KR101842668B1 - Novel Salmonella specific becteriophage SG2 and antibacterial composition comprising the same - Google Patents

Novel Salmonella specific becteriophage SG2 and antibacterial composition comprising the same Download PDF

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KR101842668B1
KR101842668B1 KR1020160133617A KR20160133617A KR101842668B1 KR 101842668 B1 KR101842668 B1 KR 101842668B1 KR 1020160133617 A KR1020160133617 A KR 1020160133617A KR 20160133617 A KR20160133617 A KR 20160133617A KR 101842668 B1 KR101842668 B1 KR 101842668B1
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김영주
연영은
최재방
한범구
김현일
박철세
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Abstract

The present invention relates to a novel bacteriophage SG2 which is specific to salmonella and has a killing function; a pharmaceutical composition, an antibiotic composition, a feed additive composition, a feed, a disinfectant, or a washing agent which is for preventing or treating infectious diseases caused by salmonella and includes the bacteriophage; and a method for preventing or treating infectious diseases caused by salmonella, wherein the method comprises a step of administrating the bacteriophage to an animal. According to the present invention, a method for preventing or treating infectious diseases caused by salmonella by using bacteriophage SG2 has very high specificity to salmonella compared to a conventional chemical material such as antibiotics or the like, and the bacteriophage SG2 is infected to salmonella to proliferate 100 times or more in the salmonella, thereby having excellent bacteriolytic activity and being strong to physiochemical stimulation. In addition, the bacteriophage SG2 of the present invention is not infected to a host, such as a human, an animal, or a plant, other than bacteria, and thus a problem of antibiotic resistant bacteria due to the abuse of antibiotics, a problem of residual antibiotics in food, and a problem of a wide range of host ranges can be solved. Accordingly, the bacteriophage SG2 of the present invention can be variously used in the fields of prevention or treatment of infectious diseases caused by salmonella, antibiotic compositions, feed additive compositions, feeds, disinfectants, or washing agents.

Description

신규한 살모넬라균 특이 박테리오파지 SG2 및 이를 포함하는 항균 조성물{Novel Salmonella specific becteriophage SG2 and antibacterial composition comprising the same}A novel Salmonella-specific bacteriophage SG2 and an antimicrobial composition comprising the Salmonella specific bacteriophage SG2 and antibacterial composition comprising the same,

본 발명은 살모넬라균에 특이적인 사멸능을 갖는 신규 박테리오파지 SG2, 상기 박테리오파지를 포함하는 살모넬라균에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제 및 상기 박테리오파지를 개체에 투여하는 단계를 포함하는 살모넬라균에 의한 감염성 질병의 예방 또는 치료 방법에 관한 것이다. The present invention relates to a novel bacteriophage SG2 having a killing activity specific to Salmonella, a pharmaceutical composition for preventing or treating an infectious disease caused by Salmonella including the bacteriophage, an antibiotic composition, a composition for feed addition, a feed, And a method for preventing or treating an infectious disease caused by Salmonella comprising the step of administering the bacteriophage to an individual.

살모넬라 (Salmonella)는 장내세균과에 속하는 그람음성의 통성혐기성 세균으로, 아포를 형성하지 않는 간균이다. 살모넬라균은 사람과 동물의 장관 내 급성 감염을 유발하는 균으로, 오염된 식품을 통하여 사람에 전염될 수 있는 인수공통전염병의 주요한 원인 중 하나이다. 살모넬라균은 혈청형이 다양한 만큼 혈청형에 따라 감염되는 동물이 다를 수 있는데, 살모넬라 티피뮤리움(Salmonella Typhimurium)이나 살모넬라 엔테리티디스(Salmonella Enteritidis)의 경우 감염되는 개체의 범위가 넓어 사람 뿐 아니라 동물에게도 동시에 감염될 수 있는 혈청형으로 알려져 있다. 상기 균들은 사람에서는 식중독을 일으키지만 닭, 돼지, 소 등 다양한 동물에서는 살모넬라증(Salmonellosis)을 유발하여 급성 또는 만성의 감염증을 유발하며, 감염 시 발열, 장염, 패혈증, 폐렴, 관절염, 유산 등이 발생할 수 있다. 한편, 가금에 특이적인 살모넬라 갈리나륨(Salmonella Gallinarum)과 살모넬라 플로럼(Salmonella Pullorum)은 가금류에게만 감염되어 티푸스, 추백리 등의 질병을 일으킨다. 살모넬라 콜레라수이스(Salmonella Choleraesuis)는 돼지에 감염되어 돼지 콜레라, 급성 패혈증을 발생시킬 수 있다. 살모넬라는 돼지의 경우 2~4개월의 어린 돼지에 다발하고, 6개월 이상이 지나면 발병이 감소한다고 알려져 있다. 또한 닭의 경우에는 다양한 계군과 일령에서 감염되어 질병을 일으킨다고 보고되고 있다. 보통 살모넬라는 건강한 돼지에도 존재하며 살모넬라 보유율은 나라, 지역, 계절에 따라 차이가 있으나 보통 10~20% 정도이다. 살모넬라가 동물에게 감염되는 경로는 경구이며, 감염원의 유입 경로는 오염된 사료, 물, 외부 요인 등이 있다. 또한 계사나 돈사 내에서는 오염된 개체들을 통해 수직 또는 수평 감염이 원인이 되어 질병을 계속해서 유발할 수 있다. Salmonella (Salmonella) is a facultative anaerobic bacteria belonging to gram-negative enteric bacteria and a bacillus which does not form a spore. Salmonella is a germ causing acute infections in humans and animals, and is one of the major causes of acquired communicable diseases that can be transmitted to humans through contaminated food. Salmonella There serotypes these animals can vary from being infected, depending on variety as serotypes, of Salmonella typhimurium (Salmonella Typhimurium) and Salmonella Entebbe utility display the range of the infected object broadened people, as well as the case of (Salmonella Enteritidis) Animals Is known to be a serotype that can be infected at the same time. Although these fungi cause food poisoning in humans, they cause salmonellosis in various animals such as chickens, pigs, and cows, leading to acute or chronic infections. Infections such as fever, enteritis, sepsis, pneumonia, arthritis, . On the other hand, poultry-specific Salmonella Gallinarum and Salmonella Pullorum are only infected by poultry, causing diseases such as typhus and shingae. Salmonella Choleraesuis can become infected with pigs and cause pig cholera and acute sepsis. Salmonella is known to be a herd of two to four months old pigs, and the disease is known to decrease after more than six months. It is also reported that chickens are infected and cause diseases in various strains and ages. Salmonella usually exists in healthy pigs, and the rate of salmonella is usually 10 to 20%, depending on the country, region and season. Salmonella is an oral route to animals, and infestation sources include contaminated feed, water, and external factors. Inside the house or pig house, contaminated objects can cause vertical or horizontal infection and continue to cause disease.

특히 살모넬라 갈리나륨(Salmonella Gallinarum)은 가금에 있어서 경구나 호흡기관을 통해서 감염, 전염되어 가금티푸스 (Fowl typhoid)를 유발하는 원인균으로 분류된다. 살모넬라 갈리나륨 세균의 경우 급성 또는 만성으로 질병을 초래할 수 있으며 감염 시 폐사율이 50% 이상이며 치사율이 거의 100%에 이르러 산란계 산업에 있어서 가장 심각한 질병이라고 할 수 있다. 살모넬라 갈리나륨에 감염되는 경우 폐사율이 높으며 산란 수가 현저히 감소하게 되므로 경제적으로도 막대한 영향을 미치고 있는 실정이다. 이 균은 양계산업에서 산란계에서 더 많은 수로 빈번하게 발병하며 오염원은 주로 감염된 사료나 물 또는 환경 등 여러 경로로 전염이 되고 있다. 이 원인균은 국내에서는 1992년 이래로 주로 갈색의 산란계 농장에 전국적으로 확산되어 있다. Especially, Salmonella Gallinarum is classified as a cause of Fowl typhoid infection in poultry through infectious disease and respiratory tract. Salmonella gallinarum bacteria can cause acute or chronic diseases, and the mortality rate is 50% or more and the mortality rate is almost 100%, which is the most serious disease in the laying hens industry. Salmonella gallinarum infection has a high mortality rate and a significant decrease in the number of eggs scattered, resulting in a great economic impact. In the poultry industry, the fungus frequently develops more in the laying hens, and the pollutants are spread by various routes, mainly infected feeds, water, or the environment. These causative organisms have spread throughout the country, mainly in brown laying hens, since 1992.

현재 살모넬라 감염증을 예방 및 치료하기 위해 항생제를 주로 사용하고 있으나 살모넬라는 동물에 감염 시 세포내에 침투하여 증식하고 감염하는 경우가 많아 항생제나 약제, 기타 생균제가 침투하여 작용하기는 어려우며, 최근 많은 분야에서 항생제 내성에 대한 문제가 발생되고 있어 이에 대한 원인을 항생제 오남용으로 규명하고 항생제를 충분히 사용하지 못하게 되었다. 이에 따라 산업동물의 배합 사료 내에 성장촉진제로 사용하는 항생제를 금지하기 시작하였고, 유럽연합에서는 2006년부터, 우리나라에서는 20011년 하반기부터 항생제 사용을 규제하기 시작하였다. 또한 항생제가 없는 안전한 먹거리에 대한 소비자의 요구가 증대되고 있어 점차 항생제 사용이 줄어들고 있다. 이에 따라 세균을 제어하는 항생제의 사용이 줄어듦에 따라 세균성 질병이 발생하고 있어 항생제를 사용하지 않고 세균성 질병을 제어하는 방법에 대한 요구가 늘어나고 있다. 이러한 연구 중 하나가 자연계에 존재하는 세균을 죽이는 바이러스인 박테리오파지를 이용하는 방법이다. Currently, antibiotics are mainly used for the prevention and treatment of Salmonella infectious diseases. However, Salmonella is infected and penetrates into cells when infected with an animal. Therefore, it is difficult for antibiotics, medicines and other probiotics to penetrate and act, There is a problem about antibiotic resistance, so the reason for this is due to abuse of antibiotics and the use of antibiotics is not enough. As a result, antibiotics used as growth promoters in industrial animal feed have begun to be banned. In the European Union, in 2006 and in Korea in the second half of 2001, antibiotics began to be regulated. Consumer demand for safe food without antibiotics is also increasing, and the use of antibiotics is gradually diminishing. As the use of antibiotics to control bacteria is reduced, bacterial diseases are occurring, and there is a growing demand for methods for controlling bacterial diseases without the use of antibiotics. One of these studies is the use of bacteriophage, a virus that kills bacteria in nature.

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

그러나, 양계 산업에 막대한 경제적 피해를 일으키고 있는 살모넬라에 의한 감염증을 예방 또는 치료하기 위한 신규한 박테리오파지에 대한 연구는 여전히 부족한 실정이다. 따라서 살모넬라에 특이적인 용균활성을 갖는 박테리오파지 및 이의 응용에 대한 기술 개발이 필요하다. However, research on new bacteriophages to prevent or treat infectious diseases caused by Salmonella, which causes enormous economic damage to the poultry industry, is still lacking. Therefore, it is necessary to develop a technology for bacteriophage having a lytic activity specific to Salmonella and its application.

이에 본 발명자들은 살모넬라균에 특이적인 용균활성을 갖는 박테리오파지를 분리하고 분리된 박테리오파지의 형태적 및 유전적 특성을 분석하여 상기 박테리오파지가 살모넬라를 선택적으로 사멸시킬 수 있음을 확인함으로써 본 발명을 완성하였다.Accordingly, the present inventors have completed the present invention by isolating bacteriophage having a lytic activity specific to Salmonella and analyzing morphological and genetic characteristics of the bacteriophage thus isolated, thereby confirming that the bacteriophage can selectively kill Salmonella.

본 발명의 목적은 살모넬라균에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 SG2를 제공하는 것이다.An object of the present invention is to provide a bacteriophage SG2 comprising the nucleotide sequence shown in SEQ ID NO: 1, which has a killing activity specific to Salmonella.

본 발명의 또 다른 목적은 상기 박테리오파지 SG2를 포함하는, 살모넬라균에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating an infectious disease caused by Salmonella, which comprises the bacteriophage SG2.

본 발명의 또 다른 목적은 상기 박테리오파지 SG2를 포함하는, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제를 제공하는 것이다.It is still another object of the present invention to provide an antibiotic composition, composition for feed addition, feed, disinfectant or detergent comprising the bacteriophage SG2.

본 발명의 또 다른 목적은 상기 박테리오파지 SG2를 개체에 투여하는 단계를 포함하는, 살모넬라균에 의한 감염성 질병의 예방 또는 치료 방법을 제공하는 것이다.Yet another object of the present invention is to provide a method for preventing or treating an infectious disease caused by Salmonella, comprising the step of administering the bacteriophage SG2 to a subject.

상기 목적을 달성하기 위하여, 본 발명은 살모넬라균에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 SG2를 제공한다.In order to achieve the above object, the present invention provides a bacteriophage SG2 comprising the nucleotide sequence shown in SEQ ID NO: 1, which has a killing activity specific to Salmonella.

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 살모넬라균에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating an infectious disease caused by Salmonella, which comprises the bacteriophage SG2.

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 항생용 조성물을 제공한다.The present invention also provides an antibiotic composition comprising the bacteriophage SG2.

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 사료 첨가용 조성물을 제공한다.Further, the present invention provides a composition for feed addition comprising the bacteriophage SG2.

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 소독제를 제공한다.The present invention also provides a disinfectant comprising the bacteriophage SG2.

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 세척제를 제공한다.The present invention also provides a cleaning agent comprising the bacteriophage SG2.

또한, 본 발명은 상기 박테리오파지 SG2를 개체에 투여하는 단계를 포함하는, 살모넬라균에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.The present invention also provides a method for preventing or treating an infectious disease caused by Salmonella, comprising the step of administering the bacteriophage SG2 to a subject.

본 발명의 박테리오파지 SG2를 이용한 살모넬라균에 의한 감염성 질병의 예방 또는 치료 방법은 기존의 항생제 등의 화학물질에 비해 살모넬라에 대한 특이성이 매우 높고, 상기 박테리오파지 SG2는 살모넬라에 감염하여 살모넬라 내에서 100배 이상 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있다. 또한 본 발명의 박테리오파지 SG2는 사람, 동물, 식물 등 세균외의 다른 숙주에는 감염되지 않으므로, 항생제의 오남용으로 인한 항생제 내성균의 문제점, 식품 내의 항생제의 잔류 문제, 광범위한 숙주범위의 문제점을 해결할 수 있는 장점이 있다. 따라서, 본 발명의 박테리오파지 SG2는 살모넬라균에 의한 감염성 질병의 예방 또는 치료, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.The method for preventing or treating infectious diseases caused by Salmonella using the bacteriophage SG2 of the present invention is highly specific for Salmonella as compared with a chemical substance such as antibiotics and the bacteriophage SG2 is more than 100 times or more as high as Salmonella in salmonella It has an advantage that it is excellent in lytic activity and strong against physicochemical stimulation by proliferation. In addition, the bacteriophage SG2 of the present invention is not infected with any host other than bacteria such as human, animal, or plant. Therefore, it is possible to solve problems of antibiotic resistant bacteria caused by misuse of antibiotics, residual antibiotics in food, have. Therefore, the bacteriophage SG2 of the present invention can be used variously in the field of preventing or treating infectious diseases caused by Salmonella, antibiotic composition, composition for feed addition, feed, disinfectant, or detergent.

도 1은 박테리오파지 SG2의 살모넬라 갈리나륨에 대한 반점 분석법 수행 결과를 나타낸 도이다.
도 2는 박테리오파지 SG2의 살모넬라 갈리나륨에 대한 용균반 형성 활성을 확인한 결과를 나타낸 도이다.
도 3은 박테리오파지 SG2의 전체 유전자를 분석한 결과를 나타낸 도이다.
도 4는 전자 현미경을 통해 박테리오파지 SG2의 형태학적 특성을 확인한 결과를 나타낸 도이다.
도 5는 박테리오파지 SG2의 단백질 패턴을 SDS-PAGE를 통해 확인한 결과를 나타낸 도이다.
도 6은 박테리오파지 SG2의 pH 안정성을 확인한 결과를 나타낸 도이다.
도 7은 박테리오파지 SG2의 열 안정성을 확인한 결과를 나타낸 도이다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing a result of performing spot analysis on gallinella galliumarium of bacteriophage SG2. FIG.
FIG. 2 is a graph showing the results of confirming the activity of the bacteriophage SG2 to form a galactomannan for gallinella gallium. FIG.
3 is a diagram showing the result of analyzing the entire gene of bacteriophage SG2.
FIG. 4 is a graph showing the morphological characteristics of the bacteriophage SG2 through an electron microscope.
FIG. 5 is a graph showing the result of SDS-PAGE analysis of the protein pattern of bacteriophage SG2.
6 is a graph showing the result of confirming the pH stability of the bacteriophage SG2.
7 is a graph showing the result of checking the thermal stability of the bacteriophage SG2.

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

본 발명은 살모넬라균에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 SG2를 제공한다.The present invention provides a bacteriophage SG2 comprising the nucleotide sequence shown in SEQ ID NO: 1, which has a killing activity specific to Salmonella.

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 " means a bacterium-specific virus that infects a specific bacterium and inhibits and inhibits the growth of the bacterium, and includes a single or double-stranded DNA or RNA as a genetic material .

본 발명에서, 상기 살모넬라균은 살모넬라 엔테리티디스(Salmonella Enteritidis), 살모넬라 티피뮤리움(Salmonella Typhimurium), 살모넬라 갈리나륨(Salmonella Gallinarum), 살모넬라 콜레라수이스(Salmonella Choleraesuis) 및 살모넬라 플로럼(Salmonella Pullorum)으로 구성된 군으로부터 선택된 1종 이상 일 수 있다.In the present invention, the Salmonella is selected from the group consisting of Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Gallinarum, Salmonella Choleraesuis, and Salmonella Pullorum. ≪ / RTI >

본 발명의 박테리오파지 SG2는 살모넬라균에 대해 용균활성이 탁월하다. 상기 박테리오파지 SG2는 150-200 nm의 크기로, 정이십면체의 머리를 가지며 꼬리가 있는 형태인 사이포비리데 (Siphoviridae)에 속함을 확인하였으며, 단백질 패턴 분석 결과 65 kDa, 45 kDa, 35 kDa, 32 kDa 그리고 16 kDa의 단백질을 포함하고, 전체 염기서열 분석 결과 33,010 bp의 크기를 갖고 GC함량이 50.1%, 전체 ORFs의 수는 47개임을 확인하였으며 그 전체 염기서열을 서열번호 1로 표시하였다. 또한 상기 박테리오파지 SG2는 열 및 pH에 대한 안정성이 우수한 특성을 가지고 있다.The bacteriophage SG2 of the present invention has excellent lytic activity against Salmonella. The bacteriophage SG2 has a size of 150-200 nm and belongs to Siphoviridae, which has a head with a regular dichroic pattern and has a tail. As a result of protein pattern analysis, 65 kDa, 45 kDa, 35 kDa, 32 kDa and 16 kDa, and the total nucleotide sequence was 33,010 bp. The GC content was 50.1% and the total number of ORFs was 47, and the entire nucleotide sequence was represented by SEQ ID NO: 1. In addition, the bacteriophage SG2 has excellent stability against heat and pH.

박테리오파지 SG2는 서열번호 1로 표시되는 염기서열을 전체 유전자의 전체 또는 일부로서 포함할 수 있다. 또한, 본 발명의 박테리오파지 SG2는 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 SG2 may contain the nucleotide sequence shown in SEQ ID NO: 1 as whole or part of the entire gene. In addition, the bacteriophage SG2 of the present invention may comprise the nucleotide sequence shown in SEQ ID NO: 1 and the functional equivalent of the nucleotide sequence. The functional equivalent means at least 70% or more, preferably 80% or more, more preferably 90% or more, still more preferably 95% or more of the nucleotide sequence shown in SEQ ID NO: 1 as a result of modification or substitution of the nucleotide sequence. Or more and having substantially the same physiological activity as the nucleotide sequence shown in SEQ ID NO: 1.

상기와 같은 살모넬라균 특이적 용균활성, 내산성, 내염기성 및 내열성은 본 발명의 박테리오파지 SG2를 살모넬라균에 의한 감염성 질병의 예방 및 치료용 조성물, 및 상기 박테리오파지 SG2를 유효성분으로 포함하는 다양한 제품에 적용함에 있어, 다양한 온도 및 pH 범위의 적용이 가능하게 한다.The salmonella-specific lytic activity, acid resistance, alkali basicity and heat resistance of the present invention are applicable to a composition for preventing and treating infectious diseases caused by Salmonella bacteria and various products containing the bacteriophage SG2 as an active ingredient This makes it possible to apply various temperature and pH ranges.

본 발명의 박테리오파지 SG2는 도계장 및 양계, 산란계 농장의 분변과 농장 부근의 하수로부터 시료를 채취하여 분리한 박테리오파지로서, 박테리오파지 SG2로 명명하고 2016년 7월 22일에 한국미생물보존센터에 수탁번호 KFCC11677P로 기탁하였다. The bacteriophage SG2 of the present invention is a bacteriophage obtained by collecting samples from feces of a slaughterhouse, a fowl farm, a laying farm and a farm, and named as bacteriophage SG2, and designated as the depository number KFCC11677P ≪ / RTI >

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 살모넬라균에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition for preventing or treating an infectious disease caused by Salmonella, which comprises the bacteriophage SG2.

본 발명에서, 상기 살모넬라균은 살모넬라 엔테리티디스(Salmonella Enteritidis), 살모넬라 티피뮤리움(Salmonella Typhimurium), 살모넬라 갈리나륨(Salmonella Gallinarum), 살모넬라 콜레라수이스(Salmonella Choleraesuis) 및 살모넬라 플로럼(Salmonella Pullorum)으로 구성된 군으로부터 선택된 1종 이상일 수 있다.In the present invention, the Salmonella is selected from the group consisting of Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Gallinarum, Salmonella Choleraesuis, and Salmonella Pullorum. ≪ / RTI >

본 발명의 박테리오파지 SG2는 살모넬라균에 특이적인 사멸능을 가지므로, 상기 균들에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물에 사용될 수 있다. 본 발명의 박테리오파지 SG2는 살모넬라균 중에서도 살모넬라 갈리나륨에 더욱 특이적이므로, 본 발명은 살모넬라 갈리나륨 감염에 의한 감염성 질병의 예방 또는 치료용 약학적 조성물을 제공할 수 있으나, 이에 제한되는 것은 아니다.Since the bacteriophage SG2 of the present invention has an ability to kill salmonella-specific, it can be used in a pharmaceutical composition for preventing or treating infectious diseases caused by the above-mentioned fungi. The bacteriophage SG2 of the present invention is more specific to Salmonella gallinarum among Salmonella, so that the present invention can provide a pharmaceutical composition for preventing or treating infectious diseases caused by Salmonella gallinarum infection, but the present invention is not limited thereto.

본 발명에서, 살모넬라균에 의한 감염성 질병은 유행성 또는 급성으로 발병하는 전염성 질환을 통칭하는 개념으로, 식중독, 장티푸스, 살모넬라증, 장염, 가금티푸스, 추백리, 돼지 콜레라, 및 패혈증으로 구성된 군으로부터 선택된 1종 이상일 수 있다.In the present invention, an infectious disease caused by Salmonella is a concept collectively referred to as an epidemic disease that occurs epidemically or acutely, and is a species selected from the group consisting of food poisoning, typhoid fever, salmonellosis, enteritis, poultry typhus, Or more.

바람직하게, 상기 살모넬라 갈리나륨에 의한 감염성 질병은 가금티푸스일 수 있으나, 이에 제한되는 것은 아니다.Preferably, the infectious disease caused by the Salmonella gallinarum may be poultry typhus, but is not limited thereto.

바람직하게, 상기 살모넬라 엔테리티디스에 의한 감염성 질병은 식중독, 장염일 수 있으나, 이에 제한되는 것은 아니다.Preferably, the infectious disease caused by Salmonella enteritidis may be food poisoning, enteritis, but is not limited thereto.

바람직하게, 상기 살모넬라 티피뮤리움에 의한 감염성 질병은 식중독, 장티푸스, 살모넬라증일 수 있으나, 이에 제한되는 것은 아니다.Preferably, the infectious disease caused by Salmonella typhimurium may be food poisoning, typhoid, or salmonellosis, but is not limited thereto.

바람직하게, 상기 살모넬라 플로럼에 의한 감염성 질병은 추백리일 수 있으나, 이에 제한되는 것은 아니다.Preferably, the infectious disease caused by the Salmonella fl rum can be, but is not limited to, shrapnel.

바람직하게, 상기 살모넬라 콜레라수이스에 의한 감염성 질병은 돼지 콜레라 또는 패혈증일 수 있으나, 이에 제한 되는 것은 아니다.Preferably, the infectious disease caused by Salmonella cholera strains is, but is not limited to, porcine cholera or sepsis.

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

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

본 발명의 용어 “치료”란 조성물의 투여로 상기 질병의 증세가 호전되거나 상기 질병의 억제 또는 경감 및 이롭게 변경되는 모든 행위를 의미한다.The term " treatment " of the present invention means all the acts of improving the symptom of the disease or inhibiting or alleviating and beneficially altering the disease upon administration of the composition.

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

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

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

상기 비강 분무의 경우 일정량의 박테리오파지 SG2를 물에 희석하여 분무기나 분무 시스템을 통해 비강으로 흡수 되도록 할 수 있다. 상기 비강 분무를 위한 비강 분무제 또는 호흡기용 제형으로는 에어로졸제 등을 들 수 있다.In the case of the nasal spray, a certain amount of bacteriophage SG2 can be diluted in water and absorbed into the nasal cavity through a sprayer or spray system. Examples of the nasal spray agent for the nasal spray or the aerosol agent for the respiratory system are mentioned.

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

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

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

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

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 항생용 조성물을 제공한다. The present invention also provides an antibiotic composition comprising the bacteriophage SG2.

본 발명에서, 용어 “항생용 조성물”은 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 " means a preparation which can be given to an animal in the form of a drug to kill the bacterium, and collectively refers to an antiseptic, a bactericide, an antibiotic and an antibacterial agent.

본 발명의 박테리오파지 SG2는 기존 항생제에 비하여 살모넬라균에 대한 특이성이 매우 높으므로, 유익균은 죽이지 않고, 특정 병원균만 사멸시킬 수 있고, 약물 내성 내지 저항성을 유도하지 않아, 기존의 항생물질에 비하여 제품수명(life cycling)이 긴 신규 항생제로서 이용될 수 있다.The bacteriophage SG2 of the present invention has a higher specificity to Salmonella than the conventional antibiotics. Therefore, the bacteriophage SG2 of the present invention can kill only specific pathogens and does not induce drug resistance or resistance, (life cycling) can be used as a new antibiotic.

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 사료 첨가용 조성물을 제공한다.Further, the present invention provides a composition for feed addition comprising the bacteriophage SG2.

축산, 수산업에서 사용되는 사료 첨가용 항생제는 질병의 예방 목적으로 사용되고 있는데, 예방 목적의 항생제 투여는 내성균 발생 가능성을 높이고 가축에 잔류하는 항생제가 사람에게 전달될 수 있어서 문제이다. 항생제가 육류를 통해 인체에 흡수되면 항생제 내성을 유발해 질병의 확산을 부를 수도 있다. 또한, 사료에 섞여 먹이는 항생제의 종류가 많고 이는 다제 내성균 발생 확률이 높아지는 문제점이 있기 때문에 좀 더 자연 친화적이면서도 기존의 항생제의 사용에서 발생한 문제를 해결할 새로운 사료 첨가용 항생물질로서 본 발명의 상기 박테리오파지 SG2를 이용할 수 있다.Antibiotics for feed additive used in animal husbandry and fisheries are used for the prevention of diseases. The administration of antibiotics for prevention is problematic because it increases the possibility of producing resistant bacteria and antibiotics remaining in the livestock can be delivered to humans. If antibiotics are absorbed into the body through meat, they may cause antibiotic resistance and spread the disease. In addition, since there are many kinds of antibiotics mixed with feed and there is a problem that the probability of occurrence of multidrug-resistant bacteria is increased, it is possible to use the bacteriophage SG2 of the present invention as a new feed additive antibiotic substance which is more natural- Can be used.

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

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

본 발명의 박테리오파지 SG2를 포함하는 사료에는 식물성으로는 곡물류, 근과류, 식품가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류, 곡물부산물류 등이 있으며, 동물성으로는 단백질류, 무기물류, 유지류, 광물성류, 유지류, 단세포 단백질, 동물성 플랑크톤류, 남은 음식물 등이 있으며 이에 제한되는 것은 아니다.Examples of the feedstuffs containing bacteriophage SG2 of the present invention include cereals, muscle roots, food processing busines, algae, fibers, pharmaceutical buses, oils, fats and oils, But are not limited to, proteins, inorganic materials, fats, oils, fats, oils, monocellular proteins, animal plankton, and remnants.

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

또한, 본 발명은 박테리오파지 SG2를 포함하는, 음용수 첨가제를 제공한다.The present invention also provides a drinking water additive comprising bacteriophage SG2.

본 발명의 음용수 첨가제는 상기 박테리오파지 SG2 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 살모넬라균의 숫자를 감소시킬 수 있는 효과가 있다.The additive for drinking water of the present invention can be used either as a method of separately preparing the bacteriophage SG2 or a composition containing the bacteriophage SG2 as a additive for drinking water, mixing the mixture with feed or drinking water, or directly adding it in the production of drinking water. As a result, the number of salmonella bacteria can be continuously reduced by mixing and supplying the drinking water.

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

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 소독제를 제공한다.The present invention also provides a disinfectant comprising the bacteriophage SG2.

살모넬라 엔테리티디스, 살모넬라 티피뮤리움, 살모넬라 갈리나륨, 살모넬라 콜레라수이스 또는 살모넬라 플로럼에 특이적 사멸능을 갖는 본 발명의 박테리오파지 SG2를 포함하는 소독제는 병원감염을 막기 위한 병원 및 보건용의 소독제로 유용하게 사용될 수 있고 일반 생활 소독제, 식품 및 조리 장소 및 설비의 소독제, 양계장, 축사 등의 건물, 축체, 음수, 깔짚, 난좌, 운반차량, 식기 등의 각종 생육 용품의 소독 등에 사용될 수 있다.Disinfectants containing the bacteriophage SG2 of the present invention, which have specific killing ability in Salmonella enteritidis, Salmonella typhimurium, Salmonella gallinarum, Salmonella cholera strain or Salmonella fl rum, And it can be used for general disinfectant, disinfectant for foods and cooking places and facilities, disinfection of various kinds of growing equipment such as poultry houses, housing buildings such as pens, shrubs, drinking water, litter, stairs, transportation vehicles and tableware.

또한, 본 발명은 상기 박테리오파지 SG2를 포함하는, 세척제를 제공한다.The present invention also provides a cleaning agent comprising the bacteriophage SG2.

본 발명의 박테리오파지 SG2는 살모넬라 엔테리티디스, 살모넬라 티피뮤리움, 살모넬라 갈리나륨, 살모넬라 콜레라수이스 또는 살모넬라 플로럼에 특이적 사멸능을 가지므로, 상기 살모넬라균에 노출되었거나 노출될 가능성이 있는 가축의 피부 표면 또는 신체 각 부위 등을 세척하는 용도로도 사용될 수 있다.The bacteriophage SG2 of the present invention has a specific killing capacity for Salmonella enteritidis, Salmonella typhimurium, Salmonella gallinarum, Salmonella cholera strain or Salmonella florum, It can also be used for cleaning skin surface or body parts.

또한, 본 발명은 상기 박테리오파지 SG2를 개체에 투여하는 단계를 포함하는, 살모넬라균에 의한 감염성 질병의 예방 또는 치료 방법을 제공한다.The present invention also provides a method for preventing or treating an infectious disease caused by Salmonella, comprising the step of administering the bacteriophage SG2 to a subject.

상기 살모넬라균은 살모넬라 엔테리티디스, 살모넬라 티피뮤리움, 살모넬라 갈리나륨, 살모넬라 콜레라수이스 또는 살모넬라 플로럼을 포함할 수 있다.The Salmonella can include Salmonella enteritidis, Salmonella typhimurium, Salmonella gallinarum, Salmonella cholera strain or Salmonella flum.

본 발명의 방법은 살모넬라균에 의한 감염성 질병, 예컨대 유행성 또는 급성으로 발병하는 전염성 질환으로, 식중독, 장티푸스, 살모넬라증, 장염, 가금티푸스, 추백리, 돼지 콜레라, 또는 패혈증이 발생한 개체에 박테리오파지 SG2를 투여하는 단계를 포함한다. The method of the present invention can be used for the treatment of infectious diseases caused by Salmonella, for example, epidemic or acute infectious diseases, in which bacteriophage SG2 is administered to individuals suffering from food poisoning, typhoid fever, salmonellosis, enteritis, poultry typhus, mulberry, pig cholera, .

본 발명에서, 용어 "개체"는 인간을 포함한 포유동물, 바람직하게는 가축류를 말하며, 가장 바람직하게는 돼지 또는 가금류를 말하나, 이에 제한되는 것은 아니다.In the present invention, the term "individual " refers to a mammal including a human, preferably a livestock, most preferably a pig or a poultry, but is not limited thereto.

상기 용어 "가축류"는 인간에 의하여 순화, 개량되어 인간과 함께 공동생활을 하는 유용한 동물을 지칭하는 개념으로, 조류, 어류, 곤충류, 포유류를 모두 포함하는 개념이며, 상기 가축류는 돼지, 소, 닭, 말, 오리 및 개로 구성된 군에서 선택될 수 있으나, 이에 제한되는 것은 아니다.The term "livestock" is a concept that refers to useful animals that are refined and improved by humans to co-live with humans, including birds, fishes, insects, and mammals. The livestock are pigs, , Horse, duck, and dog, but is not limited thereto.

상기 용어 "가금류"는, 가축 중 조류에 속하는 동물을 총칭하여 이르는 개념이며, 상기 가금류는 닭, 오리, 꿩, 메추리, 타조, 거위 및 칠면조로 구성된 군에서 선택될 수 있으나, 이에 제한되는 것은 아니다.The term "poultry" is a concept that refers collectively to animals belonging to the domestic livestock, and the poultry may be selected from the group consisting of chicken, duck, pheasant, quail, ostrich, goose and turkey, but is not limited thereto .

본 발명의 방법에서 상기 박테리오파지 SG2 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 SG2 or composition may be administered to an animal in the form of a pharmaceutical preparation, or by mixing with feed or drinking water of a livestock and feeding it.

본 발명의 방법에서 상기 박테리오파지 SG2 또는 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 경구 또는 비경구의 다양한 경로를 통하여 투여될 수 있으며, 구체적으로, 구강, 직장, 국소, 정맥 내, 복강 내, 근육 내, 동맥 내, 경피, 비측 내, 흡입 등을 통해 통상적인 방식으로 투여될 수 있다.In the method of the present invention, the administration route of the bacteriophage SG2 or composition can be administered through various routes of oral or parenteral administration, as long as it can reach the target tissues. Specifically, oral administration, rectal administration, topical administration, intravenous, Intravenous, intramuscular, intraarterial, transdermal, intranasal, inhalation, and the like.

본 발명의 방법에 투여되는 상기 박테리오파지 SG2의 적합한 총 1일 사용량은 올바른 의학적 판단범위 내에서 처치의에 의해 결정될 수 있다는 것은 당업자에게 자명한 일이다. 특정 개체에 대한 구체적인 치료적 유효량은 달성하고자 하는 반응의 종류와 정도, 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여시간, 투여 경로 및 조성물의 분비율, 치료기간, 구체적 조성물과 함께 사용되거나 동시 사용되는 약물을 비롯한 다양한 인자와 의약 분야에 잘 알려진 유사 인자에 따라 다르게 적용하는 것이 바람직하다.It will be apparent to those skilled in the art that the appropriate total daily usage of the bacteriophage SG2 administered in the methods of the present invention may be determined by treatment within the scope of sound medical judgment. The specific therapeutically effective amount for a particular individual will depend upon the type and extent of the response to be achieved, the age, weight, general health, sex and diet of the patient, the time of administration, the route of administration and the fraction of the route of administration, It is preferable to apply differently depending on various factors including drugs used or co-used and similar factors well known in the medical field.

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

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

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

1.1 박테리오파지의 스크리닝 및 단일 박테리오파지의 분리1.1 Screening of bacteriophages and isolation of single bacteriophages

살모넬라 갈리나륨(Salmonella Gallinarum)에 특이적인 박테리오파지를 스크리닝 및 분리하기 위해서 반점 분석법(spot test) 및 용균반 분석을 수행하였다. 시료는 박테리오파지가 존재할 것으로 예상되는 도계장 및 양계, 산란계 농장의 분변과 농장 부근의 하수로부터 채취하였으며, 박테리오파지 분리에 사용된 살모넬라 갈리나륨은 (주)옵티팜에서 야외 시료로부터 분리 동정된 것을 사용하였다.Spot test and sputum analysis were performed to screen and isolate bacteriophages specific to Salmonella Gallinarum. Samples were collected from the slaughterhouses and poultry fields, the feces of the laying hens farms, and the sewage near the farms where bacteriophages were expected to be present. Salmonella gallinarium used for bacteriophage isolation was obtained from Opti Farm, which was isolated from outdoor samples.

구체적으로, 채취한 시료를 균질화 한 후 살모넬라 갈리나륨과 함께 35℃에서 하룻밤 동안 진탕배양 하였다. 상기 배양액을 4,000 rpm으로 20분간 원심분리한 후, 상등액을 공극(pore) 크기가 0.45 μm인 필터를 이용하여 여과하여 시료 여과액을 얻었다. 시료 여과액에 대해 살모넬라 갈리나륨이 접종되어 있는 LB 플레이트(plate)에 전처리한 시료를 점적하여 18시간 동안 배양하여 스크리닝하는 반점 분석법(spot test)을 수행하고 그 결과를 도 1에 나타내었다. Specifically, the collected samples were homogenized and incubated with Salmonella gallinarum at 35 ° C overnight with shaking. The culture solution was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was filtered using a filter having a pore size of 0.45 μm to obtain a sample filtrate. A spot test was performed in which a sample pre-treated on an LB plate inoculated with Salmonella gallinarum was spotted and cultured for 18 hours to perform screening. The results are shown in FIG.

도 1에 나타낸 바와 같이, 용균반이 나타나는 시료를 확인하였으며, 용균반이 확인된 시료에는 살모넬라 갈리나륨을 용균할 수 있는 박테리오파지가 존재하는 것으로 추정하고 용균반 형성을 확인하였다.As shown in FIG. 1, a sample showing a sputum smear was identified, and it was assumed that a bacteriophage capable of smearing Salmonella gallinarium was present in the sample in which the sputum was confirmed, and the formation of the sputum was confirmed.

용균반 형성을 확인하기 위하여, 전배양된 살모넬라 갈리나륨 150 μL를 취해서 멸균된 튜브에 넣고 3-5 mL의 LB 탑 아가(top agar) [LB 배지(트립톤 10 g/L, 효모 추출물(yeast extract) 5 g/L, NaCl 10 g/L) + 0.6% 아가]를 첨가하여 잘 혼합한 후 미리 만들어 둔 2% LB 아가 플레이트에 중층하였다. 실온에서 30분간 방치한 후 상기 시료 여과액을 각각 10 μL씩 점적하고, 다시 실온에서 30분간 방치한 후 35℃에서 24시간 동안 배양하여 박테리오파지의 존재 유무를 확인하였다.150 μL of pre-cultured salmonella gallinarum was placed in a sterilized tube and incubated with 3-5 mL of LB top agar [LB medium (10 mg / L of tryptone, yeast extract 5 g / L of extract, 10 g / L of NaCl) + 0.6% of agar] were added to the wells and the mixture was layered on a 2% LB agar plate. After allowing to stand at room temperature for 30 minutes, the sample filtrate was dropped by 10 μL each, and the mixture was allowed to stand at room temperature for 30 minutes and then incubated at 35 ° C. for 24 hours to confirm presence or absence of bacteriophages.

박테리오파지의 존재를 확인한 시료로부터 박테리오파지를 순수 분리하기 위해서 소프트 아가 오버레이(soft agar overlay)법을 이용하였다. 박테리오파지가 존재하는 것으로 확인된 시료를 식염수로 적절히 희석하여 각 희석단의 시료 희석액 100 μL를 취하고, 대상 세균(살모넬라 갈리나륨) 배양액 150 μL에 혼합한 후 0.6% LB 탑 아가 3 mL을 첨가하여 잘 혼합한 뒤, 미리 만들어 둔 2% LB 아가 플레이트에 중층하여 방치한 후 35℃에서 18시간 배양하였다. 배양한 각 플레이트에서 형성된 용균반을 확인한 후, 이를 선택하여 400 μL의 SM 완충액 (NaCl 5.8 g/L; MgSO47H2O 2 g/L; 1M Tris-Cl (pH 7.5) 50 mL)에 현탁하고, 실온에서 4시간동안 방치하였다. 상기 박테리오파지 액 100 μL를 0.6% LB 탑 아가 및 세균 배양액 500 μL와 혼합한 뒤 150 mm 지름의 LB 플레이트에 분주하여 중층한 후 완전히 용균이 일어날 때까지 35℃에서 배양하였다. 배양이 종료된 후, LB 플레이트에 15 mL의 SM 완충액을 가하고 실온에서 4시간 동안 천천히 교반하면서 박테리오파지 용액을 회수한 후 4,000 rpm으로 20분간 원심분리하여 상등액만 취하고 이를 0.45 μm 필터로 여과하여 최종 박테리오파지 용액을 회수하였다. 이렇게 얻어진 최종 박테리오파지의 용균반을 확인한 결과를 도 2에 나타내었다. A soft agar overlay method was used to isolate the bacteriophage from the sample which confirmed the existence of the bacteriophage. A sample of which the bacteriophage was confirmed to be present was diluted appropriately with saline, 100 μL of the sample dilution of each dilution stage was taken, mixed with 150 μL of the bacterial (salmonella gallinarium) culture, and 3 mL of 0.6% LB topa After mixing, the mixture was allowed to stand in a 2% LB agar plate that had been previously prepared, followed by incubation at 35 ° C for 18 hours. The plate was incubated with a solution of 400 μL of SM buffer (NaCl 5.8 g / L; MgSO 4 7H 2 O 2 g / L; 1M Tris-Cl (pH 7.5) 50 mL) And allowed to stand at room temperature for 4 hours. 100 .mu.L of the bacteriophage solution was mixed with 500 .mu.L of 0.6% LB topagarase and bacterial culture, and the mixture was divided into 150-mm-diameter LB plates and cultured at 35.degree. After completion of the incubation, 15 mL of SM buffer was added to the LB plate, and the bacteriophage solution was recovered while slowly stirring at room temperature for 4 hours. Then, the supernatant was centrifuged at 4,000 rpm for 20 minutes to filter out the supernatant. The solution was recovered. FIG. 2 shows the result of checking the thus obtained final bacteria bacteriophage.

도 2에 나타낸 바와 같이, 3-4 mm의 크기를 가지는 비교적 크기가 큰 용균반을 형성하는 박테리오파지를 확인하였고, 상기 박테리오파지는 살모넬라 갈리나륨 특이적인 용균능을 갖는 박테리오파지임을 확인하였다. As shown in Fig. 2, a bacteriophage having a size of 3-4 mm and a relatively large size of a spore bacterium was confirmed, and it was confirmed that the bacteriophage was a bacteriophage having salmonella gallinarium specific solubility.

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

상기 실시예 1.1에서 확인한 살모넬라 갈리나륨 특이적 박테리오파지를 대량 배양하기 위하여, 1.1에서 회수한 최종 박테리오파지 배양액을 1.0x107 PFU/mL로 희석한 후 전배양된 살모넬라 갈리나륨 세균 1.0x109 CFU/mL과 혼합하여 MOI (Multiplicity of infection)를 0.01이 되도록 하여 35℃에서 120 rpm으로 9시간 배양하였다. 배양이 종료된 후 배양액 전량을 6,000 rpm으로 30분간 원심분리하여 상등액을 수득하였고, 이를 0.2 μm 필터로 여과하여 최종 박테리오파지 배양액을 회수하였다. 분리된 박테리오파지를 "박테리오파지 SG2(Bacteriophage SG2)"로 명명하고, 2016년 7월 22일에 한국미생물보존센터 (Korean Culture Center of Microorganisms, 서울, 한국)에 기탁하고, 수탁번호 KFCC11677P를 부여받았다.In order to mass-culture Salmonella Galiniarum-specific bacteriophage identified in Example 1.1 above, the final bacteriophage culture recovered at 1.1 was diluted to 1.0 x 10 7 PFU / mL, and then 1.0 x 10 9 CFU / mL of pre-cultured salmonella gallinarum bacteria And the mixture was incubated at 35 DEG C and 120 rpm for 9 hours so that MOI (multiplicity of infection) was 0.01. After the culture was completed, the whole culture was centrifuged at 6,000 rpm for 30 minutes to obtain a supernatant, which was then filtered with a 0.2 μm filter to recover the final bacteriophage culture. The isolated bacteriophage was named "Bacteriophage SG2" and deposited on July 22, 2016 at the Korean Culture Center of Microorganisms (Seoul, Korea) and received the accession number KFCC11677P.

실시예Example 2: 박테리오파지 SG2의 전체 유전자 분석 2: Whole gene analysis of bacteriophage SG2

박테리오파지 SG2의 전체 유전자를 분석하기 위해, 박테리오파지 SG2 배양액에 20 mM EDTA, 50 μg/mL의 프로테나아제 K, 0.5% (w/v) SDS를 가한 후, 50℃에서 1시간 동안 정치하였다. 동량의 페놀-클로로포름-이소프로필 알코올 (25:24:1)과 혼합한 후 실온에서 12,000 rpm으로 10분간 원심분리하여 상등액 회수하였다. 회수된 상등액을 동일 부피의 페놀-클로로포름-이소프로필 알코올 (25:24:1)과 혼합하여 실온에서 12,000 rpm으로 10분간 원심분리한 후 상등액 회수하고, 상기 상등액에 3M 소듐 아세테이트를 전체 부피의 10%(v/v)가 되도록 가한 후 혼합하였다. 여기에 2배 부피의 차가운 95% 에탄올을 가하여 혼합한 뒤 -20℃에서 1시간 동안 정치하였다. 반응이 끝난 후 0℃에서 10분간 12,000 rpm으로 원심분리하여 침전물 획득하였고, 차가운 70% 에탄올로 2회 세척한 뒤 에탄올을 완전히 제거하여 DNA 펠렛을 회수하였다. 회수한 펠렛을 최종 부피가 50 μL이 되도록 TE 완충액 (Tris-EDTA, pH8.0)에 용해시킨 후 DNA 농도를 측정하였다. 전체 유전자 분석을 위해 PacBioRSII 를 이용하여 유전자 시퀀싱을 시행하였다 (마크로젠, 한국). 시퀀싱 결과로 얻은 박테리오파지 SG2 유전자의 유전자 맵을 도 3에 나타내었다.To analyze the whole gene of bacteriophage SG2, 20 mM EDTA, 50 μg / mL proteinase K and 0.5% (w / v) SDS were added to the bacteriophage SG2 culture medium and left at 50 ° C. for 1 hour. The mixture was mixed with an equal amount of phenol-chloroform-isopropyl alcohol (25: 24: 1) and centrifuged at 12,000 rpm for 10 minutes at room temperature to recover the supernatant. The recovered supernatant was mixed with an equal volume of phenol-chloroform-isopropyl alcohol (25: 24: 1) and centrifuged at 12,000 rpm for 10 minutes at room temperature. The supernatant was recovered and 3M sodium acetate was added to the supernatant in a total volume of 10 % (v / v). The mixture was mixed with 2 volumes of cold 95% ethanol, and the mixture was allowed to stand at -20 DEG C for 1 hour. After the reaction was completed, the precipitate was collected by centrifugation at 12,000 rpm for 10 minutes at 0 ° C., washed twice with cold 70% ethanol, and then the ethanol was completely removed to recover the DNA pellet. The recovered pellet was dissolved in TE buffer (Tris-EDTA, pH 8.0) to a final volume of 50 μL, and the DNA concentration was measured. Gene sequencing was performed using PacBioRSII for whole gene analysis (Macrogen, Korea). The gene map of the bacteriophage SG2 gene obtained as a result of the sequencing is shown in Fig.

도 3에 나타낸 바와 같이, 박테리오파지 SG2의 전체 유전자의 크기는 33,010 bp이며, GC함량이 50.1%, 전체 ORFs의 수는 47개임을 확인하였다. 분석한 박테리오파지 SG2의 전체 유전자 서열을 서열번호 1로 나타내었다. As shown in FIG. 3, it was confirmed that the size of the entire gene of bacteriophage SG2 was 33,010 bp, the GC content was 50.1%, and the total number of ORFs was 47. The entire gene sequence of the bacteriophage SG2 analyzed is shown in SEQ ID NO: 1.

실시예Example 3: 박테리오파지 SG2의 형태학적 특성 3: Morphological characteristics of bacteriophage SG2

박테리오파지 SG2의 형태학적 특성을 알아보기 위해, 그리드를 고정한 뒤 0.01%의 BSA 용액 (Bovine serum albumin) 10 μL를 처리하고 10 초 후 필터 페이퍼로 제거하였다. 준비된 박테리오파지 SG2를 10 μL 취하여 그리드 위에 분주하고 1-2분간 방치 후 필터 페이퍼로 제거하는 과정을 2회 반복한 뒤, 포스포텅스텐산(phosphotungstic acid) 용액 10 μL를 샘플을 처리한 그리드 위에 처리하고 5분간 방치 한 후 필터 페이퍼로 제거하는 과정을 2회 반복하여 염색한 다음 투과전자현미경 (Carl Zeiss transmission electron microscope, 80 kV, 120,000~200,000)으로 검경하였으며 그 결과를 도 4에 나타내었다.To determine the morphological characteristics of bacteriophage SG2, 10 μL of 0.01% BSA solution (Bovine serum albumin) was applied after fixation of the grid, and after 10 seconds, it was removed with filter paper. 10 μL of the prepared bacteriophage SG2 was dispensed onto the grid, allowed to stand for 1 to 2 minutes, and then removed with filter paper twice. Then, 10 μL of phosphotungstic acid solution was treated on the sample-treated grid After leaving for 5 minutes, the filter paper was repeatedly dyed twice. The result was observed with a transmission electron microscope (Carl Zeiss transmission electron microscope, 80 kV, 120,000 ~ 200,000). The results are shown in FIG.

도 4에 나타낸 바와 같이, 박테리오파지 SG2는 정이십면체의 머리를 가지며 꼬리가 있는 형태인 사이포비리데 (Siphoviridae)에 속하며, 150-200 nm의 크기를 가지는 것으로 확인하였다.As shown in Fig. 4, the bacteriophage SG2 belongs to Siphoviridae, which has a head of regular trilobes and has a tail, and has a size of 150-200 nm.

실시예Example 4: 박테리오파지 SG2의 단백질 패턴 분석 4: Protein pattern analysis of bacteriophage SG2

박테리오파지 SG2의 단백질 패턴을 분석하기 위해, 1.0x1010 PFU/mL로 준비된 박테리오파지 SG2 40 μL를 10 μL의 5X 로딩 완충액과 혼합하고, 10분간 끓인 다음 12.5% SDS-PAGE를 수행하였다. SDS-PAGE 수행 후 단백질 전기영동 결과를 도 5에 나타내었다.To analyze the protein pattern of bacteriophage SG2, 40 μL of bacteriophage SG2 prepared to 1.0 × 10 10 PFU / mL was mixed with 10 μL of 5X loading buffer, boiled for 10 minutes and then subjected to 12.5% SDS-PAGE. The results of SDS-PAGE and protein electrophoresis are shown in FIG.

도 5에 나타낸 바와 같이, 박테리오파지 SG2는 65 kDa, 45 kDa, 35 kDa, 32 kDa 그리고 16 kDa 크기의 주요 단백질을 가지는 것을 확인하였다.As shown in Fig. 5, bacteriophage SG2 has major proteins of 65 kDa, 45 kDa, 35 kDa, 32 kDa and 16 kDa in size.

실시예Example 5: 박테리오파지 SG2의 다양한 세균에 대한 감염성 조사 5: Investigation of infectivity of various bacterium of bacteriophage SG2

박테리오파지 SG2의 다양한 세균에 대한 감염성을 조사하였다. The infectivity of bacteriophage SG2 against various bacteria was investigated.

우선, 박테리오파지 SG2의 살모넬라 갈리나륨에 대한 감수성을 소프트 아가 오버레이법을 통해 확인한 결과를 표 1에 나타내었다.First, the susceptibility of bacteriophage SG2 to Salmonella gallinarium was confirmed by soft agar overlay method. The results are shown in Table 1.

대상세균명Target bacteria name 대상 세균수Number of target bacteria 용균 활성이 확인된 세균수Number of bacterium with confirmed lytic activity 용균 활성이 확인되지 않은 세균수Number of bacteria for which lytic activity has not been confirmed 감수성 비율Susceptibility ratio 살모넬라 갈리나륨Salmonella gallinarum 15균주15 strains 12균주12 strains 3균주3 strains 86.6%86.6%

상기 표 1에 나타낸 바와 같이, 박테리오파지 SG2는 살모넬라 갈리나륨에 대한 용균활성이 탁월함을 확인하였다.As shown in Table 1 above, bacteriophage SG2 was found to be excellent in the lytic activity against gallinella gallinarium.

또한, 박테리오파지 SG2가 다양한 세균에 대해서 용균활성을 나타내는지 확인하기 위하여 ㈜옵티팜에서 분리하여 보유하고 있는 균주와 미생물 자원센터 등에서 분양받은 균주를 대상으로 소프트 아가 오버레이법을 통해서 감염성을 확인하였다. 그 결과를 표 2에 나타내었다.In order to confirm that bacteriophage SG2 exhibits lytic activity against various bacteria, the strains isolated from OptiParm Co., Ltd. and purchased from the microorganism resource center were infectious through soft agar overlay method. The results are shown in Table 2.

Figure 112016099797406-pat00001
Figure 112016099797406-pat00001

표 2에 나타낸 바와 같이, 살모넬라 갈리나륨 외에도 다른 미생물을 대상으로 감수성 검사를 진행한 결과, 박테리오파지 SG2는 살모넬라 티피뮤리움, 살모넬라 엔테리티디스, 살모넬라 플로럼, 살모넬라 콜레라수이스에도 용균활성을 가지는 것을 확인하였다. 그러나, Escherichia coli , Bacillus cereus , Lactobacillus plantarum 등 다른 속의 세균에 대해서는 용균활성이 없는 것을 확인하였다.As shown in Table 2, as a result of susceptibility testing to other microorganisms other than Salmonella gallinarum, bacteriophage SG2 has a lytic activity against Salmonella typhimurium, Salmonella enteritidis, Salmonella florum, and Salmonella cholera strain Respectively. However, Escherichia It was confirmed that there was no lytic activity for bacteria of other genus such as E. coli , Bacillus cereus , and Lactobacillus plantarum .

따라서, 본 발명의 박테리오파지 SG2는 살모넬라에 특이적으로 용균활성을 나타내는 박테리오파지임을 확인하였다.Therefore, it was confirmed that the bacteriophage SG2 of the present invention is a bacteriophage exhibiting a lytic activity specifically for Salmonella.

실시예Example 6: 박테리오파지 SG2의 pH 안정성 조사 6: Investigation of pH stability of bacteriophage SG2

박테리오파지 SG2의 pH 안정성을 확인하기 위하여, 다양한 pH 범위 (pH 2, 3, 4, 5, 6, 7, 8, 9, 10, 11)에서 박테리오파지의 수를 측정하였다. 소듐 아세테이트 완충액, 소듐 포스페이트 완충액, Tris-HCl을 이용하여 각각의 pH에 맞게 조제한 후 1 mL로 분주하여 준비하였다. 1.0x1010 PFU/mL로 준비한 박테리오파지 SG2 배양액 10 μL을 상기 용액에 첨가하여 2시간 동안 상온에서 방치하였다. 방치 후 각 반응액을 단계희석하여 소프트 아가 오버레이법에 의한 타이터(titer) 측정을 수행한 결과를 도 6에 나타내었다.The number of bacteriophages was measured in various pH ranges (pH 2, 3, 4, 5, 6, 7, 8, 9, 10, 11) to confirm pH stability of bacteriophage SG2. Sodium acetate buffer, sodium phosphate buffer, and Tris-HCl were added to each pH-adjusted solution, and the mixture was divided into 1 mL. 10 μL of the bacteriophage SG2 culture prepared to 1.0 × 10 10 PFU / mL was added to the above solution and left at room temperature for 2 hours. The reaction solution was allowed to stand, and the reaction solution was diluted stepwise, and a titer measurement was performed by soft agar overlay method. The result is shown in FIG.

도 6에 나타낸 바와 같이, pH 4-11의 범위에서는 박테리오파지 SG2가 2시간 동안 안정한 것으로 확인되었으며, pH 2에서는 박테리오파지의 수가 3 log, pH 3에서는 2 log가 감소됨을 확인하였다.As shown in FIG. 6, it was confirmed that the bacteriophage SG2 was stable for 2 hours in the range of pH 4-11, and it was confirmed that the number of bacteriophages was 3 log and 2 log was decreased at pH 3 at pH 2.

따라서, 본 발명의 박테리오파지 SG2는 pH 4-11의 범위에서 안정함을 확인하였으므로, 박테리오파지 SG2는 pH 안정성이 우수한 박테리오파지임을 알 수 있다.Therefore, it was confirmed that the bacteriophage SG2 of the present invention was stable in the pH range of 4-11, and thus it can be seen that bacteriophage SG2 is a bacteriophage having excellent pH stability.

실시예Example 7: 박테리오파지 SG2의 열 안정성 조사 7: Investigation of thermal stability of bacteriophage SG2

박테리오파지 SG2의 열 안정성을 확인하기 위하여, 다양한 온도 범위 (40℃, 50℃, 60℃, 그리고 70℃)에서 박테리오파지의 수를 측정하였다. 박테리오파지 SG2 배양액 (1.0x108 PFU/mL) 5mL을 멸균된 튜브에 분주한 후 박테리오파지를 첨가하여 2시간 동안 각 온도에서 방치하였다. 방치 후 각 반응액을 단계희석하여 소프트 아가 오버레이법에 의한 타이터 측정을 수행한 결과를 도 7에 나타내었다.In order to confirm the thermal stability of bacteriophage SG2, the number of bacteriophages was measured in various temperature ranges (40 ° C, 50 ° C, 60 ° C and 70 ° C). 5 mL of bacteriophage SG2 culture (1.0 x 10 8 PFU / mL) was dispensed into sterile tubes and bacteriophage was added and left at each temperature for 2 hours. After allowing to stand, each reaction solution was diluted stepwise, and the result of performing a tester measurement by soft agar overlay method is shown in FIG.

도 7에 나타낸 바와 같이, 40-70℃의 범위에서는 30분까지는 안정한 것을 확인하였다. As shown in Fig. 7, it was confirmed that it was stable up to 30 minutes in the range of 40-70 占 폚.

따라서, 본 발명의 박테리오파지 SG2는 내열성이 우수한 박테리오파지임을 알 수 있다.Therefore, it can be seen that the bacteriophage SG2 of the present invention is a bacteriophage having excellent heat resistance.

상기 실험으로부터 박테리오파지 SG2는 살모넬라에 대한 특이성이 매우 높고, 살모넬라균에 감염하여 살모넬라 내에서 100배 이상 증식함으로써 용균 활성이 우수하고 물리화학적 자극에 강하다는 장점을 가지고 있음을 확인하였다. 또한 본 발명의 SG2는 사람, 동물, 식물 등 세균외의 다른 숙주에는 감염되지 않으므로, 항생제의 오남용으로 인한 항생제 내성균의 문제점, 식품 내의 항생제의 잔류 문제, 광범위한 숙주범위의 문제점을 해결할 수 있는 장점이 있다. 따라서, 본 발명의 박테리오파지 SG2는 살모넬라에 의한 감염성 질병의 예방 또는 치료, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제, 또는 세척제 분야에서 다양하게 사용될 수 있다.From the above experiment, it was confirmed that bacteriophage SG2 has a high specificity to Salmonella, and has an advantage of being excellent in lytic activity and strong against physicochemical stimulation by multiplying Salmonella by 100 times or more in Salmonella. Further, since SG2 of the present invention is not infected to any host other than a bacterium such as human, animal, or plant, it has an advantage of being able to solve problems of antibiotic resistant bacteria caused by misuse of antibiotics, residual antibiotic in food, . Therefore, the bacteriophage SG2 of the present invention can be variously used in the field of preventing or treating infectious diseases caused by Salmonella, antibiotic composition, composition for feed addition, feed, disinfectant, or detergent.

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

이하 본 발명의 박테리오파지 SG2를 포함하는 감염성 질병의 예방 또는 치료용 약학적 조성물의 제제예를 설명하나, 본 발명을 한정하고자 함이 아니고 단지 이를 구체적으로 설명하고자 함이다.Hereinafter, formulation examples of pharmaceutical compositions for preventing or treating infectious diseases including bacteriophage SG2 of the present invention will be described, but the present invention is not intended to be limited thereto, but is specifically explained.

제제예Formulation example 1: 의약품의 제조 1: Manufacture of medicines

1.1 주사제의 제조1.1 Manufacture of Injection

박테리오파지 SG2 1×106 PFU/mLBacteriophage SG2 1 x 10 6 PFU / mL

주사용 멸균 증류수 적량Sterile sterilized water for injection

pH 조절제 적량pH adjuster

통상의 주사제의 제조방법에 따라 1 앰플당(2 mL) 상기의 성분 함량으 로 제조한다.(2 mL) per 1 ampoule according to the usual injection preparation method.

1.2 1.2 액제의Liquid 제조 Produce

박테리오파지 SG2 1×106 PFU/mLBacteriophage SG2 1 x 10 6 PFU / mL

설탕 20 g20 g of sugar

이성화당 20 g20 g per isomer

레몬향 적량Lemon incense quantity

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

1.3 정제의 제조1.3 Manufacture of tablets

박테리오파지 SG2 1×106 PFU/mLBacteriophage SG2 1 x 10 6 PFU / mL

옥수수전분 100 mgCorn starch 100 mg

유당 100 mgLactose 100 mg

스테아린산 마그네슘 2 mgMagnesium stearate 2 mg

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

한국미생물보존센터Korea Microorganism Conservation Center KFCC11677PKFCC11677P 2016072220160722

<110> Optipharm.CO.,LTD <120> Novel Salmonella specific bacteriophage SG2 and antibacterial composition comprising the same <130> 1_18 <160> 1 <170> KopatentIn 2.0 <210> 1 <211> 33010 <212> DNA <213> bacteriophage SG2 <400> 1 tccgtgaaca ggtagaagct caggaagacc ttatcgcatt gaagcgcgcg gcttacgaca 60 tcgataaaga agcggccaag gccgagggtc ttggtaagtt taaagtgtcg ggtaaagaca 120 acggttctac aggagattcc gcggacaatg ccgctaagaa atccgtcgac gcattcgaac 180 gtcagaagaa agccgctgag gatttctatt atcagtcaat ccaccttaac gatgacgtat 240 tccagaagat acaagctaac caagaagagc aacttactaa gctacaggag ttctacagca 300 accgtctcct tagtgaccag caatacgaaa ccgctaagac gcagattatg ctcgaggcgg 360 atacggcccg ccaggctgaa ttagataaac gcgagaaaga gcgcctggaa aaacaattct 420 ccgcagatgc ctacgtcgct cagatgcagg cccttgccga aggggaattc gccgagttag 480 atcgtcagta cgaggtcaag ctacaaaaac ttaacgactt tcatgcgcag ggtttaatcg 540 cggaagaaac ataccagcag accctaaacg caatgaatga tacttacgca ttagaccggg 600 cgaaggcaac cggcgcggct ttcggtaata tggcgagcaa catcgggtca gcactggggg 660 aagcgtccac cgcgtacaag gcatttgcca tcgcacaggc gacaatcgct acgtacacgt 720 cagcagtaga agcgtataaa tcaacggcgg ccataccggt agtcggcccg tatctggcgc 780 ctgttgctgc ggctgctgcg gtggcggctg gtttagcaaa cgtaggtaag attcgctccg 840 cccgagaaca gggtggtaac ctggccgcag ggcagatgtc cactattgca gaacgtggta 900 aaccagaagt aatcatgcct gctagcgcct cacgtgtccg gacggcggag cagatgcgac 960 agattatggg cgagaacgac gctaaatccg gcggggataa tgttactatt gtgaacaata 1020 ccaccggaag aattgattct gctgcaacag aacgcgacga tgaaggccgg ttgcgtatta 1080 taatcagtga aaccgtaagt tcagcgttgc tggatagtaa cagcgccatt tctaaatcac 1140 gtcgcgctac acgcggccaa ccaggatatt gatatgagcg attaccattt cccggcctct 1200 ttgaggccta tagtatcgaa aggctactcg atgacgcgag gtaacaacgt gtggcgagta 1260 gacctggccg gtggcggggt tcgccagggg cgtgatacat actttgatat gttcccgatt 1320 aacgttaccc tggtcgtatc accgttgggg cggcaagcat tcctcagttt catggagaag 1380 gtagacggag gggcttccag tttctggatg aaacacgact tgggtcaggg tatcgaggat 1440 taccaggtta ctctaacatc tacgtggaac gagtccaccg acgacgggaa gaactgggta 1500 ataactttca cggccacagc cgagaaatca ccattccagg aagccggcag cgcctgtctt 1560 aaccagaatc tgcccgactt atatggatgc tacggcgatt gtcttggtga atttcttaaa 1620 acttacggag tgtaccaaac tacattccct cgaatctggg acccaatgca atgagtcagg 1680 aatcagtaga agcggcatac cggcgtaagc tggcgtccaa tcccgatggc gagatggatt 1740 ttattaccct ggagatatcc caccctcttc tttcgaagcg ctggttgctt gtgcgcgggg 1800 ctaacgactt gaccgctact ctagagacgg gtgaggttgt tacattcgag ggtacgccga 1860 tggaggccaa gaacgccgcc aacaataacg atatggacca gaccgcctct ttctctttgc 1920 cggatgtgct taatatactg gacgaggaaa tggaccgcat cccttatgat aataaggaat 1980 tgcccaaatt catcttccgg cgttacgtga gcacggacct gtcataccca tgcgatgggc 2040 cggtggtata tgaattgcaa acactcacac aagagaaagg agtgttcaca gcggaaacag 2100 gtacacccat gcttaaccaa cgagctaccg gaatcctgat gacgccggag gagattcctt 2160 tacttcgagg gatactgaca tcgtgaatat taacgattac actggcctgc cgtatgactt 2220 ccgccgccgt aattgttggc accacgtccg caacgtccgg gctgacgcgg ggttatcaac 2280 tccaatgttc gatgtaacaa gcccaacggc aatagatgca gctttcgatg acggccattc 2340 agaccctaaa ggtcttaggc gagtgcttac tccgcagaat tttgacgccg ttctactcgg 2400 tgtgaaacat cgagggcgga tagtgtggca cgctggggta tattacgaag ggatggttag 2460 ccactgtgag ctggcgtcca gacaggttag actggatagt ctggaagacc ttaaagatac 2520 ttattcggag attgaatttt ggcgctagta attcactaca cacgaaacga agacggcaca 2580 tttgacgtta aacgttatcg cgataatccg atgaacttcg tcgtgaacca cgttcccgac 2640 ggggtgccgg ttcgcgtttt cattgacgaa attggagaag ataacgacgt aacagaagac 2700 ttcgaagcac tgaaagaaga cgcgactttt catattgtgg aatccgctgg tggtggcgct 2760 attaaaggcg tcatgaagat ttttagcgtt attcttaaac cgttggcgaa gctcttgtcg 2820 ccgtccgtga aaggtgcgtc ctcgaacctt gcgaactcgc aggcagactc cccaaacaac 2880 agtcttactg accgcaacaa caaggcgcgt ccgtacgagc gcagctacga tatctgcggg 2940 acggtacaaa caatacccaa taaccttatg tctacttata aggtgtttaa cgccgctggt 3000 aaaattgtag agtacggcta ctacgacgcc gggcgcggct acctcgacat ccacccggac 3060 ggtataacgg acggggatac ccgtgtatca gatataacag gtacgtcggt tgccgtgtac 3120 gcgccgtata cgtcaccaaa taatacatcc acaccacagg tcatggttgg cgacccgata 3180 gagcaaggcc tgtacatcac cgtagaatct aacgaagtag acggcgtggt tctgaaagca 3240 cctaacggcc tgggtatttc tttctcttac atgtcagggt atccgtcttt gtcaggaaac 3300 attggcacta tatacgaccc aacaggcggc tcggattttt ctggagtact ggtgcctaat 3360 gacacgtttt cgctggtgtc cgcgtggaca aatacagacg ttgacctgtc cggcggcggg 3420 tatcaggtag tcagcgtgtc cgaagggact gttaccttta tagtacctgg cggtctcatt 3480 ggtaggtggc aagaaataag accgggttcc tttttccgtg gtgacggaga ggcctcgctg 3540 caaccagaca acacgtatga gaaaacttta accgattggg tttcaataaa ccgtaccgaa 3600 gtagagcgca ttgtagccaa tatcgccgcc gcgaatggta tgtataaaga caacggcaaa 3660 tcgaaaacac tggcgtcggt caccgctgag atacagtacc agttacttga tgaaaatagt 3720 gccccttacg ggccgatata cactgcgcaa ggaaccgtgt ccggacgcac cccggactac 3780 aacggagtca ctatctatgc cgatctgccg gttgtatctc gtgtaagggt tagggcccgt 3840 agggttacgg accttgactt caattttgaa gggtctgtag tcgatgaaat aacgtatgtg 3900 aacctgtatg gacagacgcg agataacacc ccgcactacg gcaacagaac aaccgtacac 3960 tcgatgcgca agcagacacc gcgcgctgcg gaggttaagc aaccgcagtt gcgtatgatt 4020 gccaccgaaa tggtatacaa atacctcggt aatggtgttt tcgaagacac gaggacccct 4080 aatactcagg ctgtgcaatc tcttatccgc ctggcacgtg acccagatgt gggtggctta 4140 aacctgacag tgcgaaacat ggacaagtta cttgccgttc agaaagagat tgaagattat 4200 ttcggcgaca agcaagcggg ggaattttgc tacacgttcg atgactataa aaccactatg 4260 caggacatag ttagtactat agctgacgct atattctgca ccccatatcg gaaaggggcg 4320 gatatccttc ttgattttga acgccctcgc atgggcccgg agatggtgtt cacccaccga 4380 agcaaggccg gtacttctga aaaatggaca aggacattta acgatgctca ggtgttcgat 4440 agtcttaaat tctcgtacat agacccaaaa acaaacgtta aagaaacaat aacaatacct 4500 gaaactggtg gggttaaaac ggagacttat gactctaaag gtattcgcaa ctataagcag 4560 gctttctggg ccgctaatcg ccgtcaccag aagaacattt taaagaaaat ttcagtgtcg 4620 tttaccgcca cggaagaggg tatctttgct ttgccaaatc gtgccattag tgtcgttaag 4680 ggctcacgta tggctaccta cgatggctac ataaccgcag taaacgggct caccgtagaa 4740 ctatcgcaac cggttaagtt cacagcggga gatgaccatt ctttgattct caagttacgt 4800 gatggtggag tgcaaagtgt taatgtagtc cctggagcgc acgacaggca ggtgattatg 4860 acatcagtgc cgcaagaagc catttacaca ggtaatagtg ctttgaaaac tgaattttca 4920 ttcggcaacg aagcaaggca taatgctcag atgattcttg tttctacggt agaccccggc 4980 gatgacagaa cagtcaaaat aaccgggttt aactatgaca aggatttcta taagttcgac 5040 aacgtgcctc ctttcggtcg tgcgttctcc aacggattcg ataacggttt taactaagag 5100 gatagccata tgtctagtgg ttgcggtgag gtaatgtcac ttaatgattt acaggtagct 5160 aaaaaacacc agattttcga agccgaggtg atcaccggca aacagggcgg tgtagcgggt 5220 ggcgccaata tcgactacgc tactaaccag gtaaccgggc agacgcagaa gaccctgcct 5280 gcggttttac gtgatgttgg tttctccccg gcgtctttca acttcacaac cggcgggacc 5340 ctgggagctg acgacgcgga taaagcggtt ctttggccga aagaagatgg cggggacggt 5400 aactattacg catggcgtgg ccccctgccg aaagttatcc ctgcggcgtc gacacctctt 5460 acgacaggcg gcatttcgga ttccgcttgg gtcgcgttcg gagatattac ctttcgcgcg 5520 gaagcggata agaaatttaa atactccgtt aagctgtccg actttactac gttacaacaa 5580 ttggcggatg ccgccgttga tagcgttctt atcgaccgcg attacacttt cagtaataac 5640 gagaccgtta acttcagcgg gaagaccctg accatcgact gtaaagcgaa gtttatcggc 5700 gacggaaacc tggtatttac acaattaggt aaaggttcca ttgtaatagc cccctttatg 5760 gagagtgcta caacgccgtg ggtgattaaa ccgtggaccg acgataatca gtggataacc 5820 gaccccgcgg caatcgtggc cacacttaaa cagtctaaaa cagatggata ccagccgacg 5880 gtaaacgatt acgccaagtt tcctggtata gaatcccttc tccctccgga agctaaaggg 5940 caaagcatat cttctaccct ggaaattcgg gaatgtacag gcgtcgaggt tcaccgggcg 6000 agtggtctta tggcgtgttt cctgttccgc ggatgccatt tctgtaagat ggtagacgct 6060 gacaacccga gcggcggaaa agatggcgta attacctttg aaaacctgag cggcgattgg 6120 ggcaagggta actatgttat tggcgggcgc acaagttacg gttcggtaag tagcgctcaa 6180 ttcttacgaa acaatggcgg tttcgcgcgc gatggcgggg tcatcgggtt tacttcgtat 6240 cgtgcggggg aaagtggtgt taagacgtgg caaggtacgg taggttctac gacctctcgt 6300 aactacaacc tgcaattccg ggattcagca gtgctatacc ctgtatggga cggcttcgat 6360 ttaggcgcag atactgacat gaaccccgaa gatgaccgcc caggggattt ccccatttct 6420 cagtacccgg tacatatgct cccattaaac catttgatag acaatctatt tgttagaggt 6480 tcgctggggg taggtttcgg tatggacggg caaggtctgt atgtctctaa cataaccgtc 6540 gaggattgcg ctggttctgg ggcttatatt cttgcccacg aaacagtatt cactaatatc 6600 gcaataatcg acaccaatac taaaaacttc cccgcgaacc agatatatat ctcaggggcc 6660 tgccgtgtaa acggccttcg tttggtcggc atccgttcaa ctaccgaaca gggcatgacg 6720 atagacgcac ctaactccac tgtaagcgga ataacgggct tcgtggaccc ctcaaggatt 6780 aacgtagcca atttgatgga ggaaggtctt ggtaactctc gcataaacag tttcaataat 6840 ggttctgcgg cgcttcggtt tcgtattcat aaactgtcaa aaacccttga tagtgggtcc 6900 gtgtactccc acattaacgg cgggccaggt tctggttcag catggaccga aattaccgct 6960 attgcggggt ccttgcctga tgccgtgtca ttaaaaataa acaggggcga ttatcgtgct 7020 gttgagatac cggtagcggt gaccgtccta ccagacaacg ctgtcaggga taacggggct 7080 atatcactgt atctggaagg cgatagcctt aaggcgttag ttaagcgggc cgatggaagc 7140 tatacaagat taactttggc ataaatagta aaggccccgt aaggggcctt aatttatgat 7200 aacagaacca cagcagcgat aagtaaagcg accgcagcca atccgtagcc tattaaaaag 7260 catttcgcac caattgagta tttcatttag cgcccctctc tttgtcaact tcctgttgtt 7320 ctagtagcca gtcaagttgc gcgttagcag cgtctctttg ctgccgtagc cgtaaaacct 7380 cttcttcgag ttccctgata cgttttattt ctcgctttaa ttgcatagct agaagttgct 7440 cgcgggaatt aatggcgtct tttatcaatt cgttgcaagc gtcgatatca aactcattat 7500 tcatttctta cctcgtctct tcatataatt aagcaactct tcctggacag attttttctc 7560 gtccgtacgc gcggcaacga cctcatccag cgtgtcttta gcgactatgt gatacaggaa 7620 cactggacgc tcgtggccag cctgtttctg gcgtacagga cctatacgct cgacaacctg 7680 caaatagtgc tccaggttcc agccttgcga aataaacgcc agatgatgcc cgccgtcctg 7740 taaattcaaa ccatggcccg ccgatgcagg gtgcacgcac aaaatctcga tttccccgcg 7800 gttccaagat tccatctgct tattaccctt agcacctttc gcaaacgcct gcgcctgggg 7860 gaatcgccta agaatgcgct caagttcgtg cttgaactga taggccacca gcagcggcgc 7920 accctgcaac tcctcgacaa tggactctaa cgcatcgagt ttcgcgtcgt gcactttctc 7980 ccagtctttc gttgcttcac catccggccc cgacacatac acggcaccgg aagcaatctg 8040 caagcactta gccgttttag cggcggcgtt agccgcttca acttctccgc tctccagttc 8100 cgcgaataac ttctcctcca tatcgatgta ggcttgacgc gctttcttcg gcaggtcaat 8160 ctcaaccggt acaataaccg gtgcttcaca accgaaccac tcggcggcat caatggtaag 8220 gctgatatcc ttcatcttct ggtgaatctc gttatccgcg cctgggcgag catggtactc 8280 ccgcgccatc gcagatttgc ctttctgtac cgagttaaac catctatcgg taaaggccgt 8340 gtatgaagac ccaaggcgtt cacccgcgtc gatgaaccag ttctgacccc acaagtcttt 8400 gaggccgttt ggtgatggtg taccggtcag gttaatgaaa cgcttaactt taccgaacgc 8460 caccttacta agcgcctttg cccgcttgct accgccagaa cggctacgga acgatttcag 8520 cttcgtgctc tcatcggcaa cgataacggt aaaaggccag tcgtctttgc cgtagtagtc 8580 aataagccat tctataactt cgtaatttgt gcaaaccacg ttagcgtctg actccagcgc 8640 cgcgatgcgt cgcttctcag aaccagttgc atcgatgaca cgaaggcaag ggaaccccca 8700 cttagtttgt tccacaggcc acgtacccga cgcaacacgc aacggggcga ggattaatac 8760 ccggtcgtcg tcattaagtt gcccattacg gaacaggcgg ctaagcaccc acattgtcga 8820 gcttgtgttg tgcgttacgg tgaagtcacc taataaaaac cggtggtcac cgtcgatggt 8880 gaagccgtaa taatcatcta caccgaccgg ggttatagat tctatcccga cgttaagtac 8940 gtttttatta atattacgct tcggtaaatt ctggtggcga ccacgcacga aaggcacctc 9000 agaaaaatcg cctgataccg atacgcggaa gtaatccccc caaacatcag tgtttgcgca 9060 acgcttacga gttttcttct ggtaagcggc gaaaccaaga gaacggcata gatagcaaaa 9120 atcatcagcg agtcgctcgc ttaccgaaat ccagtcaaaa cctgctttag acaaatcgca 9180 gtaaccatca ctatccagaa gccctgccag caactctaaa cgctgtctcc ggtcgccgca 9240 tttgtaatta tgcggtatat gcttattgtt caggacaccc gccgatttca gcgcgtgcgt 9300 aaacccgtgc ttcttatgac ctgtgttgcc gtgagatata ctccatgtaa gtccttcttt 9360 tcgtatttgc atcccattac gtgctgcgta cgactcaaga tacgccctta tttccttttc 9420 gttctctcct gatgttatgg ccccagacga cgaagtaccg tcgcccagcc atagccccat 9480 aaggtacggg ggcaataaag cctcgtcctg ctctttcctt ggaaaatcga caggtacccg 9540 ccaaccctta agatacccat taggcccggt tacatatttc ggcaacttaa gccagtcgcg 9600 gacgcttatg tcgaaaacgg tgttatctgg ccatgaacct tttgcgatac ctgtagtagt 9660 tcgcagtgat aatatatggc tttcgtttac cgtatacgac tcgccttttc gcggtttaac 9720 ctcgtacatc atctcccggc ctctaccgag agacagtaca tttctaggcg ttgaatctgg 9780 tcccataagc acgtcaccga cgatgacgtc ttctactttc ttggtcgtac cgtcaaacat 9840 aataacttca gtcccgcgtt tcaggcactt gccgctgccc atacttgccc agatattaca 9900 gcgcgggtgt cgcagcatga acgaggtcat gagcttttga taaggccttc tagtgaattt 9960 actcatttcg ccaccagtac cagttccttc cgcccgaacg ccgtaacgtt acccgttaca 10020 tcttcgataa ccagtttacc gttcgactcg acgaacacag tatcaacggc aacaggactg 10080 cgggtcttaa cgttgaaaat catgtctcca ggtacgatgt cacgtgctgg tttgcggtca 10140 tatttgtatt tcatttctca attccttatt tctgtttggt gtgaactaaa tataatagtg 10200 ttctattaat ttatcaacct gtttcttcga cccaacaaca aaaacattcg caccgcgttt 10260 gcgcatcctc tcgtgctccc gtaactggtg cgggtccggc ttcgtgtttt cgtctttctt 10320 cacctcgacg aaccagacga tgccgccagg gagaattatc aacaggtcag gagcaccgga 10380 gcgcccctca taggaaagtt tccgaacgag gccccccagg gcctcgaatc ggtcttttgc 10440 gtatttctga atcttgcctt ccggggtcat aggcatagaa cccaagtaag gaaagcccct 10500 cctagaacgg caacgccaac aaacttaaag aacaggccat aacagaacac cgcggctatt 10560 cccgcaccga gcaacagtgc gagcatggtt acgattaccc aaaacacaaa catcgtcata 10620 atatgcaccc ctcgcgtttc gtgtgttcaa taccgcagcg aggacagatt cggcagtctt 10680 cttcccggaa ataatatata atcagtttac ttagcatacc gttttaactc cgcaccttcc 10740 gctacaagag gaaaaccttc agcccattct ggcaatgcac acattagttt ttccaattcc 10800 gctaccgtgt aataaggtgt atccggggtc tcgcatacca gttcatcatg aaccgaaaga 10860 actatagggt acccgccagc ctcaacatta agcattgcgt aggccaataa gtcacgacaa 10920 aacgcctgaa cgatgttctc acaagccttt ccgccgtgtg tgtacagggt agtccactgc 10980 cgtgttaact ggttctcgcc ctggtactta attcttacat tggtgtttac ccgcccgtct 11040 tcgtcggttt cctttgtcac gctgacgcct attcctggat atgataggat tcgccccgac 11100 ggcagctcca tgcatagcca ccatcctggg atgtttctac cggatgaatc agtttctact 11160 gttctccaga tacggatagc tctttcaccg ttgcttcgga tatttgcgcc cgcccagaaa 11220 tcgcggccag gattacgtac tgcggcaaga ataccgtctt taaggtcgca ccagaaagct 11280 acggtttgcg ggtgcgattc acgccacata cgtttaatag catcacatgt acgccatact 11340 ttcttatcga gaatatacga tggacggtca tccttctccc ccggacgcgg cggccgtttt 11400 gcctcctgta tacgcgccca ctcatacccg cgagcggtag cggcccagat atggtcgggg 11460 aaagttcctg ccattgtttt ggccatctca ataaggtcaa gacctaagtt tttggcgaat 11520 gtaacgaacg ctccgacgcc accctcatag ccgaggccaa gctcgcaagc ttttcctatc 11580 tgtctgatat ccttaaagtt tttcttaata tcatctggat ccatgccgaa catctttcct 11640 gcggttacgc agtaaatatc cagcccggcg cggaacgtat caagcgcggt ttcttcgccc 11700 gccagccatg caagcccacg gccttcgacg ttagagtaat ccgcaacaac aaacttacgc 11760 ccggcttccg ggataatgca gcttcggacg gtagacgctg ttagcttggc cacatcaaaa 11820 cggcggtgtg cacggccttt aagtaacgcg gcaatacccc tatccagttc atcgtcgtga 11880 taatacccgc gcgccaggtt ttgcggctga aaacctttcc ccgcccaccg caatgttcgc 11940 ttcgctcctc cgtattgcag gcaaccacgg cggcggtcat ctgaagaacg gcctaacagc 12000 aacggcgcgt atttcgtcga tgcggtagaa gcagccccga gacgcatctc gataatagtg 12060 cgggcgtcgt ccggtaaatc ctcatccgcc agcaaatcgt ttagtgttga cttctgtgcg 12120 ttgtgtatgc ggtgcgctgg cgctagctcc tgtaaaatag gcaagaagtc tttgccggtt 12180 agtgaaccgc catatttacg ttgggcttct tcctgtaact cttctttatg gcgcgtgact 12240 gcttcaatcg ctgcttccgc caatgccaca tcaaccttaa atcctcggtc attaatcacc 12300 tggtctaagt gcagcacgcg gtcttcgaac tcagaattcc cccacttagg cattttctta 12360 tacacttcac gcatggacgt aatgtcgctt tttgcgtatg caataaattc tgcccactct 12420 tttgggtggg tatcggcagt ataacgtcga atcttatagt tcttgggcgt aggcttactg 12480 aatcgctgta tcagcgcctt accgcgttta tccttagcca tgctggcgtc aatgtttaaa 12540 acctcgcaca acgcggctag tgacccagga agggcatggc ggaatgcaac tatcatagtg 12600 tcgatgatgt tttctatcag gagttcgaat ccccagcaat gcttaataac tggtcggtcg 12660 aacataagga agttttggcc taccagtttg aggttgctgt ctggtttttg cagctttagc 12720 atcgcccggc gcaaatcacg cggcatatca ctcccgtcgg tggcatccca tacctgcacg 12780 gggccttcat cgaaagcgta ggtgcaaata acaatttcgg tagtgggatg ttcagcgtag 12840 gcataggagc cgactttttt caaatcagct tcggagaatg tttcaatatc aagataaagc 12900 aagttcattc atttgaccct tatagaaaag gcgaccgaag ccgccttaaa tgaaatgaga 12960 atatattaac ggttacggcg acgttcgcgg cgcggggttt catcttcttc atcatcttcc 13020 aggtcatcaa cgctggcaga aaccgaagaa ccgccaaacg ctttaccttc acctgcaaat 13080 ttgatgccgt ttagttttgc gcctaaaact ttatattgct cggaaaacca aatttcgatt 13140 gagatattag ccacacaacc gctatatacc tgctcacctt caatctgctc accatcaatg 13200 ttgaagtctg gttctacctg cttttcaccc ttaaccgaag tcaggattaa tggttgttgc 13260 ttgttcttgg cctggaagta aaaaccttcc gggaagtctt cgaacggatt atcgcgctca 13320 gcgatgtcgc gcacagcgca cttgtccatg tgcttacctt cgccgtagtt ctgcttcatc 13380 catttctcag cggcggcctc tcccaatgct tccgacacta ctgcgaatac cgtatcgcga 13440 agctcttcaa tttgtgggtg ttccggagtc aggataaaag taccgttata cgtaccttta 13500 gtaacggaac cgtcattatt ttcacggtct ttagcgcgtt cgaatacgtt cagccatgcg 13560 gtttgtactt tacgaagatt aagtttaatt cccattttag ttttctcgcg ttttagagtt 13620 tatccgggga gctgcccggt cagtgattag taatataata gagtacgatt aagatgtcaa 13680 gcctctaaat cgtcttcagt aacatttttc cattctggcc gtttgtcttc gaccgttgtc 13740 acacacggcg ctcctggctt acgggttacg tgtttctcca gcgccgccca tagctcagaa 13800 cctttgtgct gcttctcggc ctcggtcggg gtcattagta cttctttatg aagcatttca 13860 gcgcccaact caaaatgtga tagcgtttcc agcagcgcat cggtatcttt ccacgcacga 13920 ttacccgggc ggccttcaac cagcttgtac cccggcactt tcttaccgga atgcaacgcg 13980 gcagccatcg ctttctcgac cttgtcgatg tgctggcgca gcagtggcaa cttctcatac 14040 tcagctacga gttgctccgg tgtaagttcc agcgcaaagt cgtcctccag ttcttccgcc 14100 agtacagagt taacggtttt tgtacgcgcg gcgcactgtt ctgagaaacg acaccactga 14160 caaccatcga ccgacggttt gaagtccgac gatttcaggt tcttcttacc gcggaaatag 14220 gcatcaagcg ctaacagcgc acgtttctgt gcgaacttag cgaacagttc cagaccttca 14280 accgagatgt cccactcgga cgccccgcca gcatacggct ggaagatgac cagacgaacc 14340 acggtgatgt tataacgtct cttgagtcgg cgataaacac cgagagcgta aagcataagc 14400 tgcttgtttt ctttcgcttc aacacgatgc cgtccggttt taaggtcgcc gataatgagc 14460 atgtgctcgt cggagttagc cagttcctga acggcaacaa ggtcagccgt tccgaatgtc 14520 tcaacgcctt cgtaacccgg atgcaatacc tcagtaagat ttacccgcat ctcgagcttg 14580 gcgtaagtcg ctacgtcgat aattggtttg cagtagtcag tgtacttgcg cacctgctca 14640 atcatgtccg ccgtaatcag taccgcgcct ttcatcgggc tgattagcgc cttaatctgg 14700 cctttaccct catccagcac gtaagcaccg acttctcgct ctagtggcag tgcagtgccg 14760 cggatatagg cgttgagatg gacctcggct atagtgtgac atgcggttcc tgtaatggcg 14820 gatttaccgg acgtgttggg gatgtctttt tcgcatgcca gtgatgcggc acagctaagc 14880 cactttttag cgccagacgg cgacagcagg gcgtgcacat cgttattgcc accgcgttcc 14940 tttaaaatca taccctgttc tcccactggt caattaaatg tcgcgtctta tggtcgcaat 15000 gcatagccca tccatacatc gactcgaaga cataaaagtc aggttttgcg aaagtcgtgc 15060 gcttaatctg cgacacgtga cggcctatat ctttaggccg tggcactttg cctaagtacg 15120 ccatctcttc catcaggtgc gcaccggacg atgcacgcaa cagccatagt gcttctgtgt 15180 tatcccgcct gtctacggcg cggtagagtt ggtaaatcat ttctttaccc tcacagaatc 15240 catccacgcg aaaacgtcat ccatagaaaa atcaccatcg aaaacttttt gtgttttatc 15300 cgacttttgt acatacgtag gacgtttgtt atccgtagcg tggaagcata caaagttatc 15360 tgtttcttct ataactcgcc ctgcaatcat aagaccactt ttcttagcta aaatagccat 15420 aattccgacc ctcagttaaa gcggcccgaa ggccgcggta tatttattct tcttcgaaat 15480 acttgttctt gattgccgtc aggcgttcca ggtactcggc caggtcttcg tctttaatcg 15540 aggcaatctt cattttctta ccggtgaact cttccagcag ttcatttgaa tcgtcacacg 15600 cggcatcgct cggaccttca ttaattgcgt cgtcgatagc cttaatctgg tcacgaagag 15660 actggtaatc tacttcttcc ttctcttctt gtactggttc ctctactttt gccttacgcg 15720 gcttacgttt tggtttctcc tcttctgccg gtttagtgcc gacgatgtcc tcaccttcga 15780 ccggaatttc ttttgctttc tcgacttctt ccagtgcttt agccagttct tttttaactg 15840 tctcgacacc ggcaacggag acgtcaacag tcatttcgct tacggatttg ttagacgtct 15900 gtttcacact attcgcagca atcagttcat gggcgactac gaaacgttca agtaatttta 15960 agaattggtc taacattatt tcttctcctg tttagttggt gataaagact ataatcgtac 16020 tctattattt atgcaagaac tttttcgtaa attgcgtcgt gtattttatt actgtactat 16080 tactgcatat ctaactaagg agtaaacata tgcagcaatc tgagctaggc gctcgcatag 16140 agcgccgccg taaagaaatc ggcatgggtc aaactgagct tgctttcaaa gctggcgtat 16200 cccaaagcct gattacccac ctggcaaccg gccgggtgtg taaggtagac tgttttaaaa 16260 tcttccacat tgccgatgct ctcggtgttg accctcgatg gttaagcttt ggtgatacag 16320 gggcctgatg gcccctttct ttttactcta aatccccttc tgttgtcttt atgttgtcgt 16380 taggttcata ccggttcttc ggtcgcttac ggtcgtctat acccatcggc aacctgtaag 16440 tattcgttac aacttcaccg ttttcgtcgc aaccaagaat caattcaccc tcgcgtagca 16500 tcttctcaag caccaggttt gtgatactgt ggtctccagc tctcgcgact atttgccgtt 16560 gtgtgaaccc tcgtcccgta tcatcggctt gctgtaaatc ctcgagagcg gacataaccg 16620 actcacgggc cgaagcgtct ttagaccgtt taaccgtatc ctttacggaa tctttacctt 16680 taccatccaa tccttcgttt cgctcttttt cctcgtccgt ttcgaacggc tggaagcccc 16740 acggcataag tacgagagct ttatgcggct ccggaaggtc gaggttgact atctccccgt 16800 atccctcacc cccagcgaat tctaccgcct ggaactcttt cggcggcggg gcctcgcgaa 16860 actgtactgg ttcaagcacc atgcctattg ttttctgctg cataccgttt ttgttcttgg 16920 tgtgcgcaac gtttatttgt ttctcggtag cgcggacaag cgttagttcc acatcgacac 16980 cggcatacag cgcaccactt ccacgcgcct tacttccacc tttcggggtg tggtggacaa 17040 cgcctacagc gcctttagta ccgtcgcgca cttctttgag catagccacg acacggccca 17100 taccgtcctg tccggctgag ttctcattga acttgtctat ccagttcccg aaggtctggt 17160 taagagtgtc gaaagcgacc atcccgacag gttcactgcc cgctatctgg cgcattttac 17220 gcaccagttt cttcgtatcc gcgaattcac cagcgtcaag cacatgcata tatcgcatac 17280 cttcatcgcc gtatttagcg gccaacgcag cgatacgggt gtgcgtgaac tccccaccct 17340 caccatcaat atagaaatga tgggctttac gggtgtctgc ccccgcgaac cgatacccgg 17400 cggcgctgat atacatcatg ccgagtgtgt agaacgattt atacgtcccg gattcaccga 17460 cgatatccca aatgcaatta gacggcatat agccctcgac gatgaagtcc gcttgtggtg 17520 ctggtggttc ttcgtcgtcc aggtcttctt cgtcgcaagt gaccccctcc gcccacccaa 17580 gcgccacctc cacacggtca aaaggtaagc cggtggccgc acaagcgtaa gcccacatat 17640 cccgtccgcc aggcttcatc ccgcccgtag ctaccaggtc ggtatcgtgg tacagggtaa 17700 cgttgggtcg ctcgaagccg tcgcgcggcc aacagaacag gaagtcgtcc tgcttaggtt 17760 cgccggtagt gtattgggcg gcgtgctccg gcgtggcggg catttgcagg ccgcggtcgg 17820 tcatacgtcc gccgaattca aaagcgaact cttcaaacag gtcggttaat tcggattgtt 17880 caccctccgg aactttataa tcagaagcac cggtaacgtt aatttcagga acttcctcca 17940 taagcttgct cgccgtaatc atgcggctac tttcagacac gatgacctgg cttcctaccg 18000 gcgggcggta cataggctgc gacagggtga acccggcgct atccacgtca cgacctttaa 18060 ggaagtgtgc caacaggccg taccggatgc ggatgatgtc gccaccggtt accggagtgc 18120 gtacaggcat aacgacgcgg tagcgcggcg cttcttcggt gtgggatgcc gtggtgtaga 18180 gcatcattgc gtaacgtgac tcgcgcacca tctcacagtc ggttgcgaac tcttccggcg 18240 tcgcgctgtc cacgtcggca taggccaggg atgaagacgt aacagacgca ttgcagcgat 18300 agaacatacc ttcggcggct tctttaccgg tagagcttac cgtggcggta caagcggcgg 18360 taatgtagcc ggggtctgtc ttggggttcc ggcgcgaacg tttaagcggt tgcataagtt 18420 caacaaactc atcccaggtg ccggaggtcg tggtgtaaac gttaatgtca gcgcgttcct 18480 cacggcgatt gcgacgcgac catgagtatg ctaaattgcc tgttgacatg tgcttttcct 18540 ttgtagtgtt taaggccctg acgttcgcgc gttggggctt ttcttttatt caaggtcttc 18600 tggggtcgcc tgggagcctt taacttcagc tactgccgaa tattttccgg ccttactttt 18660 aactgctccg tctctaacca aaacttccag cgacctctcg atgagagagg aactatagta 18720 ctgaaaataa gcgcggcgca agtcaagaac actacacgac ccttttttac gcgtaagcgc 18780 gactacagtt ctgaataccc ttttctggaa atcggtcact ttacattctc cctaatccac 18840 gcttccactt tctcccggtc aaacgtaccc ggcatccggc gacccataat ccggacacaa 18900 caatccggga acttgccgtc tcttaaccag ttattcagcg tgcggcgggt aaccccaata 18960 agctcagcta cttcattctg tgtcattctc aaatcctcca tctcagttag taaagcaagt 19020 atacccacgg cataatggga aatcaactat aaacacacct attgacattt tcattttttt 19080 cgtgtaagtt gtctcttgta acttcaagtt acttaagttt ctgtttcacc ttgtgagagc 19140 ttagcgggtc cgccctaagc ggacacgcgt taaggcttac tgaaacccgc aaacttaaaa 19200 ctcagtaact tccttcctta gtatctgaac tgccttgggc gagctatatt ctcatcggca 19260 gtatatctct taagtatctg atttccttga tgtgccgtag attcgcaacc gtaaaaatgt 19320 aacgattagt gggcaggctt aaggcctgcc ctttagttac caagtaataa gaataaaggc 19380 acaaccctgg ttgaataacc acaacggcaa cgtaactgaa atgccgaaag attcttagtc 19440 tactactcag ggattaacgc acttgcctta cggcaagcgc ttacctgacg agggacgaga 19500 ataaaggcac tctaatgggg cagggacctc cctgtaaata accccacggc aaaagattct 19560 aaagagtccg cgccggtgat tctcgggtct taaaggaggt tgccgtttat ggctagggtt 19620 ttagtggtcc ccggggccgc atccggacgc cgatgatgat gggtggatag ttcctggggt 19680 atcgtggtgg cggggccgca cctgggtgct gttaccggaa ccgcaggaag cttgtaacag 19740 tagtttagga ttttactgag tgacttacgc tgctgagtag cagcgtacca tttttgttaa 19800 tatttttcta tttcacgaaa tgcaacaatc gaagcgactg ttgcaggaga tgcaacaatg 19860 aaaccgaatg acctcgtaac ctggaccggg cgtaacggag agacccgaca cggcaaagta 19920 acatcgcttc acggcatcta tgctcgtgtc gagtggtggc gcgctcatgc taagaagcct 19980 cgtcgcattc tgacgcgtca ggatagatta atcgcgaaat agtattggca taaccaaaca 20040 taatagtgta ctattactgt atcgaaacga gaggggcggg aggaatgtac gaggatatta 20100 gcctggaaga tgcagtgtat atatggtgcg ccggagaaga tgtagtttgt gacggggacc 20160 tgaaagctat cgtatgcgag gagtgtggag atgagtagca aagacgaagt atttgaatac 20220 ctgattgacc aactacgtca gcgcgcaaat aagttcgacg ttgtagcaga aatacacaag 20280 atgagcatag accggtcgat aacaggccgc tcagggtact cggacgagga gaacgcaatt 20340 atcgacgcct acataggcca tgattctgac agcaaacaaa tcatccataa cttacaacaa 20400 cacctcgccc gtaaagacgg cgacatacgc atgctgaaag acaggttacg ccgggaggta 20460 gataaggtta aggaactacg ggatactatc gaacttatga atgccgactt ttaccgagtg 20520 actaaacgcg aagaatgtga gcactactac cgtttggttc tgtggagtag cgacttaaaa 20580 cagtgtacta aatgcggggc agtattgaac ccacaacatg agctgtacga tggttggata 20640 gaaaatccag gacgtaaggc gtgcccggtg cctagcgatt gtgaggttga ggttgaattc 20700 cgcaatggct ctatagccgt cggcgcagcc caggactacc ggtgggggat tgataacaac 20760 aactgggata tagttaagta caggataatc aaatgacaag tatacttttc atatgggttg 20820 tatccgcggg tcagctacaa cttgccggaa cagaaacgtt ttatacgtta gaggcgtgcc 20880 aggtagccgc acgcgctgcg gagaacgcgc cgttaacttt cgtagagcgt cctgttgacg 20940 ctcaggtacg cggtatttgc acactgaaac ggttatctaa gcagggggag aaatgagtct 21000 cgcaaccgat atactgaaac acgccggcat caacctggca cctccttcat cggcggtaac 21060 aaagaaggtt actcgcgtta aagaaaaggt taagcgtaaa cctaaaccgc gcgcaaagcc 21120 agttaatgag atgccggacg tgtatccgcg catccctggg gtgcaccagc ctaagtattg 21180 tgtaggcaaa ggtctgtggc gtgcgcactc ttacgacggg aagaaagtgg taaacctggg 21240 ggagttcagt agccaggcaa gggcgcatat ggcggttaaa ctgtacaaac tgtggcgtaa 21300 acgtgggtat tccgaaatcc cgcacaagcc atctattcga ctttatacgt tcaggtaatt 21360 tatatgacta cgatagcttt cgacggcaaa accatggcct gtgatacctg tgtcaccggc 21420 aattacaaat accacacgga taccaagatt tacgagaacg accactttgt tataggggta 21480 tccggggacg ccggggtggg tatgttgttg gttgccgatg atgagatact gacgcctaag 21540 cactacgact tcgacttttc ggcgctggtg tttgttaaag aggataaccg tattttcgcg 21600 gtagaattct gtagggcgtg ggacgcaccg ttaagctcgg ttataccaat tgcaggtaac 21660 gctgctgccg taggctccgg ggctccgtat gcacttacag ccatgttcat gggtgcgacg 21720 gcaacggaag cggtagaggt tgcgaagagg tttgaccctg gtaccgatgg aagcgtgata 21780 acacatcgac taggataaat ccgaattatc tgttactaaa attgtcgtat aaatgctaat 21840 ctccgaagga taaaccaact ttcggagatt ttttattgtg gacgataaat acctttggct 21900 tagtgtggcg ggcctcgccg ggggtgccgt atctcagatt aagaagcgtg aggctatttc 21960 gccatggttg cgattgtgcc atctcaccgc ttccgcctgt tgtgcagtgt acgcatcccc 22020 catcattata agttactatg aattatcaca gtctgagggt cagtacctgg ttcctttcgg 22080 ggtaggtatg ttctggctta agttattcga agctgctgac tcatccctca gcaactttaa 22140 gttaccgtgg gggagataac atgctcaact catcaattgg catcatttgc ctggttgcca 22200 tcatcgccac ctcgttaatt aatatctacg ccgactgggt tgaagacggt cttttcggcc 22260 gcctgctgta catggcgtcg gtggttaccg ccgccggagg tctcgcccat ctctttaccg 22320 gtgccattcc accgttcata attacgactc tcgtcgctat attcgcgctt aaatcggtgc 22380 gtcacatttg cgtgaaaggc gctcgctact acaaataccg gaggatgtat gtcaaaccga 22440 aacatcagta acaacggcat caaattcacc gccgcgttcg aaggtttccg cggaaccgcg 22500 tacaaggcaa cgaagagtga gaagtacctt actatcggct atggccacta cggcgcagat 22560 gtgaaagaag gccagaagat taccgaggga cagggtcttc tgctgctgca caaggatatg 22620 gctaaggccg tagctgcggt agacgctgta gcgcatccgt cgctcaatca gtcacagttc 22680 gatgcgatgt gcgacctggt gtataacgct ggcgccggtg tgattgcggc ttctaccgga 22740 acagggcagg ccctgcgtaa gggcgacgtt gccacactgc ggaataagct atctcagttc 22800 cattatcaga acggcaaatc actccttgga ttgcgtcggc gtgccgcggg tcgtgttgcg 22860 ctattcgacg gtatgccgtg gcaacaggcg gaagccatcg gccgcgaagc aaagtaggtt 22920 gacacatagg agaattccta agatactaaa cctgctcctg cttattcatc cctctagctc 22980 ctttatcccg gtaactgacc cttaccggga tttttttttt atctgtattc agaaataata 23040 gttgactagt aacattaacc ctattatatt tagttcatcg acaacgagaa cggagtagag 23100 aagatgaact tacaaagcga taaagttttt taccattgca aaccgctact ggacaccgaa 23160 gcattcaaag acgcacaact gatggcacgc attgccgtta ataacctaag tacacggatt 23220 ccagcggatg cgttctggtt cgccgcgatg cagacactga aagcagctta tgcaggagaa 23280 caaaaatgaa agtatatatc gtatccggat ggcatcacga cggggacact tacgttcttg 23340 atgtttatct cagcgaagaa aaggccgaag ccgccgccga gctggagcgc acctcacatt 23400 actatgacgg cgtagatgta ctcggatggg aggtgcagga atgagcgaca acgggcaagt 23460 aacggtaacg tttaaggtgg gcggtaaggt aagtaacacg cccttcccga cacgcgagga 23520 actgattaaa cgtaacagtt tcccaggacc ggacaagaac aagtatctca atcgaatgtg 23580 gggtaagaag aaatgactaa caacgaatat gaaaagatga tggtagaagc cgccaacggc 23640 aggtttaaga ttgaagcagt agacgcgcaa atcgaggcac accaggctgc gcttgactac 23700 cttaatgagt atcgccggga gcttatcaac cgttatgacc tgaataagtg cgagcacgat 23760 tttgttaaca atcctaaaga tatgtgcgga cagtgtgtta agtgcggggc gattaaacga 23820 tgaaacgagt atgcaaagaa acgttgcgcc aacgcatcac gtatttggag tctgtaagta 23880 agctgggcct gtcaatgaac gaagaattcc agttagaagc atacaagatg ctgctggaac 23940 gacttgagaa ggagagcgaa caggaatgat tactagtatc ccaaaactaa tagagaccca 24000 cggcacgctg gctgatacgt gccgggagac gggacttaat gagatgacat tgtccaagta 24060 ccgccatgac acaaaatgcg agcaacatgt catctataac aaccgactga tgacccatac 24120 gaaaacgtca ccggtcatct acacgaaacg tggattatcg cgcaacgacc gtatgaagct 24180 ggaggggtga tggatagttt tctcatcctc atgggtatag gattcacaac attactgctt 24240 gccggtgcta tgtacctgat gtggattttg tttctttggc catttgttga ggcggtaagt 24300 cttacgagga tgtcactagc gtatcgccgt ttgaatgctc ataaagctgg tgctggtgcc 24360 gcggtgaagc tatttgtcat gtggtacggg gaaacagtat ttggtcgtag gttcgaagcc 24420 atacatagtc gcgggtggcg atgggaagcc gtaggtaaat ggtgggttta taaagaggag 24480 agtggagaat gattgagatt ttgaaggcta ccgatgccac tggcgacctg atgttttggt 24540 ttgcctttct cagtgcgcta ataaaacaac cgactgtgtt taacagtctt ccctgatatc 24600 tgtggtggcc tatgttttta ggcttgggtg tatcaggagc ggccggcatc ctgctgtggt 24660 ttttcaaatg agattgctaa tcatcccgaa cgcctgggtt atcgccgcag ctaacgacca 24720 ttacggcggc gatggtagaa gagcactacg acacggccta tataactgac acaagcccgc 24780 ttctgcgggc ttttctgtac acaggccttc attcccctgt acaatcctta atagacgctc 24840 agggggcgtc tgatgcgcta tgatggtcat acgcatttaa gagaggattg catgaaactc 24900 aaattaaaac agcctgacgc cgaagtagtg gccgccgcgc acgaagaagc ggttagtgca 24960 aaccgtcgcc gtaaacgccc gcgtggtaaa caaagcctgt atcagtcatc cagacattcc 25020 gccgaactat gggatccgga ctattgcgac gagttaatca ggttcttcga ccgcgcgtca 25080 tgggaactcg tacccacgtc taaaggcgac gaacgcccgc ttatccagga caaaccgccg 25140 tcgctggccc gcttcgcatt acacatcggc gtaaccatcc cgattatcaa gctatggctc 25200 cgagaggttc ctgcattcgc cgaagcatat gagacggcac aagccctgga agaggcgtat 25260 ttcactgaga ccggtgcagc cgggatatcc gctacgtttg ccgcggcgaa actggggctt 25320 aataaaactg ttgtggaaga agcgcgcgac gaaccaatta gcgaagtaac tattaaggtg 25380 gtgtccggtg aacgttgata tcacagctac ggaaccgcaa ggcgcgttcc ttaatctgca 25440 ttgtaagttc ccggccttcg ttgcgggctt cggcacaggt aaatcagagg tcatgtgcaa 25500 ttccgcccta ctcgacagca tggagggcgg tagtgattca cttatcgcca tgtatgaacc 25560 gacatacgac ctggtgcgcc ttatcctcgc tccgcgtatg gaagagaagt tgtctgattg 25620 gggtattcgc tacaagtata ataaatccga caacatcatt tatacctcat ccgggcaatt 25680 cggggatttt gtccttcgca cattggataa tccagcacga attgttggct acgaatcgtt 25740 ccgcgcaaaa atcgacgagt tggacacgtt aaataaagac cacgccgagc acgcctggaa 25800 caaagttatc gcccgtaacc gtcagttgcc gcgtacatat cgtccaatta ccccgaagcc 25860 cgctaataca gtttcggtat ttacgacgcc agaaggcttc cgttttgtgc acgacagatg 25920 ggctgtaaaa aagaaaccag gctatgaaat gattcaggcc tcgacaacat ccaatccctt 25980 tctaccggaa gattatgtgc agtcgttgcg ggatacgtat ccaggccagt taattgatgc 26040 ctacatcgac ggcgaatttg tcaacctgac atccggcagc gtgtattatg cttacgaccg 26100 acgtaagaac agcagtcggg agactataca acctggagag acgctgtaca tcggtcagga 26160 cttcaacgtc gggcatatgg ctagcaccgt atacgttcag cgtgagtatg tctggcacgc 26220 ggtagccgag ctggtggata tgttcgacac cccggatgtg gtcagggaaa ttaccgagcg 26280 atgggggcga cagggccacc acatcgtcat gtatccggat gccagcggca agaaccgtaa 26340 atcgactgat gccagtacgt cagatattgc ccaattacag aacgcgggtt tcgagatacg 26400 tgcgaaatca gttaaccctg cggttaaaga ccgtgtagca tcggtgaata aagcgctgga 26460 gtctggtaga ttaatggtca acgagcaggc ttgcccggtt acagcacgtt gcctggagca 26520 acaggcttac gataaaaacg gtataccgga taagacaagc ggcaacgacc accagaacga 26580 cgcgacagga taccctatcg cctacgaaat gccgttggtt aaacctgttt cccatatccc 26640 ggttactttt gcactttaag aggattatta aatgttaaca gcaaacggcc agggttctgg 26700 cgtaaaaacc aagcaccgcg aatggctgca ctacgcgccg aaatggcaga aggtgcgcca 26760 tgcgcttgct ggagacctgg ttggctactt acgcaacgtc ggccttaatg aaccagataa 26820 agcatacggc gaagcgcgtc aggcagaata cgaggccggg ggtatcgtct acaacttcac 26880 ccggcgaacg ctgtcgggaa tggtcggtag cgtcatgcgc aaagaaccag aaatcaatat 26940 tccgaaggag ctggaatacc tgcttaaaaa tgcagatggg tctggtgtag gcctaataca 27000 gcacgcgcaa gatacactca tggagattga ctcagtaggt cgtggtggtc ttcttgttga 27060 tgctccggaa acagccgcgg ccaccgctgc cgaacaaaat gcgggcttgc ttaaccctac 27120 aatcgctttc tacactaccg agaatatcgt taactggcga ctcacgcgcg taggttctgt 27180 aaaccgggta actatggttg tgctgcgtga gacatgggag taccacgaac ctggaaacga 27240 gttcgaaact aaatacggcg agcagtaccg cgtgctggac attgacaccg atggtaatta 27300 tcgtcagcga ctgttccgtt tcgatgcgga aggcggagct caggaagagg ttgtggagat 27360 ttacccagat ttaggggagt cgttacgtgg cgtaattccg tttaccttta tcggagctac 27420 caataacgac gccaccattg acgacgctcc tttgttgcca ttggccgagc ttaatatcgg 27480 gcactaccgg aacagtgctg ataacgagga atcaagtttt gtagttggcc agcctacgct 27540 gtttatctac cccggggata accttacacc acagtcgttc aaggaagcca accccaacgg 27600 catcaaattt ggcagtcggt gcgggcataa ccttggttat ggtggtagcg ctcaacttat 27660 tcaggcgggc gaaaacaacc tggcccgcca gaatatgctg gacaaagaac agcaggctat 27720 ccagattggt gcccaactta ttacaccatc tcagcaaatt accgcagaat ccgcgcgcat 27780 ccaacgcggc gccgatacat ccgttatggc cacaatcgct cgtaacgtaa gtcaggcgta 27840 taccgatgct ttacgatggg ttgctatgat gttgggtaag ccagaagatt ctgaagtcga 27900 gttccagctt aacatggatt tcttcctgca acctatgaca gcacaggaca gggctgcgtg 27960 gatggcagac attaatgccg gattactgcc cgccactgct tattacgctg cgttgcgtaa 28020 ggcgggggtg actgactgga ccgacgagga tattctgaac gctattgaag atgcaccttt 28080 gccgtcgggt gctgttactc aggtagcggg ggagattccg caagcggcac aacaacaaca 28140 ggagtaaaga aaaaggcccc attacggggc cttagtttta gcggatatta gccgcgcatt 28200 tacgcacaac ccgcggtatc gtctctttca gctcatccca tgagttgact gcttccatat 28260 ccaacaacgt caccatagcc ttttgaatat cactccgtaa ttgattagtt ttggcgtcct 28320 gagagtccca agtatcaaaa ccggttactt cacggatatc tatagactct tttatcgcat 28380 tcgccacatg tatgaagtga ccgatacgcc caatcctacc ctccgtacgc tcaatcatcc 28440 ggctaacggc ggcattaagt tctacaaaac taacagcacc aatgttacgg agttctacct 28500 gccgttgagt taagaaacgg tctataacga ggaagtggta atagtcagaa tattgctccg 28560 caacccatat agccatatgt agacatgcgt atgttgccgc gtttttccct ctaaccacac 28620 gccacgcgct ttctatccct aattcctcac agcacacacg cgcgaacgac ctggcttgcc 28680 cggtagataa cagctgcgat aggtttttct cacccatccc attttcttca cgccacttat 28740 tcatctgctt cactaatacc gtcaggttca gctcaccttc tcgggtgaaa gttatacccc 28800 catagggaaa ttctagtgtt tgcgatttca tgtttctatc tcctttttga agtattgaat 28860 gaatactaca cctagatata gtattcgtca aatacttcta gttaacttta gaactatgat 28920 taagttgttc tactaattag ggtcgcccat agccttggga tgggttttct tatagtatgc 28980 tattaacttt agcatcacag ggtttatcta tgagcttact tacatcatta ataagccacc 29040 aggtatggtt acaacgcacc gcatccggag aggtgaaaga cctggcgcca ttcatcaagc 29100 agatgcgcga tgaggttaag cggcaggtgt tgttgtttgg tgatgacagc agaacggcgg 29160 cgagacttac caccatgctg cgggaacttg agcaagcgct gaacgggatt acgtctgggt 29220 ggtatgagaa gctactggcc gatgcccgtg agctttcaga ctatgaagtt aactggaacg 29280 taaaaaccct gtcgaccaac gttaacgcta atttcgtaac accggccgcc gaacaagtat 29340 gggccgccgc aactttcgcg cctcttgaac taagcgaaaa gccagttgat tttgtttctc 29400 taatgagcgg ctggcggcaa accgaggtaa atcgccttgt tatgggcgtt aaatctgggt 29460 tcgtacaggg gatgaccaca cgtcggatag ttaagaacgt tgtcgggccc ggtggtctgg 29520 ccgatatctc cgagcgtaac gctgcaaccg ttatccggac agcgctggcg cacgtatcca 29580 acgaagcacg gcaacaggtt tacgctcaga acggcgacat catcacgaaa tacgagtggg 29640 tatctaccct tgactccaga acctcagccg tttgtcggtc gcgggattct atgcaatacg 29700 aaataggtaa aggtccgcta ccgccagcgc atcctaactg tcggtcgagc acagcaccgg 29760 taataagtcc agaattcgac ttcctggata agggtgcaaa acgggcggcc aggggtgcag 29820 atggaggtca gcaggtaagc gcggacacga gctactacga attccttaaa caacaaccgg 29880 cgtggttcca ggacgaagca cttggccccg taaggggtaa gatttttcgt aatagtggga 29940 taacaccgga agaatttcgt gtaatatcgg tagacggatt cgggcgcccg ttaactctta 30000 aagagatggc ggagctcgat aaacgtgttg ccgattatct gaaagaggaa taaagatggg 30060 ctttttcaaa gttaaagatg tgccgtcacg tcgagtagtg cagtacgcac gtgtgtctgg 30120 tgcgggtgaa ggcgttgttt atattaaaga cgaatctgtt ctcggcgagt cggtagatga 30180 aatgccgttt gcggataaga ccggactgac cacaatctca gacggcatcc tgtacgaagt 30240 accgtatctg gatggtgcgg gagatgtgta cttcgatctg caaccggcag atgtagagct 30300 gaaagacggt tctgcaaaat taaccgtagt tgtaaaaggc ggtaaagcgc cctacgattt 30360 gcaatggttt aaagatggta aagaggtaat aaacgttcct tacgttgaag gggaactaac 30420 cgttaaagac cccggagaat atttcgtcaa ggcagtagat gccgacggtg tgtcggtggt 30480 tagcaaagcg gctaaggtct cagaacctaa gtaacgaaag gccccttgac ggggccttaa 30540 ttttaccagc gattattcca gttattcctt accaatgtaa attcaggaat actttattca 30600 aaaagtagat ttctattatt ttaaatgcta cgctaactac gctttcagcg taccgcggtg 30660 cgggtcgcgt gaagcgggcg tcacaacgcc ctagcgtaac cgcgaacgca ttcggaaggg 30720 ccgagctata ttgctttcag tatttagcta agattcgccg tatgatgttt acagatatac 30780 gagattaacg cccgacttcg tcgggcttag cgtataaaga aaggggatac tatgaattgg 30840 aaagtaaccg cggtagccac ggcagccggt atcctctcct tgtggttata cggccagtac 30900 aactacagaa gcgggtgggt agaaggccgc gccaatctcg tttcacagca acaaaagaag 30960 gcacaagctg agctgacgaa gaaaacacaa cggcagcagc agaatgatac taaggccgcc 31020 tccgccgaat cggaaggcaa agaaaattcg gaggctatca cccatgaagt catcaaatat 31080 gttatccgtc ctggccgtac tgtctgcgag tttccttctg aacgggtgtc aatcaaacga 31140 cgcgccactg agaatgctaa ttccatcccc ggatatgaca ctaatgcaac cgccgtgcaa 31200 gcttattccc ccgagcagtg acgctgatga agacctggcc atcgacgtac gtaacgcgga 31260 gtgcacacgg cagttacgtc tgaaagtgtt ccggttgcag gagtatataa ggaatattct 31320 ggaatagttg ccttagtaag tggaataatt tattcttgtt acagaaacac cgggtggccc 31380 ggtgtcctaa agtccagggg acatattgac tatgaatcgt tttttacact atccactcca 31440 ggaagaagct ggagtagaag ataaatcggg tgcaggtgat gcaccaaaaa tgtacaccgc 31500 cgaagaagtg caggccttga ttgagaaaga agtggccggg ctcaaggcaa atcaggaagc 31560 gctccttagc gagaaaaagg aagctgctcg ccgggctaaa gaagccgaag aagaacggca 31620 gcgcgcccac caggaggccc tgaaggctgc tggtaaaatg gacgaatttg aaaagacgat 31680 tcgtagtcag tatgaacccg tactgaaaga gaaagaagaa cgctacgctt ctttagccgc 31740 gcgaattctt ggaagtgaac gtaaggctgt tttgggttct ttcgccggtg atttcatcac 31800 cccggaagcg gtagaaatcc ttgctccgtt cgttaagact gaatttgaag gtgaagacgt 31860 agttactaaa ttcacggatg cggatggtaa cgtagttact accgaccctg aacagttccg 31920 caaatacctg cgcgaacata aagctttttc gcatttgatt aaagcaaatg cagcttccgg 31980 tggcggggct tccgggaata aaggcggcgg ggccgcacca gcgtttaaag atatgagtga 32040 aagtgagcgg ttggctctct ataaatctaa ccctgccgaa tttgaacggc aacttaaagc 32100 cctgaggaaa taaataatgg caatcaccac tattggtaac atcgtaactg gcaatatccc 32160 tgtactggcg tcctatatga cggaagaccc ggtagagaaa accgcgtttt tcaactccgg 32220 aattttgact ccaactccgt atgctgccga gattgcccgc ggcccgtcta acatcgctaa 32280 cctgccgttc tggaaagcca tcgatacctc tatcgaacct aactattcga acgatgtgta 32340 tcaggacatc gcgaccccgc gcgctattca gaccggtgaa atgatggccc gcgttgcgta 32400 tctgaacgaa ggtttcggcc aggcagacct gactgtcgaa ctgactagcc agaatccatt 32460 gcagtccgta gcctcccgcc tggataactt ctggcagcgc caggcccagc gtcgtctgat 32520 tgctaccgcc ctcggtctgt acaacgacaa cgtatccgct actgatgcat atcacaagca 32580 gaacgacatg gttgttgacg tttctgcaac ttcaggcttc gacgcaggtg cattcatcga 32640 tgctacccag actatgggtg atgcattgat gggcaacggc ggtgaggttc tcggtgctat 32700 cgcaatgcac agcttcgtat acgcacaggc gcgtaaagct cagcttatcg acttcatccg 32760 tgacgctgag aacaacacca tgttcgccac ctaccagggc taccgtgtta tcgtcgatga 32820 cagcatgacc gtagtaggcc agggtgcgca gcgcaagttc atctccgtca tcttcggcca 32880 gggcgctatc ggctatggcg aaggcaatcc tgagatgccg ttggaatacg agcgtgaagc 32940 atctcgcgct aacggcggcg gcgttgaaac cctatggacg cgtaaaacgt ggttgctgca 33000 tccgttggtt 33010 <110> Optipharm.CO., LTD <120> Novel Salmonella specific bacteriophage SG2 and antibacterial          composition comprising the same <130> 1_18 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 33010 <212> DNA <213> bacteriophage SG2 <400> 1 tccgtgaaca ggtagaagct caggaagacc ttatcgcatt gaagcgcgcg gcttacgaca 60 tcgataaaga agcggccaag gccgagggtc ttggtaagtt taaagtgtcg ggtaaagaca 120 acggttctac aggagattcc gcggacaatg ccgctaagaa atccgtcgac gcattcgaac 180 gtcagaagaa agccgctgag gatttctatt atcagtcaat ccaccttaac gatgacgtat 240 tccagaagat acaagctaac caagaagagc aacttactaa gctacaggag ttctacagca 300 accgtctcct tagtgaccag caatacgaaa ccgctaagac gcagattatg ctcgaggcgg 360 atacggcccg ccaggctgaa ttagataaac gcgagaaaga gcgcctggaa aaacaattct 420 ccgcagatgc ctacgtcgct cagatgcagg cccttgccga aggggaattc gccgagttag 480 atcgtcagta cgaggtcaag ctacaaaaac ttaacgactt tcatgcgcag ggtttaatcg 540 cggaagaaac ataccagcag accctaaacg caatgaatga tacttacgca ttagaccggg 600 cgaaggcaac cggcgcggct ttcggtaata tggcgagcaa catcgggtca gcactggggg 660 aagcgtccac cgcgtacaag gcatttgcca tcgcacaggc gacaatcgct acgtacacgt 720 cagcagtaga agcgtataaa tcaacggcgg ccataccggt agtcggcccg tatctggcgc 780 ctgttgctgc ggctgctgcg gtggcggctg gtttagcaaa cgtaggtaag attcgctccg 840 cccgagaaca gggtggtaac ctggccgcag ggcagatgtc cactattgca gaacgtggta 900 aaccagaagt aatcatgcct gctagcgcct cacgtgtccg gacggcggag cagatgcgac 960 agattatggg cgagaacgac gctaaatccg gcggggataa tgttactatt gtgaacaata 1020 ccaccggaag aattgattct gctgcaacag aacgcgacga tgaaggccgg ttgcgtatta 1080 taatcagtga aaccgtaagt tcagcgttgc tggatagtaa cagcgccatt tctaaatcac 1140 gtcgcgctac acgcggccaa ccaggatatt gatatgagcg attaccattt cccggcctct 1200 ttgaggccta tagtatcgaa aggctactcg atgacgcgag gtaacaacgt gtggcgagta 1260 gcctggccg gtggcggggt tcgccagggg cgtgatacat actttgatat gttcccgatt 1320 aacgttaccc tggtcgtatc accgttgggg cggcaagcat tcctcagttt catggagaag 1380 gtagacggag gggcttccag tttctggatg aaacacgact tgggtcaggg tatcgaggat 1440 taccaggtta ctctaacatc tacgtggaac gagtccaccg acgacgggaa gaactgggta 1500 ataactttca cggccacagc cgagaaatca ccattccagg aagccggcag cgcctgtctt 1560 aaccagaatc tgcccgactt atatggatgc tacggcgatt gtcttggtga atttcttaaa 1620 acttacggag tgtaccaaac tacattccct cgaatctggg acccaatgca atgagtcagg 1680 aatcagtaga agcggcatac cggcgtaagc tggcgtccaa tcccgatggc gagatggatt 1740 ttattaccct ggagatatcc caccctcttc tttcgaagcg ctggttgctt gtgcgcgggg 1800 ctaacgactt gaccgctact ctagagacgg gtgaggttgt tacattcgag ggtacgccga 1860 tggaggccaa gaacgccgcc aacaataacg atatggacca gaccgcctct ttctctttgc 1920 cggatgtgct taatatactg gacgaggaaa tggaccgcat cccttatgat aataaggaat 1980 tgcccaaatt catcttccgg cgttacgtga gcacggacct gtcataccca tgcgatgggc 2040 cggtggtata tgaattgcaa acactcacac aagagaaagg agtgttcaca gcggaaacag 2100 gtacacccat gcttaaccaa cgagctaccg gaatcctgat gacgccggag gagattcctt 2160 tacttcgagg gatactgaca tcgtgaatat taacgattac actggcctgc cgtatgactt 2220 ccgccgccgt aattgttggc accacgtccg caacgtccgg gctgacgcgg ggttatcaac 2280 tccaatgttc gatgtaacaa gcccaacggc aatagatgca gctttcgatg acggccattc 2340 agaccctaaa ggtcttaggc gagtgcttac tccgcagaat tttgacgccg ttctactcgg 2400 tgtgaaacat cgagggcgga tagtgtggca cgctggggta tattacgaag ggatggttag 2460 ccactgtgag ctggcgtcca gacaggttag actggatagt ctggaagacc ttaaagatac 2520 ttattcggag attgaatttt ggcgctagta attcactaca cacgaaacga agacggcaca 2580 tttgacgtta aacgttatcg cgataatccg atgaacttcg tcgtgaacca cgttcccgac 2640 ggggtgccgg ttcgcgtttt cattgacgaa attggagaag ataacgacgt aacagaagac 2700 ttcgaagcac tgaaagaaga cgcgactttt catattgtgg aatccgctgg tggtggcgct 2760 attaaaggcg tcatgaagat ttttagcgtt attcttaaac cgttggcgaa gctcttgtcg 2820 ccgtccgtga aaggtgcgtc ctcgaacctt gcgaactcgc aggcagactc cccaaacaac 2880 agtcttactg accgcaacaa caaggcgcgt ccgtacgagc gcagctacga tatctgcggg 2940 acggtacaaa caatacccaa taaccttatg tctacttata aggtgtttaa cgccgctggt 3000 aaaattgtag agtacggcta ctacgacgcc gggcgcggct acctcgacat ccacccggac 3060 ggtataacgg acggggatac ccgtgtatca gatataacag gtacgtcggt tgccgtgtac 3120 gcgccgtata cgtcaccaaa taatacatcc acaccacagg tcatggttgg cgacccgata 3180 gagcaaggcc tgtacatcac cgtagaatct aacgaagtag acggcgtggt tctgaaagca 3240 cctaacggcc tgggtatttc tttctcttac atgtcagggt atccgtcttt gtcaggaaac 3300 attggcacta tatacgaccc aacaggcggc tcggattttt ctggagtact ggtgcctaat 3360 gacacgtttt cgctggtgtc cgcgtggaca aatacagacg ttgacctgtc cggcggcggg 3420 tatcaggtag tcagcgtgtc cgaagggact gttaccttta tagtacctgg cggtctcatt 3480 ggtaggtggc aagaaataag accgggttcc tttttccgtg gtgacggaga ggcctcgctg 3540 caaccagaca acacgtatga gaaaacttta accgattggg tttcaataaa ccgtaccgaa 3600 gtagagcgca ttgtagccaa tatcgccgcc gcgaatggta tgtataaaga caacggcaaa 3660 tcgaaaacac tggcgtcggt caccgctgag atacagtacc agttacttga tgaaaatagt 3720 gccccttacg ggccgatata cactgcgcaa ggaaccgtgt ccggacgcac cccggactac 3780 aacggagtca ctatctatgc cgatctgccg gttgtatctc gtgtaagggt tagggcccgt 3840 agggttacgg accttgactt caattttgaa gggtctgtag tcgatgaaat aacgtatgtg 3900 aacctgtatg gacagacgcg agataacacc ccgcactacg gcaacagaac aaccgtacac 3960 tcgatgcgca agcagacacc gcgcgctgcg gaggttaagc aaccgcagtt gcgtatgatt 4020 gccaccgaaa tggtatacaa atacctcggt aatggtgttt tcgaagacac gaggacccct 4080 aatactcagg ctgtgcaatc tcttatccgc ctggcacgtg acccagatgt gggtggctta 4140 aacctgacag tgcgaaacat ggacaagtta cttgccgttc agaaagagat tgaagattat 4200 ttcggcgaca agcaagcggg ggaattttgc tacacgttcg atgactataa aaccactatg 4260 caggacatag ttagtactat agctgacgct atattctgca ccccatatcg gaaaggggcg 4320 gatatccttc ttgattttga acgccctcgc atgggcccgg agatggtgtt cacccaccga 4380 agcaaggccg gtacttctga aaaatggaca aggacattta acgatgctca ggtgttcgat 4440 agtcttaaat tctcgtacat agacccaaaa acaaacgtta aagaaacaat aacaatacct 4500 gaaactggtg gggttaaaac ggagacttat gactctaaag gtattcgcaa ctataagcag 4560 gctttctggg ccgctaatcg ccgtcaccag aagaacattt taaagaaaat ttcagtgtcg 4620 tttaccgcca cggaagaggg tatctttgct ttgccaaatc gtgccattag tgtcgttaag 4680 ggctcacgta tggctaccta cgatggctac ataaccgcag taaacgggct caccgtagaa 4740 ctatcgcaac cggttaagtt cacagcggga gatgaccatt ctttgattct caagttacgt 4800 ggggtggag tgcaaagtgt taatgtagtc cctggagcgc acgacaggca ggtgattatg 4860 acatcagtgc cgcaagaagc catttacaca ggtaatagtg ctttgaaaac tgaattttca 4920 ttcggcaacg aagcaaggca taatgctcag atgattcttg tttctacggt agaccccggc 4980 gatgacagaa cagtcaaaat aaccgggttt aactatgaca aggatttcta taagttcgac 5040 aacgtgcctc ctttcggtcg tgcgttctcc aacggattcg ataacggttt taactaagag 5100 gatagccata tgtctagtgg ttgcggtgag gtaatgtcac ttaatgattt acaggtagct 5160 aaaaaacacc agattttcga agccgaggtg atcaccggca aacagggcgg tgtagcgggt 5220 ggcgccaata tcgactacgc tactaaccag gtaaccgggc agacgcagaa gaccctgcct 5280 gcggttttac gtgatgttgg tttctccccg gcgtctttca acttcacaac cggcgggacc 5340 cggggagctg acgacgcgga taaagcggtt ctttggccga aagaagatgg cggggacggt 5400 aactattacg catggcgtgg ccccctgccg aaagttatcc ctgcggcgtc gacacctctt 5460 acgacaggcg gcatttcgga ttccgcttgg gtcgcgttcg gagatattac ctttcgcgcg 5520 gaagcggata agaaatttaa atactccgtt aagctgtccg actttactac gttacaacaa 5580 ttggcggatg ccgccgttga tagcgttctt atcgaccgcg attacacttt cagtaataac 5640 gagaccgtta acttcagcgg gaagaccctg accatcgact gtaaagcgaa gtttatcggc 5700 gacggaaacc tggtatttac acaattaggt aaaggttcca ttgtaatagc cccctttatg 5760 gagagtgcta caacgccgtg ggtgattaaa ccgtggaccg acgataatca gtggataacc 5820 gaccccgcgg caatcgtggc cacacttaaa cagtctaaaa cagatggata ccagccgacg 5880 gtaaacgatt acgccaagtt tcctggtata gaatcccttc tccctccgga agctaaaggg 5940 caaagcatat cttctaccct ggaaattcgg gaatgtacag gcgtcgaggt tcaccgggcg 6000 agtggtctta tggcgtgttt cctgttccgc ggatgccatt tctgtaagat ggtagacgct 6060 gacacccga gcggcggaaa agatggcgta attacctttg aaaacctgag cggcgattgg 6120 ggcaagggta actatgttat tggcgggcgc acaagttacg gttcggtaag tagcgctcaa 6180 ttcttacgaa acaatggcgg tttcgcgcgc gatggcgggg tcatcgggtt tacttcgtat 6240 cgtgcggggg aaagtggtgt taagacgtgg caaggtacgg taggttctac gacctctcgt 6300 aactacaacc tgcaattccg ggattcagca gtgctatacc ctgtatggga cggcttcgat 6360 ttaggcgcag atactgacat gaaccccgaa gatgaccgcc caggggattt ccccatttct 6420 cagtacccgg tacatatgct cccattaaac catttgatag acaatctatt tgttagaggt 6480 tcgctggggg taggtttcgg tatggacggg caaggtctgt atgtctctaa cataaccgtc 6540 gaggattgcg ctggttctgg ggcttatatt cttgcccacg aaacagtatt cactaatatc 6600 gcaataatcg acaccaatac taaaaactc cccgcgaacc agatatatat ctcaggggcc 6660 tgccgtgtaa acggccttcg tttggtcggc atccgttcaa ctaccgaaca gggcatgacg 6720 atagacgcac ctaactccac tgtaagcgga ataacgggct tcgtggaccc ctcaaggatt 6780 aacgtagcca atttgatgga ggaaggtctt ggtaactctc gcataaacag tttcaataat 6840 ggttctgcgg cgcttcggtt tcgtattcat aaactgtcaa aaacccttga tagtgggtcc 6900 gtgtactccc acattaacgg cgggccaggt tctggttcag catggaccga aattaccgct 6960 attgcggggt ccttgcctga tgccgtgtca ttaaaaataa acaggggcga ttatcgtgct 7020 gttgagatac cggtagcggt gaccgtccta ccagacaacg ctgtcaggga taacggggct 7080 atatcactgt atctggaagg cgatagcctt aaggcgttag ttaagcgggc cgatggaagc 7140 tatakaagat taactttggc ataaatagta aaggccccgt aaggggcctt aatttatgat 7200 aacagaacca cagcagcgat aagtaaagcg accgcagcca atccgtagcc tattaaaaag 7260 catttcgcac caattgagta tttcatttag cgcccctctc tttgtcaact tcctgttgtt 7320 ctagtagcca gtcaagttgc gcgttagcag cgtctctttg ctgccgtagc cgtaaaacct 7380 cttcttcgag ttccctgata cgttttattt ctcgctttaa ttgcatagct agaagttgct 7440 cgcgggaatt aatggcgtct tttatcaatt cgttgcaagc gtcgatatca aactcattat 7500 tcatttctta cctcgtctct tcatataatt aagcaactct tcctggacag attttttctc 7560 gtccgtacgc gcggcaacga cctcatccag cgtgtcttta gcgactatgt gatacaggaa 7620 cactggacgc tcgtggccag cctgtttctg gcgtacagga cctatacgct cgacaacctg 7680 caaatagtgc tccaggttcc agccttgcga aataaacgcc agatgatgcc cgccgtcctg 7740 taaattcaaa ccatggcccg ccgatgcagg gtgcacgcac aaaatctcga tttccccgcg 7800 gttccaagat tccatctgct tattaccctt agcacctttc gcaaacgcct gcgcctgggg 7860 gaatcgccta agaatgcgct caagttcgtg cttgaactga taggccacca gcagcggcgc 7920 accctgcaac tcctcgacaa tggactctaa cgcatcgagt ttcgcgtcgt gcactttctc 7980 ccagtctttc gttgcttcac catccggccc cgacacatac acggcaccgg aagcaatctg 8040 caagcactta gccgttttag cggcggcgtt agccgcttca acttctccgc tctccagttc 8100 cgcgaataac ttctcctcca tatcgatgta ggcttgacgc gctttcttcg gcaggtcaat 8160 ctcaaccggt acaataaccg gtgcttcaca accgaaccac tcggcggcat caatggtaag 8220 gctgatatcc ttcatcttct ggtgaatctc gttatccgcg cctgggcgag catggtactc 8280 ccgcgccatc gcagatttgc ctttctgtac cgagttaaac catctatcgg taaaggccgt 8340 gtatgaagac ccaaggcgtt cacccgcgtc gatgaaccag ttctgacccc acaagtcttt 8400 gaggccgttt ggtgatggtg taccggtcag gttaatgaaa cgcttaactt taccgaacgc 8460 caccttacta agcgcctttg cccgcttgct accgccagaa cggctacgga acgatttcag 8520 cttcgtgctc tcatcggcaa cgataacggt aaaaggccag tcgtctttgc cgtagtagtc 8580 aataagccat tctataactt cgtaatttgt gcaaaccacg ttagcgtctg actccagcgc 8640 cgcgatgcgt cgcttctcag aaccagttgc atcgatgaca cgaaggcaag ggaaccccca 8700 cttagtttgt tccacaggcc acgtacccga cgcaacacgc aacggggcga ggattaatac 8760 ccggtcgtcg tcattaagtt gcccattacg gaacaggcgg ctaagcaccc acattgtcga 8820 gcttgtgttg tgcgttacgg tgaagtcacc taataaaaac cggtggtcac cgtcgatggt 8880 gaagccgtaa taatcatcta caccgaccgg ggttatagat tctatcccga cgttaagtac 8940 gtttttatta atattacgct tcggtaaatt ctggtggcga ccacgcacga aaggcacctc 9000 agaaaaatcg cctgataccg atacgcggaa gtaatccccc caaacatcag tgtttgcgca 9060 acgcttacga gttttcttct ggtaagcggc gaaaccaaga gaacggcata gatagcaaaa 9120 atcatcagcg agtcgctcgc ttaccgaaat ccagtcaaaa cctgctttag acaaatcgca 9180 gtaaccatca ctatccagaa gccctgccag caactctaaa cgctgtctcc ggtcgccgca 9240 tttgtaatta tgcggtatat gcttattgtt caggacaccc gccgatttca gcgcgtgcgt 9300 aaacccgtgc ttcttatgac ctgtgttgcc gtgagatata ctccatgtaa gtccttcttt 9360 tcgtatttgc atcccattac gtgctgcgta cgactcaaga tacgccctta tttccttttc 9420 gttctctcct gatgttatgg ccccagacga cgaagtaccg tcgcccagcc atagccccat 9480 aaggtacggg ggcaataaag cctcgtcctg ctctttcctt ggaaaatcga caggtacccg 9540 ccaaccctta agatacccat taggcccggt tacatatttc ggcaacttaa gccagtcgcg 9600 gcgcttatg tcgaaaacgg tgttatctgg ccatgaacct tttgcgatac ctgtagtagt 9660 tcgcagtgat aatatatggc tttcgtttac cgtatacgac tcgccttttc gcggtttaac 9720 ctcgtacatc atctcccggc ctctaccgag agacagtaca tttctaggcg ttgaatctgg 9780 tcccataagc acgtcaccga cgatgacgtc ttctactttc ttggtcgtac cgtcaaacat 9840 aataacttca gtcccgcgtt tcaggcactt gccgctgccc atacttgccc agatattaca 9900 gcgcgggtgt cgcagcatga acgaggtcat gagcttttga taaggccttc tagtgaattt 9960 actcatttcg ccaccagtac cagttccttc cgcccgaacg ccgtaacgtt acccgttaca 10020 tcttcgataa ccagtttacc gttcgactcg acgaacacag tatcaacggc aacaggactg 10080 cgggtcttaa cgttgaaaat catgtctcca ggtacgatgt cacgtgctgg tttgcggtca 10140 tatttgtatt tcatttctca attccttatt tctgtttggt gtgaactaaa tataatagtg 10200 ttctattaat ttatcaacct gtttcttcga cccaacaaca aaaacattcg caccgcgttt 10260 gcgcatcctc tcgtgctccc gtaactggtg cgggtccggc ttcgtgtttt cgtctttctt 10320 cacctcgacg aaccagacga tgccgccagg gagaattatc aacaggtcag gagcaccgga 10380 gcgcccctca taggaaagtt tccgaacgag gccccccagg gcctcgaatc ggtcttttgc 10440 gtatttctga atcttgcctt ccggggtcat aggcatagaa cccaagtaag gaaagcccct 10500 cctagaacgg caacgccaac aaacttaaag aacaggccat aacagaacac cgcggctatt 10560 cccgcaccga gcaacagtgc gagcatggtt acgattaccc aaaacacaaa catcgtcata 10620 atatgcaccc ctcgcgtttc gtgtgttcaa taccgcagcg aggacagatt cggcagtctt 10680 cttcccggaa ataatatata atcagtttac ttagcatacc gttttaactc cgcaccttcc 10740 gctacaagag gaaaaccttc agcccattct ggcaatgcac acattagttt ttccaattcc 10800 gctaccgtgt aataaggtgt atccggggtc tcgcatacca gttcatcatg aaccgaaaga 10860 actatagggt acccgccagc ctcaacatta agcattgcgt aggccaataa gtcacgacaa 10920 aacgcctgaa cgatgttctc acaagccttt ccgccgtgtg tgtacagggt agtccactgc 10980 cgtgttaact ggttctcgcc ctggtactta attcttacat tggtgtttac ccgcccgtct 11040 tcgtcggttt cctttgtcac gctgacgcct attcctggat atgataggat tcgccccgac 11100 ggcagctcca tgcatagcca ccatcctggg atgtttctac cggatgaatc agtttctact 11160 gttctccaga tacggatagc tctttcaccg ttgcttcgga tatttgcgcc cgcccagaaa 11220 tcgcggccag gattacgtac tgcggcaaga ataccgtctt taaggtcgca ccagaaagct 11280 acggtttgcg ggtgcgattc acgccacata cgtttaatag catcacatgt acgccatact 11340 ttcttatcga gaatatacga tggacggtca tccttctccc ccggacgcgg cggccgtttt 11400 gcctcctgta tacgcgccca ctcatacccg cgagcggtag cggcccagat atggtcgggg 11460 aaagttcctg ccattgtttt ggccatctca ataaggtcaa gacctaagtt tttggcgaat 11520 gtaacgaacg ctccgacgcc accctcatag ccgaggccaa gctcgcaagc ttttcctatc 11580 tgtctgatat ccttaaagtt tttcttaata tcatctggat ccatgccgaa catctttcct 11640 gcggttacgc agtaaatatc cagcccggcg cggaacgtat caagcgcggt ttcttcgccc 11700 gccagccatg caagcccacg gccttcgacg ttagagtaat ccgcaacaac aaacttacgc 11760 ccggcttccg ggataatgca gcttcggacg gtagacgctg ttagcttggc cacatcaaaa 11820 cggcggtgtg cacggccttt aagtaacgcg gcaatacccc tatccagttc atcgtcgtga 11880 taatacccgc gcgccaggtt ttgcggctga aaacctttcc ccgcccaccg caatgttcgc 11940 ttcgctcctc cgtattgcag gcaaccacgg cggcggtcat ctgaagaacg gcctaacagc 12000 aacggcgcgt atttcgtcga tgcggtagaa gcagccccga gacgcatctc gataatagtg 12060 cgggcgtcgt ccggtaaatc ctcatccgcc agcaaatcgt ttagtgttga cttctgtgcg 12120 ttgtgtatgc ggtgcgctgg cgctagctcc tgtaaaatag gcaagaagtc tttgccggtt 12180 agtgaaccgc catatttacg ttgggcttct tcctgtaact cttctttatg gcgcgtgact 12240 gcttcaatcg ctgcttccgc caatgccaca tcaaccttaa atcctcggtc attaatcacc 12300 tggtctaagt gcagcacgcg gtcttcgaac tcagaattcc cccacttagg cattttctta 12360 tacacttcac gcatggacgt aatgtcgctt tttgcgtatg caataaattc tgcccactct 12420 tttgggtggg tatcggcagt ataacgtcga atcttatagt tcttgggcgt aggcttactg 12480 aatcgctgta tcagcgcctt accgcgttta tccttagcca tgctggcgtc aatgtttaaa 12540 acctcgcaca acgcggctag tgacccagga agggcatggc ggaatgcaac tatcatagtg 12600 tcgatgatgt tttctatcag gagttcgaat ccccagcaat gcttaataac tggtcggtcg 12660 aacataagga agttttggcc taccagtttg aggttgctgt ctggtttttg cagctttagc 12720 atcgcccggc gcaaatcacg cggcatatca ctcccgtcgg tggcatccca tacctgcacg 12780 gggccttcat cgaaagcgta ggtgcaaata acaatttcgg tagtgggatg ttcagcgtag 12840 gcataggagc cgactttttt caaatcagct tcggagaatg tttcaatatc aagataaagc 12900 aagttcattc atttgaccct tatagaaaag gcgaccgaag ccgccttaaa tgaaatgaga 12960 atatattaac ggttacggcg acgttcgcgg cgcggggttt catcttcttc atcatcttcc 13020 aggtcatcaa cgctggcaga aaccgaagaa ccgccaaacg ctttaccttc acctgcaaat 13080 ttgatgccgt ttagttttgc gcctaaaact ttatattgct cggaaaacca aatttcgatt 13140 gagatattag ccacacaacc gctatatacc tgctcacctt caatctgctc accatcaatg 13200 ttgaagtctg gttctacctg cttttcaccc ttaaccgaag tcaggattaa tggttgttgc 13260 ttgttcttgg cctggaagta aaaaccttcc gggaagtctt cgaacggatt atcgcgctca 13320 gcgatgtcgc gcacagcgca cttgtccatg tgcttacctt cgccgtagtt ctgcttcatc 13380 catttctcag cggcggcctc tcccaatgct tccgacacta ctgcgaatac cgtatcgcga 13440 agctcttcaa tttgtgggtg ttccggagtc aggataaaag taccgttata cgtaccttta 13500 gtaacggaac cgtcattatt ttcacggtct ttagcgcgtt cgaatacgtt cagccatgcg 13560 gtttgtactt tacgaagatt aagtttaatt cccattttag ttttctcgcg ttttagagtt 13620 tatccgggga gctgcccggt cagtgattag taatataata gagtacgatt aagatgtcaa 13680 gcctctaaat cgtcttcagt aacatttttc cattctggcc gtttgtcttc gaccgttgtc 13740 acacacggcg ctcctggctt acgggttacg tgtttctcca gcgccgccca tagctcagaa 13800 cctttgtgct gcttctcggc ctcggtcggg gtcattagta cttctttatg aagcatttca 13860 gcgcccaact caaaatgtga tagcgtttcc agcagcgcat cggtatcttt ccacgcacga 13920 ttacccgggc ggccttcaac cagcttgtac cccggcactt tcttaccgga atgcaacgcg 13980 gcagccatcg ctttctcgac cttgtcgatg tgctggcgca gcagtggcaa cttctcatac 14040 tcagctacga gttgctccgg tgtaagttcc agcgcaaagt cgtcctccag ttcttccgcc 14100 agtacagagt taacggtttt tgtacgcgcg gcgcactgtt ctgagaaacg acaccactga 14160 caaccatcga ccgacggttt gaagtccgac gatttcaggt tcttcttacc gcggaaatag 14220 gcatcaagcg ctaacagcgc acgtttctgt gcgaacttag cgaacagttc cagaccttca 14280 accgagatgt cccactcgga cgccccgcca gcatacggct ggaagatgac cagacgaacc 14340 acggtgatgt tataacgtct cttgagtcgg cgataaacac cgagagcgta aagcataagc 14400 tgcttgtttt ctttcgcttc aacacgatgc cgtccggttt taaggtcgcc gataatgagc 14460 atgtgctcgt cggagttagc cagttcctga acggcaacaa ggtcagccgt tccgaatgtc 14520 tcaacgcctt cgtaacccgg atgcaatacc tcagtaagat ttacccgcat ctcgagcttg 14580 gcgtaagtcg ctacgtcgat aattggtttg cagtagtcag tgtacttgcg cacctgctca 14640 atcatgtccg ccgtaatcag taccgcgcct ttcatcgggc tgattagcgc cttaatctgg 14700 cctttaccct catccagcac gtaagcaccg acttctcgct ctagtggcag tgcagtgccg 14760 cggatatagg cgttgagatg gacctcggct atagtgtgac atgcggttcc tgtaatggcg 14820 gatttaccgg acgtgttggg gatgtctttt tcgcatgcca gtgatgcggc acagctaagc 14880 cactttttag cgccagacgg cgacagcagg gcgtgcacat cgttattgcc accgcgttcc 14940 tttaaaatca taccctgttc tcccactggt caattaaatg tcgcgtctta tggtcgcaat 15000 gcatagccca tccatacatc gactcgaaga cataaaagtc aggttttgcg aaagtcgtgc 15060 gcttaatctg cgacacgtga cggcctatat ctttaggccg tggcactttg cctaagtacg 15120 ccatctcttc catcaggtgc gcaccggacg atgcacgcaa cagccatagt gcttctgtgt 15180 tatcccgcct gtctacggcg cggtagagtt ggtaaatcat ttctttaccc tcacagaatc 15240 catccacgcg aaaacgtcat ccatagaaaa atcaccatcg aaaacttttt gtgttttatc 15300 cgacttttgt acatacgtag gacgtttgtt atccgtagcg tggaagcata caaagttatc 15360 tgtttcttct ataactcgcc ctgcaatcat aagaccactt ttcttagcta aaatagccat 15420 aattccgacc ctcagttaaa gcggcccgaa ggccgcggta tatttattct tcttcgaaat 15480 acttgttctt gattgccgtc aggcgttcca ggtactcggc caggtcttcg tctttaatcg 15540 aggcaatctt cattttctta ccggtgaact cttccagcag ttcatttgaa tcgtcacacg 15600 cggcatcgct cggaccttca ttaattgcgt cgtcgatagc cttaatctgg tcacgaagag 15660 actggtaatc tacttcttcc ttctcttctt gtactggttc ctctactttt gccttacgcg 15720 gcttacgttt tggtttctcc tcttctgccg gtttagtgcc gacgatgtcc tcaccttcga 15780 ccggaatttc ttttgctttc tcgacttctt ccagtgcttt agccagttct tttttaactg 15840 tctcgacacc ggcaacggag acgtcaacag tcatttcgct tacggatttg ttagacgtct 15900 gtttcacact attcgcagca atcagttcat gggcgactac gaaacgttca agtaatttta 15960 agaattggtc taacattatt tcttctcctg tttagttggt gataaagact ataatcgtac 16020 tctattattt atgcaagaac tttttcgtaa attgcgtcgt gtattttatt actgtactat 16080 tactgcatat ctaactaagg agtaaacata tgcagcaatc tgagctaggc gctcgcatag 16140 agcgccgccg taaagaaatc ggcatgggtc aaactgagct tgctttcaaa gctggcgtat 16200 cccaaagcct gattacccac ctggcaaccg gccgggtgtg taaggtagac tgttttaaaa 16260 tcttccacat tgccgatgct ctcggtgttg accctcgatg gttaagcttt ggtgatacag 16320 gggcctgatg gcccctttct ttttactcta aatccccttc tgttgtcttt atgttgtcgt 16380 taggttcata ccggttcttc ggtcgcttac ggtcgtctat acccatcggc aacctgtaag 16440 tattcgttac aacttcaccg ttttcgtcgc aaccaagaat caattcaccc tcgcgtagca 16500 tcttctcaag caccaggttt gtgatactgt ggtctccagc tctcgcgact atttgccgtt 16560 gtgtgaaccc tcgtcccgta tcatcggctt gctgtaaatc ctcgagagcg gacataaccg 16620 actcacgggc cgaagcgtct ttagaccgtt taaccgtatc ctttacggaa tctttacctt 16680 taccatccaa tccttcgttt cgctcttttt cctcgtccgt ttcgaacggc tggaagcccc 16740 acggcataag tacgagagct ttatgcggct ccggaaggtc gaggttgact atctccccgt 16800 atccctcacc cccagcgaat tctaccgcct ggaactcttt cggcggcggg gcctcgcgaa 16860 actgtactgg ttcaagcacc atgcctattg ttttctgctg cataccgttt ttgttcttgg 16920 tgtgcgcaac gtttatttgt ttctcggtag cgcggacaag cgttagttcc acatcgacac 16980 cggcatacag cgcaccactt ccacgcgcct tacttccacc tttcggggtg tggtggacaa 17040 cgcctacagc gcctttagta ccgtcgcgca cttctttgag catagccacg acacggccca 17100 taccgtcctg tccggctgag ttctcattga acttgtctat ccagttcccg aaggtctggt 17160 taagagtgtc gaaagcgacc atcccgacag gttcactgcc cgctatctgg cgcattttac 17220 gcaccagttt cttcgtatcc gcgaattcac cagcgtcaag cacatgcata tatcgcatac 17280 cttcatcgcc gtatttagcg gccaacgcag cgatacgggt gtgcgtgaac tccccaccct 17340 caccatcaat atagaaatga tgggctttac gggtgtctgc ccccgcgaac cgatacccgg 17400 cggcgctgat atacatcatg ccgagtgtgt agaacgattt atacgtcccg gattcaccga 17460 cgatatccca aatgcaatta gacggcatat agccctcgac gatgaagtcc gcttgtggtg 17520 ctggtggttc ttcgtcgtcc aggtcttctt cgtcgcaagt gaccccctcc gcccacccaa 17580 gcgccacctc cacacggtca aaaggtaagc cggtggccgc acaagcgtaa gcccacatat 17640 cccgtccgcc aggcttcatc ccgcccgtag ctaccaggtc ggtatcgtgg tacagggtaa 17700 cgttgggtcg ctcgaagccg tcgcgcggcc aacagaacag gaagtcgtcc tgcttaggtt 17760 cgccggtagt gtattgggcg gcgtgctccg gcgtggcggg catttgcagg ccgcggtcgg 17820 tcatacgtcc gccgaattca aaagcgaact cttcaaacag gtcggttaat tcggattgtt 17880 caccctccgg aactttataa tcagaagcac cggtaacgtt aatttcagga acttcctcca 17940 taagcttgct cgccgtaatc atgcggctac tttcagacac gatgacctgg cttcctaccg 18000 gcgggcggta cataggctgc gacagggtga acccggcgct atccacgtca cgacctttaa 18060 ggaagtgtgc caacaggccg taccggatgc ggatgatgtc gccaccggtt accggagtgc 18120 gtacaggcat aacgacgcgg tagcgcggcg cttcttcggt gtgggatgcc gtggtgtaga 18180 gcatcattgc gtaacgtgac tcgcgcacca tctcacagtc ggttgcgaac tcttccggcg 18240 tcgcgctgtc cacgtcggca taggccaggg atgaagacgt aacagacgca ttgcagcgat 18300 agaacatacc ttcggcggct tctttaccgg tagagcttac cgtggcggta caagcggcgg 18360 taatgtagcc ggggtctgtc ttggggttcc ggcgcgaacg tttaagcggt tgcataagtt 18420 caacaaactc atcccaggtg ccggaggtcg tggtgtaaac gttaatgtca gcgcgttcct 18480 cacggcgatt gcgacgcgac catgagtatg ctaaattgcc tgttgacatg tgcttttcct 18540 ttgtagtgtt taaggccctg acgttcgcgc gttggggctt ttcttttatt caaggtcttc 18600 tggggtcgcc tgggagcctt taacttcagc tactgccgaa tattttccgg ccttactttt 18660 aactgctccg tctctaacca aaacttccag cgacctctcg atgagagag aactatagta 18720 ctgaaaataa gcgcggcgca agtcaagaac actacacgac ccttttttac gcgtaagcgc 18780 gactacagtt ctgaataccc ttttctggaa atcggtcact ttacattctc cctaatccac 18840 gcttccactt tctcccggtc aaacgtaccc ggcatccggc gacccataat ccggacacaa 18900 caatccggga acttgccgtc tcttaaccag ttattcagcg tgcggcgggt aaccccaata 18960 agctcagcta cttcattctg tgtcattctc aaatcctcca tctcagttag taaagcaagt 19020 atacccacgg cataatggga aatcaactat aaacacacct attgacattt tcattttttt 19080 cgtgtaagtt gtctcttgta acttcaagtt acttaagttt ctgtttcacc ttgtgagagc 19140 ttagcgggtc cgccctaagc ggacacgcgt taaggcttac tgaaacccgc aaacttaaaa 19200 ctcagtaact tccttcctta gtatctgaac tgccttgggc gagctatatt ctcatcggca 19260 gtatatctct taagtatctg atttccttga tgtgccgtag attcgcaacc gtaaaaatgt 19320 aacgattagt gggcaggctt aaggcctgcc ctttagttac caagtaataa gaataaaggc 19380 acaaccctgg ttgaataacc acaacggcaa cgtaactgaa atgccgaaag attcttagtc 19440 tactactcag ggattaacgc acttgcctta cggcaagcgc ttacctgacg agggacgaga 19500 ataaaggcac tctaatgggg cagggacctc cctgtaaata accccacggc aaaagattct 19560 aaagagtccg cgccggtgat tctcgggtct taaaggaggt tgccgtttat ggctagggtt 19620 ttagtggtcc ccggggccgc atccggacgc cgatgatgat gggtggatag ttcctggggt 19680 atcgtggtgg cggggccgca cctgggtgct gttaccggaa ccgcaggaag cttgtaacag 19740 tagtttagga ttttactgag tgcttacgc tgctgagtag cagcgtacca tttttgttaa 19800 tatttttcta tttcacgaaa tgcaacaatc gaagcgactg ttgcaggaga tgcaacaatg 19860 aaaccgaatg acctcgtaac ctggaccggg cgtaacggag agacccgaca cggcaaagta 19920 acatcgcttc acggcatcta tgctcgtgtc gagtggtggc gcgctcatgc taagaagcct 19980 cgtcgcattc tgacgcgtca ggatagatta atcgcgaaat agtattggca taaccaaaca 20040 taatagtgta ctattactgt atcgaaacga gaggggcggg aggaatgtac gaggatatta 20100 gcctggaaga tgcagtgtat atatggtgcg ccggagaaga tgtagtttgt gacggggacc 20160 tgaaagctat cgtatgcgag gagtgtggag atgagtagca aagacgaagt atttgaatac 20220 ctgattgacc aactacgtca gcgcgcaaat aagttcgacg ttgtagcaga aatacacaag 20280 atgagcatag accggtcgat aacaggccgc tcagggtact cggacgagga gaacgcaatt 20340 atcgacgcct acataggcca tgattctgac agcaaacaaa tcatccataa cttacaacaa 20400 cacctcgccc gtaaagacgg cgacatacgc atgctgaaag acaggttacg ccgggaggta 20460 gataaggtta aggaactacg ggatactatc gaacttatga atgccgactt ttaccgagtg 20520 actaaacgcg aagaatgtga gcactactac cgtttggttc tgtggagtag cgacttaaaa 20580 cagtgtacta aatgcggggc agtattgaac ccacaacatg agctgtacga tggttggata 20640 gaaaatccag gacgtaaggc gtgcccggtg cctagcgatt gtgaggttga ggttgaattc 20700 cgcaatggct ctatagccgt cggcgcagcc caggactacc ggtgggggat tgataacaac 20760 aactgggata tagttaagta caggataatc aaatgacaag tatacttttc atatgggttg 20820 tatccgcggg tcagctacaa cttgccggaa cagaaacgtt ttatacgtta gaggcgtgcc 20880 aggtagccgc acgcgctgcg gagaacgcgc cgttaacttt cgtagagcgt cctgttgacg 20940 ctcaggtacg cggtatttgc acactgaaac ggttatctaa gcagggggag aaatgagtct 21000 cgcaaccgat atactgaaac acgccggcat caacctggca cctccttcat cggcggtaac 21060 aaagaaggtt actcgcgtta aagaaaaggt taagcgtaaa cctaaaccgc gcgcaaagcc 21120 agttaatgag atgccggacg tgtatccgcg catccctggg gtgcaccagc ctaagtattg 21180 tgtaggcaaa ggtctgtggc gtgcgcactc ttacgacggg aagaaagtgg taaacctggg 21240 ggagttcagt agccaggcaa gggcgcatat ggcggttaaa ctgtacaaac tgtggcgtaa 21300 acgtgggtat tccgaaatcc cgcacaagcc atctattcga ctttatacgt tcaggtaatt 21360 tatatgacta cgatagcttt cgacggcaaa accatggcct gtgatacctg tgtcaccggc 21420 aattacaaat accacacgga taccaagatt tacgagaacg accactttgt tataggggta 21480 tccggggacg ccggggtggg tatgttgttg gttgccgatg atgagatact gacgcctaag 21540 cactacgact tcgacttttc ggcgctggtg tttgttaaag aggataaccg tattttcgcg 21600 gtagaattct gtagggcgtg ggacgcaccg ttaagctcgg ttataccaat tgcaggtaac 21660 gctgctgccg taggctccgg ggctccgtat gcacttacag ccatgttcat gggtgcgacg 21720 gcaacggaag cggtagaggt tgcgaagagg tttgaccctg gtaccgatgg aagcgtgata 21780 acacatcgac taggataaat ccgaattatc tgttactaaa attgtcgtat aaatgctaat 21840 ctccgaagga taaaccaact ttcggagatt ttttattgtg gacgataaat acctttggct 21900 tagtgtggcg ggcctcgccg ggggtgccgt atctcagatt aagaagcgtg aggctatttc 21960 gccatggttg cgattgtgcc atctcaccgc ttccgcctgt tgtgcagtgt acgcatcccc 22020 catcattata agttactatg aattatcaca gtctgagggt cagtacctgg ttcctttcgg 22080 ggtaggtatg ttctggctta agttattcga agctgctgac tcatccctca gcaactttaa 22140 gttaccgtgg gggagataac atgctcaact catcaattgg catcatttgc ctggttgcca 22200 tcatcgccac ctcgttaatt aatatctacg ccgactgggt tgaagacggt cttttcggcc 22260 gt; gtgccattcc accgttcata attacgactc tcgtcgctat attcgcgctt aaatcggtgc 22380 gtcacatttg cgtgaaaggc gctcgctact acaaataccg gaggatgtat gtcaaaccga 22440 aacatcagta acaacggcat caaattcacc gccgcgttcg aaggtttccg cggaaccgcg 22500 tacaaggcaa cgaagagtga gaagtacctt actatcggct atggccacta cggcgcagat 22560 gtgaaagaag gccagaagat taccgaggga cagggtcttc tgctgctgca caaggatatg 22620 gctaaggccg tagctgcggt agacgctgta gcgcatccgt cgctcaatca gtcacagttc 22680 gatgcgatgt gcgacctggt gtataacgct ggcgccggtg tgattgcggc ttctaccgga 22740 acagggcagg ccctgcgtaa gggcgacgtt gccacactgc ggaataagct atctcagttc 22800 cattatcaga acggcaaatc actccttgga ttgcgtcggc gtgccgcggg tcgtgttgcg 22860 ctattcgacg gtatgccgtg gcaacaggcg gaagccatcg gccgcgaagc aaagtaggtt 22920 gacacatagg agaattccta agatactaaa cctgctcctg cttattcatc cctctagctc 22980 ctttatcccg gtaactgacc cttaccggga tttttttttt atctgtattc agaaataata 23040 gttgactagt aacattaacc ctattatatt tagttcatcg acaacgagaa cggagtagag 23100 aagatgaact tacaaagcga taaagttttt taccattgca aaccgctact ggacaccgaa 23160 gcattcaaag acgcacaact gatggcacgc attgccgtta ataacctaag tacacggatt 23220 ccagcggatg cgttctggtt cgccgcgatg cagacactga aagcagctta tgcaggagaa 23280 caaaaatgaa agtatatatc gtatccggat ggcatcacga cggggacact tacgttcttg 23340 atgtttatct cagcgaagaa aaggccgaag ccgccgccga gctggagcgc acctcacatt 23400 actatgacgg cgtagatgta ctcggatggg aggtgcagga atgagcgaca acgggcaagt 23460 aacggtaacg tttaaggtgg gcggtaaggt aagtaacacg cccttcccga cacgcgagga 23520 actgattaaa cgtaacagtt tcccaggacc ggacaagaac aagtatctca atcgaatgtg 23580 gggtaagaag aaatgactaa caacgaatat gaaaagatga tggtagaagc cgccaacggc 23640 aggtttaaga ttgaagcagt agacgcgcaa atcgaggcac accaggctgc gcttgactac 23700 cttaatgagt atcgccggga gcttatcaac cgttatgacc tgaataagtg cgagcacgat 23760 tttgttaaca atcctaaaga tatgtgcgga cagtgtgtta agtgcggggc gattaaacga 23820 tgaaacgagt atgcaaagaa acgttgcgcc aacgcatcac gtatttggag tctgtaagta 23880 agctgggcct gtcaatgaac gaagaattcc agttagaagc atacaagatg ctgctggaac 23940 gacttgagaa ggagagcgaa caggaatgat tactagtatc ccaaaactaa tagagaccca 24000 cggcacgctg gctgatacgt gccgggagac gggacttaat gagatgacat tgtccaagta 24060 ccgccatgac acaaaatgcg agcaacatgt catctataac aaccgactga tgacccatac 24120 gaaaacgtca ccggtcatct acacgaaacg tggattatcg cgcaacgacc gtatgaagct 24180 ggaggggtga tggatagttt tctcatcctc atgggtatag gattcacaac attactgctt 24240 gccggtgcta tgtacctgat gtggattttg tttctttggc catttgttga ggcggtaagt 24300 cttacgagga tgtcactagc gtatcgccgt ttgaatgctc ataaagctgg tgctggtgcc 24360 gcggtgaagc tatttgtcat gtggtacggg gaaacagtat ttggtcgtag gttcgaagcc 24420 atacatagtc gcgggtggcg atgggaagcc gtaggtaaat ggtgggttta taaagaggag 24480 agtggagaat gattgagatt ttgaaggcta ccgatgccac tggcgacctg atgttttggt 24540 ttgcctttct cagtgcgcta ataaaacaac cgactgtgtt taacagtctt ccctgatatc 24600 tgtggtggcc tatgttttta ggcttgggtg tatcaggagc ggccggcatc ctgctgtggt 24660 ttttcaaatg agattgctaa tcatcccgaa cgcctgggtt atcgccgcag ctaacgacca 24720 ttacggcggc gatggtagaa gagcactacg acacggccta tataactgac acaagcccgc 24780 ttctgcgggc ttttctgtac acaggccttc attcccctgt acaatcctta atagacgctc 24840 agggggcgtc tgatgcgcta tgatggtcat acgcatttaa gagaggattg catgaaactc 24900 ggttagtgca aaccgtcgcc gtaaacgccc gcgtggtaaa caaagcctgt atcagtcatc cagacattcc 25020 gccgaactat gggatccgga ctattgcgac gagttaatca ggttcttcga ccgcgcgtca 25080 tgggaactcg tacccacgtc taaaggcgac gaacgcccgc ttatccagga caaaccgccg 25140 tcgctggccc gcttcgcatt acacatcggc gtaaccatcc cgattatcaa gctatggctc 25200 cgagaggttc ctgcattcgc cgaagcatat gagacggcac aagccctgga agaggcgtat 25260 ttcactgaga ccggtgcagc cgggatatcc gctacgtttg ccgcggcgaa actggggctt 25320 aataaaactg ttgtggaaga agcgcgcgac gaaccaatta gcgaagtaac tattaaggtg 25380 gtgtccggtg aacgttgata tcacagctac ggaaccgcaa ggcgcgttcc ttaatctgca 25440 ttgtaagttc ccggccttcg ttgcgggctt cggcacaggt aaatcagagg tcatgtgcaa 25500 ttccgcccta ctcgacagca tggagggcgg tagtgattca cttatcgcca tgtatgaacc 25560 gacatacgac ctggtgcgcc ttatcctcgc tccgcgtatg gaagagaagt tgtctgattg 25620 gggtattcgc tacaagtata ataaatccga caacatcatt tatacctcat ccgggcaatt 25680 cggggatttt gtccttcgca cattggataa tccagcacga attgttggct acgaatcgtt 25740 ccgcgcaaaa atcgacgagt tggacacgtt aaataaagac cacgccgagc acgcctggaa 25800 caaagttatc gcccgtaacc gtcagttgcc gcgtacatat cgtccaatta ccccgaagcc 25860 cgctaataca gtttcggtat ttacgacgcc agaaggcttc cgttttgtgc acgacagatg 25920 ggctgtaaaa aagaaaccag gctatgaaat gattcaggcc tcgacaacat ccaatccctt 25980 tctaccggaa gattatgtgc agtcgttgcg ggatacgtat ccaggccagt taattgatgc 26040 ctacatcgac ggcgaatttg tcaacctgac atccggcagc gtgtattatg cttacgaccg 26100 acgtaagaac agcagtcggg agactataca acctggagag acgctgtaca tcggtcagga 26160 cttcaacgtc gggcatatgg ctagcaccgt atacgttcag cgtgagtatg tctggcacgc 26220 ggtagccgag ctggtggata tgttcgacac cccggatgtg gtcagggaaa ttaccgagcg 26280 atgggggcga cagggccacc acatcgtcat gtatccggat gccagcggca agaaccgtaa 26340 atcgactgat gccagtacgt cagatattgc ccaattacag aacgcgggtt tcgagatacg 26400 tgcgaaatca gttaaccctg cggttaaaga ccgtgtagca tcggtgaata aagcgctgga 26460 gtctggtaga ttaatggtca acgagcaggc ttgcccggtt acagcacgtt gcctggagca 26520 acaggcttac gataaaaacg gtataccgga taagacaagc ggcaacgacc accagaacga 26580 cgcgacagga taccctatcg cctacgaaat gccgttggtt aaacctgttt cccatatccc 26640 ggttactttt gcactttaag aggattatta aatgttaaca gcaaacggcc agggttctgg 26700 cgtaaaaacc aagcaccgcg aatggctgca ctacgcgccg aaatggcaga aggtgcgcca 26760 tgcgcttgct ggagacctgg ttggctactt acgcaacgtc ggccttaatg aaccagataa 26820 agcatacggc gaagcgcgtc aggcagaata cgaggccggg ggtatcgtct acaacttcac 26880 ccggcgaacg ctgtcgggaa tggtcggtag cgtcatgcgc aaagaaccag aaatcaatat 26940 tccgaaggag ctggaatacc tgcttaaaaa tgcagatggg tctggtgtag gcctaataca 27000 gcacgcgcaa gatacactca tggagattga ctcagtaggt cgtggtggtc ttcttgttga 27060 tgctccggaa acagccgcgg ccaccgctgc cgaacaaaat gcgggcttgc ttaaccctac 27120 aatcgctttc tacactaccg agaatatcgt taactggcga ctcacgcgcg taggttctgt 27180 aaaccgggta actatggttg tgctgcgtga gacatgggag taccacgaac ctggaaacga 27240 gttcgaaact aaatacggcg agcagtaccg cgtgctggac attgacaccg atggtaatta 27300 tcgtcagcga ctgttccgtt tcgatgcgga aggcggagct caggaagagg ttgtggagat 27360 ttacccagat ttaggggagt cgttacgtgg cgtaattccg tttaccttta tcggagctac 27420 caataacgac gccaccattg acgacgctcc tttgttgcca ttggccgagc ttaatatcgg 27480 gcactaccgg aacagtgctg ataacgagga atcaagtttt gtagttggcc agcctacgct 27540 gtttatctac cccggggata accttacacc acagtcgttc aaggaagcca accccaacgg 27600 catcaaattt ggcagtcggt gcgggcataa ccttggttat ggtggtagcg ctcaacttat 27660 tcaggcgggc gaaaacaacc tggcccgcca gaatatgctg gacaaagaac agcaggctat 27720 ccagattggt gcccaactta ttacaccatc tcagcaaatt accgcagaat ccgcgcgcat 27780 ccaacgcggc gccgatacat ccgttatggc cacaatcgct cgtaacgtaa gtcaggcgta 27840 taccgatgct ttacgatggg ttgctatgat gttgggtaag ccagaagatt ctgaagtcga 27900 gttccagctt aacatggatt tcttcctgca acctatgaca gcacaggaca gggctgcgtg 27960 gatggcagac attaatgccg gattactgcc cgccactgct tattacgctg cgttgcgtaa 28020 ggcgggggtg actgactgga ccgacgagga tattctgaac gctattgaag atgcaccttt 28080 gccgtcgggt gctgttactc aggtagcggg ggagattccg caagcggcac aacaacaaca 28140 ggagtaaaga aaaaggcccc attacggggc cttagtttta gcggatatta gccgcgcatt 28200 tacgcacaac ccgcggtatc gtctctttca gctcatccca tgagttgact gcttccatat 28260 ccaacaacgt caccatagcc ttttgaatat cactccgtaa ttgattagtt ttggcgtcct 28320 gagagtccca agtatcaaaa ccggttactt cacggatatc tatagactct tttatcgcat 28380 tcgccacatg tatgaagtga ccgatacgcc caatcctacc ctccgtacgc tcaatcatcc 28440 ggctaacggc ggcattaagt tctacaaaac taacagcacc aatgttacgg agttctacct 28500 gccgttgagt taagaaacgg tctataacga ggaagtggta atagtcagaa tattgctccg 28560 caacccatat agccatatgt agacatgcgt atgttgccgc gtttttccct ctaaccacac 28620 gccacgcgct ttctatccct aattcctcac agcacacacg cgcgaacgac ctggcttgcc 28680 cggtagataa cagctgcgat aggtttttct cacccatccc attttcttca cgccacttat 28740 tcatctgctt cactaatacc gtcaggttca gctcaccttc tcgggtgaaa gttatacccc 28800 catagggaaa ttctagtgtt tgcgatttca tgtttctatc tcctttttga agtattgaat 28860 gaatactaca cctagatata gtattcgtca aatacttcta gttaacttta gaactatgat 28920 taagttgttc tactaattag ggtcgcccat agccttggga tgggttttct tatagtatgc 28980 tattaacttt agcatcacag ggtttatcta tgagcttact tacatcatta ataagccacc 29040 aggtatggtt acaacgcacc gcatccggag aggtgaaaga cctggcgcca ttcatcaagc 29100 agatgcgcga tgaggttaag cggcaggtgt tgttgtttgg tgatgacagc agaacggcgg 29160 cgagacttac caccatgctg cgggaacttg agcaagcgct gaacgggatt acgtctgggt 29220 ggtatgagaa gctactggcc gatgcccgtg agctttcaga ctatgaagtt aactggaacg 29280 taaaaaccct gtcgaccaac gttaacgcta atttcgtaac accggccgcc gaacaagtat 29340 gggccgccgc aactttcgcg cctcttgaac taagcgaaaa gccagttgat tttgtttctc 29400 taatgagcgg ctggcggcaa accgaggtaa atcgccttgt tatgggcgtt aaatctgggt 29460 tcgtacaggg gatgaccaca cgtcggatag ttaagaacgt tgtcgggccc ggtggtctgg 29520 ccgatatctc cgagcgtaac gctgcaaccg ttatccggac agcgctggcg cacgtatcca 29580 acgaagcacg gcaacaggtt tacgctcaga acggcgacat catcacgaaa tacgagtggg 29640 tatctaccct tgactccaga acctcagccg tttgtcggtc gcgggattct atgcaatacg 29700 aaataggtaa aggtccgcta ccgccagcgc atcctaactg tcggtcgagc acagcaccgg 29760 taataagtcc agaattcgac ttcctggata agggtgcaaa acgggcggcc aggggtgcag 29820 atggaggtca gcaggtaagc gcggacacga gctactacga attccttaaa caacaaccgg 29880 cgtggttcca ggacgaagca cttggccccg taaggggtaa gatttttcgt aatagtggga 29940 taacaccgga agaatttcgt gtaatatcgg tagacggatt cgggcgcccg ttaactctta 30000 aagagatggc ggagctcgat aaacgtgttg ccgattatct gaaagaggaa taaagatggg 30060 ctttttcaaa gttaaagatg tgccgtcacg tcgagtagtg cagtacgcac gtgtgtctgg 30120 tgcgggtgaa ggcgttgttt atattaaaga cgaatctgtt ctcggcgagt cggtagatga 30180 aatgccgttt gcggataaga ccggactgac cacaatctca gacggcatcc tgtacgaagt 30240 accgtatctg gatggtgcgg gagatgtgta cttcgatctg caaccggcag atgtagagct 30300 gaaagacggt tctgcaaaat taaccgtagt tgtaaaaggc ggtaaagcgc cctacgattt 30360 gcaatggttt aaagatggta aagaggtaat aaacgttcct tacgttgaag gggaactaac 30420 cgttaaagac cccggagaat atttcgtcaa ggcagtagat gccgacggtg tgtcggtggt 30480 tagcaaagcg gctaaggtct cagaacctaa gtaacgaaag gccccttgac ggggccttaa 30540 ttttaccagc gattattcca gttattcctt accaatgtaa attcaggaat actttattca 30600 aaaagtagat ttctattatt ttaaatgcta cgctaactac gctttcagcg taccgcggtg 30660 cgggtcgcgt gaagcgggcg tcacaacgcc ctagcgtaac cgcgaacgca ttcggaaggg 30720 ccgagctata ttgctttcag tatttagcta agattcgccg tatgatgttt acagatatac 30780 gagattaacg cccgacttcg tcgggcttag cgtataaaga aaggggatac tatgaattgg 30840 aaagtaaccg cggtagccac ggcagccggt atcctctcct tgtggttata cggccagtac 30900 aactacagaa gcgggtgggt agaaggccgc gccaatctcg tttcacagca acaaaagaag 30960 gcacaagctg agctgacgaa gaaaacacaa cggcagcagc agaatgatac taaggccgcc 31020 tccgccgaat cggaaggcaa agaaaattcg gaggctatca cccatgaagt catcaaatat 31080 gttatccgtc ctggccgtac tgtctgcgag tttccttctg aacgggtgtc aatcaaacga 31140 cgcgccactg agaatgctaa ttccatcccc ggatatgaca ctaatgcaac cgccgtgcaa 31200 gcttattccc ccgagcagtg acgctgatga agacctggcc atcgacgtac gtaacgcgga 31260 gtgcacacgg cagttacgtc tgaaagtgtt ccggttgcag gagtatataa ggaatattct 31320 ggaatagttg ccttagtaag tggaataatt tattcttgtt acagaaacac cgggtggccc 31380 ggtgtcctaa agtccagggg acatattgac tatgaatcgt tttttacact atccactcca 31440 ggaagaagct ggagtagaag ataaatcggg tgcaggtgat gcaccaaaaa tgtacaccgc 31500 cgaagaagtg caggccttga ttgagaaaga agtggccggg ctcaaggcaa atcaggaagc 31560 gctccttagc gagaaaaagg aagctgctcg ccgggctaaa gaagccgaag aagaacggca 31620 gcgcgcccac caggaggccc tgaaggctgc tggtaaaatg gacgaatttg aaaagacgat 31680 tcgtagtcag tatgaacccg tactgaaaga gaaagaagaa cgctacgctt ctttagccgc 31740 gcgaattctt ggaagtgaac gtaaggctgt tttgggttct ttcgccggtg atttcatcac 31800 cccggaagcg gtagaaatcc ttgctccgtt cgttaagact gaatttgaag gtgaagacgt 31860 agttactaaa ttcacggatg cggatggtaa cgtagttact accgaccctg aacagttccg 31920 caaatacctg cgcgaacata aagctttttc gcatttgatt aaagcaaatg cagcttccgg 31980 tggcggggct tccgggaata aaggcggcgg ggccgcacca gcgtttaaag atatgagtga 32040 aagtgagcgg ttggctctct ataaatctaa ccctgccgaa tttgaacggc aacttaaagc 32100 cctgaggaaa taaataatgg caatcaccac tattggtaac atcgtaactg gcaatatccc 32160 tgtactggcg tcctatatga cggaagaccc ggtagagaaa accgcgtttt tcaactccgg 32220 aattttgact ccaactccgt atgctgccga gattgcccgc ggcccgtcta acatcgctaa 32280 cctgccgttc tggaaagcca tcgatacctc tatcgaacct aactattcga acgatgtgta 32340 tcaggacatc gcgaccccgc gcgctattca gaccggtgaa atgatggccc gcgttgcgta 32400 tctgaacgaa ggtttcggcc aggcagacct gactgtcgaa ctgactagcc agaatccatt 32460 gcagtccgta gcctcccgcc tggataactt ctggcagcgc caggcccagc gtcgtctgat 32520 tgctaccgcc ctcggtctgt acaacgacaa cgtatccgct actgatgcat atcacaagca 32580 gaacgacatg gttgttgacg tttctgcaac ttcaggcttc gacgcaggtg cattcatcga 32640 tgctacccag actatgggtg atgcattgat gggcaacggc ggtgaggttc tcggtgctat 32700 cgcaatgcac agcttcgtat acgcacaggc gcgtaaagct cagcttatcg acttcatccg 32760 tgacgctgag aacaacacca tgttcgccac ctaccagggc taccgtgtta tcgtcgatga 32820 cagcatgacc gtagtaggcc agggtgcgca gcgcaagttc atctccgtca tcttcggcca 32880 gggcgctatc ggctatggcg aaggcaatcc tgagatgccg ttggaatacg agcgtgaagc 32940 atctcgcgct aacggcggcg gcgttgaaac cctatggacg cgtaaaacgt ggttgctgca 33000 tccgttggtt 33010

Claims (12)

살모넬라 갈리나륨(Salmonella Gallinarum), 살모넬라 콜레라수이스(Salmonella Choleraesuis) 및 살모넬라 플로럼(Salmonella Pullorum)으로 구성된 군으로부터 선택된 1종 이상인 살모넬라균에 특이적인 사멸능을 갖는, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 SG2 (기탁번호 KFCC11677P).
1, which has a killing activity specific to Salmonella, which is at least one selected from the group consisting of Salmonella Gallinarum, Salmonella Choleraesuis and Salmonella Pullorum. Bacteriophage SG2 (Accession No. KFCC11677P).
삭제delete 삭제delete 삭제delete 삭제delete 삭제delete 제1항의 박테리오파지 SG2를 포함하는, 항생용 조성물.
An antibiotic composition comprising the bacteriophage SG2 of claim 1.
제1항의 박테리오파지 SG2를 포함하는, 사료 첨가용 조성물.
A composition for feed addition comprising the bacteriophage SG2 of claim 1.
제8항의 사료 첨가용 조성물을 포함하는, 사료.
A feed comprising the feed additive composition of claim 8.
제1항의 박테리오파지 SG2를 포함하는, 소독제.
A disinfectant comprising bacteriophage SG2 of claim 1.
제1항의 박테리오파지 SG2를 포함하는, 세척제.
A cleaning agent comprising the bacteriophage SG2 of claim 1.
제1항의 박테리오파지 SG2를 인간을 제외한 동물에 투여하는 단계를 포함하는, 살모넬라 갈리나륨(Salmonella Gallinarum), 살모넬라 콜레라수이스(Salmonella Choleraesuis) 및 살모넬라 플로럼(Salmonella Pullorum)으로 구성된 군으로부터 선택된 1종 이상인 살모넬라균에 의한 감염성 질병의 예방 또는 치료 방법.A method for producing a bacteriophage SG2 according to claim 1, which comprises administering the bacteriophage SG2 to an animal other than a human, comprising at least one selected from the group consisting of Salmonella Gallinarum, Salmonella Choleraesuis, and Salmonella Pullorum, A method for preventing or treating an infectious disease caused by Salmonella.
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KR102193477B1 (en) * 2019-06-26 2020-12-22 주식회사 옵티팜 Novel Salmonella specific bacteriophage ST20 and antibacterial composition comprising the same

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US8148131B2 (en) 2008-12-02 2012-04-03 Cj Cheiljedang Corporation Bacteriophage and antibacterial composition comprising the same

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US8148131B2 (en) 2008-12-02 2012-04-03 Cj Cheiljedang Corporation Bacteriophage and antibacterial composition comprising the same

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Title
Appl. Environ. Microbiol. 2013, vol. 79, no. 6, pp. 1956-1968.*
Salmonella phage wksl3, complete genome, GenBank: JX202565.1, 2013.03.08.*

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102193477B1 (en) * 2019-06-26 2020-12-22 주식회사 옵티팜 Novel Salmonella specific bacteriophage ST20 and antibacterial composition comprising the same

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