttcgtaatcc tggtcgtgaa ttttgggctg gggcacactt gcgccggaat ggtgaaaggg 9420 ctatccgcat atggcgtacc gtagaatttg attcgtcggg caggaaggtt cctggctggt 9480 ggttgtgtat ggagttgccg tcaggccgta tcctatcgta tccgggaatc ggcgttagtg 9540 tgacaaagga aacagacgaa gacgggtgga taaacaccag tgtaagaatt aagtaccagg 9600 gtgagaacca gttaacacgg caatggacca ccctgtacac acacggcgga aaggcttgtg 9660 aaaacattgt tcaggcgttg tgccgtgact tattggccta tgcgatgctt aatgtagaag 9720 ccggtgggta tccaatagtt ctttctgttc acgatgaact ggtatgcgag actccggata 9780 catcagatta cacggtagct gaactggaaa aactaatgtg tgcattgccg gagtgggctg 9840 atggttttcc tcttgtagcg gaaggtgcgg agttaaaacg gtatgctaag taaactgatt 9900 atttattatt tcagggaaga agactgtcga atctgtccgc gctgtgggat taagcacacg 9960 aaacgtgagg ggtgcataga tgagtacgcc tgagggccgc gtccagaaat acgcaaaaga 10020 ccgatttgag gccctggggg gcctcgtacg caaattgtcg tacgaaaata gagtgggcgc 10080 tcctgacctg ctggtgattc tccctggtgg catcatctgg ttcgtcgagg tgaagaaaga 10140 cgaaaacacg aagccagacc cgcaccagtt gcgcgagcac gagaggatgc gtaaacgcgg 10200 cgcaaatgtt tttgttgttg ggtcgaagaa acaggttgat aaattaatag aacactatta 10260 tatttagtta acaccaaaca gaaataagga attgagaaat gaaatatgaa tatgaccgca 10320 aaccagcacg tgacatcgta ccaggcgata tgattttcaa cgttaaaacc cgccagccgg 10380 ttgccgttga tactgtgttc gtcgagtcga atggcaaact ggttatcgaa gatgtaacgg 10440 gtaacgttac ggcgttcggg cggaaggaac tggtactggt ggcgaaatga gtaaattcac 10500 tagaaggcct tatcaaaagc tcatgacctc gttcatgctg cgacacccgc gctgtaatat 10560 ctgggcaagt atgggcagcg gcaagtgcct gaaacgcggg actgaagtta ttatgtttga 10620 cggtacgacc aagaaagtag aagacgtcat cgtcggtgac gtgcttatgg gaccagattc 10680 aacgcctaga aatgtactgt ctctcggtag aggccgggag atgatgtacg aggttaaacc 10740 gcgaaaaggc gagtcgtata cggtaaacga aagccatata ttatcactgc gaactactac 10800 aggtatcgca aaaggttcat ggccagataa caccgttttc gacataagcg tccgcgactg 10860 gcttaagttg ccgaaatatg taaccgggcc taatgggtat cttaagggtt ggcgggtacc 10920 tgtcgatttt ccaaggaaag agcaggacga ggctttattg cccccgtacc ttatggggct 10980 atggctgggc gacggtactt cgtcgtctgg ggccataaca tcaggagaga acgaaaagga 11040 aataagggcg tatcttgagt cgtacgcagc acgtaatggg atgcaaatac gaaaagaagg 11100 acttacatgg agtatatctc acggcaacac aggtcataag aagcacgggt ttacgcacgc 11160 gctgaaatcg gcgggtgtcc tgaacaataa gcatataccg cataattaca aatgcggcga 11220 ccggagacag cgtttagagt tgctggcagg gcttctggat agtgatggtt actgcgattt 11280 gtctaaagca ggttttgact ggatttcggt aagcgagcga ctcgctgatg atttttgcta 11340 tctatgccgt tctcttggtt tcgccgctta caagaagaaa actcgtaagc gttgcgcaaa 11400 cactgatgtt tggggggatt acttccgcgt atcggtatca ggcgattttt ctgaggtgcc 11460 tttcgtgcgt ggtcgccacc agaatttacc gaagcgtaat attaataaaa acgtacttaa 11520 cgtcgggata gaatctataa ccccggtcgg tgtagatgat tattacggct tcaccatcga 11580 cggtgaccac cggtttttat taggtgactt caccgtaacg cacaacacaa gctcgacaat 11640 gtgggtcctc aaccgactgt tccgtaatgg gcaacttaat gacgacgacc gagtgttaat 11700 tctggcccct ttacgtgttg cgtcaggtac gtggccggca gaacaagaga aatggaactt 11760 cccgtgtctg agtgtagtag atgcaactgg ttctgagaag cgacgcatcg cggcgctgga 11820 gtcagacgct aacgtggtat gcactaacta cgaggttatc gaatggctta ttgactacta 11880 cggcaaagac gactggcctt ttaccgttat agttgccgat gaaagcacga agcttaaatc 11940 gttccgcagc cgttcgggcg ggagcaaacg ggcaagggcg cttagcaagg ttgcgttcgg 12000 taaggttaag cgtttcatta accttaccgg gacaccatcg ccaaacggct taaaagattt 12060 gtggggtcag aactggttta tcgacgcagg tgagcgcctt ggttcttcgt acaccgcatt 12120 caccgaccgc tggtttaatt cagtacagaa aggaaagtcg gcgatggcac gggagtacca 12180 cgcccgcccc ggcgcggata acgagattca ccagaagatg aaggatatca gcctgaccat 12240 tgatgctgcc gagtggttcg gttgtgaagc gccggttatc gtaccggttg agattgacct 12300 gcctaagaaa gcacgccagg cttacatcga tatggaggag aagttattcg cggagctaga 12360 aagcggagaa gtcgaagcgg ctaatgccgc tgctaaaacg tcgaaatgcc tccagattgc 12420 ctccggtgct gtatacgtat cagggccaga tggcgaagca accaaagact gggagaaagt 12480 gcacgacacg aaactggacg cactggagtc aattgtggag gagttgcagg gcgcgccgtt 12540 gctggtggcg tatcagttta agcacgaact tgagcgcatt cttaagcgtt tcccacaggc 12600 gcaggcgttc gccaaaggtg ccaagggcaa taagcagatg gaagcgtgga accgcggaga 12660 gatacctatg ctcctgatac acccagcctc cgcgggtcac gggctgtcgc tacaacacgg 12720 cgggcaccac atggccttta tgagtcaggg gtggaacctg gaacattatc tgcaggttat 12780 cgagcgcata ggtccggtaa ggcagatgca atctgggtac aaacggtctg tattcattta 12840 taaaataata gcccgagata ctcttgatga agttgtcgcc gctcgtacgg acgagaaaag 12900 gtctgtacag gacgaactac ttaattatat gaaacgtagg agtaagaaat gagcaaggta 12960 tcttacgacc ctgaaaccgg tattttcaca tgggtagaaa caaaacgtag gggttttatt 13020 ggaaagaaag cgggatgctt ttgtccgagg gatgggcaca tttgtatccg aataaacgga 13080 aaactagaac tagcacatcg gttagcatgg ggtgcctgca atcccgataa ccaacttaca 13140 ccacaggagc agatagacca cattaaccat attcgaactg ataaccgaat atgcaacccg 13200 cggaaagtta ccaacactga aaacagccgc aacgcatcaa taggcgtaaa caatacgagc 13260 ggcgcactcg gagtctggtt cgagaagcga cggaataagt gggtcgcaga aataaaggta 13320 gaccgtaaaa agatacatct cggtcaattt gaaaagtttt ccgatgctgt agcagagcgt 13380 aaagctgccg aagttaaata cggattccat gaaaaccacg gcaagaagag aggtgtgtga 13440 tgggaactta cattggtaaa tcttggaaag atttatacga tagaatcaag gaactcgaag 13500 ccgagaacgc cagactacgg cagcagcgag acgcggcgaa cgcgcaatta gactggctac 13560 ttgagcaacc ggaagttgac aaagagaggg gcgctaaatg aaatactcaa ttggtgtgaa 13620 atgcttttta ataggctacg gattggccgc ggtcgcttta cttatcgctg ctgtggatct 13680 gttatcataa attaaggccc cttacggggc ctttactatt tatgccaaag ttaatcttgt 13740 atagcttccg tcggcccgct taactaacgc tttaaggtta tcgccctcca gatacagtga 13800 tatagacccg ttatccctga cagcgttgtc tggtaggacg gtcgtcgcta ccggtatctc 13860 aacagcacga taatcgcccc tatttatttt taatgatacg gcatcgggca agctccccgc 13920 aatagcggta atttcggtcc atgctgagcc agaacccggc ccgccgttaa tgtgggagta 13980 cacggaccca ctatcaaggg tttttgacag tttatgaata cgaaaccgaa gcgccgcaga 14040 atcattatcg aaactgttta tgcgagagtt accaagacct ccctccatca aatcggctac 14100 gttaatcctt gaggggtcca cgaagcccgt tagtccgctt acagtggagt taggtgcgtc 14160 taccgtcatg ccctgttcgg tagttgaacg gatgccgacc aaacgaaggc cgtttacacg 14220 acaggccccc gagatatata tctggttcgc agggaagttt ttagtattgg tgtcgattat 14280 tgcgatatta gtgaatactg tttcgtgggc aagaatataa gccccagaac cagcgcaatc 14340 ctcgacggtt atgttagaga catacagacc ttgcccgtcc ataccgaatc ctacccccag 14400 cgaacctcta acaaatagat tgtctatcaa atggtttaaa gggagcatat gtaccgggta 14460 ctgagaaatg gggaaatccc ctgggcggtc atcttcgggg ttcatgtcag tatctgcgcc 14520 taaatcgaag ccgtcccata cagggtacag taccgccgaa tcccggaatt gcaggttgta 14580 gttacgagat gtcgtagaac ctaccgtacc ttgccacgtc ttaacaccac tttcccctgc 14640 acgatacgag gtaaacccga tgaccccgcc atcgcgcgcg aaaccgccat tgttgcgtaa 14700 gaattgcgcg ctacttaccg aaccgtaact tgtgcgcccg ccgataacat agttaccttt 14760 gccccaatct ccgcttaagt tttcgaaggt gattacgccg tgtgcaccgc cgctcgggtt 14820 gtcagcgtct accatcttac agaaatggca tccacggaac aggaaacacg ccataagacc 14880 actcgcccgg tgaacctcga cgcctgtaca ttcccgaatt tccagggtag aagatatgct 14940 ttgcccttta gcttccggag ggagaaggga ttctatgcca gggaacttgg cgtaatcgtt 15000 taccgtcggc tggtatccat ctgttttaga ctgtttaagt gtggccacga ttgccgcggg 15060 gtcggttatc cattgattat cgtcagtcca cggtttaatc acccacggcg ttgtagcact 15120 ctccataaag ggggctatta caatggaacc tttacctaat tgcgtaaata ccaggtttcc 15180 gtcgccgata aacttcgctt tacagtcgat ggtcagggtc ttcccgctga agttaacggt 15240 ctcgttatta ctgaaagtgt aatcgcggtc gataagaacg ctatcaacgg cggcatccgc 15300 caattgttgt aacgtagtaa agtcggacag cttaacggag tatttaaatt tcttatccgc 15360 ttccgcgcga aaggtaatat ctccgaacgc gacccaagcg gaatacgaaa tgccgcctgt 15420 cgtaaggggt gtcgacgccg cagggataac tttcggcagg gagccacgcc atgcgtagta 15480 gttaccgtct ccaccatctt ctttcggcca aagaaccgct ttattggcgt cgtttacgcc 15540 gagagtaccc ccggtcgtaa agttaaaaga tgccggggag aaaccagcat cacgtaagac 15600 tgcgggaagc gtcttctgcg tctgcccggt tacctggtta gtggcgtagt cgatatccgc 15660 accaccagct acaccgcctt gtttaccggt gatcacctcg gcttcgaaaa tctggtgttt 15720 tttagctatt tgtaaatcat taagtgacaa tacatcaccg caaccgctgg acatatggct 15780 atcctcttag ttaaaaccgt tatcgaatcc gttagagaac gcacgaccga aaggaggcac 15840 gttgtcaaac ttatagaaat ccttgtcata gttaaacccg gttattttga ctgttctgtc 15900 atcgccaggg tctaccgtag aaacaagaat catctgagca ttatgccttg cttcgttgcc 15960 gaatgaaaat tcagttttca gggcgctatt acccgtgtaa atagcttcct gcggcaccga 16020 cgtcataatt acttgtcggt catgtgcacc agggacgaca cgaacacttt ggactccgcc 16080 atcacgtaac tttagaacca aagaatggtc atctccggat gtgaacttaa ccggctggga 16140 tagctctacg gtaagaccgt taaccgcggt tacgtagccg tcgtaagtag ccatacgcga 16200 acccttaacc acactaacgg cacgattcgg aagggcaaaa ataccctctt cagtggcagt 16260 aaacgacacc gaaattttct ttaaaatgtt cttctggtgg cgacggtttg ccgcccagaa 16320 agcctgctta tagttgcgga tcccttttga gtcataagtc tccgttttaa ggcccccggt 16380 ttcaggtatt gttatggttt ctttaacgtt tgttttaggg tctatgtacg agaatttaag 16440 actatcgaac acctgagcat cgttaaaggt tctggtccat ttttcggaag taccagcctt 16500 gcttcggtgg gtgaacacca tctcaggacc catgcgaggg cgttcaaaat cgagaaggat 16560 atccgcccca cgcctatatg gagtacagaa gatagcgtcg gctatagtac taactatatc 16620 ctgcatagtg gttttatagt catcaaacgt gtaacaaaat tctccagcct gtttgtcgcc 16680 aaaatacgct tcaacctcgt tctgcacggc aagtaactta tccatgttgc gtactgtcag 16740 gtttaaaccg ccaacatccg ggtcacgcgc caggcggatg agggattgta cagcctgtgt 16800 attaggtgtc atcgtatctt cgaaaacacc gttaccgagg tatttgtaca ccatttcagt 16860 agcaatcata cgcaactgcg gttgctttac ttccgcagca cgtggggtct gcttgcgcat 16920 cgagtgtact gttgttctgt tgccgtagtg tggggtgttg tcacgtgttt gcccgtataa 16980 gttaacgtac gttatctcat caactacaga cccctcgaaa ttaaagtcca ggtctgttac 17040 ccttctggcg cgcacccgca cccgtgacac aaccggcagg tcggcgtaaa tagtgacgcc 17100 gttgtagtcc ggtgtacgcc cggacacagt tccttgcgca gtatatatcg gtccgtaagg 17160 agtgctgttt tcatcaagca attgatactg tatttcggcg gtgactgacg ccagtgtttt 17220 cgatttgccg ttgtctttat acatgccgtt cgcggcggcg atattggcga ctatacgctc 17280 aacctcggta cggtttattg aaacccaatc ggttaatgtt ttctcatacg tgttgtctgg 17340 ttgcagcgag gcctctccat caccgcggaa aaatgaaccg ggtcttattt cttgccatcg 17400 cccgataagg ccgccaggca cgataaaggt aacggtccct tcggatacgc ttaacacctg 17460 atacccgccg ccggacaagt cgacgtccgt atttgtccac gctgacacca acgaaaacgt 17520 gtcattaggc accaacactc cagaaaaatc cgaaccacct gttgggtcgt atatcgtgcc 17580 gatgttcccg gacaaagacg gatacccgga catgtaagag aaagaaatac ccagaccgtt 17640 gggtgcttta agaaccacac catcgacttc gttagattct acggtaatat acaagccctg 17700 ctctatcggg tctccgacca tgacctgggg tgtagacgtg ttattgggtg acgtatacgg 17760 cgcgtacacg gcaaccgacg tgcctgttat atccgataca cgggtgtccc cgtcagtgat 17820 atcttcaggg cgtatgtcga gatagccgcg accggcgtcg taataaccgt actctacaat 17880 tttaccggcg gcgttaaaca ccttgtaagt agtcataagg ttattgggga tggtttgcac 17940 cgtcccgcag atgtcgtaac tgcgctcgta cgggcgcgcc ttgttgttgc ggtcggtgag 18000 actgttgttc ggggaatccg cctgcgagtt cgccaggtta gaggacgccc ctttcacgga 18060 tggtgatagt agtttcgcca gcggtttaag gataacgcta aaaaccttca tgacgccttt 18120 aatagccccc ccaccggcag attctacaat gtggaaagtc gcgttttctt tcagtgcttc 18180 gaagtcttct gttacgtcgt tatcttctcc gatttcgtca atgaaaacac gtaccggaac 18240 cccgtccgga acatggttca cgacgaagtt catcggatta tcgtgataac gtttagcgtc 18300 aaatgtgccg tcttcgttgc gggtgtagtg gattactagc gccaaaattc aatctccgaa 18360 taagtatctt taaggtcttc caggctatcc agtctgacct gtctggacgc cagctcacag 18420 tggctaacca ttccttcgta atatacccca gcatgccaca ctattcgccc cctatgtttc 18480 actcctagta gaaccgcgtc aaaattctgc ggtgtgaccg ctcgtaccag acctttcgga 18540 ttagagtggc catcgtcgaa ggcggcgtct attgccgttg ggctggtgac gtcaaacatt 18600 ggagttgata acccagcatc cgcgcggacg ttgcggacgt gatgccagca attacgacgg 18660 cgaaagtcat acggcaagcc agtgtaatca ttaatattca tgatgtcagt atcccgcgaa 18720 gtaaaggaat ctcctccggt gtcatcaaaa taccggtagc ccgctggtta agcatcggcg 18780 taccggtttc cgccgtaaat acaccttttt cttgtgtgag tgtttgcaac tcataaacca 18840 ccgggccgtc gcatggatag gataggtccg tactcacata gcgacggaag atgaatttag 18900 gcagttcctt gttgtcgtaa ggtatacggt ccatttcctc atccagtatg ttaagcacat 18960 ccggaagcga aaaggacgcg gtctggtcca tatcgttatt gttggcggcg ttcttagcct 19020 ccatcggcgt accttcaaat gtcactacct ccccggtctc aagagttgcg gttaagtctt 19080 taactccgcg cacaagtaac caccgtttcg aaagaagtgg gtggtatatc tcaagagtaa 19140 taaaatccat ttcaccgtct ggattggatg ccagtttacg tcggtatgcc gcttctactg 19200 attcctgact cattgcatag ggtcccagat tctcgggaat gtagtctgat acactccata 19260 ggttttaagg aattcgccaa ggcaatcgcc atagcacccg tacaaatcgg gtaggttctg 19320 gttaaggcaa gcgttgttgg cttcctggaa tggcgacttc tcggcggtgg ccgtgaaggt 19380 gattacccaa ttctttccgt catctgtgga ctcgttccac gtggacgtta acgttacctg 19440 gtaatcgtca ataccctggc caaggtcgtg tttcatccag aaactggaag cccctccgtc 19500 taccttctcc atgaaactga ggaatgcttg tctacccagt ggcgacacga ccagcgtaac 19560 gctaatcggg aacacatcaa agtatgtatc acgcccctgg cgaactccgc caccggccag 19620 gtctacccgc cacacgttgt tgccgcgagt catcgagtat cctttcgata ctatcggcct 19680 caaagaggcc gggaaatggt aatcgctcat atcaatatcc tggttggccg cgtgtagcgc 19740 gacgtgcctt agaaatagcg ctgttactat cctgtagcgc ggaacttacg gtttcactga 19800 ttataatacg taaacgacct tcgtcgtcgc gttctgttgc agccgaatca attcttccag 19860 tggtgttgtt tacaatagta acattatccc cacctgattt agtgccattc tcgcccataa 19920 tctgtcgcat ctgctccgcc gtccggacac gggaggcgct agcgggcatg attacttctg 19980 gtttaccacg ttctgcaata gtggatatct gccctgcggc caggttacca ccctgttctc 20040 gggcggagcg aatcttacct acgtttgcta aaccagccgc caccgcagca gccgcagcaa 20100 caggcgccag atacgggccg actaccggta tggctgctgt tgatttatac gcttcgaccg 20160 ctgatgtgta cgtagctatg gtcgcctgag caattgcaaa tgctttgtat gccgtggaag 20220 cctctcctag cgcagccccg atgttactag ccatgttacc gaaagccgcg ccggtcgcct 20280 tcactcggtc aagggagtaa gattcatcca tggcgcttaa cgtttgttgg taagtttcat 20340 cggcgatcaa accctgcgca tgaaaatcgt taagtttttg tagcttaacc tcgtactggc 20400 ggtctaactc cgcgaactct ccttcagcaa gggcttgcat ctgagcgacg tacgcgtctg 20460 cggagaactg tttttccagg cgctctttct cgcgtttatc taattcagcc tggcgggccg 20520 tatccgcctc gagcataatc tgcgtcttag cggtttcgta ttgctggtca ctaaggagac 20580 ggttgctgta gaactcctgt agctttgtaa gttgctcttc ttggttagct tgtatcttct 20640 ggaatacgtc gtcgttaagg tggattgact gataatagaa atcctcagcg gctttcttct 20700 ggcgttcgaa tgcgtcgatg gatttcttgg cggccttgtc tgcggaatca cctgtagaac 20760 cgctgtcctt acctgatacc ttaaacttac ctaacccctc ggccttagct gcttctttat 20820 cgatgtcgta agccgctcgc ttcagcgcga taaggtcttc ctgagcttct acttgttcgc 20880 ggaaactttt ccgtgtcagg tcagccacac ccctttgagt atccatagca agcgctaact 20940 ggaagttaag gttcttggcc tcagctaccg actctcccgg cgtcaatgca gcctttacaa 21000 gacgccccgc atcctgagca cctttaacca tatcttcaaa aaaccgctcc attgctgtaa 21060 attcctgggc aatgaagtcc agcgcatccg ccgctgactc agctatcgta gatgcggtct 21120 cctctccgct tccggtaatg ctttcactgg tgtaagtcca ggcggagttc aaagcgttta 21180 acgcgtctgc aaccatccca aacgccgcgt cgaatgaggc ctctatcgca cccccaacat 21240 cggatgccat gtcaccccac atccgcatct cggctacgaa ctcaccggat gcgacctgag 21300 cgtttatctc ctgtatcagg tcgtcaatat aacgtaaagg ctccgccagc gaccctacat 21360 ccagactagt atcgagagtc atcttcaact gggaccatga gtcctccgca ctagcgatag 21420 ctccgtccag ggtattagcc tggtcagcca tcgctccggc gaagttaacg ttacctatgt 21480 tgagaagata tcgctcaata tcggcagcgt ttttcttaac tacagtagtc gtcccctgga 21540 aggtaaattc gatatcactt tggttctgct tggccttaat gccgaattct ttcagacgtt 21600 cgaattcaaa ggtgctcgca tccgcgacgg cctcaatcat ctggttaagg tctttaccca 21660 tagccgatgc ggtgttgcca taggagcgga gcgcttcttc tgatggggtg aggcctagag 21720 ctaccagttt acggaaccct tctaccgcct gttcaaggcc gtaaggtgtg tcacgggcga 21780 agtcctgtag tatactcagg gcctgttttg caccttgtgt gcttccggtt agtgttttca 21840 ggctggccga catcttatcc agttgccgtt gtgactcgac tagttcctga gccgccttat 21900 aggctgcggc ggctacaacg gcagccagtc cggtgaccgc ggcccccgca acttttgcag 21960 atttagagag gttatcaaga cggtctgatg tggccttcgc gccttgttcg gttaccttta 22020 ctactaagct agctacatca gccatcgttt ctaccctcaa aaatggcgtc taagcccatg 22080 ataatttcag attcaaacat ccctatattc tgacctgaaa tagtgctata cgccactaaa 22140 tctgaccaac taagttgctc tcttgggtat agctttacgc cgtcgtcatc gacgcggcgg 22200 gtaaatttaa catcacgata cttctcaaaa gtggttagta gttcaggggg gcattcaggc 22260 ccggcgtcct gcgttgcttc ttccgcgtct tctattaccc ccatagcgat aagcgcggct 22320 ttgtgcccgt cggctatact gtcaaacttg cgcctttcgt ggcgggttat gaagttccaa 22380 cgggcgaact gaaacaacgc gtctactttt cctgcaattg gcgcaactgt tcgtggtgga 22440 atacgactac atgttccgcc aggcgcttat attgggtgag aagagtctca agattttcct 22500 tagtgaactc atcatcgaga ctccaaccat tcaccaattc gagagccaat tgccggttta 22560 ggtctcctgc cgcgtcttcc atttttaaat tgtattctga gaagtctttt tgctcttcgc 22620 atttatctcg aagcggtttt aatttaccta ccgccgctcg gtacgccagg gtgaaggcgc 22680 gcatagcttt aaccgcaata tcggcctccg gagagaccac gttcaaccat tcccctgaat 22740 ctgtaccatc ttttaaagga atcggcatgc gcgcgccttt ctcggcttct gcttcgtagt 22800 aaaaatcgct aagtttcata gtaatgccct ttggttagtc ggttactggt tggatagttg 22860 cgtcaggcgg gaaccacccg cttttctggc gcgaccgtag acgctaaata agaatagcaa 22920 aatagcttgc gcaaccgtta atagtgtcct attatctcta cgtcaactaa acaatatgag 22980 gttaagaaca tgagcgtttt tatcggtatt atcgcgttta cctatttcat cccgttcctg 23040 gtagccttgc tacgtaatca taaagcgaaa ttaggaatat tcgtatgtaa tcttttactg 23100 gggtggattc ttctgccgtg ggttggggct ctcatctggg cctgtaactc taatgttaat 23160 ggtaaataat ctggctaaat taaacttacg ttattagaat cagatttatc ttaaacagtc 23220 aggaaacaag catggtcact cgcaaaataa cagatgaaca gttgcagcaa gagctaaatg 23280 ctggcctcgg ccctactgaa atagccaaaa agtataatat gtcccgccgt aatgttcagc 23340 ttcgttccgc gcgcctggca aagaaaggag ttggccacgg gcgcgacgta agccaccttg 23400 tgccagatgg gtataagatt aagggtacgt cgtcactggt agatgagttc ggtaacacga 23460 aacttcagtg ggttaagact gacgccgacg ccgagcgtca ggtcgagttg atgcgagccg 23520 taatagatgg gatgaagtcc gatattacgc cagtttcttc ggtccctcgg cctaaaaagc 23580 gactaaacga aaagttgcta aatctttaca cggtttctga cttccattta ggtatgttgg 23640 cctgggcgga tgagagtggc gacgactggg atatgaagat tgcagaagac ctgttctcga 23700 gatggttcga cgcggcgttt cagaaggcgc ctgatgccgg tgtgggtgtt attaacctat 23760 taggggactt tgcgcatttt gacagccttg atgccgttac gcctgccagt ggccatgtac 23820 tggacgcgga cacgcgttac cagaaactgg tgcgctacat gattcgtatg gtcaggcgcg 23880 tggttaatat ggcgcttgtt aagcataaaa atgttcatct cctaattgtg caaggaaacc 23940 atgacgagtc aggcatgatt tggttggctg aaatgtttaa cactctttac gataatgaac 24000 ctcgtgtttt tgtagataca tcggctgatg tctacaagat ggtccaacac ggtaagacga 24060 cccttttctt tcaccatggg cataaggcga gattcgatgc tatcgaacca gttatgatcg 24120 ccaagttccg taaggcgttc ggagagagcg tttacagtta cgcccatgtg ggccaccttc 24180 accatcagaa gattgtggaa agccgtaaca tgattgttga acaacaccgc actctcgcgg 24240 cgaaagatgc ttacgcatca cgtggtggat ggatgtcagg ccgcagcgcg aatgttatta 24300 cttacagtgc cgaatacggc gaagtcgcgc gtttaactat ttcaccggag atgcttggat 24360 gactaacaaa tacaatcgca caatgacaaa tactgacgga gatagcatta cctgtgatgt 24420 gtacgacgtt ctgagggcat ttgatatccg cgacccggca ctacagcatg cgttgaagaa 24480 actgttgtgt atgggcttgc gggggcacaa ggacacagga accgacttag cagaagcaat 24540 tgaaagtctg gagaagttac ggaaataccg tagtaatatt gatgagtgag aaaaaggccc 24600 cgaaaggggc cttttttttt atcgtcacga tatagaatct gatttgaacg ataaggatgt 24660 tcgaagttaa atatatctac accaacaaga ggggatgaaa ccatgaacgg attaatgaat 24720 gttagtgaag cacaaacaat gtccagccgt gagattgcgg aactgacagg taaggagcat 24780 aaaaatgttc tggctgatat ccgcaaaatg ttgtctgaga ttcaatcggc tgaaaagtca 24840 gccgattaca aagacagcaa ggggcgcacg tataaaatgc ttcttttgga taaagaggaa 24900 acgttaatcc tgatttcagg gtacagcatc aaaatgcgcg ctgccatcat ccgccgttgg 24960 caggaactgg agtcacaagc gcgcaaacca tccttaccgg tgcccaagac aatgggggag 25020 gctttaaggt tggctgcgga cttgtgggag gagaaagaac gccttgcgct tgagaacaag 25080 gaaatggcac caaaagctga tgtctacgac cgcatcatcg acagaaataa cttgtacaac 25140 gcgacgcagg ttgcccagaa gttcggtcaa tctgccgtgt ggatgaataa acaacttgaa 25200 caatttggcg tatacaatcg ctccgtaaaa cgcggtcgtg tcttccagca atggttcatc 25260 gataagggtt acggcattat gcgtgaaaca gaaactgggc attcacaggc tatgttcttc 25320 gctgaaggtg aaatgtggat tatcggtaag ttaacggcag aaggcttgat ttaaaactaa 25380 ggcccctttc ggggcctttt tcttaggcgt acttaatgcg ctggataaca attgaagact 25440 ggaactggtt gccggttgcc tggccctcaa gagacagggt aatagactcc gggccgccga 25500 tttctggggt tgcggacgta agtgaagcac gtttgagtgt aaagctcata gcgccatccg 25560 gaccatcaag aaccgaagat acctccattt ctgtttcatc caggaattta tttaacagcg 25620 tcaggtcgta cagtttacct gccagcgaga aggtgttggc cgcacgactg cgctcaacaa 25680 atgccacgct gttatttccc aactcgaact gtgcggatgc cccgttgtcg ttagtaatag 25740 tgaacgtgtc aataagttta agcggtgcag caccgtcgta aacagataca tcaaccgagg 25800 caaacgggcg ggcgctaaaa ttggtcgtga agtctgaacc actcggcggt gcttgtaaga 25860 tttcctgatt caagccgatg aacgggaaag aaccggtgac cattgcgtta acagcctgtt 25920 cgatagtgaa cccggtaaac tcaacgcctc tggttacgat atatgaatcc gggtttccgc 25980 acttaccttt caaccacgta agaatcgaat aggtcttaca caagttaccg gtttccagtt 26040 tatctgcgat tcgcaaatcg gcttgggcgt ctgcttgggc ggttagagtg tgctggatgc 26100 cagcaccggt tacaaccgta gcggttaccg cggtaacgag gaaagcttta tcgttattac 26160 cggacagacc atcgaattgc accaggtcac ctacttcaac accatcggtt acaaagctac 26220 cggtagcccg tgtaaaagtt ttcgctaccg ggtctactgt gacgccgatt gccgatgcgg 26280 tggaacccgc tacccatgaa ctagccattg cgcctgctaa cagctcgtct tggctttccg 26340 cacttagctc aatagcgtac tcacctgtta cctgacggtt gccagtgcgg atagctgatg 26400 tttcgttgct gccgtctagc tcattagaga tgagggcgtc tcgggttacc gcggggatgc 26460 cgcctgtgtt gcggagtggc tgccataccg ggttcgctgg cgtcaccccc ggcgtcgtct 26520 caagaacgta aaattgcgcg gtcatcgcgc ctttgtatgg ttgtaacgcc attatctaat 26580 cctcgctgta aaggctatga aattaattga gagaggcctc ttcgcccatc cattctcaac 26640 aataagagga ccgaggctta ctgattgtac ctcagcacag atttcgttac gactaaaaca 26700 gttaccggcg gcgaaggcgg cgtttagctt atcagctaat cggttaatag gagcgctacc 26760 caaggccgac ccaacgttaa tgtccacctg ataaacacct gcccgttgtt ccgtccagaa 26820 caagtcggct tgttccgtat cagacaatag catataactc gcaagatacg gtgtatcagt 26880 agatgtcggc gcatcgatgt tctctaacgc aaccttggta ccgttgtcca tgccgaatgt 26940 caccagtgct gtgtcaaatg ctttcgttaa gtcctcaaaa tatgttgcca ttatttcacc 27000 ttagaggctt cttcatttag tagttgctgg aacctgctga cattaaccct aacgaagcct 27060 tgaggcgcct gttgcgacca accatactca aggcgctgcg catacggcaa attgttagtt 27120 agagtaaacg tgtgccagtc agcggcattt agcacgaaat tagcggcatt actggtcgcc 27180 gtggcgccag ctttatcggt agcgtccgta gttccgtcgg cgggagtacc gccggacgcc 27240 atccagttca tacgaaagcg acctgtatct acaggactcg ctttgataat cgcagaaaac 27300 aacttgatag atacctgacg aattaccttt tcaggattct tcttagcctt ttccacgaac 27360 ttggacacat caagagcaaa actcattttc tcacctgaat aaagtacgct acaacgtcat 27420 cgttaaccat cttcttctcg atggcgacca cagaccactg ttcaccattg aattgtactt 27480 tatcatccat cttaggaatt acgctgtaat cggccttaac caccatatcc cctgcctgaa 27540 tggtggttcc gtttaccaga ccagcattaa ccggtacagg aacggattta agagggaccg 27600 tcgtgtccac cccccaaacg tactcaccta aaacagggtc ccatgttttc tggcctttgc 27660 gcaccaaagt taccgtgctg ccgtattttg acagtaggcg agtacctacg ccttgcatcc 27720 gtttactaaa agcggtgctc atcacacacc ctccagtctt gaaacgacca gtagtgcgga 27780 tggggccaca ccccaggacg taaccgttgc cacttgcggg tataacccgc cgaagttgga 27840 accggcggca tcacgcataa tctctaccgt aaaagtctgc ccggctgcgg cattcacaac 27900 aacacgagat tcaatgggaa cagtaacatc ggtgctagcc agcttagcga cggcgggtga 27960 accgaattga gcgccggcta gcagaacccg cgacagaagg atggatgttc cgctcgcccc 28020 ggtgcgaccg gcctgtaact taacacggat agcgtagtta ccggcgacgt tgaacgtaac 28080 taccccggaa gcgctaatca ttacagggtc ggcgcttgtt ttctgcgcac caccaaaggc 28140 tacagttaat gctgtgccgg ttgtagtagg cgcctggata tctgtagacg acgcacgaag 28200 tacctccact tccttagccc cgtacagcat agaatccgcc acctgagtgg taacttcgcg 28260 caacttttcc ggtgtaatta gcccggactg gttatcaggg aagtttgccc caattaaggc 28320 gaatatttcc gatttagtct tagccatgtt tagcccctaa aaacattaaa ggagtaggca 28380 ttattgctac cgcataagag aggtcgtagc gcatcatcgg ctgccgtgat gcttactgtg 28440 ccgcctgagt agccattctt aaagtaggac accgttactg caccctcaac ccgttcggtc 28500 tgaacctctc gcccgtctgt agaccctcga acgtcggtac cagcgccata ctcgaccgcg 28560 gccattacct gagcttgaat aactaacgat ggaattacat tggatggctg ggggaacccg 28620 tgtaaagtaa caccggttcg agggaacgcc agcgcctggt ttgcggatac acgacggccg 28680 cacatctggg attcaaaaag cccaacgtat accgcgccgt tgcgtagcgc cgcttcggcc 28740 ttagcatcat cttctggcag ctcaagaccg tacttagagg ccatggacct cgcgtcttcc 28800 aggctgacgt aagagtcagc attcgggatg ccttgaccgg tttccacgat aagcggcata 28860 aattattcct ctacgctttt gcggcgacga cgctgcttag gctctccgcc accgttgtaa 28920 tgcggttctt cggccactgg ctgagtttcg attaatgctt ccgggttcgc ttcttcgcgc 28980 atcgggacta actgcccgtc aacctcgacc acacccttgt attgttcccg tactacgtaa 29040 ttatctgtca tgtctaatcc ttaaagcggc ccggaggccg cgtgtattta acttacggta 29100 actaccgtac tatctgaaat gatattaccg taaccgtcgt gcgctacgac tttgtaagta 29160 ccggaatccg ccgctgcgga gttggcctta gtgtaagtca aagcggttgc accggagatt 29220 gcgttattat ctttatacca ttgaactgtt ttagggtcca ttccgtcagc tagtacaacg 29280 gtcagggtga tagcctgacc tgcggttacc gcagtagtat catttagacc agtagagaaa 29340 cgcaacggca tcacgttttc catatcaatc tctacctgcc cgtcctgcgg gctgtcatca 29400 gataccccaa caatacgacg tttaattaca tcaaccattt ttatcctcct caggatacag 29460 taccagcgtt tttaagggcg gttaacagat tagtaacggc agtgcgcaat gaggttacat 29520 cggtgcgcaa cttattgtag ttagacacca acgcgtcaaa ctcttctttc gtaggagcgg 29580 ctgctgcggc tgcaccacta ccggcactaa tcgctggcgg tgcggctact gtagcggact 29640 ttttaacccc tccgattgtc gttgtggtag cggcgggaat atcgaattcc gcgttaacca 29700 ggctgcgggg taacccttta cctgtctttg acattatgta gtctccgtat aaatgaaaag 29760 agggaccgaa gtccctcaag agtataccct acgattaagc acctacgcca gttaccagga 29820 acgcaattgg cacatgctta cggtcgacta cacggttcca gttggcggcg ttagccaggt 29880 cctgccagct agcggagcgg gcggtagtct cggtgccgtt gccggtgatt acagcgctgg 29940 tgaagctgta accaaacgga tgcagcaacc acgttttacg agtccacagg gtttcaacac 30000 caccaccatt agcgcgagat gcttcacgtt cgtattccag cggcatctca ggattgcctt 30060 cgccgtaacc gatagcaccc tggccgaaga tgatggagat gaatttacgg ctagtgtcct 30120 gaccaactac ggtcatgctg tcatcgacga tgacgcggta accctggtag gtggcgaaca 30180 tggtgttgtt ctcagcatca cggatgaagt cgataagctg ggctttacgg gcctgtgcgt 30240 atacgaagct gtgcattgcg atagcaccga gaacctcacc gccgttgccc atcaatgcat 30300 cacccatagt ctgggtagca tcgatgaatg cacctgcgtc gaagcctgaa gttgcagaaa 30360 cgtcaacaac catgtcgttc tgctcgtgat acgcatcagt agcggatacg ttgtcgttgt 30420 acaggccgag tgcagttgca ataaggcgac gctgcgcctg acgctgccag aagttatcca 30480 gacgggatgc gacggactgc aacggattct ggctggtaag ttctacagtc aggtctgcct 30540 ggccgaaacc ttcgttcaga tatgcaacgc gggccatcat ttcaccggtc tgaatagcgc 30600 gcggggtcgc gatgtcctgg tatacgtcat tcgagtagtt aggctcgata gaagtatcga 30660 tagctttcca gaacggcagg ttagcgatgt tagacggtcc gcgggcaatc tcggcagcat 30720 acggagttgg ggtcaaaatt ccggagttga aaaacgcggt tttctctacc gggtcttccg 30780 tcatatagga cgccagtaca gggatattgc cagttacgat gttaccaata gtggtgattg 30840 ccattattta tttcctcagg gctttaagtt gccgttcaaa ttcggcaggg ttagatttat 30900 agagagccaa ccgctcactt tcactcatat ctttaaacgc tggtgcggcc ccgccgcctt 30960 tattcccgga agccccgcca ccggaagctg catttgcttt aatcaaatgc gaaaaagctt 31020 tatgttctcg caggtatttg cggaactgtt cagggtcggt agtaactacg ttaccatccg 31080 catccgtgaa tttagtaact acgtcttcac cttcaaattc agtcttaacg aacggagcaa 31140 ggatttctac cgcttccggg gtgatgaaat caccggcgaa agaacccaaa acagccttac 31200 gttcacttcc aagaattcgc gcggctaaag aagcgtagcg ttcttctttc tctttcagta 31260 cgggttcata ctgactacga atcgtctttt caaattcgtc cattttacca gcagccttca 31320 gggcctcctg gtgggcgcgc tgccgttctt cttcggcttc tttagcccgg cgagcagctt 31380 cctttttctc gctaaggagc gcttcctgat ttgccttgag cccggccact tctttctcaa 31440 tcaaggcctg cacttcttcg gcggtgtaca tttttggtgc atcacctgca cccgatttat 31500 cttctacccc agcttcttcc tggagtggat agtgtaaaaa acgattcata gtcaatatgt 31560 cccctggact ttaggacacc gggccacccg gtgtttctgt aacaagaata aattattcca 31620 cttactaagg caactattcc agaatattcc ttatgtactc ctgcaaccgg aacactttca 31680 gacgtaactg ccgtgtgcac tccgcgttac gtacgtcaat ggccaggtct tcatcagcgt 31740 cactgctcgg tggaataagc ttgcacggcg gctgcattag tgtcatatcc agggatggaa 31800 ttagcattct cggcggcgcg tcgtttgatt gacacccgtt caggaggaaa ctcacagaca 31860 gtacggccag gacgggtaac gtatttgata acttcacggg tgatagcctc cgaattttct 31920 ttgccttccg attcggcgga ggcggcctta gtatcattct gctgctgccg ttgtgttttc 31980 ttcgtcaact cagcttgtgc cttcttttgt tgttgtgaaa caagattctc tcggccttct 32040 acccacccgc ttctgtagtt gtactggccg tataaccaca aggaaaggat gccggctgcc 32100 gtgattactg cggttacttt ccaattcata gtatcccctt tctttatacg ctaagcccgg 32160 cgtaagccgg gcgttaaact cgtatatctg taaacatcat acggcgaatc ttagataaat 32220 actgaaagca atatagctcg gcccttccga atgcgttcgc ggttacgcta gggcgctgtg 32280 gcgcccgctt cacgcgtccg caccgcggta cgctgaaagc gtaggtagcg taacataaaa 32340 aataatagaa gtctactttt tgaataaagt attcctgaat ttacattggt aaggaataac 32400 tggaataatg gtgtgtaaaa ttaaggcccc ttggcggggc ctttcgttac ttaggttctg 32460 agaccttagc cgctttactt accaccgata tcccgtcggc atctaccgcc ctgacaaaat 32520 actcccccgg gtctttaaca gttagttcac cctcaacgta cgggacgttg atcacctctt 32580 taccattttt aaaccattgc agatcgtaag gagccttacc acctttcacg actacggtta 32640 gtttagcgga accgtctttc agttttgtgt ctaccggttg tgtatcaaag tacacgtcac 32700 ccgctccgtc aagatacggc acctcgtaca ggacaccgtc tgagattgca gtcagcccgg 32760 tcttatcggc gaacggcatt tcatctaccg actcaccaag aacagattca tctttaatgt 32820 acacgacacc ttcgccagca ccagacacac gtgagtactg aacaacccga cgcgccggca 32880 catctttaac tttgaaaaag cccatcttta ttcctctttc aggtatttgg caacacgctt 32940 atcgagctcc gccatctctt taagggttaa cggacgcccg aatccatcta ctgatattac 33000 acgaaattct tccggggtta tcccactatt gcgaaaaatc ttacccctga caggaccaag 33060 tgcttcatcc tggaaccacg ccggctgttg tttaaggaac tcgtagtagg tcgtgcccgc 33120 gtttacttgc tggcctcctt cagcacccct ggccgcccgt tttgcaccct tatccaggaa 33180 gtcgaattct ggactgatta ctggtgccgt ggtcgaccga cagttagggt gcgcgggcgg 33240 tagcgggcct ttgcctattt cgtactgcat cgaatcccgc gaccgacaaa cggctgaggt 33300 tctggagtca agggtagata cccactcgta tttcgtgatg atatcgtcat tccgcgcgta 33360 aacctgttgc cgtgcttcat tagatacgtg cgccagggct gtacggataa ccgtcgccgc 33420 attacgttcg gaaatatccg ccagtccacc aggcccgaca acgttcttaa ctatctgacg 33480 tgtggtcatc ccctgtacga acccagattt aacgcccata acaaggcgat ttacctcggt 33540 ttgccgccag ccactcatta gagaaacaaa atcaactggc ttttcgctta gttcaagagg 33600 cgcgaaagtt gcggcggccc atacttgttc ggcggctggg gttacgaaat tagcgttaac 33660 gttggtcgac agggttttta cgttccagtt aatttcatag tcagaaagct cacgagcgtc 33720 agccagtaat ttctcgtacc atcctgatgt aatcacgttc agttcttgct caagttcccg 33780 cagcatgatg gtaagtctcg ctgccgttct gctgtcatca ccaaacaaca acacctgtcg 33840 cttaacctca tcgcacatct gcttgatgaa tggcgccagg tctttcacct ctccggatgc 33900 ggtgcgttgt aaccatacct ggtggctgat taatgatgta agtagtggca tatatccctc 33960 ctgtttgcgt tgttgatgca tcttagcatg gcataggtta ttcctacaca aacacctaaa 34020 aaagtctaat ttgctataat tttgcgtatt aatgtttgcg taaacaaaga aattaggaat 34080 ataatttaaa cgatacaatt aagaggagaa aaacagatga atgtgcagac tttagaattt 34140 ccgtatggcg aggtggaatt tacacgtgat gggtatatag atgcctcaac attaaccatc 34200 cagatgaaca gatggaggtt cgataacgga ctacaaccaa gaaatcttgc gcagatacta 34260 aaaaccgatg actgcaaaaa ctttatgcgt gtgtgcgagg aggagacagg tattgttccg 34320 ttaaaaacat caagagggag gaacgggaag acgtggttgt gcctgcacct ggccgtgtac 34380 attgcagaac agtatagtgc gtattttcac ttcctggtta tagaccgatt tattacgcaa 34440 cggcaggtag agctgcgcaa cattggtgcg gtgagctttg tcgagcttaa cgctgccgtt 34500 agtcgaatga ttgagcgcac ggagggtagg attgggcata gtggtcactt catacaggtg 34560 gctaaagcca taaaggaatc tgtagatata cgtgagataa ccgggtttga tacatgggac 34620 tcccaggatg caaaaactaa tcaattacgg agtgatattc aaaaggctat ggtgacgttg 34680 ttggatatgg aagcagtcaa ctcatgggat gagctgaaag agacgatacc gcgggttgta 34740 cgtaaatgcg cggctaatat ccgctaaaac taaggccccg taatggggcc tttttcttta 34800 ctcctgttgt tgttgtgccg cttgcggaat ctcccccgct acctgagtaa cagcacccga 34860 cggcaaaggt gcatcttcaa tagcgttcag aataccctcg tcggtccagt cagtgacacc 34920 ggccctacgc aacgcagcgt aataagcagt ggcgggcagt aatccggcat taatgtctgc 34980 catccacgca gccctgtcct gtgctgtcat aggttgcagg aagaaatcca tgttaatctg 35040 gaactcgact tcagtatctt ctggcttacc caacatcata gcaacccatc gtaaagcatc 35100 ggtatacgcc tgacttacgt tacgagcgat tgtggccata acggatgtat cggcgccgcg 35160 ttggatgcgc gcggattctg cggtaatttg ctgagatggt gtaataagtt gtgcgccaat 35220 ctggatagcc tgctgttctt tgtccagcat attctggcgg gccaggttgt tttcgcctgc 35280 ctgaataagt tgagcgctac cgccataacc aaggttatgc ccacaccgac taccaaattt 35340 gatcccattg gggttcgctt ccttgaacga ttgcggggtt aagttctcac cgggatagat 35400 aaacagggtc ggctggccga ctacgaagct ggactcctcg ttatcagcgc tattgcggta 35460 atgacctatg ttaagctctg ccagcggcaa caaaggcgcg tcgtcgatgg tggcgtcgtt 35520 attggtcgcc ccgataaagg taaacggaat tacccctcgt aatgattccc ctaagtccgg 35580 gtaaatctcc acgacgtctt cctgcgcacc accttccgca tcgaaacgaa aaagacgttg 35640 acggtagttg ccgtcgctat cgatgtccag aacgcgatac tgttcgccgt atttcgtttc 35700 gaactcgttt cccggttcgt tgtactccca tgtctcacgc agcacgacca tagttacgcg 35760 gtttacggaa cctacgcgcg tgagtcgcca gttaacgata ttctcggtag tgtagaaagc 35820 gattgtaggg ttgagcagac ccgcgttctg ttcggccgct gtagctgcgc cggtttccgg 35880 ggcgtcaaca agaagaccgc cacgacctac tgagtcaatc tccatgagcg tgtcctgcgc 35940 atgctgtatt aggcctacac cggacccatc tgcgttttta agcaagtatt ccagctcctt 36000 cggaatattt atttcaggtt ctttgcgcat gacgctacca accatccctg acagcgtgcg 36060 ccgggtgaag ttgtatacga tgccgcccgc ctcgtattct gcctgacgcg cttcgccgta 36120 ggctttatct ggctcattca ggccgacgtt acgtaagtag ccaaccaggt ctccggcaag 36180 cgcatggcgc accttctgcc atttcggtgc gtagtgcagc cattcgcggt gcttggtttt 36240 tacgccagaa ccctggccgt ttgctgttaa catttaataa tcctcttaaa gtgcaaaagt 36300 aaccgggata tgggaaacag gtttaaccaa cggcatttca taagcaatag ggtatcctgt 36360 cgcgtcgttc tggtggtcgt tgccgcttgt cttatccggt ataccgtttt tatcgtaagc 36420 ctgttgctcc aggcaacgtg ctgtaaccgg gcaagcctgc tcgttgacca ttaatctacc 36480 agactccagc gctttattca ccgatgctac acggtcttta acagcagggt taaccgattt 36540 cacccgtatc tcgaaaccag cgttctgtaa ttgggcaata tccgacatac tggcatcagt 36600 cgatttacgg ttcttgccgc tggcgtccgg gtacatgacg atgtggtggc cgtgtcgttt 36660 ccaccgctcg gtaatctccc tgaccacatc cggggtatcg cgcatatcta ctaactcggc 36720 taccgcgtgc cagacgtaat cgcgctgaac gtatatcgta gaggccatat gtccgacgtt 36780 gaagtcctga ccgatataca aagtttcacc aggttgtata gtctcccggc tgctgtgttt 36840 tttgcggaca taagcgtaat aaacactgcc cgatgtcagg ttaacaaatt tgccgtcgat 36900 gtaggcgtca attaactggc ccggatatgt atcccgcaac gactgcacat aatcttccgg 36960 tagaaaggga ttggatgttg tcgaggcctg aatcatttca tagcctggtt gcttttttac 37020 ggcccatctg tcgtgcacaa aacggaagcc ttctggcgtc gtaaacaccg atactgtatt 37080 agctggcttc ggggtaattg gacgatatgt acacggcaac tgacggttac gggcaataac 37140 tttattccag gcgtgctcgg cgtggtcttt atttaacgtg tccaactcgt cgatttttgc 37200 gcgaaaagat tcgtaaccga cgattcgcgc cgggttatcc aatgtacgca ggacaaaatc 37260 cccgaattgc ccggatgatg tatagattat gttgtctgat ttattgtact tgtaacgaat 37320 accccaatca gacaacttct cttccatacg cggagcgagg ataaggcgca ccaggtcgta 37380 tgtcggttca tacatggcga taagtgaatc actaccacct tccatgctgt cgagtagggc 37440 ggagttgcac atgacctccg atttacctgt gccgaatcca gcgacgaagg ccgggaattt 37500 acaatgcaga ttaaggaacg cgccttgcgg ttccgtagcc gttatatcaa cgttcaccgg 37560 acaccacctt aatagttact tcgctaattg gttcgtcgcg cgcttcttcc acaacagttt 37620 tattaagccc aagttttgct gcggcgaacg tagcagatat cccggcagca ccggtctcag 37680 tgaaatacgc ctcttccaag gcctgcgccg tttcgtatgc ttcagcgaag gcaggaacct 37740 ctcgcagcca cagcttgata atcggaacgg ttacgcctat gtgtagtgcg aaacgggcca 37800 gcgacggcgg tttgtcctgg ataagcgggc gttcgtcgcc tttagacgtg ggtacgagtt 37860 cccacgacgt gcggtcgaag aaccggatta actcgtcaca atagtccggg tcccacagtt 37920 cggcggaatg cctggatgac tggtacagac tttgtttgcc acgcgggcgc ttgcgtcgac 37980 ggtttgcgtt aaccgcttct tcgtgcgcgg cggccactac ttcggcgtca ggctgtttta 38040 atttgagttt catgcaatcc tctcttaaat gcgtatgacc atcatagcgc atcagacacc 38100 ccctgagcgt ctattaagga ttgtataggg aaagataggc ctgtgtacag aaaagcccgc 38160 atatgcgggc ttgtgtcagt tatataggcc gtgtcgtagt gctcttctac catcgccgcc 38220 gtaatggtcg ttagctgcgg cgataaccca ggcgttcggg atgattagca atctcatttg 38280 aaaaaccaca gcaggatgcc ggccgctcct gatacaccca agcctaaaaa cataggccac 38340 cacagatatc ggggaagact gttaaacaca gtcggttgtt ttattagcgc actgagaaag 38400 gcgaaccaaa acatcaggtc gccagtggca tcggtagcct tcaaaatctc aatcattctc 38460 cactctcctc tttataaacc caccatttac ctactgcttc ccatcgccac ccgcgactat 38520 gtatggcttc gaacctacga ccaaatactg tttcagcata ccacgtaaag aacaaacgta 38580 tcgtacgcca ggcacctcgc ttgcgcccat taatacgcca atactctctc gtcatccggg 38640 taagactgac agcttcgaca aacggccaaa gaaaaaaaat ccacatcagg tatatagcac 38700 cggcaagcag taatgttgtg aatcctatac ccatgagaat gagaaaacta tccatcactc 38760 ctccagcttc atacggtcgt tgcgcgataa tccacgtttc gtgtagatga ccggtgacgt 38820 tttcgtatgg gtcatcagtc ggttgttata gatgacatgt tgctcgcatt ttgtgtcatg 38880 gcggtacttg gacaatgtca tctcattaag tcccgtctcc cggcacgtat ccgctaatgt 38940 gccgtgtgtc tctattagtt ttgggatact ggtaatcatt cctgttcgct ctccttctca 39000 agtcgttcag gcagcatctt gtatgcttct aaatggaatt cttcgttcat tgacaggccc 39060 agcttactta cagaccccaa atacataatt cgctggcgta aagtttctta acatacgcga 39120 ttcatttatt ctccccacct acgtatttgt taaggtcgta gcggttaata agctcacgcc 39180 ggtgttcgtt aaggtgatca agcgcggctt ggtgtgcctt aatctgcgcg tcaatggctt 39240 cgattttaaa cctgccgttt acagcctcta ccatcatctt ttcatagtcg cggtcaacca 39300 tcatttacgc ttcccccaca tgcgattgag atacttgttc tggtcaggac ctgggaaact 39360 attgcgtttt aacagttcct cgcgtgttgg gaacggggta ttgcctactt tcccgcccac 39420 ctttaacgtt tctgctacct gtccgttgtc gctcattcct gcacctccca ctcgcatacc 39480 tttacaccat cataaatgcc tttagctcgt tcctcgtcgg catcagcatt agcttttcct 39540 tcgctacgat aaactccgag aatgttactg ccttcatagt gataaaatcc gtatacaata 39600 tataccttca tttcttctct cctgcataag ctgctttcag tgtctgcatc gcggcaaacc 39660 agaacgcgtc cgccgggatg ctgttgctca ggttactgac ggcaatgcgc gccatcagtt 39720 gtgcgtcttt gaatgcttct gtatctgtga ttttcatctt ctctactccg ttctcgttgt 39780 cgatgagcta aatatagtag tgttatggtt actcgtcaac tattatccca ggatatagat 39840 aaaaaaaaat cccggtaagg gtcagttacc gggataaagg agctagaggg atgaataagc 39900 aggagcaggt ttagtatctt aggaattctc ctatgtgtca acctactttg cttcgcggcc 39960 gatggcttcc gcctgttgcc acggcatacc gtcgaatagc gcaacacgac ccgcggcacg 40020 ccgacgcaat ccaaggagtg atttgccgtt ctgataatgg aactgagtta acttattccg 40080 cagtgtggca acgtcgccct tacgcagggc ctgccctgtt ccggtagaag ccgcaatcac 40140 accggcacca gcgttataca ccaggtcgca catcgcatcg aactgtgact gattgagcga 40200 cggatgcgca acggcgtcta ccgcggctac agccttagcc atgtccctgt tcagtagcaa 40260 aagaccctgg ccttcggtaa ttttctggcc ctctttcaca tctgcgccgt agtggccgta 40320 gccaattgtc aggtacttct cgttcttcgt tgccttgtac gcagttccgc ggaacccctc 40380 gaacgcggcg gtgaagtgta gcccgttatc actgatgttt cggtttgaca tacatcctcc 40440 ggtatttgta gtagcgagcg cctttcacgc aaatatgacg caccgattta agcgcgaata 40500 cggcgacgag agtcgtaatt atgaatggcg ggatgccacc ggtaaagaga tgagctagcc 40560 caccggcggc ggtaacaacg gacgccatat acagcagccg cccgaaaaga ccgtcttcaa 40620 tccagtgagc gtagatatta attaacgagg tgatgatgat tgcgaccaga caaatggcgc 40680 cgatttgtga gttgagcatg ttattttccc cacggtaact taaagttgct gagggatgag 40740 tcagcagctt cgaataactt aagccagaac atacctaccc cgaaaggaac caggtactga 40800 ccctcagact gtgataattc atagtaactt ataatgatgg gggatgcgta caccgcacaa 40860 caggcggagg caaccaggtg gcacaatcgc aaccaaggcg aaatagcctc acgcttctta 40920 atctgggata cggcgccccc ggcgaggccc gccacactaa gccaaaggta tctatcgtcc 40980 acaataaaaa atctccgaaa gttggtttac ccttcggaga ttaacattta tacgacaatt 41040 ttagtaacag ataattcgga tttatctgaa cgtataaagt ctgatggatg gcttgtgcgg 41100 gatatctgag tgcccgcgtt tacgccacag tttgtacagt ttaaccgcca tatgcgcccg 41160 cgcctggcta ctgaactcac caaggtttac cactttcttc ccgtcgtaag agtgcgcacg 41220 ccacagacct ttgcctacac aatacttagg ctggtgcacc ccagggatgc gcggatacac 41280 gtccggcatc tcattaactg gctttgcgcg cggtttaggt ttacgcttaa ccttttcttt 41340 aacgcgagta accttctttg ttaccgccga tgaaggaggt gccaggttga tgccggcgtg 41400 tttcagtata tcggttgcga gactcataat cacttctcct gtttaccgag tcgcttaggc 41460 gaacagatag cgcgtacttc tgaatcattt ggcttgtcgc cctgaaacag gaagtgcgca 41520 ttctcggcgg cgcgtgccgc tgaccggcac gcctctaatg tgtaaaacgt ttccgatgcc 41580 gcgagttgca tctggcctgc ggacagtacc cagataaata agatactggt catttcttta 41640 acccgcatat tctgcacatc atcgacgcac ttgtatcgta aaagtaccaa tcgtgctcac 41700 acccctgagc atcttcttta cggaaaccag attgctccgg cccgtgttca cctttgcgcg 41760 caagtttgta cgcctccacg tcgcccgtag cttttaccgg cgccatacgc aactccttaa 41820 cttgccctgt caggtgtgat atgtgctcag atttcttaat gttgtcttcc tgtaactcag 41880 ccaccagcac cgacacatca cgtaactgat tcttaagtga ctgcacctcc tgcatcaggt 41940 cttcacactg gttgctgttt acccgctgcc gtagctggtc aatcaggtac tctaatacct 42000 cgtctttgtt attcaccccc acactcctct atctcgtttc gataaagtaa tagtacccta 42060 ttatgttttg ttacgtcaat actattttac gattaattta tcctgacaca ccgtaatgta 42120 gcggggcttc ttcgaatgag cgcgccacca ctcaacacgg gcgtagatgc cgtggagcga 42180 tattactttg ccgtgtcggg tctctccgtt acgcccggtc caggttacaa ggtcattcgg 42240 tttcattgtt gcacctcctg caatagt 42267 <110> Optipharm.CO.,LTD <120> Novel Salmonella specific bacteriophage ST20 and antibacterial composition comprising the same <130> optipharm1-84P <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 42267 <212> DNA <213> Unknown <220> <223> Bacteriophage ST20 <400> 1 cgcttcgatt gttgcgtttg gcgaaataga aaaatattaa caaaaatggt acgctgctac 60 tcagcagcgt aagtcactca gtaaaatcct aaactactgt tacaagcttc ctgcggttcc 120 gataacagcg cccaggtgcg gccccgccac cacgataccc caggaactac ccacccacca 180 tcatcggcgt ccggatgcgg ccccggggac cactaaaacc ctagccataa acggcaacct 240 cctttaagac ccgagaatca ccggcgcgga ctctttagaa tcttttgccg tggggtattt 300 acagggaggt ccccgccccc attagagtgc ctttattccc gtccctcgtc aggtaagcgc 360 ttgccgcaag gcaagtgcgt taattcatga gtagtagact aagaatcttt cggcatacaa 420 gagaacagtt acgtttgccg ttgtccagtg ttgtgccttt attcctatta tctggtaact 480 aaagggcagg tctaaaacct gcccgctaat cgttacattt tacggttgcg aatctacggc 540 acatcaagga attcagatac ttaagagata tgctgccgat gagaatatag ctcgcccagg 600 gcagttcaga tactaaggaa gggaagttac tgagttttaa gtttgcgggt ttcagtaagc 660 cttaacgcgt atccgcttgg ggcggacccg ctaagctctc acaaagtgaa acagaaactt 720 aagtaacttg aagttacaag agacaactta cacgaaaaaa atgaaaatgt caataggtgt 780 gtttatagtt gatttcccat tatgccgtgg gtatacttgc tttactaact gagatggagg 840 atttgagaat gacacagaat gaagtagctg agcttattgg ggttacccgc cgcacgctga 900 ataactggct aagagacggc aagttcccgg attgttgtgt ccggattatg ggtcgcagga 960 tgccgggtac gtttgaccgg gagaaagtgg aagcgtggat tagggagaat gtaaagtgac 1020 cgatttccag aaaagggtat tcagaacggt agtcgcgctt acgtgtaaaa aagggtcgtg 1080 tagtgttctt gacttgcgcc gcacttattt tcagtactat agttcctctc tcatcgagaa 1140 cacgttgaag gttctggtta gagacggagc agttaaaagt aaggccggaa aatattcggc 1200 agtagctgaa gttaaaggct cccaggcgac cccagaagac cttgaataaa agaaaagccc 1260 caacgcgcga acgtcagggc cttaaacact acaaaggaaa agcacatgtc aacaggtaat 1320 ttagcatact catggtcgcg tcgcaatcgc cgtgaggaac gcgctgacat taacgtttac 1380 accacaactt ccggcacctg ggatgagttt gtcgaactga tgcagccgct taaacgttcg 1440 cgccggaatc ccaagacgga cccgggttac attaccgccg cttgtactgc cacggtaagc 1500 tctaccggta aagaagccgc cgaaggtatg ttctaccgct gcaatgcatc tgttacgtct 1560 tcatccctgg cttacgccga tgtggatagt gcgactccgg aagagtttgc caccgactgt 1620 gaaatggtgc gcgagtcacg tttcgctatg atgctctaca ccacagcatc gcacactgcg 1680 gaagcgccgc gctatcgcgt cgttatgccc gtgcgtacac cggtcactgg tggcgacgtt 1740 atccgcatcc ggtacggcct gttgacccac ttccttaaag gacgtgacgt agatagcgcc 1800 gggttcaccc tgtctcagcc gatgtaccgc ccgcctgtcg ggagtcaggt catcgtgtcc 1860 gaaagtagcc gcatgattac ggccagtaag cttatggagg gagttcctga aattaacgtt 1920 acaggcgctt ccgattacaa agtgcccgag ggtgagcaat ccgaattaac tgacctgttt 1980 gaagagttcg ccttcgagtt cggcggccgt atgaccgacc gcggcctgca aatgcccgcc 2040 acgccggagc acgccgccca atacactacc ggtgaaccta agcaggatga cttcctgttc 2100 tgttggccgc gcgacggctt cgagcgaccc aacgttacca tgtaccacga taccgacctg 2160 gtagctacgg gcggtatgaa gcctggcgga cgggatatgt gggcttacgc ttgtgcggcc 2220 accggcttac cttttgaccg tgtggaggtg gcgcttgggt gggcggaggg ggtcgcttgt 2280 gacgaagaag acctggccga cgaagaacca ccagcaccac aagcggactt catcgtcgaa 2340 ggatatatgc cgtctaactg catctgggat atcgtcggtg aatccgggac gtacaaatcg 2400 ttctacacac tcggcatgat gtatatcagt gctgccgggt atcggttcgc gggggcagac 2460 acccgtaaag cccatcattt ctatattgat ggtgagggtg gggagttcac gcacacccgt 2520 atcgctgcgt tggccgctaa atacggcgat gaaggtatgc ggtatatgca tgtgcttgac 2580 gctggtgaat tcgcggatac gaagaaactg gtgcgtaaaa tgcgccagat agcgggcagt 2640 gaacctgtcg ggatggtcgc tttcgacact cttaaccaga ccttcgggaa ctggatagac 2700 aagttcaatg agaactcagc cgggcaagac gggatgggcc gtgtcgtggc tatgcttaaa 2760 gaagtgcgcg acggtactaa aggcgctgta ggcgttgtcc accacacccc gaaaggtggg 2820 agtaaggcgc gcggaagtgg ggcgctgtat gctggtgtcg acgtcgaact tacgcttgtc 2880 cgagcgaccg agaaacaaat aaacgttgcg cacaccaaga acaaaaacgg catgcagcag 2940 aaaacgatag gtatggtgct tgaaccggta cagtttcgtg aagctccacc gccgaaagag 3000 ttccaggctg tagaattcgt tggaggtgag ggatacggtg aaatcgttaa cctcgacctg 3060 ccggaaccac ataaagctct tgtactcatg ccgtggggct tccagccgtt cgaaacggat 3120 gaagaaaaag aacgcaacga aggactcgat ggtaaaggta aagattccgt aaagggtact 3180 gttaaacggt ctaaagatgc ttccgcccgc gaatcagtta tgtctgctct cgaagattta 3240 cagcaagccg atgatacagg acgtggcttc acacaacggc aaatagtcgc aagagcgagg 3300 gaccacagca tcacaaacct ggtacttgag aagatgctac gtgagggtga attgatgctt 3360 ggttgcgatg aaaacggtga agttgtaacg aatacttaca ggttgccgac gggtatagat 3420 gaccgtaagc ggccaaagaa ccggtatgaa cctaacgaca acataaagac aacagaagtg 3480 gatttagagt aaaaagaaag gggccatcag gcccctgtat caccaaagct taaccatcga 3540 gggtcaacac cgagagcatc ggcaatgtgg aagattttaa aacagtctac cttacatacc 3600 cgaccagtcg ccaagtgggt aatcaggctt tgggataccc cggctttgaa ggcaagctca 3660 gtttgaccca tgccgatttc tttgcgacgg cgttctacac gagcgcctag ctcagattgc 3720 tgcatatgtt tactccttag ttagatatgt agtaatagta cagtaataaa atacacggtg 3780 caatttacga aaaagtgctt gcataaataa tagagtacga ttatagttct tatcaccaac 3840 taaacaggag aacaaataat gttagagaaa ttcttggtat tacttgaacg tttcgtagtt 3900 gcccatgaac tgattgctgc gaatagcgcg aaacagactg tgtaccaact taaaagtgat 3960 ataaccacgg taaacaaaga gttgacagaa aaagcggtag aaatggcgat gcaggataat 4020 tataaggcta gttctgcggt acgcaaagca tcagaaaaag aactcccggt agaaggcgaa 4080 agcgtcgttg acactaaacc ggcggaagaa gagaagccga aacgtaagac gcgtaaagct 4140 aaagtagagg agccggaaga agagaaagaa ggagttgact accagtctct tcgcgaccag 4200 attcaggcta tcgacgatgc gattagcgaa ggtcctagcg atgacgctta cgatgattcc 4260 aatgaactgc tgggagagtt caccggtaag aagatgaagc ttgccgcgat taaggacgaa 4320 gacctggccg aatacctgga acgcctgaca gcgatcaaga acaagtattt cgaagaagaa 4380 gaataactat cccgcggcct tcgggccgct ttacatggag attgacggat gaaattaaaa 4440 gaacgcggtg gcaacaacga tgtgcattcc ctgctgtcgc cgtctggggc caaaaagtgg 4500 ctggcgtgca gtgcatcact ggcttgtgaa aaagatatac ccaacacgtc cggtaaagcc 4560 gcggtgttag gaacggcgat gcacacactt gccgagattc atcttaacca gtacatcaaa 4620 ggcacttctt tgccgttaga gcgagatgta ggggcatatg tgctggaaga aggtaaaggc 4680 gcggttaagg ccctaattaa gccgatgaaa ggcgcggtgc tggttacaga tgacatggtc 4740 gagcaggtgc gaaagtacac cgactactgt aaagcggtta tcgatgtagc ggcctacgcc 4800 aagcttgaaa tgcgcgttaa tcttagtgag gtgttgcacc ccggatacga aggcgttgag 4860 acgttcggga ccgctgacct tgttgccgta caggaattgg ctaacactga cgatcacatg 4920 ctcgttattg gcgacctgaa aacagggcgc catagagtag aggcgaaaga aaacaagcaa 4980 cttatgctct atgctctcgg tgtttaccgc cgcctaaaga gacgctataa cataacagct 5040 gttcgtctgg ttatctttca accgtacacc ggtggcgcgt cggagtggga tatctcagtt 5100 gaaggtcttg aactattcgc taagttcgcg cagaaacgtg cggtagccgc acttgacgcg 5160 tatcgacgtg gtaagaaaaa ccttaaacca tcggacttca agccgtcggt tgatggttgc 5220 caatggtgcc gtttctcaga acaatgtgcc gcgcgtacaa aaaccgttaa ttctgtactg 5280 gcggaagaac tggaagacga tacggatgta tatcgtacag ccgtagaacg tgtagctaag 5340 ttctgcaacg acgcggacta tatgagcgtt tgggaggatg ttcagccgag cgatattctt 5400 aagttgctgc ctgatggccc agatttacct aagaagccag tgagagggga cgactttgca 5460 ctggagctta cgccggagca actcgtcgct gagtatgaga agttgccgtt gttgcgccag 5520 cacatcgaca aagtagaaaa agctatggca gccgctttgc attccggtaa gaaagttccc 5580 ggttacaaac tggttgaggg ccgtcctggt aatcgcacgt ggaaagatga agatgctgta 5640 aataaattta gggaaaacca tgtaaacggc tatctgctgg ataagactgt tcccgtatcg 5700 ccgaccgaag cggaaaagat catcggcaag gaagaccccg aactttgggc tgagctatct 5760 aaactcgtaa ctcgtaagcc gggtgtgcca tgtgttgcat cggcggatga caggcgacct 5820 gattggaaaa atattacgga ggaggatttg gaatgaagaa attgacgtgg tggctgcttg 5880 ccaccccact aataccggtg gcttttatat actatctaat gagagcgtta gagaaagttg 5940 gtgaaacgtc ggcgatttta gctgacagga tgctagactc aaaaatgagg atgaaatatt 6000 tagcatggcg taatcgaaaa tttggttgac aagttaatag tactctatta tattcttaat 6060 cactgacctg gcagcttcct ggataaactc taaaacgcga gaaaactaaa atgggaatca 6120 aacttaatct tcgtaaagta caaactgcat ggttgaacgt attcgaacgc gctaaagacc 6180 gtgaaaataa tgacggttcc gttactaaag gtacgtataa cggtactttt atcctgactc 6240 cggaacaccc gcaaattgaa gaacttcgcg atacggtatt cgctgtagtg tcggaagcac 6300 tgggagaggc cgccgcccag aaatggatga agcagaacta cggtgaaggg aagcatatgg 6360 ataaatgcgc cgtgcgcgac atcgccgaac gtgataaccc gttcgaagac ttcccggaag 6420 gtttttactt ccaggcaaag aacaagcaac agccattaat cctgacttcg gttaagggcg 6480 aaaagcaggt agaaccagac ttcaatattg atggtgagca gattgaaggt aagcaggtat 6540 atagcggttg tgtagctaat atatcaatcg aaatttggtt ttccgagcaa tataaagttt 6600 taggcgcaaa actaaacggc atcaaatttg caggtgaagg aaaggcattc ggtggttctg 6660 cggtttctgc cagcgttgac gacctggaag atgatgaaga cgaaacaccg cgtcgcgaac 6720 gccgtcgtaa ccgttaatgt attttctttt caattaaggc ggcttcggtc gccttttcta 6780 taagggtcaa ataaatgaat ctgctttatc tcgatactga aacattctca gaagccgatt 6840 tgaaaaaagt cggttcctat gcctatgccg aacatccgac tactgaaatt gttatctgca 6900 cttatgcttt cgatgaaggc cccgtgcaag tatgggacgc caccgacggc agcgatatgc 6960 cgagagattt gcgcagggcg atgctaaagc tgcaaaaacc agacagcaac ctcaaactgg 7020 taggccaaaa cttccttatg ttcgaccgac cagttattaa gcattgctgg ggattcgaac 7080 tcctggtaga aaacattata gacactatga tagtcgcgtt ccgacatgcc ctcccgggtt 7140 cactggccgc gctgtgtgag gttttaaaca ttgacgcaag catggctaag gataaacgcg 7200 gcaaggcgct catacagcgg ttcagtaagc ctacgcctaa gaactataag attcgacgat 7260 ataccgccaa tacccaccca aaagagtggg cagaatttat cgcatacgca aaaagcgaca 7320 ttacgtctat gcgtgaagtg tataagaaaa tgccgaagtg ggggaattct gagttcgaag 7380 accgcgtgct gtggttagac caagtaatta acgaccgcgg atttaaggtt gacgttgccc 7440 tcgctgaggc cgcgattgaa gcagtgacgc gccacaagga agagctacag gaagaagccc 7500 aacgcaaata cggcggttca ctaaccggca aagacttctt gcctatttta caggaactgg 7560 cgccggcgca ccgcattcac aacgcacaga agtcaacact aaacgatttg ctggcggatg 7620 aggatttacc ggacgacgcc cgcactatta tcgagatgcg tctcggggct gcttctactg 7680 catcgacgaa atacgcgccg ttgctgttag gccgttcttc agatgaccgc cgccgtggtt 7740 gcctgcaata cggaggagcg aagcgaacat tgcggtgggc ggggaaaggt tttcagccgc 7800 aaaacctggc gcgcgggtat tatcacgacg atgaactgga taggggtatt gccgcgttac 7860 ttaaaggccg tgcacaccgc cgttttgacg tggcaaaact aacagcgtct accgtccgaa 7920 gctgcattat cccggaagcc gggcgtaagt ttgttgttgc ggattactct aacgtcgaag 7980 gccgtgggct tgcatggctg gcgggcgaag aaaccgcgct tgatacgttc cgcgccgggc 8040 tggatattta ctgcgtaacc gcaggtaaga tgtttggcat ggacccagat tacattaaga 8100 aagaacgtaa agatttacgc cagattggta aagcctgcct tcaccgccat acccaagttt 8160 tgaccaatgg cggatttaag gatattatgg cggtcacatc gacagataaa gtatggagtg 8220 gtgaaaaatg ggtgaataca aaaggcgcgc accttatggg gtggaaaccg gtgataaatg 8280 tggacggggt gctaatgacc gaagaccaca aaatcttgac gcattcctgg aagcaggcaa 8340 agcaactcgt ttcaaacaaa tatatgatgg accgcgccct ggagataggt atggacgcct 8400 ggttatcctg cgcgagttac caaaacgaca aggccaaaga caactactcg tccaatgtga 8460 ttgtggagag atgcctgggc gggtctatta tgacaatgtc agggcgggta aaaccactca 8520 atgcaaccgc tgcgcccttg aagcaacaaa aaaatatcgt aaacagtatt tctgctacga 8580 agacgcaatg ccggatgatg cgcacagaac gcgattactc aaccggctgt ccgcggcaat 8640 cgtgcggaca acaagccccg gaaacaagag ttacaaaaac tatggggcta ggggtatcac 8700 ggtatttgac cagtggaggg ccgataaaag gtcgtttctt agatatgttc aaactttgga 8760 aggctgggat gacccaaacc tcgaaatgga ccgaatcgac acagacggta actacgagcc 8820 tggcaacatt cggtttgtca gccgttctga aaactgccgt aacagacgtc gaatacctga 8880 gcttcaacga aagtatgatg cagccgttgc ccgcattgct gagcttgaac gggaaattaa 8940 cctactgcga gccagtttac gacctgattg atgtggaaga cggcaaccgt tttcttatag 9000 cctcagactc gggctttctt gtggcgcata actgcgaact gggcctcggc tacgagggag 9060 gtgtcggagc attcgttacg tttgctaaaa acctgggtct tgaccttatt gagatggcaa 9120 aaacaatgga cgggactttc cccgaccaca tctgggctgc tactgcacgt gggtatgagt 9180 gggcgcgtat tcaggaagcc aagagaccac cgcgccctgg cgaaaaggat gacaggccat 9240 cgtatattct ggataagaaa gtgtggcgta cctgtgatgc catcaagcgc atgtggcgcg 9300 agtcacaccc tgaaacagta gcgttctggc gcgaccttaa agacggaatt ttagccgctg 9360 ttcgtaatcc tggtcgtgaa ttttgggctg gggcacactt gcgccggaat ggtgaaaggg 9420 ctatccgcat atggcgtacc gtagaatttg attcgtcggg caggaaggtt cctggctggt 9480 ggttgtgtat ggagttgccg tcaggccgta tcctatcgta tccgggaatc ggcgttagtg 9540 tgacaaagga aacagacgaa gacgggtgga taaacaccag tgtaagaatt aagtaccagg 9600 gtgagaacca gttaacacgg caatggacca ccctgtacac acacggcgga aaggcttgtg 9660 aaaacattgt tcaggcgttg tgccgtgact tattggccta tgcgatgctt aatgtagaag 9720 ccggtgggta tccaatagtt ctttctgttc acgatgaact ggtatgcgag actccggata 9780 catcagatta cacggtagct gaactggaaa aactaatgtg tgcattgccg gagtgggctg 9840 atggttttcc tcttgtagcg gaaggtgcgg agttaaaacg gtatgctaag taaactgatt 9900 atttattatt tcagggaaga agactgtcga atctgtccgc gctgtgggat taagcacacg 9960 aaacgtgagg ggtgcataga tgagtacgcc tgagggccgc gtccagaaat acgcaaaaga 10020 ccgatttgag gccctggggg gcctcgtacg caaattgtcg tacgaaaata gagtgggcgc 10080 tcctgacctg ctggtgattc tccctggtgg catcatctgg ttcgtcgagg tgaagaaaga 10140 cgaaaacacg aagccagacc cgcaccagtt gcgcgagcac gagaggatgc gtaaacgcgg 10200 cgcaaatgtt tttgttgttg ggtcgaagaa acaggttgat aaattaatag aacactatta 10260 tatttagtta acaccaaaca gaaataagga attgagaaat gaaatatgaa tatgaccgca 10320 aaccagcacg tgacatcgta ccaggcgata tgattttcaa cgttaaaacc cgccagccgg 10380 ttgccgttga tactgtgttc gtcgagtcga atggcaaact ggttatcgaa gatgtaacgg 10440 gtaacgttac ggcgttcggg cggaaggaac tggtactggt ggcgaaatga gtaaattcac 10500 tagaaggcct tatcaaaagc tcatgacctc gttcatgctg cgacacccgc gctgtaatat 10560 ctgggcaagt atgggcagcg gcaagtgcct gaaacgcggg actgaagtta ttatgtttga 10620 cggtacgacc aagaaagtag aagacgtcat cgtcggtgac gtgcttatgg gaccagattc 10680 aacgcctaga aatgtactgt ctctcggtag aggccgggag atgatgtacg aggttaaacc 10740 gcgaaaaggc gagtcgtata cggtaaacga aagccatata ttatcactgc gaactactac 10800 aggtatcgca aaaggttcat ggccagataa caccgttttc gacataagcg tccgcgactg 10860 gcttaagttg ccgaaatatg taaccgggcc taatgggtat cttaagggtt ggcgggtacc 10920 tgtcgatttt ccaaggaaag agcaggacga ggctttattg cccccgtacc ttatggggct 10980 atggctgggc gacggtactt cgtcgtctgg ggccataaca tcaggagaga acgaaaagga 11040 aataagggcg tatcttgagt cgtacgcagc acgtaatggg atgcaaatac gaaaagaagg 11100 acttacatgg agtatatctc acggcaacac aggtcataag aagcacgggt ttacgcacgc 11160 gctgaaatcg gcgggtgtcc tgaacaataa gcatataccg cataattaca aatgcggcga 11220 ccggagacag cgtttagagt tgctggcagg gcttctggat agtgatggtt actgcgattt 11280 gtctaaagca ggttttgact ggatttcggt aagcgagcga ctcgctgatg atttttgcta 11340 tctatgccgt tctcttggtt tcgccgctta caagaagaaa actcgtaagc gttgcgcaaa 11400 cactgatgtt tggggggatt acttccgcgt atcggtatca ggcgattttt ctgaggtgcc 11460 tttcgtgcgt ggtcgccacc agaatttacc gaagcgtaat attaataaaa acgtacttaa 11520 cgtcgggata gaatctataa ccccggtcgg tgtagatgat tattacggct tcaccatcga 11580 cggtgaccac cggtttttat taggtgactt caccgtaacg cacaacacaa gctcgacaat 11640 gtgggtcctc aaccgactgt tccgtaatgg gcaacttaat gacgacgacc gagtgttaat 11700 tctggcccct ttacgtgttg cgtcaggtac gtggccggca gaacaagaga aatggaactt 11760 cccgtgtctg agtgtagtag atgcaactgg ttctgagaag cgacgcatcg cggcgctgga 11820 gtcagacgct aacgtggtat gcactaacta cgaggttatc gaatggctta ttgactacta 11880 cggcaaagac gactggcctt ttaccgttat agttgccgat gaaagcacga agcttaaatc 11940 gttccgcagc cgttcgggcg ggagcaaacg ggcaagggcg cttagcaagg ttgcgttcgg 12000 taaggttaag cgtttcatta accttaccgg gacaccatcg ccaaacggct taaaagattt 12060 gtggggtcag aactggttta tcgacgcagg tgagcgcctt ggttcttcgt acaccgcatt 12120 caccgaccgc tggtttaatt cagtacagaa aggaaagtcg gcgatggcac gggagtacca 12180 cgcccgcccc ggcgcggata acgagattca ccagaagatg aaggatatca gcctgaccat 12240 tgatgctgcc gagtggttcg gttgtgaagc gccggttatc gtaccggttg agattgacct 12300 gcctaagaaa gcacgccagg cttacatcga tatggaggag aagttattcg cggagctaga 12360 aagcggagaa gtcgaagcgg ctaatgccgc tgctaaaacg tcgaaatgcc tccagattgc 12420 ctccggtgct gtatacgtat cagggccaga tggcgaagca accaaagact gggagaaagt 12480 gcacgacacg aaactggacg cactggagtc aattgtggag gagttgcagg gcgcgccgtt 12540 gctggtggcg tatcagttta agcacgaact tgagcgcatt cttaagcgtt tcccacaggc 12600 gcaggcgttc gccaaaggtg ccaagggcaa taagcagatg gaagcgtgga accgcggaga 12660 gatacctatg ctcctgatac acccagcctc cgcgggtcac gggctgtcgc tacaacacgg 12720 cgggcaccac atggccttta tgagtcaggg gtggaacctg gaacattatc tgcaggttat 12780 cgagcgcata ggtccggtaa ggcagatgca atctgggtac aaacggtctg tattcattta 12840 taaaataata gcccgagata ctcttgatga agttgtcgcc gctcgtacgg acgagaaaag 12900 gtctgtacag gacgaactac ttaattatat gaaacgtagg agtaagaaat gagcaaggta 12960 tcttacgacc ctgaaaccgg tattttcaca tgggtagaaa caaaacgtag gggttttatt 13020 ggaaagaaag cgggatgctt ttgtccgagg gatgggcaca tttgtatccg aataaacgga 13080 aaactagaac tagcacatcg gttagcatgg ggtgcctgca atcccgataa ccaacttaca 13140 ccacaggagc agatagacca cattaaccat attcgaactg ataaccgaat atgcaacccg 13200 cggaaagtta ccaacactga aaacagccgc aacgcatcaa taggcgtaaa caatacgagc 13260 ggcgcactcg gagtctggtt cgagaagcga cggaataagt gggtcgcaga aataaaggta 13320 gaccgtaaaa agatacatct cggtcaattt gaaaagtttt ccgatgctgt agcagagcgt 13380 aaagctgccg aagttaaata cggattccat gaaaaccacg gcaagaagag aggtgtgtga 13440 tgggaactta cattggtaaa tcttggaaag atttatacga tagaatcaag gaactcgaag 13500 ccgagaacgc cagactacgg cagcagcgag acgcggcgaa cgcgcaatta gactggctac 13560 ttgagcaacc ggaagttgac aaagagaggg gcgctaaatg aaatactcaa ttggtgtgaa 13620 atgcttttta ataggctacg gattggccgc ggtcgcttta cttatcgctg ctgtggatct 13680 gttatcataa attaaggccc cttacggggc ctttactatt tatgccaaag ttaatcttgt 13740 atagcttccg tcggcccgct taactaacgc tttaaggtta tcgccctcca gatacagtga 13800 tatagacccg ttatccctga cagcgttgtc tggtaggacg gtcgtcgcta ccggtatctc 13860 aacagcacga taatcgcccc tatttatttt taatgatacg gcatcgggca agctccccgc 13920 aatagcggta atttcggtcc atgctgagcc agaacccggc ccgccgttaa tgtgggagta 13980 cacggaccca ctatcaaggg tttttgacag tttatgaata cgaaaccgaa gcgccgcaga 14040 atcattatcg aaactgttta tgcgagagtt accaagacct ccctccatca aatcggctac 14100 gttaatcctt gaggggtcca cgaagcccgt tagtccgctt acagtggagt taggtgcgtc 14160 taccgtcatg ccctgttcgg tagttgaacg gatgccgacc aaacgaaggc cgtttacacg 14220 acaggccccc gagatatata tctggttcgc agggaagttt ttagtattgg tgtcgattat 14280 tgcgatatta gtgaatactg tttcgtgggc aagaatataa gccccagaac cagcgcaatc 14340 ctcgacggtt atgttagaga catacagacc ttgcccgtcc ataccgaatc ctacccccag 14400 cgaacctcta acaaatagat tgtctatcaa atggtttaaa gggagcatat gtaccgggta 14460 ctgagaaatg gggaaatccc ctgggcggtc atcttcgggg ttcatgtcag tatctgcgcc 14520 taaatcgaag ccgtcccata cagggtacag taccgccgaa tcccggaatt gcaggttgta 14580 gttacgagat gtcgtagaac ctaccgtacc ttgccacgtc ttaacaccac tttcccctgc 14640 acgatacgag gtaaacccga tgaccccgcc atcgcgcgcg aaaccgccat tgttgcgtaa 14700 gaattgcgcg ctacttaccg aaccgtaact tgtgcgcccg ccgataacat agttaccttt 14760 gccccaatct ccgcttaagt tttcgaaggt gattacgccg tgtgcaccgc cgctcgggtt 14820 gtcagcgtct accatcttac agaaatggca tccacggaac aggaaacacg ccataagacc 14880 actcgcccgg tgaacctcga cgcctgtaca ttcccgaatt tccagggtag aagatatgct 14940 ttgcccttta gcttccggag ggagaaggga ttctatgcca gggaacttgg cgtaatcgtt 15000 taccgtcggc tggtatccat ctgttttaga ctgtttaagt gtggccacga ttgccgcggg 15060 gtcggttatc cattgattat cgtcagtcca cggtttaatc acccacggcg ttgtagcact 15120 ctccataaag ggggctatta caatggaacc tttacctaat tgcgtaaata ccaggtttcc 15180 gtcgccgata aacttcgctt tacagtcgat ggtcagggtc ttcccgctga agttaacggt 15240 ctcgttatta ctgaaagtgt aatcgcggtc gataagaacg ctatcaacgg cggcatccgc 15300 caattgttgt aacgtagtaa agtcggacag cttaacggag tatttaaatt tcttatccgc 15360 ttccgcgcga aaggtaatat ctccgaacgc gacccaagcg gaatacgaaa tgccgcctgt 15420 cgtaaggggt gtcgacgccg cagggataac tttcggcagg gagccacgcc atgcgtagta 15480 gttaccgtct ccaccatctt ctttcggcca aagaaccgct ttattggcgt cgtttacgcc 15540 gagagtaccc ccggtcgtaa agttaaaaga tgccggggag aaaccagcat cacgtaagac 15600 tgcgggaagc gtcttctgcg tctgcccggt tacctggtta gtggcgtagt cgatatccgc 15660 accaccagct acaccgcctt gtttaccggt gatcacctcg gcttcgaaaa tctggtgttt 15720 tttagctatt tgtaaatcat taagtgacaa tacatcaccg caaccgctgg acatatggct 15780 atcctcttag ttaaaaccgt tatcgaatcc gttagagaac gcacgaccga aaggaggcac 15840 gttgtcaaac ttatagaaat ccttgtcata gttaaacccg gttattttga ctgttctgtc 15900 atcgccaggg tctaccgtag aaacaagaat catctgagca ttatgccttg cttcgttgcc 15960 gaatgaaaat tcagttttca gggcgctatt acccgtgtaa atagcttcct gcggcaccga 16020 cgtcataatt acttgtcggt catgtgcacc agggacgaca cgaacacttt ggactccgcc 16080 atcacgtaac tttagaacca aagaatggtc atctccggat gtgaacttaa ccggctggga 16140 tagctctacg gtaagaccgt taaccgcggt tacgtagccg tcgtaagtag ccatacgcga 16200 acccttaacc acactaacgg cacgattcgg aagggcaaaa ataccctctt cagtggcagt 16260 aaacgacacc gaaattttct ttaaaatgtt cttctggtgg cgacggtttg ccgcccagaa 16320 agcctgctta tagttgcgga tcccttttga gtcataagtc tccgttttaa ggcccccggt 16380 ttcaggtatt gttatggttt ctttaacgtt tgttttaggg tctatgtacg agaatttaag 16440 actatcgaac acctgagcat cgttaaaggt tctggtccat ttttcggaag taccagcctt 16500 gcttcggtgg gtgaacacca tctcaggacc catgcgaggg cgttcaaaat cgagaaggat 16560 atccgcccca cgcctatatg gagtacagaa gatagcgtcg gctatagtac taactatatc 16620 ctgcatagtg gttttatagt catcaaacgt gtaacaaaat tctccagcct gtttgtcgcc 16680 aaaatacgct tcaacctcgt tctgcacggc aagtaactta tccatgttgc gtactgtcag 16740 gtttaaaccg ccaacatccg ggtcacgcgc caggcggatg agggattgta cagcctgtgt 16800 attaggtgtc atcgtatctt cgaaaacacc gttaccgagg tatttgtaca ccatttcagt 16860 agcaatcata cgcaactgcg gttgctttac ttccgcagca cgtggggtct gcttgcgcat 16920 cgagtgtact gttgttctgt tgccgtagtg tggggtgttg tcacgtgttt gcccgtataa 16980 gttaacgtac gttatctcat caactacaga cccctcgaaa ttaaagtcca ggtctgttac 17040 ccttctggcg cgcacccgca cccgtgacac aaccggcagg tcggcgtaaa tagtgacgcc 17100 gttgtagtcc ggtgtacgcc cggacacagt tccttgcgca gtatatatcg gtccgtaagg 17160 agtgctgttt tcatcaagca attgatactg tatttcggcg gtgactgacg ccagtgtttt 17220 cgatttgccg ttgtctttat acatgccgtt cgcggcggcg atattggcga ctatacgctc 17280 aacctcggta cggtttattg aaacccaatc ggttaatgtt ttctcatacg tgttgtctgg 17340 ttgcagcgag gcctctccat caccgcggaa aaatgaaccg ggtcttattt cttgccatcg 17400 cccgataagg ccgccaggca cgataaaggt aacggtccct tcggatacgc ttaacacctg 17460 atacccgccg ccggacaagt cgacgtccgt atttgtccac gctgacacca acgaaaacgt 17520 gtcattaggc accaacactc cagaaaaatc cgaaccacct gttgggtcgt atatcgtgcc 17580 gatgttcccg gacaaagacg gatacccgga catgtaagag aaagaaatac ccagaccgtt 17640 gggtgcttta agaaccacac catcgacttc gttagattct acggtaatat acaagccctg 17700 ctctatcggg tctccgacca tgacctgggg tgtagacgtg ttattgggtg acgtatacgg 17760 cgcgtacacg gcaaccgacg tgcctgttat atccgataca cgggtgtccc cgtcagtgat 17820 atcttcaggg cgtatgtcga gatagccgcg accggcgtcg taataaccgt actctacaat 17880 tttaccggcg gcgttaaaca ccttgtaagt agtcataagg ttattgggga tggtttgcac 17940 cgtcccgcag atgtcgtaac tgcgctcgta cgggcgcgcc ttgttgttgc ggtcggtgag 18000 actgttgttc ggggaatccg cctgcgagtt cgccaggtta gaggacgccc ctttcacgga 18060 tggtgatagt agtttcgcca gcggtttaag gataacgcta aaaaccttca tgacgccttt 18120 aatagccccc ccaccggcag attctacaat gtggaaagtc gcgttttctt tcagtgcttc 18180 gaagtcttct gttacgtcgt tatcttctcc gatttcgtca atgaaaacac gtaccggaac 18240 cccgtccgga acatggttca cgacgaagtt catcggatta tcgtgataac gtttagcgtc 18300 aaatgtgccg tcttcgttgc gggtgtagtg gattactagc gccaaaattc aatctccgaa 18360 taagtatctt taaggtcttc caggctatcc agtctgacct gtctggacgc cagctcacag 18420 tggctaacca ttccttcgta atatacccca gcatgccaca ctattcgccc cctatgtttc 18480 actcctagta gaaccgcgtc aaaattctgc ggtgtgaccg ctcgtaccag acctttcgga 18540 ttagagtggc catcgtcgaa ggcggcgtct attgccgttg ggctggtgac gtcaaacatt 18600 ggagttgata acccagcatc cgcgcggacg ttgcggacgt gatgccagca attacgacgg 18660 cgaaagtcat acggcaagcc agtgtaatca ttaatattca tgatgtcagt atcccgcgaa 18720 gtaaaggaat ctcctccggt gtcatcaaaa taccggtagc ccgctggtta agcatcggcg 18780 taccggtttc cgccgtaaat acaccttttt cttgtgtgag tgtttgcaac tcataaacca 18840 ccgggccgtc gcatggatag gataggtccg tactcacata gcgacggaag atgaatttag 18900 gcagttcctt gttgtcgtaa ggtatacggt ccatttcctc atccagtatg ttaagcacat 18960 ccggaagcga aaaggacgcg gtctggtcca tatcgttatt gttggcggcg ttcttagcct 19020 ccatcggcgt accttcaaat gtcactacct ccccggtctc aagagttgcg gttaagtctt 19080 taactccgcg cacaagtaac caccgtttcg aaagaagtgg gtggtatatc tcaagagtaa 19140 taaaatccat ttcaccgtct ggattggatg ccagtttacg tcggtatgcc gcttctactg 19200 attcctgact cattgcatag ggtcccagat tctcgggaat gtagtctgat acactccata 19260 ggttttaagg aattcgccaa ggcaatcgcc atagcacccg tacaaatcgg gtaggttctg 19320 gttaaggcaa gcgttgttgg cttcctggaa tggcgacttc tcggcggtgg ccgtgaaggt 19380 gattacccaa ttctttccgt catctgtgga ctcgttccac gtggacgtta acgttacctg 19440 gtaatcgtca ataccctggc caaggtcgtg tttcatccag aaactggaag cccctccgtc 19500 taccttctcc atgaaactga ggaatgcttg tctacccagt ggcgacacga ccagcgtaac 19560 gctaatcggg aacacatcaa agtatgtatc acgcccctgg cgaactccgc caccggccag 19620 gtctacccgc cacacgttgt tgccgcgagt catcgagtat cctttcgata ctatcggcct 19680 caaagaggcc gggaaatggt aatcgctcat atcaatatcc tggttggccg cgtgtagcgc 19740 gacgtgcctt agaaatagcg ctgttactat cctgtagcgc ggaacttacg gtttcactga 19800 ttataatacg taaacgacct tcgtcgtcgc gttctgttgc agccgaatca attcttccag 19860 tggtgttgtt tacaatagta acattatccc cacctgattt agtgccattc tcgcccataa 19920 tctgtcgcat ctgctccgcc gtccggacac gggaggcgct agcgggcatg attacttctg 19980 gtttaccacg ttctgcaata gtggatatct gccctgcggc caggttacca ccctgttctc 20040 gggcggagcg aatcttacct acgtttgcta aaccagccgc caccgcagca gccgcagcaa 20100 caggcgccag atacgggccg actaccggta tggctgctgt tgatttatac gcttcgaccg 20160 ctgatgtgta cgtagctatg gtcgcctgag caattgcaaa tgctttgtat gccgtggaag 20220 cctctcctag cgcagccccg atgttactag ccatgttacc gaaagccgcg ccggtcgcct 20280 tcactcggtc aagggagtaa gattcatcca tggcgcttaa cgtttgttgg taagtttcat 20340 cggcgatcaa accctgcgca tgaaaatcgt taagtttttg tagcttaacc tcgtactggc 20400 ggtctaactc cgcgaactct ccttcagcaa gggcttgcat ctgagcgacg tacgcgtctg 20460 cggagaactg tttttccagg cgctctttct cgcgtttatc taattcagcc tggcgggccg 20520 tatccgcctc gagcataatc tgcgtcttag cggtttcgta ttgctggtca ctaaggagac 20580 ggttgctgta gaactcctgt agctttgtaa gttgctcttc ttggttagct tgtatcttct 20640 ggaatacgtc gtcgttaagg tggattgact gataatagaa atcctcagcg gctttcttct 20700 ggcgttcgaa tgcgtcgatg gatttcttgg cggccttgtc tgcggaatca cctgtagaac 20760 cgctgtcctt acctgatacc ttaaacttac ctaacccctc ggccttagct gcttctttat 20820 cgatgtcgta agccgctcgc ttcagcgcga taaggtcttc ctgagcttct acttgttcgc 20880 ggaaactttt ccgtgtcagg tcagccacac ccctttgagt atccatagca agcgctaact 20940 ggaagttaag gttcttggcc tcagctaccg actctcccgg cgtcaatgca gcctttacaa 21000 gacgccccgc atcctgagca cctttaacca tatcttcaaa aaaccgctcc attgctgtaa 21060 attcctgggc aatgaagtcc agcgcatccg ccgctgactc agctatcgta gatgcggtct 21120 cctctccgct tccggtaatg ctttcactgg tgtaagtcca ggcggagttc aaagcgttta 21180 acgcgtctgc aaccatccca aacgccgcgt cgaatgaggc ctctatcgca cccccaacat 21240 cggatgccat gtcaccccac atccgcatct cggctacgaa ctcaccggat gcgacctgag 21300 cgtttatctc ctgtatcagg tcgtcaatat aacgtaaagg ctccgccagc gaccctacat 21360 ccagactagt atcgagagtc atcttcaact gggaccatga gtcctccgca ctagcgatag 21420 ctccgtccag ggtattagcc tggtcagcca tcgctccggc gaagttaacg ttacctatgt 21480 tgagaagata tcgctcaata tcggcagcgt ttttcttaac tacagtagtc gtcccctgga 21540 aggtaaattc gatatcactt tggttctgct tggccttaat gccgaattct ttcagacgtt 21600 cgaattcaaa ggtgctcgca tccgcgacgg cctcaatcat ctggttaagg tctttaccca 21660 tagccgatgc ggtgttgcca taggagcgga gcgcttcttc tgatggggtg aggcctagag 21720 ctaccagttt acggaaccct tctaccgcct gttcaaggcc gtaaggtgtg tcacgggcga 21780 agtcctgtag tatactcagg gcctgttttg caccttgtgt gcttccggtt agtgttttca 21840 ggctggccga catcttatcc agttgccgtt gtgactcgac tagttcctga gccgccttat 21900 aggctgcggc ggctacaacg gcagccagtc cggtgaccgc ggcccccgca acttttgcag 21960 atttagagag gttatcaaga cggtctgatg tggccttcgc gccttgttcg gttaccttta 22020 ctactaagct agctacatca gccatcgttt ctaccctcaa aaatggcgtc taagcccatg 22080 ataatttcag attcaaacat ccctatattc tgacctgaaa tagtgctata cgccactaaa 22140 tctgaccaac taagttgctc tcttgggtat agctttacgc cgtcgtcatc gacgcggcgg 22200 gtaaatttaa catcacgata cttctcaaaa gtggttagta gttcaggggg gcattcaggc 22260 ccggcgtcct gcgttgcttc ttccgcgtct tctattaccc ccatagcgat aagcgcggct 22320 ttgtgcccgt cggctatact gtcaaacttg cgcctttcgt ggcgggttat gaagttccaa 22380 cgggcgaact gaaacaacgc gtctactttt cctgcaattg gcgcaactgt tcgtggtgga 22440 atacgactac atgttccgcc aggcgcttat attgggtgag aagagtctca agattttcct 22500 tagtgaactc atcatcgaga ctccaaccat tcaccaattc gagagccaat tgccggttta 22560 ggtctcctgc cgcgtcttcc atttttaaat tgtattctga gaagtctttt tgctcttcgc 22620 atttatctcg aagcggtttt aatttaccta ccgccgctcg gtacgccagg gtgaaggcgc 22680 gcatagcttt aaccgcaata tcggcctccg gagagaccac gttcaaccat tcccctgaat 22740 ctgtaccatc ttttaaagga atcggcatgc gcgcgccttt ctcggcttct gcttcgtagt 22800 aaaaatcgct aagtttcata gtaatgccct ttggttagtc ggttactggt tggatagttg 22860 cgtcaggcgg gaaccacccg cttttctggc gcgaccgtag acgctaaata agaatagcaa 22920 aatagcttgc gcaaccgtta atagtgtcct attatctcta cgtcaactaa acaatatgag 22980 gttaagaaca tgagcgtttt tatcggtatt atcgcgttta cctatttcat cccgttcctg 23040 gtagccttgc tacgtaatca taaagcgaaa ttaggaatat tcgtatgtaa tcttttactg 23100 gggtggattc ttctgccgtg ggttggggct ctcatctggg cctgtaactc taatgttaat 23160 ggtaaataat ctggctaaat taaacttacg ttattagaat cagatttatc ttaaacagtc 23220 aggaaacaag catggtcact cgcaaaataa cagatgaaca gttgcagcaa gagctaaatg 23280 ctggcctcgg ccctactgaa atagccaaaa agtataatat gtcccgccgt aatgttcagc 23340 ttcgttccgc gcgcctggca aagaaaggag ttggccacgg gcgcgacgta agccaccttg 23400 tgccagatgg gtataagatt aagggtacgt cgtcactggt agatgagttc ggtaacacga 23460 aacttcagtg ggttaagact gacgccgacg ccgagcgtca ggtcgagttg atgcgagccg 23520 taatagatgg gatgaagtcc gatattacgc cagtttcttc ggtccctcgg cctaaaaagc 23580 gactaaacga aaagttgcta aatctttaca cggtttctga cttccattta ggtatgttgg 23640 cctgggcgga tgagagtggc gacgactggg atatgaagat tgcagaagac ctgttctcga 23700 gatggttcga cgcggcgttt cagaaggcgc ctgatgccgg tgtgggtgtt attaacctat 23760 taggggactt tgcgcatttt gacagccttg atgccgttac gcctgccagt ggccatgtac 23820 tggacgcgga cacgcgttac cagaaactgg tgcgctacat gattcgtatg gtcaggcgcg 23880 tggttaatat ggcgcttgtt aagcataaaa atgttcatct cctaattgtg caaggaaacc 23940 atgacgagtc aggcatgatt tggttggctg aaatgtttaa cactctttac gataatgaac 24000 ctcgtgtttt tgtagataca tcggctgatg tctacaagat ggtccaacac ggtaagacga 24060 cccttttctt tcaccatggg cataaggcga gattcgatgc tatcgaacca gttatgatcg 24120 ccaagttccg taaggcgttc ggagagagcg tttacagtta cgcccatgtg ggccaccttc 24180 accatcagaa gattgtggaa agccgtaaca tgattgttga acaacaccgc actctcgcgg 24240 cgaaagatgc ttacgcatca cgtggtggat ggatgtcagg ccgcagcgcg aatgttatta 24300 cttacagtgc cgaatacggc gaagtcgcgc gtttaactat ttcaccggag atgcttggat 24360 gactaacaaa tacaatcgca caatgacaaa tactgacgga gatagcatta cctgtgatgt 24420 gtacgacgtt ctgagggcat ttgatatccg cgacccggca ctacagcatg cgttgaagaa 24480 actgttgtgt atgggcttgc gggggcacaa ggacacagga accgacttag cagaagcaat 24540 tgaaagtctg gagaagttac ggaaataccg tagtaatatt gatgagtgag aaaaaggccc 24600 cgaaaggggc cttttttttt atcgtcacga tatagaatct gatttgaacg ataaggatgt 24660 tcgaagttaa atatatctac accaacaaga ggggatgaaa ccatgaacgg attaatgaat 24720 gttagtgaag cacaaacaat gtccagccgt gagattgcgg aactgacagg taaggagcat 24780 aaaaatgttc tggctgatat ccgcaaaatg ttgtctgaga ttcaatcggc tgaaaagtca 24840 gccgattaca aagacagcaa ggggcgcacg tataaaatgc ttcttttgga taaagaggaa 24900 acgttaatcc tgatttcagg gtacagcatc aaaatgcgcg ctgccatcat ccgccgttgg 24960 caggaactgg agtcacaagc gcgcaaacca tccttaccgg tgcccaagac aatgggggag 25020 gctttaaggt tggctgcgga cttgtgggag gagaaagaac gccttgcgct tgagaacaag 25080 gaaatggcac caaaagctga tgtctacgac cgcatcatcg acagaaataa cttgtacaac 25140 gcgacgcagg ttgcccagaa gttcggtcaa tctgccgtgt ggatgaataa acaacttgaa 25200 caatttggcg tatacaatcg ctccgtaaaa cgcggtcgtg tcttccagca atggttcatc 25260 gataagggtt acggcattat gcgtgaaaca gaaactgggc attcacaggc tatgttcttc 25320 gctgaaggtg aaatgtggat tatcggtaag ttaacggcag aaggcttgat ttaaaactaa 25380 ggcccctttc ggggcctttt tcttaggcgt acttaatgcg ctggataaca attgaagact 25440 ggaactggtt gccggttgcc tggccctcaa gagacagggt aatagactcc gggccgccga 25500 tttctggggt tgcggacgta agtgaagcac gtttgagtgt aaagctcata gcgccatccg 25560 gaccatcaag aaccgaagat acctccattt ctgtttcatc caggaattta tttaacagcg 25620 tcaggtcgta cagtttacct gccagcgaga aggtgttggc cgcacgactg cgctcaacaa 25680 atgccacgct gttatttccc aactcgaact gtgcggatgc cccgttgtcg ttagtaatag 25740 tgaacgtgtc aataagttta agcggtgcag caccgtcgta aacagataca tcaaccgagg 25800 caaacgggcg ggcgctaaaa ttggtcgtga agtctgaacc actcggcggt gcttgtaaga 25860 tttcctgatt caagccgatg aacgggaaag aaccggtgac cattgcgtta acagcctgtt 25920 cgatagtgaa cccggtaaac tcaacgcctc tggttacgat atatgaatcc gggtttccgc 25980 acttaccttt caaccacgta agaatcgaat aggtcttaca caagttaccg gtttccagtt 26040 tatctgcgat tcgcaaatcg gcttgggcgt ctgcttgggc ggttagagtg tgctggatgc 26100 cagcaccggt tacaaccgta gcggttaccg cggtaacgag gaaagcttta tcgttattac 26160 cggacagacc atcgaattgc accaggtcac ctacttcaac accatcggtt acaaagctac 26220 cggtagcccg tgtaaaagtt ttcgctaccg ggtctactgt gacgccgatt gccgatgcgg 26280 tggaacccgc tacccatgaa ctagccattg cgcctgctaa cagctcgtct tggctttccg 26340 cacttagctc aatagcgtac tcacctgtta cctgacggtt gccagtgcgg atagctgatg 26400 tttcgttgct gccgtctagc tcattagaga tgagggcgtc tcgggttacc gcggggatgc 26460 cgcctgtgtt gcggagtggc tgccataccg ggttcgctgg cgtcaccccc ggcgtcgtct 26520 caagaacgta aaattgcgcg gtcatcgcgc ctttgtatgg ttgtaacgcc attatctaat 26580 cctcgctgta aaggctatga aattaattga gagaggcctc ttcgcccatc cattctcaac 26640 aataagagga ccgaggctta ctgattgtac ctcagcacag atttcgttac gactaaaaca 26700 gttaccggcg gcgaaggcgg cgtttagctt atcagctaat cggttaatag gagcgctacc 26760 caaggccgac ccaacgttaa tgtccacctg ataaacacct gcccgttgtt ccgtccagaa 26820 caagtcggct tgttccgtat cagacaatag catataactc gcaagatacg gtgtatcagt 26880 agatgtcggc gcatcgatgt tctctaacgc aaccttggta ccgttgtcca tgccgaatgt 26940 caccagtgct gtgtcaaatg ctttcgttaa gtcctcaaaa tatgttgcca ttatttcacc 27000 ttagaggctt cttcatttag tagttgctgg aacctgctga cattaaccct aacgaagcct 27060 tgaggcgcct gttgcgacca accatactca aggcgctgcg catacggcaa attgttagtt 27120 agagtaaacg tgtgccagtc agcggcattt agcacgaaat tagcggcatt actggtcgcc 27180 gtggcgccag ctttatcggt agcgtccgta gttccgtcgg cgggagtacc gccggacgcc 27240 atccagttca tacgaaagcg acctgtatct acaggactcg ctttgataat cgcagaaaac 27300 aacttgatag atacctgacg aattaccttt tcaggattct tcttagcctt ttccacgaac 27360 ttggacacat caagagcaaa actcattttc tcacctgaat aaagtacgct acaacgtcat 27420 cgttaaccat cttcttctcg atggcgacca cagaccactg ttcaccattg aattgtactt 27480 tatcatccat cttaggaatt acgctgtaat cggccttaac caccatatcc cctgcctgaa 27540 tggtggttcc gtttaccaga ccagcattaa ccggtacagg aacggattta agagggaccg 27600 tcgtgtccac cccccaaacg tactcaccta aaacagggtc ccatgttttc tggcctttgc 27660 gcaccaaagt taccgtgctg ccgtattttg acagtaggcg agtacctacg ccttgcatcc 27720 gtttactaaa agcggtgctc atcacacacc ctccagtctt gaaacgacca gtagtgcgga 27780 tggggccaca ccccaggacg taaccgttgc cacttgcggg tataacccgc cgaagttgga 27840 accggcggca tcacgcataa tctctaccgt aaaagtctgc ccggctgcgg cattcacaac 27900 aacacgagat tcaatgggaa cagtaacatc ggtgctagcc agcttagcga cggcgggtga 27960 accgaattga gcgccggcta gcagaacccg cgacagaagg atggatgttc cgctcgcccc 28020 ggtgcgaccg gcctgtaact taacacggat agcgtagtta ccggcgacgt tgaacgtaac 28080 taccccggaa gcgctaatca ttacagggtc ggcgcttgtt ttctgcgcac caccaaaggc 28 140 tacagttaat gctgtgccgg ttgtagtagg cgcctggata tctgtagacg acgcacgaag 28200 tacctccact tccttagccc cgtacagcat agaatccgcc acctgagtgg taacttcgcg 28260 caacttttcc ggtgtaatta gcccggactg gttatcaggg aagtttgccc caattaaggc 28320 gaatatttcc gatttagtct tagccatgtt tagcccctaa aaacattaaa ggagtaggca 28380 ttattgctac cgcataagag aggtcgtagc gcatcatcgg ctgccgtgat gcttactgtg 28440 ccgcctgagt agccattctt aaagtaggac accgttactg caccctcaac ccgttcggtc 28500 tgaacctctc gcccgtctgt agaccctcga acgtcggtac cagcgccata ctcgaccgcg 28560 gccattacct gagcttgaat aactaacgat ggaattacat tggatggctg ggggaacccg 28620 tgtaaagtaa caccggttcg agggaacgcc agcgcctggt ttgcggatac acgacggccg 28680 cacatctggg attcaaaaag cccaacgtat accgcgccgt tgcgtagcgc cgcttcggcc 28740 ttagcatcat cttctggcag ctcaagaccg tacttagagg ccatggacct cgcgtcttcc 28800 aggctgacgt aagagtcagc attcgggatg ccttgaccgg tttccacgat aagcggcata 28860 aattattcct ctacgctttt gcggcgacga cgctgcttag gctctccgcc accgttgtaa 28920 tgcggttctt cggccactgg ctgagtttcg attaatgctt ccgggttcgc ttcttcgcgc 28980 atcgggacta actgcccgtc aacctcgacc acacccttgt attgttcccg tactacgtaa 29040 ttatctgtca tgtctaatcc ttaaagcggc ccggaggccg cgtgtattta acttacggta 29100 actaccgtac tatctgaaat gatattaccg taaccgtcgt gcgctacgac tttgtaagta 29160 ccggaatccg ccgctgcgga gttggcctta gtgtaagtca aagcggttgc accggagatt 29220 gcgttattat ctttatacca ttgaactgtt ttagggtcca ttccgtcagc tagtacaacg 29280 gtcagggtga tagcctgacc tgcggttacc gcagtagtat catttagacc agtagagaaa 29340 cgcaacggca tcacgttttc catatcaatc tctacctgcc cgtcctgcgg gctgtcatca 29400 gataccccaa caatacgacg tttaattaca tcaaccattt ttatcctcct caggatacag 29460 taccagcgtt tttaagggcg gttaacagat tagtaacggc agtgcgcaat gaggttacat 29520 cggtgcgcaa cttattgtag ttagacacca acgcgtcaaa ctcttctttc gtaggagcgg 29580 ctgctgcggc tgcaccacta ccggcactaa tcgctggcgg tgcggctact gtagcggact 29640 ttttaacccc tccgattgtc gttgtggtag cggcgggaat atcgaattcc gcgttaacca 29700 ggctgcgggg taacccttta cctgtctttg acattatgta gtctccgtat aaatgaaaag 29760 agggaccgaa gtccctcaag agtataccct acgattaagc acctacgcca gttaccagga 29820 acgcaattgg cacatgctta cggtcgacta cacggttcca gttggcggcg ttagccaggt 29880 cctgccagct agcggagcgg gcggtagtct cggtgccgtt gccggtgatt acagcgctgg 29940 tgaagctgta accaaacgga tgcagcaacc acgttttacg agtccacagg gtttcaacac 30000 caccaccatt agcgcgagat gcttcacgtt cgtattccag cggcatctca ggattgcctt 30060 cgccgtaacc gatagcaccc tggccgaaga tgatggagat gaatttacgg ctagtgtcct 30120 gaccaactac ggtcatgctg tcatcgacga tgacgcggta accctggtag gtggcgaaca 30180 tggtgttgtt ctcagcatca cggatgaagt cgataagctg ggctttacgg gcctgtgcgt 30240 atacgaagct gtgcattgcg atagcaccga gaacctcacc gccgttgccc atcaatgcat 30300 cacccatagt ctgggtagca tcgatgaatg cacctgcgtc gaagcctgaa gttgcagaaa 30360 cgtcaacaac catgtcgttc tgctcgtgat acgcatcagt agcggatacg ttgtcgttgt 30420 acaggccgag tgcagttgca ataaggcgac gctgcgcctg acgctgccag aagttatcca 30480 gacgggatgc gacggactgc aacggattct ggctggtaag ttctacagtc aggtctgcct 30540 ggccgaaacc ttcgttcaga tatgcaacgc gggccatcat ttcaccggtc tgaatagcgc 30600 gcggggtcgc gatgtcctgg tatacgtcat tcgagtagtt aggctcgata gaagtatcga 30660 tagctttcca gaacggcagg ttagcgatgt tagacggtcc gcgggcaatc tcggcagcat 30720 acggagttgg ggtcaaaatt ccggagttga aaaacgcggt tttctctacc gggtcttccg 30780 tcatatagga cgccagtaca gggatattgc cagttacgat gttaccaata gtggtgattg 30840 ccattattta tttcctcagg gctttaagtt gccgttcaaa ttcggcaggg ttagatttat 30900 agagagccaa ccgctcactt tcactcatat ctttaaacgc tggtgcggcc ccgccgcctt 30960 tattcccgga agccccgcca ccggaagctg catttgcttt aatcaaatgc gaaaaagctt 31020 tatgttctcg caggtatttg cggaactgtt cagggtcggt agtaactacg ttaccatccg 31080 catccgtgaa tttagtaact acgtcttcac cttcaaattc agtcttaacg aacggagcaa 31140 ggatttctac cgcttccggg gtgatgaaat caccggcgaa agaacccaaa acagccttac 31200 gttcacttcc aagaattcgc gcggctaaag aagcgtagcg ttcttctttc tctttcagta 31260 cgggttcata ctgactacga atcgtctttt caaattcgtc cattttacca gcagccttca 31320 gggcctcctg gtgggcgcgc tgccgttctt cttcggcttc tttagcccgg cgagcagctt 31380 cctttttctc gctaaggagc gcttcctgat ttgccttgag cccggccact tctttctcaa 31440 tcaaggcctg cacttcttcg gcggtgtaca tttttggtgc atcacctgca cccgatttat 31500 cttctacccc agcttcttcc tggagtggat agtgtaaaaa acgattcata gtcaatatgt 31560 cccctggact ttaggacacc gggccacccg gtgtttctgt aacaagaata aattattcca 31620 cttactaagg caactattcc agaatattcc ttatgtactc ctgcaaccgg aacactttca 31680 gacgtaactg ccgtgtgcac tccgcgttac gtacgtcaat ggccaggtct tcatcagcgt 31740 cactgctcgg tggaataagc ttgcacggcg gctgcattag tgtcatatcc agggatggaa 31800 ttagcattct cggcggcgcg tcgtttgatt gacacccgtt caggaggaaa ctcacagaca 31860 gtacggccag gacgggtaac gtatttgata acttcacggg tgatagcctc cgaattttct 31920 ttgccttccg attcggcgga ggcggcctta gtatcattct gctgctgccg ttgtgttttc 31980 ttcgtcaact cagcttgtgc cttcttttgt tgttgtgaaa caagattctc tcggccttct 32040 acccacccgc ttctgtagtt gtactggccg tataaccaca aggaaaggat gccggctgcc 32 100 gtgattactg cggttacttt ccaattcata gtatcccctt tctttatacg ctaagcccgg 32160 cgtaagccgg gcgttaaact cgtatatctg taaacatcat acggcgaatc ttagataaat 32220 actgaaagca atatagctcg gcccttccga atgcgttcgc ggttacgcta gggcgctgtg 32280 gcgcccgctt cacgcgtccg caccgcggta cgctgaaagc gtaggtagcg taacataaaa 32340 aataatagaa gtctactttt tgaataaagt attcctgaat ttacattggt aaggaataac 32400 tggaataatg gtgtgtaaaa ttaaggcccc ttggcggggc ctttcgttac ttaggttctg 32460 agaccttagc cgctttactt accaccgata tcccgtcggc atctaccgcc ctgacaaaat 32520 actcccccgg gtctttaaca gttagttcac cctcaacgta cgggacgttg atcacctctt 32580 taccattttt aaaccattgc agatcgtaag gagccttacc acctttcacg actacggtta 32640 gtttagcgga accgtctttc agttttgtgt ctaccggttg tgtatcaaag tacacgtcac 32700 ccgctccgtc aagatacggc acctcgtaca ggacaccgtc tgagattgca gtcagcccgg 32760 tcttatcggc gaacggcatt tcatctaccg actcaccaag aacagattca tctttaatgt 32820 acacgacacc ttcgccagca ccagacacac gtgagtactg aacaacccga cgcgccggca 32880 catctttaac tttgaaaaag cccatcttta ttcctctttc aggtatttgg caacacgctt 32940 atcgagctcc gccatctctt taagggttaa cggacgcccg aatccatcta ctgatattac 33000 acgaaattct tccggggtta tcccactatt gcgaaaaatc ttacccctga caggaccaag 33060 tgcttcatcc tggaaccacg ccggctgttg tttaaggaac tcgtagtagg tcgtgcccgc 33 120 gtttacttgc tggcctcctt cagcacccct ggccgcccgt tttgcaccct tatccaggaa 33180 gtcgaattct ggactgatta ctggtgccgt ggtcgaccga cagttagggt gcgcgggcgg 33240 tagcgggcct ttgcctattt cgtactgcat cgaatcccgc gaccgacaaa cggctgaggt 33300 tctggagtca agggtagata cccactcgta tttcgtgatg atatcgtcat tccgcgcgta 33360 aacctgttgc cgtgcttcat tagatacgtg cgccagggct gtacggataa ccgtcgccgc 33420 attacgttcg gaaatatccg ccagtccacc aggcccgaca acgttcttaa ctatctgacg 33480 tgtggtcatc ccctgtacga acccagattt aacgcccata acaaggcgat ttacctcggt 33540 ttgccgccag ccactcatta gagaaacaaa atcaactggc ttttcgctta gttcaagagg 33600 cgcgaaagtt gcggcggccc atacttgttc ggcggctggg gttacgaaat tagcgttaac 33660 gttggtcgac agggttttta cgttccagtt aatttcatag tcagaaagct cacgagcgtc 33720 agccagtaat ttctcgtacc atcctgatgt aatcacgttc agttcttgct caagttcccg 33780 cagcatgatg gtaagtctcg ctgccgttct gctgtcatca ccaaacaaca acacctgtcg 33840 cttaacctca tcgcacatct gcttgatgaa tggcgccagg tctttcacct ctccggatgc 33900 ggtgcgttgt aaccatacct ggtggctgat taatgatgta agtagtggca tatatccctc 33960 ctgtttgcgt tgttgatgca tcttagcatg gcataggtta ttcctacaca aacacctaaa 34020 aaagtctaat ttgctataat tttgcgtatt aatgtttgcg taaacaaaga aattaggaat 34080 ataatttaaa cgatacaatt aagaggagaa aaacagatga atgtgcagac tttagaattt 34 140 ccgtatggcg aggtggaatt tacacgtgat gggtatatag atgcctcaac attaaccatc 34200 cagatgaaca gatggaggtt cgataacgga ctacaaccaa gaaatcttgc gcagatacta 34260 aaaaccgatg actgcaaaaa ctttatgcgt gtgtgcgagg aggagacagg tattgttccg 34320 ttaaaaacat caagagggag gaacgggaag acgtggttgt gcctgcacct ggccgtgtac 34380 attgcagaac agtatagtgc gtattttcac ttcctggtta tagaccgatt tattacgcaa 34440 cggcaggtag agctgcgcaa cattggtgcg gtgagctttg tcgagcttaa cgctgccgtt 34500 agtcgaatga ttgagcgcac ggagggtagg attgggcata gtggtcactt catacaggtg 34560 gctaaagcca taaaggaatc tgtagatata cgtgagataa ccgggtttga tacatgggac 34620 tcccaggatg caaaaactaa tcaattacgg agtgatattc aaaaggctat ggtgacgttg 34680 ttggatatgg aagcagtcaa ctcatgggat gagctgaaag agacgatacc gcgggttgta 34740 cgtaaatgcg cggctaatat ccgctaaaac taaggccccg taatggggcc tttttcttta 34800 ctcctgttgt tgttgtgccg cttgcggaat ctcccccgct acctgagtaa cagcacccga 34860 cggcaaaggt gcatcttcaa tagcgttcag aataccctcg tcggtccagt cagtgacacc 34920 ggccctacgc aacgcagcgt aataagcagt ggcgggcagt aatccggcat taatgtctgc 34980 catccacgca gccctgtcct gtgctgtcat aggttgcagg aagaaatcca tgttaatctg 35040 gaactcgact tcagtatctt ctggcttacc caacatcata gcaacccatc gtaaagcatc 35 100 ggtatacgcc tgacttacgt tacgagcgat tgtggccata acggatgtat cggcgccgcg 35 160 ttggatgcgc gcggattctg cggtaatttg ctgagatggt gtaataagtt gtgcgccaat 35220 ctggatagcc tgctgttctt tgtccagcat attctggcgg gccaggttgt tttcgcctgc 35280 ctgaataagt tgagcgctac cgccataacc aaggttatgc ccacaccgac taccaaattt 35340 gatcccattg gggttcgctt ccttgaacga ttgcggggtt aagttctcac cgggatagat 35400 aaacagggtc ggctggccga ctacgaagct ggactcctcg ttatcagcgc tattgcggta 35460 atgacctatg ttaagctctg ccagcggcaa caaaggcgcg tcgtcgatgg tggcgtcgtt 35520 attggtcgcc ccgataaagg taaacggaat tacccctcgt aatgattccc ctaagtccgg 35580 gtaaatctcc acgacgtctt cctgcgcacc accttccgca tcgaaacgaa aaagacgttg 35640 acggtagttg ccgtcgctat cgatgtccag aacgcgatac tgttcgccgt atttcgtttc 35700 gaactcgttt cccggttcgt tgtactccca tgtctcacgc agcacgacca tagttacgcg 35760 gtttacggaa cctacgcgcg tgagtcgcca gttaacgata ttctcggtag tgtagaaagc 35820 gattgtaggg ttgagcagac ccgcgttctg ttcggccgct gtagctgcgc cggtttccgg 35880 ggcgtcaaca agaagaccgc cacgacctac tgagtcaatc tccatgagcg tgtcctgcgc 35940 atgctgtatt aggcctacac cggacccatc tgcgttttta agcaagtatt ccagctcctt 36000 cggaatattt atttcaggtt ctttgcgcat gacgctacca accatccctg acagcgtgcg 36060 ccgggtgaag ttgtatacga tgccgcccgc ctcgtattct gcctgacgcg cttcgccgta 36120 ggctttatct ggctcattca ggccgacgtt acgtaagtag ccaaccaggt ctccggcaag 36180 cgcatggcgc accttctgcc atttcggtgc gtagtgcagc cattcgcggt gcttggtttt 36240 tacgccagaa ccctggccgt ttgctgttaa catttaataa tcctcttaaa gtgcaaaagt 36300 aaccgggata tgggaaacag gtttaaccaa cggcatttca taagcaatag ggtatcctgt 36360 cgcgtcgttc tggtggtcgt tgccgcttgt cttatccggt ataccgtttt tatcgtaagc 36420 ctgttgctcc aggcaacgtg ctgtaaccgg gcaagcctgc tcgttgacca ttaatctacc 36480 agactccagc gctttattca ccgatgctac acggtcttta acagcagggt taaccgattt 36540 cacccgtatc tcgaaaccag cgttctgtaa ttgggcaata tccgacatac tggcatcagt 36600 cgatttacgg ttcttgccgc tggcgtccgg gtacatgacg atgtggtggc cgtgtcgttt 36660 ccaccgctcg gtaatctccc tgaccacatc cggggtatcg cgcatatcta ctaactcggc 36720 taccgcgtgc cagacgtaat cgcgctgaac gtatatcgta gaggccatat gtccgacgtt 36780 gaagtcctga ccgatataca aagtttcacc aggttgtata gtctcccggc tgctgtgttt 36840 tttgcggaca taagcgtaat aaacactgcc cgatgtcagg ttaacaaatt tgccgtcgat 36900 gtaggcgtca attaactggc ccggatatgt atcccgcaac gactgcacat aatcttccgg 36960 tagaaaggga ttggatgttg tcgaggcctg aatcatttca tagcctggtt gcttttttac 37020 ggcccatctg tcgtgcacaa aacggaagcc ttctggcgtc gtaaacaccg atactgtatt 37080 agctggcttc ggggtaattg gacgatatgt acacggcaac tgacggttac gggcaataac 37140 tttattccag gcgtgctcgg cgtggtcttt atttaacgtg tccaactcgt cgatttttgc 37200 gcgaaaagat tcgtaaccga cgattcgcgc cgggttatcc aatgtacgca ggacaaaatc 37260 cccgaattgc ccggatgatg tatagattat gttgtctgat ttattgtact tgtaacgaat 37320 accccaatca gacaacttct cttccatacg cggagcgagg ataaggcgca ccaggtcgta 37380 tgtcggttca tacatggcga taagtgaatc actaccacct tccatgctgt cgagtagggc 37440 ggagttgcac atgacctccg atttacctgt gccgaatcca gcgacgaagg ccgggaattt 37500 acaatgcaga ttaaggaacg cgccttgcgg ttccgtagcc gttatatcaa cgttcaccgg 37560 acaccacctt aatagttact tcgctaattg gttcgtcgcg cgcttcttcc acaacagttt 37620 tattaagccc aagttttgct gcggcgaacg tagcagatat cccggcagca ccggtctcag 37680 tgaaatacgc ctcttccaag gcctgcgccg tttcgtatgc ttcagcgaag gcaggaacct 37740 ctcgcagcca cagcttgata atcggaacgg ttacgcctat gtgtagtgcg aaacgggcca 37800 gcgacggcgg tttgtcctgg ataagcgggc gttcgtcgcc tttagacgtg ggtacgagtt 37860 cccacgacgt gcggtcgaag aaccggatta actcgtcaca atagtccggg tcccacagtt 37920 cggcggaatg cctggatgac tggtacagac tttgtttgcc acgcgggcgc ttgcgtcgac 37980 ggtttgcgtt aaccgcttct tcgtgcgcgg cggccactac ttcggcgtca ggctgtttta 38040 atttgagttt catgcaatcc tctcttaaat gcgtatgacc atcatagcgc atcagacacc 38 100 ccctgagcgt ctattaagga ttgtataggg aaagataggc ctgtgtacag aaaagcccgc 38 160 atatgcgggc ttgtgtcagt tatataggcc gtgtcgtagt gctcttctac catcgccgcc 38220 gtaatggtcg ttagctgcgg cgataaccca ggcgttcggg atgattagca atctcatttg 38280 aaaaaccaca gcaggatgcc ggccgctcct gatacaccca agcctaaaaa cataggccac 38340 cacagatatc ggggaagact gttaaacaca gtcggttgtt ttattagcgc actgagaaag 38400 gcgaaccaaa acatcaggtc gccagtggca tcggtagcct tcaaaatctc aatcattctc 38460 cactctcctc tttataaacc caccatttac ctactgcttc ccatcgccac ccgcgactat 38520 gtatggcttc gaacctacga ccaaatactg tttcagcata ccacgtaaag aacaaacgta 38580 tcgtacgcca ggcacctcgc ttgcgcccat taatacgcca atactctctc gtcatccggg 38640 taagactgac agcttcgaca aacggccaaa gaaaaaaaat ccacatcagg tatatagcac 38700 cggcaagcag taatgttgtg aatcctatac ccatgagaat gagaaaacta tccatcactc 38760 ctccagcttc atacggtcgt tgcgcgataa tccacgtttc gtgtagatga ccggtgacgt 38820 tttcgtatgg gtcatcagtc ggttgttata gatgacatgt tgctcgcatt ttgtgtcatg 38880 gcggtacttg gacaatgtca tctcattaag tcccgtctcc cggcacgtat ccgctaatgt 38940 gccgtgtgtc tctattagtt ttgggatact ggtaatcatt cctgttcgct ctccttctca 39000 agtcgttcag gcagcatctt gtatgcttct aaatggaatt cttcgttcat tgacaggccc 39060 agcttactta cagaccccaa atacataatt cgctggcgta aagtttctta acatacgcga 39120 ttcatttatt ctccccacct acgtatttgt taaggtcgta gcggttaata agctcacgcc 39180 ggtgttcgtt aaggtgatca agcgcggctt ggtgtgcctt aatctgcgcg tcaatggctt 39240 cgattttaaa cctgccgttt acagcctcta ccatcatctt ttcatagtcg cggtcaacca 39300 tcatttacgc ttcccccaca tgcgattgag atacttgttc tggtcaggac ctgggaaact 39360 attgcgtttt aacagttcct cgcgtgttgg gaacggggta ttgcctactt tcccgcccac 39420 ctttaacgtt tctgctacct gtccgttgtc gctcattcct gcacctccca ctcgcatacc 39480 tttacaccat cataaatgcc tttagctcgt tcctcgtcgg catcagcatt agcttttcct 39540 tcgctacgat aaactccgag aatgttactg ccttcatagt gataaaatcc gtatacaata 39600 tataccttca tttcttctct cctgcataag ctgctttcag tgtctgcatc gcggcaaacc 39660 agaacgcgtc cgccgggatg ctgttgctca ggttactgac ggcaatgcgc gccatcagtt 39720 gtgcgtcttt gaatgcttct gtatctgtga ttttcatctt ctctactccg ttctcgttgt 39780 cgatgagcta aatatagtag tgttatggtt actcgtcaac tattatccca ggatatagat 39840 aaaaaaaaat cccggtaagg gtcagttacc gggataaagg agctagaggg atgaataagc 39900 aggagcaggt ttagtatctt aggaattctc ctatgtgtca acctactttg cttcgcggcc 39960 gatggcttcc gcctgttgcc acggcatacc gtcgaatagc gcaacacgac ccgcggcacg 40020 ccgacgcaat ccaaggagtg atttgccgtt ctgataatgg aactgagtta acttattccg 40080 cagtgtggca acgtcgccct tacgcagggc ctgccctgtt ccggtagaag ccgcaatcac 40140 accggcacca gcgttataca ccaggtcgca catcgcatcg aactgtgact gattgagcga 40200 cggatgcgca acggcgtcta ccgcggctac agccttagcc atgtccctgt tcagtagcaa 40260 aagaccctgg ccttcggtaa ttttctggcc ctctttcaca tctgcgccgt agtggccgta 40320 gccaattgtc aggtacttct cgttcttcgt tgccttgtac gcagttccgc ggaacccctc 40380 gaacgcggcg gtgaagtgta gcccgttatc actgatgttt cggtttgaca tacatcctcc 40440 ggtatttgta gtagcgagcg cctttcacgc aaatatgacg caccgattta agcgcgaata 40500 cggcgacgag agtcgtaatt atgaatggcg ggatgccacc ggtaaagaga tgagctagcc 40560 caccggcggc ggtaacaacg gacgccatat acagcagccg cccgaaaaga ccgtcttcaa 40620 tccagtgagc gtagatatta attaacgagg tgatgatgat tgcgaccaga caaatggcgc 40680 cgatttgtga gttgagcatg ttattttccc cacggtaact taaagttgct gagggatgag 40740 tcagcagctt cgaataactt aagccagaac atacctaccc cgaaaggaac caggtactga 40800 ccctcagact gtgataattc atagtaactt ataatgatgg gggatgcgta caccgcacaa 40860 caggcggagg caaccaggtg gcacaatcgc aaccaaggcg aaatagcctc acgcttctta 40920 atctgggata cggcgccccc ggcgaggccc gccacactaa gccaaaggta tctatcgtcc 40980 acaataaaaa atctccgaaa gttggtttac ccttcggaga ttaacattta tacgacaatt 41040 ttagtaacag ataattcgga tttatctgaa cgtataaagt ctgatggatg gcttgtgcgg 41100 gatatctgag tgcccgcgtt tacgccacag tttgtacagt ttaaccgcca tatgcgcccg 41160 cgcctggcta ctgaactcac caaggtttac cactttcttc ccgtcgtaag agtgcgcacg 41220 ccacagacct ttgcctacac aatacttagg ctggtgcacc ccagggatgc gcggatacac 41280 gtccggcatc tcattaactg gctttgcgcg cggtttaggt ttacgcttaa ccttttcttt 41340 aacgcgagta accttctttg ttaccgccga tgaaggaggt gccaggttga tgccggcgtg 41400 tttcagtata tcggttgcga gactcataat cacttctcct gtttaccgag tcgcttaggc 41460 gaacagatag cgcgtacttc tgaatcattt ggcttgtcgc cctgaaacag gaagtgcgca 41520 ttctcggcgg cgcgtgccgc tgaccggcac gcctctaatg tgtaaaacgt ttccgatgcc 41580 gcgagttgca tctggcctgc ggacagtacc cagataaata agatactggt catttcttta 41640 acccgcatat tctgcacatc atcgacgcac ttgtatcgta aaagtaccaa tcgtgctcac 41700 acccctgagc atcttcttta cggaaaccag attgctccgg cccgtgttca cctttgcgcg 41760 caagtttgta cgcctccacg tcgcccgtag cttttaccgg cgccatacgc aactccttaa 41820 cttgccctgt caggtgtgat atgtgctcag atttcttaat gttgtcttcc tgtaactcag 41880 ccaccagcac cgacacatca cgtaactgat tcttaagtga ctgcacctcc tgcatcaggt 41940 cttcacactg gttgctgttt acccgctgcc gtagctggtc aatcaggtac tctaatacct 42000 cgtctttgtt attcaccccc acactcctct atctcgtttc gataaagtaa tagtacccta 42060 ttatgttttg ttacgtcaat actattttac gattaattta tcctgacaca ccgtaatgta 42120 gcggggcttc ttcgaatgag cgcgccacca ctcaacacgg gcgtagatgc cgtggagcga 42180 tattactttg ccgtgtcggg tctctccgtt acgcccggtc caggttacaa ggtcattcgg 42240 tttcattgtt gcacctcctg caatagt 42267
Claims (12)
Bacteriophage ST20 (Accession No. KFCC11828P) having a killing ability specific to Salmonella bacteria.
The method of claim 1, wherein the bacteriophage ST20 is a bacteriophage ST20 (accession number KFCC11828P), characterized in that consisting of a nucleotide sequence represented by SEQ ID NO: 1.
The method of claim 1, wherein the Salmonella bacteria are any one selected from the group consisting of Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Gallinarum, and Salmonella Pullorum. Lee Sang-in, Bacteriophage ST20 (Accession No. KFCC11828P).
The bacteriophage ST20 (accession number KFCC11828P) according to claim 1, wherein the bacteriophage ST20 is stable in the range of pH 4 to pH 11.
The bacteriophage ST20 (accession number KFCC11828P) according to claim 1, wherein the bacteriophage ST20 is stable in the range of 40°C to 70°C.
A composition for antibiotics comprising the bacteriophage ST20 of claim 1.
A composition for feed addition, comprising the bacteriophage ST20 of claim 1.
A feed comprising the composition for feed addition of claim 7.
A disinfectant comprising the bacteriophage ST20 of claim 1.
A detergent comprising the bacteriophage ST20 of claim 1.
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| KR20230164829A (en) * | 2022-05-26 | 2023-12-05 | 전북대학교산학협력단 | Composition for inhibiting proliferation of Salmonella enteritidis comprising bacteriophage |
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| KR20090030385A (en) * | 2007-09-20 | 2009-03-25 | 주식회사 인트론바이오테크놀로지 | Novel bacteriophage having killing activity specific to salmonella selected from infectious bacteriophages of salmonella enteritidis |
| KR20120070533A (en) * | 2010-12-21 | 2012-06-29 | 씨제이제일제당 (주) | Novel bacteriophage and antibacterial composition comprising the same |
| KR101842668B1 (en) * | 2016-10-14 | 2018-03-27 | 주식회사 옵티팜 | Novel Salmonella specific becteriophage SG2 and antibacterial composition comprising the same |
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| KR20090030385A (en) * | 2007-09-20 | 2009-03-25 | 주식회사 인트론바이오테크놀로지 | Novel bacteriophage having killing activity specific to salmonella selected from infectious bacteriophages of salmonella enteritidis |
| KR20120070533A (en) * | 2010-12-21 | 2012-06-29 | 씨제이제일제당 (주) | Novel bacteriophage and antibacterial composition comprising the same |
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| KR20230164829A (en) * | 2022-05-26 | 2023-12-05 | 전북대학교산학협력단 | Composition for inhibiting proliferation of Salmonella enteritidis comprising bacteriophage |
| KR102673134B1 (en) | 2022-05-26 | 2024-06-10 | 전북대학교산학협력단 | Composition for inhibiting proliferation of Salmonella enteritidis comprising bacteriophage |
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