KR20190069901A - Novel Vibrio specific bacteriophage VP2 and antibacterial composition comprising the same - Google Patents

Novel Vibrio specific bacteriophage VP2 and antibacterial composition comprising the same Download PDF

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KR20190069901A
KR20190069901A KR1020170170253A KR20170170253A KR20190069901A KR 20190069901 A KR20190069901 A KR 20190069901A KR 1020170170253 A KR1020170170253 A KR 1020170170253A KR 20170170253 A KR20170170253 A KR 20170170253A KR 20190069901 A KR20190069901 A KR 20190069901A
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bacteriophage
vibrio
present
bacteria
feed
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김영주
이종석
한범구
김현일
박철세
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주식회사 옵티팜
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/195Antibiotics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66Microorganisms or materials therefrom
    • A61K35/76Viruses; Subviral particles; Bacteriophages
    • CCHEMISTRY; METALLURGY
    • C11ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11DDETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16Organic compounds
    • C11D3/38Products with no well-defined composition, e.g. natural products
    • C11D3/381Microorganisms
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    • C12N2795/00Bacteriophages
    • C12N2795/00011Details
    • C12N2795/00021Viruses as such, e.g. new isolates, mutants or their genomic sequences
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2795/00Bacteriophages
    • C12N2795/00011Details
    • C12N2795/00031Uses of virus other than therapeutic or vaccine, e.g. disinfectant
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2795/00Bacteriophages
    • C12N2795/00011Details
    • C12N2795/00032Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention relates to a novel vibrio-specific bacteriophage VP2, and more specifically, to an antibacterial composition comprising the vibrio-specific bacteriophage VP2. According to the present invention, the bacteriophage VP2 is very highly specific to vibrio, is infected to vibrio to grow in the vibrio, thereby having an excellent lytic activity. In addition, the bacteriophage VP2 is strong against physiochemical stimulus and is not infected to other hosts such as humans, animals, plants, except bacteria, thereby solving problems of antibiotic-resistant bacteria for abuse and misuse of antibiotics, residual antibiotics in foods and a wide host range. Accordingly, the present invention may be used in antibiotic compositions, feed additive compositions, feed, and disinfectants or detergents fields.

Description

신규한 비브리오균 특이 박테리오파지 VP2 및 이를 포함하는 항균 조성물{Novel Vibrio specific bacteriophage VP2 and antibacterial composition comprising the same}[0001] The present invention relates to novel Vibrio bacteriophage VP2 and antibacterial composition comprising the same,

본 발명은 신규한 비브리오균 특이 박테리오파지 VP2에 관한 것으로서, 더욱 상세하게는 상기 비브리오균 특이 박테리오파지 VP2를 포함하는 항균 조성물에 관한 것이다.The present invention relates to a novel Vibrio bacteria-specific bacteriophage VP2, and more particularly to an antimicrobial composition comprising the Vibrio bacterium-specific bacteriophage VP2.

장염비브리오균(Vibrio parahaemolyticus)은 살모넬라균, 황색포도상구균과 함께 식중독을 일으킬 수 있는 3대 식중독균 가운데 하나로, 해수 또는 기수에서 서식한다. 야생종은 바닷물에 널리 분포하고 있으며, 사람이나 어류에서 병원성을 나타내며, 특히 해산물을 날 것이나 덜 익힌 상태로 섭취하였을 경우, 또는 장염비브리오균에 오염된 기구를 통해 조리되었거나 오염된 식품을 섭취하였을 경우 감염이 되어 장염을 일으킬 수 있다. 장염비브리오균은 사람에서 식중독을 유발할 뿐만 아니라, 항생제 내성 비브리오균의 출현으로 인해 어패류 양식 분야의 생산성 저하를 초래한다.Enteritis Vibrio parahaemolyticus ) is one of the three food poisoning bacteria that can cause food poisoning with Salmonella and Staphylococcus aureus, and lives in seawater or in water. Wild species are widely distributed in seawater and exhibit pathogenicity in humans and fish, especially when they are consumed with raw or undercooked seafood, or when they have been cooked or contaminated with food contaminated with enteritis Vibrio infection And can cause enteritis. Not only does Vibrio parahaemolyticus cause food poisoning in humans, but also the productivity of fish and shellfish production is deteriorated due to the emergence of antibiotic resistant Vibrio bacteria.

한편, 박테리오파지(bacteriophage)는 특정 세균을 감염시켜 감염된 세균의 성장을 억제하고 저해하는 세균 특이적 바이러스를 의미한다. 박테리오파지는 박테리아에 감염(infection)한 후 박테리아 세포 내부에서 증식을 하고, 증식 후 자손 박테리오파지들이 박테리아 밖으로 나올 때 숙주인 박테리아의 세포벽을 파괴하는 방식으로 박테리아를 사멸시키는 능력을 갖고 있다. 박테리오파지의 박테리아 감염 방식은 매우 특이성이 높아서 특정 박테리아에 감염할 수 있는 박테리오파지의 종류는 일부로 한정된다. 즉, 특정 박테리오파지는 특정 범주의 박테리아에만 감염할 수 있고 이로 인하여 특정 박테리오파지는 특정 박테리아만을 사멸시키며 다른 박테리아에는 영향을 주지 않는다. 따라서 최근 세균성 질환의 대처 방안으로 박테리오파지의 활용이 크게 주목을 받고 있으며, 자연 친화적 방식의 선호로 인하여 박테리오파지에 대한 관심은 어느 때보다 높다고 할 수 있다. 기존 항생제의 대체 물질로의 개발 가능성이 부각되면서 다시 박테리오파지가 항-박테리아제로 주목을 받고 있다. 이러한 박테리오파지가 여러 세균성 질병을 대상으로 활발히 연구되고 있으며, 박테리오파지를 이용한 장염비브리오균의 제어는 안전한 수산물 생산과 장염비브리오에 의한 질병의 위험을 감소할 수 있는 방안이 될 수 있을 것으로 판단된다.On the other hand, bacteriophage refers to a bacterium-specific virus that infects a specific bacterium to inhibit and inhibit the growth of the infected bacterium. Bacteriophages have the ability to kill bacteria by infecting bacteria, multiplying inside bacterial cells, and destroying the cell walls of the host bacteria when the progeny bacteriophages come out of the bacteria after proliferation. The bacteriophage infection method of bacteriophage is highly specific, so the type of bacteriophage that can infect a specific bacterium is limited to some. That is, certain bacteriophages can infect only a specific category of bacteria, which causes certain bacteriophages to kill only certain bacteria and not other bacteria. Recently, the use of bacteriophage has attracted much attention as a countermeasure against bacterial diseases, and interest in bacteriophage is higher than ever due to preference of nature friendly method. Bacteriophage is attracting attention as an anti-bacterial agent again with the possibility of development as a substitute for existing antibiotics. These bacteriophages have been actively studied for various bacterial diseases, and control of Vibrio parahaemolyticus by bacteriophage may be a way to reduce the risk of disease caused by safe aquatic products and enteritis vibrio.

이에 본 발명자들은 박테리오파지를 기존 항생제의 대체 물질로 활용하기 위하여, 해수로부터 비브리오균 특이 박테리오파지를 분리하였고, 분리된 박테리오파지의 형태적 및 유전학적 특성 분석을 통해 장염비브리오균을 선택적으로 사멸시킬 수 있음을 확인함으로써 본 발명을 완성하게 되었다.Therefore, in order to utilize bacteriophage as a substitute for existing antibiotics, the present inventors have isolated the bacteriophage specific bacteriophage from seawater and can selectively kill the enteric bacterium by analyzing the morphological and genetic characteristics of the isolated bacteriophage The present invention has been completed.

따라서 본 발명의 목적은 비브리오균(Vibrio)에 특이적인 사멸능을 갖고, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 VP2를 제공하는 것이다.Accordingly, an object of the present invention is to provide a bacteriophage VP2 having the nucleotide sequence shown in SEQ ID NO: 1 and having an extermination activity specific to Vibrio .

본 발명의 다른 목적은 상기 박테리오파지 VP2를 포함하는 항생용 조성물을 제공하는 것이다.Another object of the present invention is to provide an antibiotic composition comprising the bacteriophage VP2.

본 발명의 또 다른 목적은 상기 박테리오파지 VP2를 포함하는 사료 첨가용 조성물을 제공하는 것이다.Yet another object of the present invention is to provide a composition for feed addition comprising the bacteriophage VP2.

본 발명의 또 다른 목적은 상기 박테리오파지 VP2를 포함하는 소독제를 제공하는 것이다.It is still another object of the present invention to provide a disinfectant containing the bacteriophage VP2.

본 발명의 또 다른 목적은 상기 박테리오파지 VP2를 포함하는 세척제를 제공하는 것이다.It is still another object of the present invention to provide a cleaning agent comprising the bacteriophage VP2.

본 발명의 또 다른 목적은 상기 박테리오파지 VP2를 인간을 제외한 개체에 투여하는 단계를 포함하는 비브리오균에 의한 감염성 질병의 예방 또는 치료 방법을 제공하는 것이다.It is still another object of the present invention to provide a method for preventing or treating infectious diseases caused by Vibrio bacteria comprising the step of administering the bacteriophage VP2 to a subject other than a human.

상기 목적을 달성하기 위하여, 본 발명은 비브리오균에 특이적인 사멸능을 갖고, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 VP2를 제공한다.In order to accomplish the above object, the present invention provides a bacteriophage VP2 having the nucleotide sequence shown in SEQ ID NO: 1 and having a killing activity specific to Vibrio germ.

또한 본 발명은 상기 박테리오파지 VP2를 포함하는 항생용 조성물을 제공한다.The present invention also provides an antibiotic composition comprising the bacteriophage VP2.

또한 본 발명은 상기 박테리오파지 VP2를 포함하는 사료 첨가용 조성물을 제공한다.The present invention also provides a composition for adding a feed comprising the bacteriophage VP2.

또한 본 발명은 상기 박테리오파지 VP2를 포함하는 소독제를 제공한다.The present invention also provides a disinfectant comprising the bacteriophage VP2.

또한 본 발명은 상기 박테리오파지 VP2를 포함하는 세척제를 제공한다.The present invention also provides a detergent comprising the bacteriophage VP2.

또한 본 발명은 상기 박테리오파지 VP2를 인간을 제외한 개체에 투여하는 단계를 포함하는 비브리오균에 의한 감염섬 질병의 예방 또는 치료방법을 제공한다.The present invention also provides a method of preventing or treating an infectious islet disease caused by Vibrio bacteria comprising the step of administering the bacteriophage VP2 to a subject other than a human.

본 발명에 따른 박테리오파지 VP2는 비브리오균에 대한 특이성이 매우 높고, 비브리오균에 감염하여 비브리오균 내에서 증식함으로써 용균 활성이 우수하며, 물리화학적 자극에 강하다는 장점을 가지고 있다. 또한 본 발명의 박테리오파지 VP2는 사람, 동물, 식물 등 세균 외의 다른 숙주에는 감염되지 않으므로, 항생제의 오남용으로 인한 항생제 내성균의 문제점, 식품 내의 항생제 잔류문제, 광범위한 숙주범위의 문제점을 해결할 수 있는바, 항생용 조성물, 사료 첨가용 조성물, 사료, 소독제 또는 세척제 분야에서 다양하게 활용될 수 있다.The bacteriophage VP2 according to the present invention has an extremely high specificity to Vibrio sp., And has an advantage of being strong against physicochemical stimuli because of its excellent lytic activity by proliferation in Vibrio spp. Since the bacteriophage VP2 of the present invention is not infected with any host other than bacteria such as human, animal, and plant, problems of antibiotic resistant bacteria caused by misuse of antibiotics, antibiotic residues in foods, and problems in a wide host range can be solved. A composition for feed addition, a feed, a disinfectant or a detergent.

도 1은 본 발명에 따른 박테리오파지 VP2의 장염비브리오균에 대한 용균반 형성 활성을 확인한 결과를 나타낸 도이다.
도 2는 본 발명에 따른 박테리오파지 VP2의 전체 유전자를 분석한 결과를 나타낸 도이다.
도 3은 전자 현미경을 통해 본 발명에 따른 박테리오파지 VP2의 형태학적 특성을 확인한 결과를 나타낸 도이다.
도 4는 SDS-PAGE를 통해 본 발명에 따른 박테리오파지 VP2의 단백질 패턴을 확인한 결과를 나타낸 도이다.
도 5는 본 발명에 따른 박테리오파지 VP2의 pH 안정성을 확인한 결과를 나타낸 도이다.
도 6은 본 발명에 따른 박테리오파지 VP2의 열 안정성을 확인한 결과를 나타낸 도이다.
도 7은 본 발명에 따른 박테리오파지 VP2의 염 안정성을 확인한 결과를 나타낸 도이다.
FIG. 1 is a graph showing the results of confirming the activity of the bacteriophage VP2 according to the present invention for the preparation of a microorganism for Vibrio parahaemolyticus.
FIG. 2 is a diagram showing the result of analyzing the entire gene of bacteriophage VP2 according to the present invention. FIG.
FIG. 3 is a graph showing the morphological characteristics of bacteriophage VP2 according to the present invention through an electron microscope.
FIG. 4 is a diagram showing the result of confirming the protein pattern of bacteriophage VP2 according to the present invention through SDS-PAGE. FIG.
5 is a graph showing the result of confirming the pH stability of bacteriophage VP2 according to the present invention.
6 is a graph showing the results of confirming the thermal stability of the bacteriophage VP2 according to the present invention.
7 is a graph showing the results of confirming the salt stability of bacteriophage VP2 according to the present invention.

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

본 발명의 양태에 따르면, 본 발명은 비브리오균(Vibrio)에 특이적인 사멸능을 갖고, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 VP2를 제공한다.According to an aspect of the present invention, the present invention provides a bacteriophage VP2 having the nucleotide sequence shown in SEQ ID NO: 1 and having a killing activity specific to Vibrio .

본 발명에 있어서, “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, " 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 발명의 박테리오파지 VP2는 27 내지 32 nm의 크기로, 정이십면체의 머리를 가지며 짧은 꼬리가 있는 형태인 포도비리대(Podoviridae)에 속함을 확인하였다.The bacteriophage VP2 of the present invention has a size of 27 to 32 nm and belongs to Podoviridae, which has a head of regular twenty-sided and a short tail.

본 발명의 박테리오파지 VP2의 단백질 패턴 분석 결과 72, 38, 35 및 32 kDa 크기의 단백질을 포함한다는 것을 확인하였으며, 상기 박테리오파지 VP2의 전체 염기서열 분석 결과, 전체 유전자 크기는 43,069 bp이고, GC 함량은 49%, 전체 ORFs의 수는 46개임을 확인하였다. 또한 상기 박테리오파지 VP2는 열, pH 및 염에 대한 안정성이 우수한 특성을 가지고 있다.As a result of analyzing the protein pattern of the bacteriophage VP2 of the present invention, it was confirmed that it contained 72, 38, 35 and 32 kDa proteins. As a result of the whole base sequence analysis of bacteriophage VP2, the total gene size was 43,069 bp and the GC content was 49 %, And the total number of ORFs was 46. In addition, the bacteriophage VP2 has excellent stability against heat, pH and salt.

본 발명의 박테리오파지 VP2는 서열번호 1로 표시되는 염기서열을 전체유전자의 전체 또는 일부로서 포함하는 것이 바람직하다. 또한 상기 염기서열의 변이체가 본 발명의 범위 내에 포함된다. 구체적으로, 상기 유전자는 서열번호 1의 염기서열과 70% 이상, 더욱 바람직하게는 80% 이상, 더 더욱 바람직하게는 90% 이상, 가장 바람직하게는 95% 이상의 서열 상동성을 가지는 것으로, 서열번호 1로 표시되는 염기서열과 실질적으로 동질의 생리활성을 나타내는 서열을 의미한다. 폴리뉴클레오티드에 대한 "서열 상동성의 %"는 두 개의 최적으로 배열된 서열과 비교 영역을 비교함으로써 확인되며, 비교 영역에서의 폴리뉴클레오티드 서열의 일부는 두 서열의 최적 배열에 대한 참고 서열(추가 또는 삭제를 포함하지 않음)에 비해 추가 또는 삭제(즉, 갭)를 포함할 수 있다.The bacteriophage VP2 of the present invention preferably contains the nucleotide sequence shown in SEQ ID NO: 1 as a whole or a part of the entire gene. Variants of the above base sequences are also included within the scope of the present invention. Specifically, the gene has a sequence homology of at least 70%, more preferably at least 80%, even more preferably at least 90%, most preferably at least 95% with the nucleotide sequence of SEQ ID NO: 1, 1 < / RTI > of the nucleotide sequence shown in SEQ ID NO: 1. "% Of sequence homology to polynucleotides" is ascertained by comparing the comparison region with two optimally aligned sequences, and a portion of the polynucleotide sequence in the comparison region is the reference sequence for the optimal alignment of the two sequences (I. E., A gap) relative to the < / RTI >

본 발명의 바람직한 구체예에 따르면, 본 발명의 박테리오파지 VP2는 해수로부터 시료를 채취하여 분리한 박테리오파지로서, 박테리오파지 VP2로 명명하고 2017년 9월 13일에 한국미생물보존센터에 기탁하여 수탁번호 KFCC11727P를 부여받았다.According to a preferred embodiment of the present invention, the bacteriophage VP2 of the present invention is a bacteriophage isolated from bacteriophage VP2 by collecting a sample from seawater and named as bacteriophage VP2, deposited on September 13, 2017 with the Korean Microorganism Conservation Center and assigned accession number KFCC11727P received.

본 발명의 바람직한 구체예에 따르면, 본 발명의 박테리오파지 VP2는 비브리오균(Vibrio)에 우수한 용균활성을 나타내며, 상기 비브리오균의 예로는 콜레라균(V. cholerae), 장염비브리오균(V. parahaemolyticus), 비브리오 앵길라룸(V. anguillarum), 비브리오 피셰리(V. fischeri), 비브리오 코스티콜라(V. costicola) 및 비브리오 미미쿠스(V. mimicus) 등이 있다.According to a preferred embodiment of the invention, the bacteriophage VP2 of the present invention exhibit excellent activity in lysed parahaemolyticus (Vibrio), examples of the Vibrio bacteria is Vibrio cholerae (V. cholerae), Vibrio parahaemolyticus (V. parahaemolyticus), V. anguillarum , V. fischeri , V. costicola and V. mimicus , and the like.

본 발명의 다른 양태에 따르면, 본 발명은 상기 박테리오파지 VP2를 포함하는 항생용 조성물을 제공한다.According to another aspect of the present invention, there is provided an antibiotic composition comprising the bacteriophage VP2.

본 발명에 있어서, “항생용 조성물”은 약제 형태로 동물에게 제공되어 균을 사멸시킬 수 있는 제제를 의미하며, 방부제, 살균제, 항생제 및 항균제를 총칭하는 것이다.In the present invention, " antibiotic composition " means a preparation which can be given to an animal in the form of a drug to kill bacteria, and includes a preservative, a bactericide, an antibiotic and an antibacterial agent.

본 발명의 박테리오파지 VP2는 기존 항생제에 비하여 비브리오균에 대한 특이성이 매우 높으므로, 유익균은 죽이지 않고, 특정 병원균만 사멸시킬 수 있으며, 약물 내성 및 저항성을 유도하지 않아, 기존의 항생물질에 비하여 제품수명(life cycling)이 긴 신규 항생제로서 이용될 수 있다.Since the bacteriophage VP2 of the present invention is highly specific for Vibrio than the existing antibiotics, it does not kill useful bacteria and can kill only specific pathogens, and does not induce drug resistance and resistance, (life cycling) can be used as a new antibiotic.

본 발명의 또 다른 양태에 따르면, 본 발명은 상기 박테리오파지 VP2를 포함하는 사료 첨가용 조성물을 제공한다.According to still another aspect of the present invention, there is provided a feed composition comprising the bacteriophage VP2.

축산, 수산업, 반려동물 산업에서 사용되는 사료 첨가용 항생제는 질병의 예방 목적으로 사용되고 있는데, 예방 목적의 항생제 투여는 항생제 내성균 발생 가능성을 높이고, 동물에 잔류하는 항생제가 사람에게 전달될 수 있는 문제가 있다. 항생제가 인체에 흡수되면 항생제 내성을 유발해 질병의 확산을 초래할 수 있다. 또한 사료에 첨가되는 항생제의 종류가 많을 경우, 이는 다제내성균 발생 확률을 높인다는 문제가 있다. 따라서 종래 항생제 사용으로 발생한 문제를 해결할 새로운 사료 첨가용 항생물질로서, 본 발명의 박테리오파지 VP2를 이용할 수 있다.The use of antibiotics for feed addiction in animal husbandry, fisheries and companion animal industries is used for the prevention of diseases. The administration of antibiotics for prevention increases the possibility of the occurrence of antibiotic resistant bacteria and the problem that antibiotics remaining in animals can be transmitted to humans have. If antibiotics are absorbed into the body, they can cause antibiotic resistance and spread the disease. In addition, when there are many kinds of antibiotics added to the feed, this raises the probability of occurrence of multidrug resistant bacteria. Therefore, the bacteriophage VP2 of the present invention can be used as a new feed additive antibiotic substance that solves the problems caused by the use of antibiotics in the past.

본 발명의 사료 첨가용 조성물에 포함된 박테리오파지 VP2는 액상 또는 건조 상태일 수 있으며, 바람직하게는 건조된 분말형태이다. 박테리오파지 VP2의 건조 방법은 통풍건조, 자연건조, 분무건조 및 동결건조가 가능하며, 이에 제한되는 것은 아니다.The bacteriophage VP2 contained in the feed composition of the present invention may be in a liquid or dry state, preferably in the form of a dried powder. Drying methods of bacteriophage VP2 can be, but not limited to, air drying, natural drying, spray drying and freeze drying.

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), 소의 위와 같은 혐기적 조건에서 생리적 활성 및 유기물 분해능이 있는 락토바실러스 균주(Lactobacillus sp.), 동물의 체중을 증가시키며 우유의 산유량을 늘리고 사료의 소화 흡수율을 높이는 효과를 보여주는 아스퍼질러스 오리자에(Aspergillus oryzae) 및 사카로미세스 세레비지에(Saccharomyces cerevisiae)로 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 Bacillus subtilis capable of producing proteolytic enzymes, lipolytic enzymes and glycosyltransferases, Lactobacillus strains having physiological activity and organic degradation ability under anaerobic conditions such as those above, sp.), increases the weight of the animal (Aspergillus in Aspergillus duck, showing the effect of increasing the milk production of the milk to increase the absorption of digested food party oryzae , and Saccharomyces cerevisiae .

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.

본 발명의 일 구체예에서는, 상기 사료 첨가용 조성물을 포함하는 사료를 제공한다. 상기 사료는 박테리오파지 VP2를 사료 첨가제 형태로 제조하여 사료에 혼합하거나, 사료 제조 시 박테리오파지 VP2를 직접 첨가하여 제조할 수 있다. 본 발명의 사료 내 박테리오파지 VP2는 액상 또는 건조 상태일 수 있으며, 바람직하게는 건조된 분말형태이다. 상기 분말 형태의 박테리오파지 VP2는 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 one embodiment of the present invention, there is provided a feed comprising the feed composition. The feed can be prepared by preparing bacteriophage VP2 in the form of a feed additive and mixing it with feed, or by directly adding bacteriophage VP2 in the production of feed. The bacteriophage VP2 in the feed of the present invention may be in a liquid or dry state, preferably in the form of a dried powder. The bacteriophage VP2 in powder form may be mixed at 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 feeds may further include conventional additives that can enhance the preservability of the feed.

본 발명이 박테리오파지 VP2를 포함하는 사료는 식물성 사료 및 동물성사료인 것이 바람직하고, 구체적으로 상기 식물성 사료는 곡물류, 근과류, 식품가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류 및 곡물부산물류 등이 있고, 상기 동물성 사료는 단백질류, 무기물류, 유지류, 광물성류, 단세포 단백질, 동물성 플랑크톤류 및 남은 음식물 등이 있으며, 이에 제한되는 것은 아니다.The feed containing the bacteriophage VP2 of the present invention is preferably a vegetable feed and an animal feed. Specifically, the vegetable feed is selected from the group consisting of grains, muscle roots, food processing Busan logistics, algae, fibrous materials, , Pulses and grain by-products, and the animal feeds include, but are not limited to, proteins, inorganic materials, oils, minerals, single cell proteins, zooplankton and residual foods.

본 발명의 또 다른 양태에 따르면, 본 발명은 상기 박테리오파지 VP2를 포함하는 소독제를 제공한다.According to still another aspect of the present invention, there is provided a disinfectant comprising the bacteriophage VP2.

본 발명의 박테리오파지 VP2를 포함하는 소독제는 비브리오균에 탁월한 용균활성을 나타내므로, 병원감염을 막기 위한 병원 및 보건용의 소독제로 유용하게 사용될 수 있다. 또한 상기 소독제는 일반 생활 소독제, 식품, 조리 장소·설비의 소독제, 양계장 및 축사 등의 건물, 축체, 음수, 깔짚, 난좌, 운반차량, 식기 등의 각종 생육 용품의 소독제로 사용될 수 있다.The disinfectant containing the bacteriophage VP2 of the present invention exhibits excellent lytic activity in Vibrio germs, and thus can be effectively used as a disinfectant for hospitals and healthcare to prevent hospital infection. The disinfectant may be used as a disinfectant for various kinds of growth products such as general household disinfectants, foods, disinfectants for cooking places and facilities, buildings such as poultry houses and barns, shavings, water, litter, stairs, transportation vehicles and tableware.

본 발명의 또 다른 양태에 따르면, 본 발명은 상기 박테리오파지 VP2를 포함하는 세척제를 제공한다.According to another aspect of the present invention, the present invention provides a detergent comprising the bacteriophage VP2.

본 발명의 박테리오파지 VP2는 비브리오균에 특이적 사멸능을 가지므로, 상기 비브리오균에 노출되었거나 노출될 가능성이 있는 동물의 피부 표면 또는 신체 각 부위 등을 세척하는 용도로 사용될 수 있다.Since the bacteriophage VP2 of the present invention has a specific killing ability for Vibrio germs, it can be used for washing the skin surface or body parts of an animal which is exposed to the Vibrio germ or is likely to be exposed.

본 발명의 또 다른 양태에 따르면, 본 발명은 상기 박테리오파지 VP2를 인간을 제외한 개체에 투여하는 단계를 포함하는 비브리오균에 의한 감염성 질병의 예방 또는 치료방법을 제공한다.According to another aspect of the present invention, the present invention provides a method for preventing or treating an infectious disease caused by Vibrio bacteria, comprising the step of administering the bacteriophage VP2 to a subject other than a human.

본 발명에 있어서, “개체”는 인간을 포함한 동물, 바람직하게는 인간, 가축류 또는 반려동물을 말하며, 가장 바람직하게는 인간, 개, 고양이, 말 또는 새를 말하나, 이에 제한되는 것은 아니다.In the present invention, " an individual " refers to an animal including a human, preferably a human, a livestock or companion animal, and most preferably a human, a dog, a cat, a horse or a bird.

상기 "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" refers to useful animals that are refined and improved by humans to co-live with humans, and is a concept including all birds, fishes, insects, and mammals, and the livestock includes pigs, cows, Horse, duck, and dog, but is not limited thereto.

본 발명의 비브리오균에 의한 감염성 질병의 예방 또는 치료방법은 박테리오파지 VP2를 약학적 조성물의 형태로 개체에 직접 투여하거나, 개체의 사료 또는 음용수에 혼합하여 이를 섭식시키는 방법을 통해 수행될 수 있다.The method for preventing or treating infectious diseases caused by Vibrio bacteria according to the present invention can be carried out by directly administering bacteriophage VP2 to a subject in the form of a pharmaceutical composition or mixing the subject with feed or drinking water of an individual and feeding them.

본 발명의 비브리오균에 의한 감염성 질병의 예방 또는 치료방법에서 상기 박테리오파지 VP2의 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 for preventing or treating infectious diseases caused by Vibrio bacteria according to the present invention, the administration route of the bacteriophage VP2 may be administered through various oral or parenteral route as long as it can reach the target tissues. Specifically, Can be administered in a conventional manner via topical, intravenous, intraperitoneal, intramuscular, intraarterial, transdermal, intranasal, inhalation and the like.

본 발명의 방법에 투여되는 상기 박테리오파지 VP2의 적합한 총 1 일 사용량은 올바른 의학적 판단범위 내에서 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 VP2 administered in the methods of the present invention can be determined 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 발명의 일 구체예에서, 비브리오균에 의한 질병은 장염비브리오성 식중독, 콜레라, 비브리오 패혈증 및 어류 비브리오 감염증인 것이 바람직하나, 이에 제한되지 않는다.In one embodiment of the present invention, the disease caused by Vibrio sp. Is preferably, but not limited to, Vibrio parahaemolyticus, cholera, Vibrio sepsis and fish Vibrio infection.

이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these 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

장염비브리오균(Vibrio parahaemolyticus)에 특이적인 박테리오파지를 스크리닝 및 분리하기 위해서 반점 용균반 분석을 수행하였다. 시료는 박테리오파지가 존재할 것으로 예상되는 전남 고흥 지역의 해수로부터 시료를 채취하였으며, 박테리오파지 분리에 사용된 장염비브리오균은 한국미생물보존센터에서 분양받은 장염비브리오균 KCCM11965를 사용하였다.Enteritis Vibrio To investigate and isolate bacteriophage specific to parahaemolyticus , spot analysis was performed. Samples were collected from seawater in Goheung area, Jeonnam province where bacteriophages are expected to be present. Vibrio parahaemolyticus, which was used for bacteriophage isolation, was obtained from KCM11965.

구체적으로, 채취한 시료를 균질화 한 후 장염비브리오균과 함께 35℃에서 하룻밤 동안 진탕배양 하였다. 상기 배양액을 4,000 rpm으로 20분간 원심분리한 후, 상등액을 공극(pore) 크기가 0.2 μm인 필터를 이용하여 여과하여 시료 여과액을 얻었다. Specifically, the collected samples were homogenized and incubated with the Vibrio parahaemolyticus at 35 ° C overnight with shaking. The culture was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was filtered using a filter having a pore size of 0.2 μm to obtain a sample filtrate.

전배양된 장염비브리오균 150 μL을 취해서 멸균된 튜브에 넣고, 3 mL의 2.5% 염화나트륨이 첨가된 트립틱 소이 탑 아가[(트립톤 17.0 g, 소이톤 3.0 g, 포도당 2.5 g, 염화나트륨 5.0 g, 인산수소칼륨 2.5 g) + 0.6% 아가]를 첨가하여 잘 혼합한 후 미리 만들어 둔 2.5% 염화나트륨이 첨가된 TSB 아가 플레이트에 중층하였다. 실온에서 30분간 방치한 후 준비된 샘플 여과액을 각각 10 uL씩 점적한 후 실온에서 30분 동안 방치하였고, 이를 35℃에서 하룻밤 동안 배양한 후 박테리오파지의 유무를 확인하였다.150 μL of the pre-cultured enterovirus Vibrio was taken in a sterilized tube, and 3 mL of 2.5% sodium chloride-added tryptic soytopyridine [(tryptone 17.0 g, soytone 3.0 g, glucose 2.5 g, sodium chloride 5.0 g, 2.5 g of potassium hydrogenphosphate + 0.6% of agar] was added thereto, and the mixture was layered on a TSB agar plate supplemented with 2.5% sodium chloride previously prepared. After incubation at room temperature for 30 minutes, 10 μL of the prepared sample filtrate was added to each well, and the mixture was allowed to stand at room temperature for 30 minutes. After incubation at 35 ° C. overnight, bacteriophage was observed.

박테리오파지의 존재가 확인된 샘플에서 박테리오파지를 분리하기 위해서 소프트 아가 오버레이법을 이용하였다. 박테리오파지가 있다고 확인된 샘플을 식염수로 적절히 희석하여 각 희석단의 샘플 희석액 100 uL을 대상 세균(장염비브리오균) 배양액 150 uL와 혼합한 후 TSA 탑 아가(0.6% 아가 + TSB + 2.5% 염화나트륨) 3 ml 첨가하여 잘 혼합한 뒤, 미리 만들어 둔 TSA 아가 플레이트(2% 아가 + TSB + 2.5% 염화나트륨)에 중층하여 방치한 후 35℃에서 24시간 동안 배양하였다. 배양한 각 플레이트에서 형성된 용균반을 확인하였고, 확인된 용균반 중 하나를 선택하여 400 uL의 SM 완충액(NaCl 5.8 g/L; MgSO47H2O 2 g/L; 1 M Tris-Cl (pH7.5) 50 mL/L)에 현탁하고, 실온에서 4시간 동안 방치하였다. 상기 박테리오파지 반응액 100 uL를 채취하여 2.5%의 염화나트륨이 첨가된 트립틱 소이 탑 아가 (0.6% 아가 농도)와 장염비브리오균 배양액 500 uL와 혼합한 뒤 150 mm 지름의 TSB 플레이트(Tryptic soy agar + 2.5% NaCl)에 분주하여 중층하였고, 완전히 용균 될 때까지 35 ℃에서 배양하였다. 배양이 종료된 후, 상기 TSB 플레이트에 SM 완충액 15 mL을 가하고 실온에서 4시간동안 천천히 교반하여 박테리오파지 용액을 회수하였다. 회수된 박테리오파지 용액을 4,000 rpm으로 20분간 원심분리하여 상등액만 취했고, 0.2 μm 필터로 여과하여 최종 박테리오파지 용액을 회수하였다. 이렇게 얻어진 최종 박테리오파지의 용균반을 확인한 결과를 도 1에 나타내었다.The soft agar overlay method was used to isolate the bacteriophage from the sample in which the presence of the bacteriophage was identified. The samples were confirmed to have bacteriophage and diluted appropriately with saline. 100 μL of sample dilution at each dilution was mixed with 150 μL of the bacterial culture medium (0.6% agar + TSB + 2.5% sodium chloride) ml, well mixed, and then allowed to stand in a TSA agar plate (2% agar + TSB + 2.5% sodium chloride) which had been previously prepared, followed by incubation at 35 ° C for 24 hours. One of the identified wells was selected, and 400 μL of SM buffer (NaCl 5.8 g / L; MgSO 4 7H 2 O 2 g / L; 1 M Tris-Cl (pH 7 5) 50 mL / L), and left at room temperature for 4 hours. 100 .mu.L of the bacteriophage reaction solution was collected and mixed with 500 .mu.L of a culture medium containing 0.5% of sodium chloride and 0.6% agar dilution of the culture medium. Then, a TSB plate (Tryptic soy agar + 2.5 % NaCl) and incubated at 35 ° C until completely lysed. After completion of the incubation, 15 mL of SM buffer was added to the TSB plate, and the bacteriophage solution was recovered by slowly stirring at room temperature for 4 hours. The recovered bacteriophage solution was centrifuged at 4,000 rpm for 20 minutes to collect only the supernatant, and the final bacteriophage solution was recovered by filtration through a 0.2 μm filter. FIG. 1 shows the result of checking the thus obtained final bacteria bacterium.

도 1에 나타낸 바와 같이, 5 내지 7 mm의 크기를 가지는 용균반을 형성하는 박테리오파지를 확인하였고, 상기 박테리오파지는 장염비브리오균에 특이적인 용균능을 갖는 박테리오파지임을 확인하였다.As shown in FIG. 1, a bacteriophage forming a fungus having a size of 5 to 7 mm was confirmed, and it was confirmed that the bacteriophage is a bacteriophage having a specific ability to enter the enteritis Vibrio sp.

1-2. 박테리오파지의 대량 배양 및 정제1-2. Bulk culture and purification of bacteriophages

상기 실시예 1-1에서 확인한 장염비브리오균 특이적 박테리오파지를 대량 배양하기 위하여, 실시예 1-1에서 회수한 최종 박테리오파지 배양액을 1.0x107 PFU/mL로 희석한 후 전배양된 장염비브리오균 KCCM11965 1.0x109 CFU/mL과 혼합하여 MOI(Multiplicity of infection)가 0.01이 되도록 35℃에서 120 rpm으로 9시간 동안 배양하였다. 배양이 종료된 후 배양액 전량을 6,000 rpm으로 30분간 원심분리하여 상등액을 수득하였고, 이를 0.2 μm 필터로 여과하여 최종 박테리오파지 배양액을 회수하였다. 분리된 박테리오파지를 "박테리오파지 VP2(Bacteriophage VP2)"로 명명하고, 2017년 9월 13일 한국미생물보존센터 (Korean Culture Center of Microorganisms, 서울, 한국)에 기탁하여 기탁번호 KFCC11727P를 부여받았다.In order to mass-culture the V. vulnificus-specific bacteriophage identified in Example 1-1, the final bacteriophage culture recovered in Example 1-1 was diluted to 1.0 x 10 < 7 > PFU / mL and then the pre-cultured Vibrio parahaemolyticus KCCM11965 1.0 x 10 < 9 > CFU / mL and cultured 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 VP2" and deposited with the Korean Culture Center of Microorganisms (Seoul, Korea) on September 13, 2017 and received the deposit number KFCC11727P.

실시예Example 2. 박테리오파지 VP2의 전체 유전자 분석 2. Whole gene analysis of bacteriophage VP2

박테리오파지 VP2의 전체 유전자를 분석하기 위해, 박테리오파지 VP2 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 3 M 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, pH 8.0)에 용해시킨 후 DNA 농도를 측정하였다. 전체 유전자 분석을 위해 PacBioRSII(마크로젠, 한국)를 이용하여 유전자 시퀀싱을 시행하였다. 시퀀싱 결과로 얻은 박테리오파지 VP2의 유전자 맵은 도 2에 나타내었다.To analyze the entire gene of bacteriophage VP2, 20 mM EDTA, 50 μg / mL proteinase K and 0.5% (w / v) SDS were added to the culture medium of bacteriophage VP2 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 3 M sodium acetate was added to the supernatant 10% (v / v). The mixture was mixed with 2 volumes of cold 95% ethanol, and the mixture was allowed to stand at 20 占 폚 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. Genetic sequencing was performed using PacBioRSII (Macrogen, Korea) for total gene analysis. The gene map of bacteriophage VP2 obtained as a result of sequencing is shown in Fig.

도 2에 나타낸 바와 같이, 박테리오파지 VP2의 전체 유전자의 크기는 43,069 bp이며, GC 함량은 49%, 전체 ORFs의 수는 46개임을 확인하였다. 분석한 박테리오파지 VP2의 전체 유전자 서열을 서열번호 1로 나타내었다. As shown in FIG. 2, it was confirmed that the total gene of bacteriophage VP2 was 43,069 bp, the GC content was 49%, and the total number of ORFs was 46. The entire gene sequence of the bacteriophage VP2 analyzed is shown in SEQ ID NO: 1.

실시예Example 3. 박테리오파지 VP2의 형태학적 특성 3. Morphological characteristics of bacteriophage VP2

박테리오파지 VP2의 형태학적 특성을 알아보기 위해, 박테리오파지 VP2의 순수배양액을 0.01% 젤라틴 용액에 희석하여 2.5% 글루타르알데하이드(glutaraldehyde) 용액으로 고정하였다. 이를 탄소-코팅 운모 플레이트(carbon-coated mica plate(ca. 2.5 x 2.5 mm))에 적가한 후, 10분간 적응시킨 후 멸균증류수로 세척하였다. 이를 구리 격자(copper grid)에 끼워 2% 우라닐 아세테이트(Uranyl acetate)에서 30 내지 60초 동안 염색하고 건조시킨 후, 투과전자현미경(Tecnai G2 Spirit Twin, Bio-Transmission electron microscope, 120 kV, 120,000~400,000)으로 검경하였으며, 그 결과를 도 3에 나타내었다.To examine the morphological characteristics of bacteriophage VP2, the pure culture medium of bacteriophage VP2 was diluted in 0.01% gelatin solution and fixed with 2.5% glutaraldehyde solution. This was applied to a carbon-coated mica plate (ca. 2.5 x 2.5 mm), followed by a 10-minute adaptation and washing with sterile distilled water. The resultant was stained with 2% Uranyl acetate for 30 to 60 seconds, and dried using a transmission electron microscope (Tecnai G2 Spirit Twin, Bio-Transmission electron microscope, 120 kV, 400,000), and the results are shown in FIG.

도 3에 나타낸 바와 같이, 박테리오파지 VP2는 27 내지 32 nm의 크기로, 정이십면체의 머리를 가지며 짧은 꼬리가 있는 형태인 포도비리대(Podoviridae)에 속함을 확인하였다.As shown in Fig. 3, bacteriophage VP2 was 27 to 32 nm in size, and it belongs to Podoviridae, which has a short twisted head with a regular dodecahedron.

실시예Example 4. 박테리오파지 VP2의 단백질 패턴 분석 4. Protein pattern analysis of bacteriophage VP2

박테리오파지 VP2의 단백질 패턴을 분석하기 위해, 1.0x1010 PFU/mL로 준비된 박테리오파지 VP2 40 μL를 10 μL의 5X 로딩 완충액과 혼합하고, 10분간 끓인 다음 12.5% SDS-PAGE를 수행하였다. SDS-PAGE 수행 후 단백질 전기영동 결과를 도 4에 나타내었다.To analyze the protein pattern of bacteriophage VP2, 40 μL of bacteriophage VP2 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 4에 나타낸 바와 같이, 박테리오파지 VP2는 72, 38, 35 및 32 kDa 크기의 단백질을 가지는 것을 확인하였다.As shown in FIG. 4, it was confirmed that bacteriophage VP2 has proteins of 72, 38, 35 and 32 kDa in size.

실시예Example 5. 박테리오파지 VP2의 다양한 세균에 대한 감염성 조사 5. Investigation of infectivity of bacteriophage VP2 against various bacteria

박테리오파지 VP2가 장염비브리오균 외의 비브리오 속 세균 및 다른 종의 어류 질병 원인 세균들에 대해 감염성을 나타내는지 확인하기 위하여, 한국미생물보존센터와 한국생명공학연구원 미생물자원센터로부터 분양받은 균주를 대상으로 소프트 아가 오버레이법을 통해 감염성을 확인하였다. 박테리오파지 VP2의 감염성 확인 결과는 표 1에 나타내었다.In order to determine whether bacteriophage VP2 is infectious to bacteria of the genus Vibrio other than Vibrio parahaemolyticus and other species of fish diseases, the strains purchased from the Korean Microorganism Conservation Center and the Korea Research Institute of Bioscience and Biotechnology, The infectivity was confirmed by overlay method. The results of the infectivity test of bacteriophage VP2 are shown in Table 1.

Figure pat00001
Figure pat00001

표 1에 나타낸 바와 같이, 박테리오파지 VP2는 장염비브리오균에 용균활성을 나타내고, 장염비브리오균 외의 비브리오 속 세균 및 다른 종의 어류 질병 원인 세균에 대해서는 용균활성을 나타내지 않음을 확인하였다. 상기 감염성 확인 결과는 박테리오파지 VP2가 장염비브리오균에 특이적으로 용균활성을 나타낸다는 것을 의미한다.As shown in Table 1, it was confirmed that bacteriophage VP2 exhibited lytic activity in enteritis Vibrio bacteria, and did not exhibit lytic activity against bacteria of the genus Vibrio other than Vibrio parahaemolyticus and other species of fish diseases. The result of the infectivity confirmation means that bacteriophage VP2 shows lytic activity specifically against Vibrio parahaemolyticus.

실시예Example 6. 박테리오파지 VP2의 pH 안정성 조사 6. pH stability of bacteriophage VP2

박테리오파지 VP2의 pH 안정성을 확인하기 위하여, 다양한 pH 범위(pH2, 3, 4, 5, 6, 7, 8, 9, 10, 11)에서 박테리오파지의 수를 측정하였다. 소듐 아세테이트 완충액, 소듐 포스페이트 완충액, Tris-HCl을 이용하여 각각의 pH에 맞게 조제한 후 990 μL씩 분주하여 준비하였다. 1.0x108 PFU/mL로 준비한 박테리오파지 VP2 배양액 10 μL을 상기 용액에 첨가하여 2시간 동안 상온에서 방치하였다. 방치 후 각 반응액을 단계희석하여 소프트 아가 오버레이법에 의한 타이터(titer) 측정을 수행하였고, 그 결과를 도 5에 나타내었다.In order to confirm the pH stability of bacteriophage VP2, the number of bacteriophages was measured in various pH ranges (pH 2, 3, 4, 5, 6, 7, 8, 9, 10, 11). Sodium acetate buffer, sodium phosphate buffer, and Tris-HCl were added to each pH, and 990 μL aliquots were prepared. 10 μL of the bacteriophage VP2 culture prepared to 1.0 × 10 8 PFU / mL was added to the above solution and left at room temperature for 2 hours. After allowing to stand, each reaction solution was diluted stepwise and titer measurement was performed by soft agar overlay method. The results are shown in FIG.

도 5에 나타낸 바와 같이, pH3 내지 10의 범위에서는 박테리오파지 VP2가 2시간 동안 안정한 것을 확인하였다. 따라서 본 발명의 박테리오파지 VP2는 pH3 내지 10의 범위에서 안정함을 확인하였으므로, 박테리오파지 VP2는 pH 안정성이 우수한 박테리오파지임을 알 수 있다.As shown in Fig. 5, it was confirmed that bacteriophage VP2 was stable for 2 hours in the range of pH 3 to 10. Therefore, it was confirmed that the bacteriophage VP2 of the present invention is stable in the range of pH 3 to 10, so that bacteriophage VP2 is a bacteriophage having excellent pH stability.

실시예Example 7. 박테리오파지 VP2의 열 안정성 조사 7. Investigation of thermal stability of bacteriophage VP2

박테리오파지 VP2의 열 안정성을 확인하기 위하여, 다양한 온도 범위(40℃, 50℃, 60℃, 70℃)에서 박테리오파지의 수를 측정하였다. 박테리오파지 VP2 배양액 (1.0x108 PFU/mL) 5 mL을 멸균된 튜브에 분주한 후 2시간 동안 각 온도에서 방치하였다. 방치 후 각 반응액을 단계 희석하여 소프트 아가 오버레이법에 의한 타이터 측정을 수행한 결과를 도 6에 나타내었다.In order to confirm the thermal stability of bacteriophage VP2, the number of bacteriophages was measured in various temperature ranges (40 ° C, 50 ° C, 60 ° C and 70 ° C). 5 mL of bacteriophage VP2 culture (1.0 x 10 8 PFU / mL) was dispensed into sterile tubes 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 6에 나타낸 바와 같이, 박테리오파지 VP2는 40 내지 50℃에서 10분 동안 안정한 것을 확인하였고, 특히 40℃에서는 60분 후에도 안정하였다. 따라서 본 발명의 박테리오파지 VP2는 내열성이 우수한 박테리오파지임을 알 수 있다.As shown in Fig. 6, the bacteriophage VP2 was found to be stable at 40 to 50 DEG C for 10 minutes, and particularly stable at 40 DEG C after 60 minutes. Therefore, the bacteriophage VP2 of the present invention is a bacteriophage having excellent heat resistance.

실시예Example 8. 박테리오파지 VP2의 염에 대한 안정성 조사 8. Investigation of the stability of bacteriophage VP2 against salt

박테리오파지 VP2의 염에 대한 안정성을 확인하기 위하여, 다양한 염 농도 범위(NaCl 3%, 2%, 1%)에서 시험하였다. 염화나트륨을 이용하여 각각의 농도에 맞게 염 용액을 조제한 후 990 μL로 준비하였다. 농도가 1.0x108 PFU/mL인 박테리오파지 VP2 배양액 10 μL를 상기 염 용액에 첨가하여 24시간 동안 상온에서 방치하였다. 방치 후 각 반응액을 단계희석하여 소프트 아가 오버레이법에 의한 역가 측정을 시행하였고, 그 결과는 도 7에 나타내었다.In order to confirm the stability of bacteriophage VP2 to salt, it was tested in various salt concentration ranges (NaCl 3%, 2%, 1%). Sodium chloride was used to prepare salt solutions for each concentration, and then 990 μL was prepared. 10 [mu] L of bacteriophage VP2 culture having a concentration of 1.0 x 10 < 8 > PFU / mL was added to the salt solution and left at room temperature for 24 hours. After allowing to stand, each reaction solution was diluted stepwise and titration was carried out by soft agar overlay method. The results are shown in FIG.

도 7에 나타낸 바와 같이, 박테리오파지 VP2는 해수와 동일한 3% 염농도에서 24시간 동안 안정한 것으로 확인되었다. 따라서 본 발명의 박테리오파지 VP2는 내염성(salt tolerance)이 우수한 박테리오파지임을 알 수 있다.As shown in Fig. 7, bacteriophage VP2 was found to be stable for 24 hours at the same 3% salt concentration as sea water. Therefore, it can be understood that the bacteriophage VP2 of the present invention is a bacteriophage having excellent salt tolerance.

종합적으로 본 발명자들은 박테리오파지 VP2가 장염비브리오균에 대한 특이성이 매우 높고, 장염비브리오균에 감염하여 장염비브리오균 내에서 증식함으로써 용균 활성이 우수하며, 물리화학적 자극에 강하다는 장점을 가지고 있음을 확인하였다. 또한 본 발명의 박테리오파지 VP2는 사람, 동물, 식물 등 세균 외의 다른 숙주에는 감염되지 않으므로, 항생제의 오남용으로 인한 항생제 내성균의 문제점, 식품 내의 항생제의 잔류문제, 광범위한 숙주범위의 문제점을 해결할 수 있는 장점이 있다. 따라서 본 발명의 박테리오파지 VP2는 항생용 조성물, 사료첨가용 조성물, 사료, 소독제 또는 세척제 분야에서 다양하게 활용될 수 있다.Taken together, the inventors of the present invention confirmed that bacteriophage VP2 is highly specific for Vibrio parahaemolyticus and is highly resistant to physicochemical stimuli by proliferation in Vibrio parahaemolyticus by infection with Vibrio parahaemolyticus . In addition, since bacteriophage VP2 of the present invention is not infected by any host other than bacteria such as human, animal, and plant, it can solve problems of antibiotic resistant bacteria caused by abuse of antibiotics, residual antibiotics in food, have. Therefore, the bacteriophage VP2 of the present invention can be used in various fields in antibiotic composition, composition for feed addition, feed, disinfectant or detergent.

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

제제예Formulation example 1. 사료 첨가용 조성물 1. Composition for Feed Additive

박테리오파지 VP2 1x107 PFU/mLBacteriophage VP2 1 x 10 7 PFU / mL

비타민 믹스 100 mgVitamin mix 100 mg

미네랄 믹스 100 mgMineral mix 100 mg

스테아린산 마그네슘 2 mgMagnesium stearate 2 mg

통상의 사료 첨가제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.The above components are mixed and prepared according to a conventional feed additive manufacturing method.

제제예Formulation example 2. 소독제 조성물 2. Disinfectant composition

박테리오파지 VP2 1x107 PFU/mLBacteriophage VP2 1 x 10 7 PFU / mL

에탄올 적량Ethanol quantity

멸균 증류수 적량Sterilized distilled water suitable amount

pH 조절제 적량pH adjuster

통상의 소독제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.The above components are mixed and prepared according to a conventional disinfectant preparation method.

제제예Formulation example 3. 세척제 조성물 3. Cleaner composition

박테리오파지 VP2 1x107 PFU/mLBacteriophage VP2 1 x 10 7 PFU / mL

에탄올 적량Ethanol quantity

효소 복합물 적량Enzyme complex

멸균 증류수 적량Sterilized distilled water suitable amount

통상의 세척제 제조방법에 따라 상기의 성분들을 혼합하여 제조한다.The above components are mixed and prepared according to a conventional method for producing a detergent.

이상, 본 발명내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적인 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다.Having described specific portions of the present invention in detail, those skilled in the art will appreciate that these specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

한국미생물보존센터Korea Microorganism Conservation Center KFCC11727PKFCC11727P 2017091320170913

<110> Optipharm.CO.,LTD <120> Novel Vibrio specific bacteriophage VP2 and antibacterial composition comprising the same <130> 1.59P <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 43069 <212> DNA <213> Unknown <220> <223> Bacteriophage VP2 <400> 1 cccctacggg ggaaacgcgc gcgtggatgg gggagaaccc cctcgcctga gtataacatt 60 tttcttgtgc agggttttac acgctttccg ctaggaggcc aatggctttc acaggctttc 120 cccatgtcac cactggcttt ccattagctt tccactctct acacatggac acttcgtgtc 180 agaggcttaa ccctcgtaca ccgcgttgtg ttcagcttgc ttcgtttcac ttcgctgcgc 240 ttcaccacga actgccattg ctaagaatgt acgctctttc tctagctcag cctcatacag 300 cttctgcttc atcaggtacg cttcaagagg ccacactttc ttcatggcat ccaccttagc 360 ccaatgctct ccaaccgcct tgtcgaagtt ctctgggtca acacaagctg attcacctgt 420 gacagtgaaa ccattctcca gtgttaacac acagaacgtc agagtagtac ccggcacaac 480 cacgtactgc tcagacttaa cctttgactc tacgagttcc tgtgttacct tatgttcaga 540 catccatctt ccccattact tctttgtatt tatcagagcg gtaatcccat cgggctttct 600 caccgcgtgt atcaacgtgt acaccccagt cgtacagacc gacgccaccg ttagggttca 660 gagctaaagc cgtgtcatac agcttctgga gcttctggaa gttccctcgg tctttgttca 720 gcaagtgcaa gtccactgca tcaccatgta agtgacgaga gttctccgca ccacctacgg 780 ctttattgtg tactgggcag cgataaccgg attcaacctt tagcggctct ccgaagtgct 840 cccgtagcat ttgtaccagc ataagtaggg ctaggtctag tttcaaagcc ccacagtgct 900 gacacgcgaa cttcacaatc ttgaaattcg cagtgtcttt aattagcata attactccta 960 ggctttattg ccaattaggt tgaactcaac aggttgacca atgttgtcct ctagtagcgc 1020 atacgcagca gcagtgaaag cctcaccctc tgcgttctta ggggtttccc aacggttagc 1080 acctgtcttg ataagtgtga tttcctgtgt acgaccaata gtgaatgtgg tgaacaggtc 1140 taagttatca gccagcgtga ggcgtaggaa cttagataca ctgttaacct caagggcgaa 1200 cacattagcg tttgtagtgc cgtctgtgac gtaactaggc gtaagactac cgttgctagg 1260 gttcatacct agaacacctg ttacaatctg tgcagggacg attgtgtggg tgtacgtaac 1320 accagcatcg actgtcaatg cggtactctc agtaacagag ccacctagtg tttggatagt 1380 acactcaatg ttagctgtac ctgctgccac accagtaacc agtccggtac ttgacacagt 1440 aacaaccgca gggttagtgc tattccatgt cttagtgcca actacggctc caaccggaga 1500 tatatccgta atcgtagctt gagcggttgc accaacctcg ataggtgtgt ctagcacgac 1560 ggagaagctc actagcggta ctaaagggtc aacgcctagg ataagctcta cctgcccatc 1620 acggagcact gggtaagcct cacgacattc ttggataagc tgtgtgagga cgtcttgtgg 1680 gattgggtct gggaagtccc aacgcagcaa ctcgttagcg cgggtacgca cagcagcacg 1740 cacccacgga atcactggta atcccggtac tcgatacgga attacaattt gcatacaatc 1800 tcctacattg cagctagctt agtagcgacg ttaacggcgg tctctaagag cttaccgtct 1860 ccacccacta ggtacactgc acccacacct gctaaaaagt acagggtctt tttaacaact 1920 ggggcaactt ccttaaaggt cttcaacatt tcagcggaat cttcacgcat ctgcttcaaa 1980 ctgtccgcaa gttctttctg tgtctgttcc atagcgaata accgtcgttc gtggtcgagc 2040 aggatttctc gggtagtttt atcctcaaag tcctgcatga agacagcctc tttatctgcc 2100 atcacctact ccttacattt tatttttcct tgcgacgaag cacaaagtcg aagattgtac 2160 gtagaagacc gaaccagtct tcgcgtttaa atgagtctaa tcgtttgttc atcatagtct 2220 acctcgtttg cggtgtcggg agttcaagcg tgatagtgtg cctgtaccta ctccgatacc 2280 agttcgttta tgaagtggct tgccagctac cgataggcgg cttgcagtgc ctttgttgcg 2340 tgggcgagtg tctacgcctt ggtaagccat tgggttcaga atgaactcgg ctacacgcat 2400 ttgtagtgct ttctcagcgg ctgaacgttc gtcctcaact aagtgggcat tcagttcttg 2460 aaccagcata gcgatagcat ctgcgcggtc atccttagcc agacagcctt tgtcgtacgt 2520 gatgtcctgt aattgtagga acgcactgta ctgccaacgg cggtcacgtg gatacgacat 2580 gcagcactcg atgtccatgt ctaatgcaga cgtatggaca actaagcggt gacgacgagt 2640 aaccggagag atagtgtcga tgatacgacg ctctttctgc gtactgttac gtaagtcgcg 2700 cacgccaata tgtgcaatgt cacgttgcgc tagggcgttc tggaacagca ttgtaactgt 2760 accgtggccc atgttgctct cgataacgat gtccttaacc tcgaagtcct cggctaagtc 2820 aatcaagcgg ttcatgtttt cctcagacac gccaccttgg aaaccacctg tgccgaacaa 2880 gtgcacgtac gcactagcag caccaccaat tgcaaaggag atttcatcac caccacaacc 2940 agcagggtca attaccatga ttttgtgctt gtacgggata atctcagtcc caatgctcaa 3000 aggagtgtac aagcgtgcgc caaggacacc atcatgcacc tctgggtaaa gggcacgttt 3060 atccgctgtc caactgaaca tgtccggtgc agagttcgaa tcaccagcat ggataatcat 3120 atcagatagt ttgatacgcg tacgcatcgc gtctgatagg gttgtatcca gcatgtactg 3180 taatgcgaaa ccctctgggc cgaagtccag ctctttctct atcaacgcgt cctcgtcgta 3240 acgctgaggg tcagtagttt ctccaagcgt accatcaacg ccgaagccag tgcgtcgtgc 3300 gccctgctca atcagcatct tgatgtacgg ggctaatgtg tcgccgtact tttcctcttg 3360 ctctacagac gggatacgtc cacaccacac tcgtacttca aaaccacgcg agggtagtgt 3420 cttatagata ctgtctttag tctgaggtgt acccagataa agcgtatcac cgtgcgtgtt 3480 gattgccgca aagtctttcg agattgtcaa caagtgctca cgctgtgttt gagtcagacc 3540 gttcttggtc gtctcaatat catcggggat tagcaagtca gcacgcttac cctgtagctg 3600 agcggtgata cccacacaag ccacggatgg tgctttctct agcggtttca agtcacagtg 3660 aacatcgtaa ccctcgtatg aggtacggtc gccacgagca gggtcggcac gtagccaaca 3720 cagcaagtcc catgtctcaa tcaaacggat aacgagggtt gcaacctcgg atgcctgttt 3780 ctcaccaccg gatacaatta gtacacgtgt gctctggtct tggataagac gccacacgct 3840 gtacagagca gcaagcgtac tctttgcctc accacgctgt gcacacacca tcttcttacg 3900 agggccgtgc tgcatgtact gggcaatgtc tgcttgcatg tctgttaagt cgaacccaag 3960 gaatcgcata ccaacgtacg cgaagtcctc gaagttcttg aatgtgattg cgaacatcaa 4020 cgccagctcc tgacgtgtgt ccactgggat gttacgtgga ttttcagcaa gctttttaca 4080 gcgttgtgct aacaacttta atcgtgtaat cacctgtaca gataaaccca cgataacctc 4140 cttagataat acctgctaat tcatcatcgt ccccatcttg gagacggcct agcagttctt 4200 gtttacgtgc ctctcggcgt tgcgcgagtt catcctcgaa ctcgtcggct agttcaccca 4260 tagcctcatc atctgcgtcg gcagaaatgt tgttatcttt gaggaactta gcaatcactg 4320 acttatcggc ggctggaagt ggaatttctt cctcacgcga ttgcttcaat tcctcaatca 4380 aagcgtcagt gaatgcagcg tgcaacgccg ctagacgcga acgactagca gcttttgcca 4440 tgtttactcc ttaaatacgc tctaaccgga tgtcagtcag accctcgtat ttattgtttg 4500 ttgtgttacg acgcaaggat acccgttgtt ggctctgcgc aatgaaagtc aactcatagg 4560 tgtcaccctc gaacattggg atgtcgatta catcattgcc cggctcgaac gtcaccaact 4620 gcaaaccagc ggcttgcgtt gtagtacgtg ccaccttaat tgtcatcttg taacgagcgc 4680 cttgcactaa gttcaagaac tggaattcac cgtttcctga gccttgcaat ttgcggtcat 4740 tacgccactc caatcggtac ggtgtgatgt actgtggtgg gttcttacca gcggctagat 4800 acgacgttgg caaccacgtt ccatcggcat ctttcggcat gtcatcgtca tatggcttag 4860 gtagctcata tggcggtggc agctcgttat agaaaccgga gaagaagttc tcccacttcc 4920 agtctcgcca tacgtcagca gggtatgttt cacctaactg gcgcactggt tcatctggga 4980 acgggatgta gtggaagtag tcaacgtcaa tcgagtcata aatgtagtcg gcactatgtc 5040 cagcccaacg gttgcccggt gcgcgtggat accaaagggc aatccataga cgacccggaa 5100 tgtcagggat gtgtgtggtg ttagtccatt tcaatacacc gtcaacgtag aagtccacac 5160 gtttcgcagg tttccctgct ggtgggttct gttctccagt atgccagtcg aaacgaatct 5220 cgtgccactt accgtcactc aacgcctcat ccacccactt ctcgaagaag tcagtgtatt 5280 cagagaagta gttcgggtca ctctctggta aatcccaagt acgtagctca ccctcccaag 5340 tgttcagacg cgctttatca gccgatacat cctccatatc cacagcatct ttcaatgcag 5400 tcgggtactc gatgtcaatc tcgtggttac gcacaatgta gtaaccatcg tcctcattac 5460 cttgagcgtg taagccctcg tccaagaagc tctgccaacg tgggtcgttc tcgtagattt 5520 cctcgtagtg gaatgtccac aaggctgtcg ctgcaccttt ccggtacgga gagcgcaact 5580 tacagcggaa actgcccggc cctaagtact cacgtgtaac cagtgctgaa cctacctcag 5640 tcttacgacc atcaggggca gggtgtccga tacggtcaac accgatgaca tcaccatcgt 5700 agtacgtacc atgaccacgt agacgtaaca cacctttcac agcgtttgct acatcagagt 5760 agtcagggta aacctcaacg ttctcacgca ccacaccacc gttcagtagc gcggtttggt 5820 ttcccacaac gccagtacca ccccattgct tatgtgcaac gagccagttc tcacgcattt 5880 caatcaagtc agagaagtcg gaatacaact cagtcttgcg gctaatgtca cctttgatgc 5940 gcactggcag cgtgttactc acaccgtagt tggtcttgta ccggataaag tactgaccgt 6000 caggcacatc aggcagcgtg taaatacaac ggcggttatc ctgctcatgc ggccagttca 6060 ccagagaact cgatatatcc tgccacgtgc tgctgccctc agcttgaatc tcaatgttca 6120 agtaaggctg ctcgtagtag cgcggaatca cgttctcgtt gaacgtacca tgcgtgtcga 6180 tgtagaaacg acggatgtca atgttaccaa tccacgactt gtcgtaaaca tcagtaatag 6240 ggacgccagt cgtgcccaca cactcagggc cacagttgaa taatacactg tcaaactgtt 6300 taccccacag cttcaatgag cacgaagctg actgttcatt cggcgtacct ttatcaactg 6360 taatcgtgca gttgtggttc gcgtcctgcg ttagttgcac ttgatgccaa ccactcagac 6420 ttgctaattg cacacgtgcg tacacgtaag tactgttgaa tcgacggtag tacatcaact 6480 caccagagcg tgatacatgc gcaccaaagc taatctgagc actcttttgt tgaccagtgc 6540 cttgttctgt ccaagttaca ttagagccac catacagcag ccacatgcgg tcttgtccat 6600 caacgaagtc acccaatcgg aattgtagtg accaaatgtt cgggtagatt gcagtcggga 6660 ttgtacccac gaatgcaccc acaccccagt tataaccctc ttgaccgggg cgagtaactg 6720 tgtaaagata ctgagcattt gcttgtcgca gcttacacat acggtaaatg ttatcaatac 6780 cgtaagggtt cgctgtaccc tcgtaaccga agttaacata gttctcaagc gtgtcagcac 6840 cattactgta aagcttgtat tgtgagcctg tgaagaatgg gagtgggtct ggtagttctt 6900 tcagaccatc accaaacaag gctacgtcgc cacgataagg cgcacctgtg aagtaacgcg 6960 cgttatcaag gctcactgtg cctacagagt caggattatt gctataggct aagtaccagc 7020 ggttgttgtt accaatccag cggtaaagca cttcgcggta ctggtacact gggtcgtcgt 7080 cttcaccgac attcactgca atgtactgtt tgccattttc gtaccaaacc tgattctgac 7140 cgtggtaatg cttaggggct aaaccctcta gtacatcagc catagatacc tctactcttg 7200 gtctttatcg ttagatgagt taacaaacgc accattgtcc acatcaccaa tcatcaccca 7260 ttcctcgtac tgaaccggaa cggtgtcagg ggatttagtg gagatagtga ttgggcctcg 7320 gcggttcgta cccttaatgt tctcaattgc ggttacacgg tctgcaactt ctttcaattg 7380 gaggaacgtt acagcgtctc ggttctccgt accttgtgcc atgttacgca ctcggtagcg 7440 gtgcatatcc aagtcaccat agaaatcgct aatgccagaa ccctcagtca tttcttgact 7500 taggtacagc aattgcgtgt gcgcgtcgtc taagttagct cgggtgaact cagcaccctc 7560 agagaagata tgacgcagcg cagtgcggtt ggtacgccgt gtaagcgtaa cctctacccc 7620 atcagctaac ggggcgttgg taaagttaat ttgtgttcga ccctgccaag tgtaatctgt 7680 accctcgatt gcaggtgtct catcaagcat aacgaaaatg tcacgcgtgt caaagaattc 7740 aatgctgagg acgatgttgg tcatagtacc gtcgctcgta gcacgttgta cacttaatgc 7800 cataaagcct ctttaaaggg ggattactcc cccatgttgt tgataactgc acgtagcggc 7860 atgaactctt gcgcgaacgg aagaatcttg ctcatacgct gtgtgtccat acgaccatcg 7920 ctgaccatat cctgcaccgc cccgatgctc tgcgtcacgt aacctagtga ggtcagtgag 7980 tgtcgcggag attcaccaag gaatagcggt gacagcaaag tgataccacc aatagctgac 8040 atgttcatcg cagtgttagt gattagttca cgtgggctct gtgcctcttt accgtccaac 8100 gcaagcttag catgggtagc cataagcatg agagggaact ggtacatcat aagcatcgca 8160 agaccagcgt aatcaccatt ctgcgtgtat cggcgcagta gtttgttagt agccgcaatc 8220 gcaaaggatt ggtagccaat cacaatctta cctaccggac tgaattgtgc aaagtgtgac 8280 gtctcaccag tacggatgtt ctgcatcact gagtccatag cacgtgtcga tacaatctcc 8340 aactcacgtt gaatctcgta cgggaacacg ccagtcgggt tctcagcata cgcagtacgt 8400 agcttctgtg tcaactcagg tgtcagacca tgctcagtta gagcgcggct tgtaccacct 8460 ttcaacacgc gttcgacttc atcacaaatg ataccagcgt tcaagttcgc cattaggcgg 8520 tgaaccatgc tgaaaccgtt tgctagtcga gctgcttgcg acaggttctg agttgtctgg 8580 aatgcagcac ttgtgtttgt caagtcaagg ttatcgtcag catatgtgtg gatgtgacgg 8640 tatcgtgctt cgttctggta cgcaccattc aagatgctgt acagacgctc acgagagcca 8700 gtatccgcaa caactgcgtc agccttacgt aacagaccag tgcttgctaa accacgcagt 8760 acacgcgcta aaccgaactc tttcattgcg atactaatgt cagtcatctg gtataaaccg 8820 gagttcttta gcatcacagt gttagcaagg ttgcttgcac cacgcatcag ctcagggatg 8880 tcagtaccga ctgggtagcc tagcaggtgg tcgattgtgt cgtccactgc tttgccccat 8940 gattctgcac cctcacccat acctagtcga gtctcagtga ttgaacggtc aagtgcacgt 9000 aaatcagtga taccacgatt cgcaaatgcc acacgacctg acatacggtt tgtgtaacct 9060 tgcatcaagc caagtgtgtt agtctctaga atgtcagata aacgtagcac atcaccatca 9120 atgtcgaact ctttgttcaa gttcaagtca agacggttac gcaagttctt cactggcgca 9180 gtgctgcctg tcttcggtac gttcacacgt aggaactcat tgattttgtc gtcttcaacg 9240 ccagcatcac gcagagccat aaacaactca tcgttatcca taccttgcat caagttcttc 9300 cattgtgggc caccgttacc ggatacgcca ttcacaatgc ggttgtacca agagtcggct 9360 acacgtgcag ctagcttacc gtccatgtta gtgtaagtat cctgcaacgc agagcgcagc 9420 agtttcttaa ctttaggagg ttcgatgtta cgcactttct cataactata ttgacgaggg 9480 atgtagtact tcgagcgtgg gaactcgtct gcactaatag taccggattt cacaaggtgc 9540 tcatgccaag actcagccca accggaatcg gtgtaggctt tcaccaaacg ctcaatgtcg 9600 gctggtggtg ttgggatttc gcgccctgct ttctcagcac tgtatgcttc atgtaggtac 9660 ttagtcagct tgtcttcttc cacactacga gctgtacgag ccttagcacg gttaaagaac 9720 atatcaaagg tgttaacgcc gtgcttctca cgcatagcag ccacaatcgc atcttcgacc 9780 gcagcagcac gtaggttctg ctcaagagtc aagttacgtt tatggtctac cacactggta 9840 gcacgttcac ccatacgggt agcatcagat acaagtaggt tcgctaggtc ttcactacca 9900 cgagcaatgt tgtcccagag tgcgaacgac tgcttcaaag atttacggac actctgctca 9960 agacgtttag cacgctcaac cgcagtagca ccacggttcg catcatctgc aaggtcagcc 10020 atgctatcag agaagaaagc attcacttta tctaggttat cagcttggta gttagcaacc 10080 ttaacaaagg catcaaccgc ttcatcaaga gctgagtctg tacctttgaa gcctaacatt 10140 ttcagaatct tctcagccac agcttgcaaa gcattctgac ctgcacggac acccggtaac 10200 ttcgctaggt ctttcaccat ctccggctta gacgccaact ctgctaggaa ctcacgattg 10260 ttagtcatag cgtagctgta cttacgagag aactcacggt tgtttttcag tgaggcatgt 10320 aaagcatcaa ggtcacggat agcctctgca atctcaggag caacattacc tttacttgct 10380 tggaataaag tcttagatgt cgcagcatgg ataagctcat gcacagcaac gtctgcatct 10440 acgtggttca acatatcgcc cacggtctta aacactgtac cgtctttcgc tgtagagcgt 10500 agtttcagaa actcagtgtt agcaccaaag gtgtaagatg agcgacgacc agtagcattc 10560 attaaacgca cttccatacc agagatagta tctggtagtg catccataag agcacgagct 10620 gagtctgaca aactatccat agtacggagg tgtgtcacta agtcttgtgc tttaatttga 10680 actgtcttac gtccttgtcg agaggcaaca attcgaccct caacaggtgt gtcctgtaat 10740 gggaaacttc gttccccctt acctgtacgt gggactttta ccacctctgg cacttcattt 10800 acaggtgcgt cagggtcagg cattcgctgt gcagcgacct ctggttttac cacatcaggc 10860 acgatttctg gagcatcact taggacaccg ccagtctgtg ctttacgtgc taaacctgtc 10920 tcagcggtga acacgtgctc agttgcagaa cccaaagcgc ggctctcaac ctctgctgca 10980 ttcgctgctg cacggacacc gcgagttgca cgcgatacgt cgtacagttg gtcaacaccg 11040 attagacctg cggcaaccgc tgtaacccac tcagcctgac ctagttggtc ttgggcgtag 11100 taagtagttg ctaactcacc agcacgcaat gcagcacgca cagccatacc agtacgacca 11160 accacacctg ctgcacccat aggagcaagt aggaacggcg cgtcacctgc taagttacca 11220 gcgataccag cgaacatgtt ctctgcttgt acaccatcac gcttgcgttg gtcttccaca 11280 gcctctttgc ggtactggta ctcatcaagg ctacgtgagc caccgatgaa gttcaattct 11340 tcttctgagt accctagaat gcgcagagag gtatccgctg tcatagtagt accaaggtca 11400 aaggatgtgt cctctgggaa gtccggcatc gttgctttac gcacagccgc acctacgata 11460 ctgttaccaa cacctgctgc gaatgattcg aggccagttg ctttcggtgt ttcaaggcgt 11520 ttaaggttcg cctcacgctc cgcagcgttt aaacctaaat cggtagtgtc gtcccatgct 11580 agtgcaccag cgtccatatc tacgccctta gcattagact ctaagccagc accctgtgaa 11640 gatgggcgac ggttctcaaa aatagccata gtaattcctc aaagtataat gataagggag 11700 gggcaaatgc cccaccactt agtaataggt tgggtacgga ttcttagcag gttgattctg 11760 ttgaagccaa cttgtcaagt cagtacgtaa ttgctcttgt cgagtctttc ctgattgctt 11820 gtacgcagga gtttgcatca tagcgttcat cactagcata cggttagtga ggttgtcgaa 11880 ttggctcaca cccttagcgt aatcagccat agccttagcc acagggttcg cgctaccacc 11940 accaccatga tacgcaaggt caaccagcac agggtacgca gcatcactca actcaagacc 12000 agcagactta gctgctgact taacaattgg gatgtacgta tcttctaggt cgcccttgag 12060 cataccgata gcttgcggga gtgttacctc atcagcaatg gccttaccgg agttagtgtg 12120 acgtccgaag cctacagtct cgccaccacc tgctttcttc ttcttgccaa ggaagccctc 12180 agattccatc agattagcgg ttagcttgcc gaacacaaac gggtcgatac cagacttgtt 12240 cacaccagtc acgttcattg tcacagcgcg gttattcgtt gcgtcataga acgtagcagc 12300 ggttaggcca ctttgttgtg aaccaagacc tgcggcagtc ttaacctcgt catcgtacat 12360 cgtagttgca gcctgttgca gagtgctgcc tgagatactg taacgagtca caataccgtc 12420 ggagtcttcc acagctacaa ccatacccat agcgtcacca cctgctgcac cgatgtcaac 12480 agttacaccg ctcacatcag tctgaatgct tgacagcaca gaagccactt ggttgttcat 12540 agctttaacg aaagtctcct gagagccttt gtatgagcca tagaagtcgt ctagcttctt 12600 acccacagga acgtacgtgt ggatgttgtc accgcccacc tgcaactcca cctgacgcgt 12660 acgggcatct gcataagcag ctttctccat agcgtccaag tcgttagtca gctctgggcg 12720 tttctgtgca atgtacttcc agctctgtgc cacagcagta gcaccagcag tcacctgacg 12780 gtcactttct gtgccgaagt caagccagtt gttcagttta gtaccatcaa tcaaagcgtt 12840 aggcattgtc gcgttctggg cgttgtactt accagaagca atagcctgag cattgcgtcg 12900 caggtcgtcg aacaggatgt tactcgcgtt ctcagggttc tgtaatgcag cattacccat 12960 cgggccacgt aggttctccg gtaggctctg gagcatcgtc tgcttaccaa tggcagtact 13020 tttgctgtac atcatcatcc atgtggcaac ctgcgcctgt tgttcctcac tgacgacatt 13080 atccttgcct gtgaagtcga ttgtcgctaa gacctgacca gtgttcgtac tcatacgttt 13140 agccgctagg gacgccatgt atgagtcgtt agcttgtgaa gcatatgcca tcattgccaa 13200 gttaccttga gcagtgtctg ggaagttctt ctctagctct ttacgagtct tgtctaaatc 13260 cataccgagc tgaccagcta ggacagttga cgggatagca ttgctcagtg cgagttggaa 13320 agtgttcgct tgacgcgctt tcttctccgc ctcgttcaga cgcttcacat acgaaccagc 13380 agtaccagcc tcgataatac cctgctgcgc cttgtcgtcc acgaagttac gtagtcgagt 13440 caactcagcc gcttgttcat ccggtgatag tgctgctaga ccgttgatgt ccgtttcgaa 13500 actcaccata gcctgtgcag cttgctggtt tccggcacgg ttgtactcag tgcgtaacga 13560 cttaataacc gtagggctta gcacgcccat ttccttatca accacgcctt gcagcatgtt 13620 gataatactt ggctcatcag tgcgctgtgc aactgtctgt acgtacgctg tggcacgttg 13680 gattttgtca tcctttgaca agtgtgtaga caggttaatg ctgcgaagac ctttcgcaat 13740 actagcctgt gccatttctg gtgcacccga gtccacaaag ctgtagaact ccgctgctgt 13800 gctgtctagg ttgctgtcta acgcacggtc agcagctacg ttagccattg cagcggagtc 13860 tttctggaac ttgttacgag ccgctacaga cgtttcctgt aggtcttgca acaatgccat 13920 tgccgattgc ttagggatgt gtgggaggta cttactcatc tgactggcga acgcagcgtt 13980 gcgctcttgg ctcagcttgt cgaagtactc taagtcctta ccctgagata cggcattatt 14040 gatttggtcg atttgctcct gacggaactt gcccatttcc tgattcactg ctgcactcaa 14100 gtagcccgat tggtaggact ccttgaatag catgttcacc ttgctgagct tttcatcttg 14160 cttgctcatg gcgtcagtcg tggcagtggc gtctagcgca ccttgaattg cggcatctgc 14220 tgcggcttga ttgtacgcac gttccactgc tggcacggct gtttgcagga atccagcaat 14280 cagcgcacca cttgcgtcag gctcagcctg tacgttctcc atacgtggag aggccgcacc 14340 aatcaacagg ttgccagttg cagctccgag ttgtagcgga ttgctttgga ttggcatatt 14400 atctccttac caacttgtag ttattggttt atccgaataa ctgaaattaa ggtcaatttg 14460 gtcaaaggca ctacgtcccc accaatcgta atcagcgtcg ttctttccta cacttgggac 14520 ttcttgtggc tgtgtctgac tttgtggggt taactggctg aaaccgtagt tcgcagcagc 14580 gcccaccatg ataccagcag cctgcccgta tgcagcgttc atgatttgtt caccggatga 14640 tgccacacca ctttgtagag cattactgcc ctcgtcggta gtcttacgca acatcatgtt 14700 taaaccctcg ttcgtgatta catgttggtt ctcaactgta ccttgagcac ggtcagcgtt 14760 gaccatgact gattgcgcag cgtcacgtgc actaacgccc acagtgtccg tggcagcagc 14820 catagcacgt gtttggttag cagcagcgtt cttctgcgca ttgatattaa atagcgcact 14880 agcggtcttc tggttctcaa tagagcgttg caagttaatc tgcgacacag actgagctgt 14940 gctgagcatg actttcttgt tgtactccat gatagcctga tttcgagcta cggtggcttt 15000 gttctccgcc ttagctgctt gaccaccagt gaacagtgct gacgccgcac ctactacagc 15060 accgatagca gcaccccaag ggccacctac cgatgcaccc atagcagcac cagacgtagc 15120 accagtggcg acaccgccac cacttgcacc tgccatatta actccttata cgcgacgtcg 15180 gcgttggttg taacgaatga tatattcaat ttccagaatg ttcatatcct gtgtaccatc 15240 agtactcaag tacatttcag tagactgcgc attgcttcga cacggcacag ggacgcgtgc 15300 taggtcagca agtaacgcca tatctggctg aacgctatca gtaccaaaca caaggctagt 15360 tgtgctatct gagtaatcca atccgatact tgggtcaacg atgcgtacat cgaactcacc 15420 agtatttcga gttgttaact cataacgcaa cagacgaacc ggagcagtac tgattagagt 15480 atcgttctgg tctttcagca ttggaggtgt tggtgcgaac atagactcgt accggaaacc 15540 tagaaagatt tcttcatgtg ttgagcctga ttcaacagtg aactcccaag tgatttcgtc 15600 tacgtcaata gccacttcct cagaggcacg tgcaccagtc gctacagcac cagtgagctt 15660 gccctccgct acccaaggac gcatatagat tggcacgata cctttaccct cagtcgtaat 15720 aggtacacgg ctcattgcgt ctagatgtgg taatcggtca acatcaaacg tcacaccgcc 15780 ctgtttaggg tcgatagttg tgatagtgat tttgtcggta ctaccgtcgt catctgcggc 15840 aaacaacaca acacggtcac gagcaaagtg cagactagcg acacggtacg gaaagagcca 15900 tttatgccat gcttggtgta ctttaccttg gttcgtgaag tgatactcgt gtgcaatcac 15960 ttgacggttg tcaccagtag ttgccattag cacgatgtta gcagcacttg cactctgcat 16020 gaaacgagcc tcgccctcaa tgtaacgggg gatatgagtc gtaacgtctt gagatacgta 16080 ctgagacgat gtgtattgtg atggtacaag ctccagaact gcactgaatg cttcgcttcg 16140 aggtgcaggg tacatcaggg tctgtgaggt tgtaacaggt gctacaaacg cattacaggc 16200 caaatctgag gtcaacacca cactagcgtt atcaggcgca agtaactgtt gcagggatgg 16260 tactaccgcc tgtgtactgt caccaagcag aatcaagtcc ttgttgaact gcaacgcttg 16320 gcggaacact gagttctgag ccgaaccgga tgcaatatca atacggtctg ttgggtcaag 16380 ggaactcacg gtagagcgga agaaacggtc tggttcgcca gtggcactca tacagacgta 16440 tgcaccagat agcagcacta agcggccttg gaacgtaccg ataccagtga tgcggcgttc 16500 ttcaaggaat gtcggtgcag ggttagttaa gtcgtcacct gcgagacgcc cctccatgat 16560 atgttgctct atgttgtcat cgacaatctt atagggcaca tccacgctaa tagcagtcac 16620 ggagttgtag tcaccagatt ccaaccacac acccttttcg gcgctgtaac gataccacac 16680 aagtgcacgc tctgattgac ccacagcaca caacgcacca tcagcagacg ggtgcagacg 16740 cgcagggagg tcagtctcta ggtttacctg tgactggttc gagtagccaa cgtaaggtga 16800 acctgacgta ctggtgattt taaggtcagt acctgtaatt aattcaaagt agatgtacgg 16860 gccgacacgc acagcgaagt ccacacccac agcaattaac ttctcgacca gcttacgtgc 16920 aattgcttct ggcacagatt ggtctgcgtc acctgctgtt gtaccgtcag gtgtagtata 16980 tgtcacggtt tggctaccga gcgaccacac tacggacagg tcgtactctt tactgaacgc 17040 accactcttg acgaagtaga aacctgttgt cttcgggtct ttcttgtcag agttgtcagt 17100 cgttgcagac ggacgtttct cagtgttcag aatgtacgtg ataccaccca tactggtagt 17160 ctggatacta cgacggtcag ccgctaggag gtatgacaag ttaccctcac tcacgatgtt 17220 cttagcctcg cggtctaaga gccaccagcc gcctgtgttg gtgttgatta ctaggtgtcg 17280 tccatcagca cctcgttcga gatactgtgt gtacagcgca tcaccctcac caatcgggcc 17340 gaaatcgtgc acactcacga acaaactacc gggacgacgg cggatgcttg ttactgggtc 17400 ggacagcata ttaacttgtg cagagcattg tccagccaca cgcgcacgtg ggacttgttg 17460 cgacacgcct tgcagcaggt cattcaatgc accctcaaat ggtcgagcca tgattcacct 17520 ctatgtatat agtgcagagc ggatacgacg gtaacgccct gtcttgcttg tactgtagcg 17580 gcggttacgt aggttctgct tgtgcagcac tcggaacgcc tcttgcgact gacggtctag 17640 gttctggtac tggttgtcca cacctaaatc gtttgcataa acttgcatag ccgcttcata 17700 cgcgacgatg tgttgggcca tttctggcat gtgttcaaat ggtgtatcaa acacgatacg 17760 caatagcttg gactcatcga atacgatgtt attggtggcg taatccaaca gcaatccgtc 17820 ttcattctcg ccgtacatca catcactagc cgcatcgtaa atgtcgatgg ttgcgctagg 17880 taggtagata tgcccctcaa catcagggta cagcgtcatt tcacgctcgt tgaaccacca 17940 accatcgctc agcaatagtt tgcgtttagt ttcaagagtg tccttgacga taccaatggt 18000 agggttgcga acagtgtcgg atgtgatgcg gtactcacca atcgcagtca acgagatatt 18060 gattgcatct agtaaggtca tattttaagc actccaaaca aaaaagggga gcagccgaag 18120 ccactcccct tgggtattat tgctgattag ctctcagtct ctgttttctc tggtgcacca 18180 gcggctacag gtttgttctg acgagcgtca cgcttctgct tttgacgcgc tgtgtccggt 18240 gtccacttag cgcgtaggtc gattacttta gccatttatt tgctccttat actgctgctg 18300 gttcttcgaa cttagtaacg attgcagtgt ctggacgacg ctgaccaact gtgtacatag 18360 cgtaacagtc aagtacgtta cagaactctt tctcgtcatc ccagatacga gttgtgaatg 18420 ggtgtgcttc cacagtcaca agtgtacgtg atttagagaa agtaaccatt tgacatttag 18480 catcttccgc agatacgttg aacgaagtgc caagtgggtg agcagcgatt gcaccagttg 18540 ggaactcggt acactcaact actggtacac cgttcatacg aacgacacga cgaccgccgt 18600 agtcagacat atcaacgttg aactcttggt tcagcagttt tggatgctct actagcgcag 18660 agtaaatgtc tggtgataca agggtaattt ggtcagtcag aggaacttta cgtttcacta 18720 gctcagtcac accagccttg tgagcttcgt tgatagagat agcgttcgct tcaagttctt 18780 cttgagaaac cgcagcagcc ttgtaagttg ccagtacttc tacaccgtca ctgaaagcag 18840 gtttcaagtg gtcaggcgcg tcccaagtac gacccttgat gatttggata aggtgcgctt 18900 ggtcgaagac ctcagcgaac tcagagccgt tgttctgacc catttcggtt aggaagtcag 18960 gtgcagtcca gtcgtcttgg tagtcgattg ggttacggat gtacagcact gtatccacga 19020 cgataatcat tttatcgtta cgtacaggcg tgttatccag agtttcaccg gaacgacgac 19080 ctttcactgt tgaagtgttc aagcggtcaa tacggtaagt gttagagcgt tcagcagttg 19140 aacgttgtgc agacaggcca cggaacatag ctgcgtactg gaaacgagtg tcaacttcgt 19200 tttggtacac ttctaggtgg atgtcttggt cggaagcagc accaccccaa tgtgggcgag 19260 ttagacctgc tttgtatgaa gtatcagcca ttttaaatcc ttatggttga aaagtttaaa 19320 tctggagttt cagacgccag aaacgcaaaa aggggcttat cgccccagtt gtttgccacg 19380 tgcgcggcgt gcaagcagat tctgatacgc ttgaccagca gggccacttt caaaagaagt 19440 gttaccaaac ttggtacgta aatccgcaag ttctttcgag aactcagcag ccgacaacgc 19500 agtgttgcca gcttgaccac ccattccatg aatgtgttgg ccttgagttg gtaaagcacc 19560 agactgacgg gcgtagtcta gtaccatctg agctgcgtcc ttggctagac cgctgtttgc 19620 catagtcttg accgcagtct gaatatgagc aggagcgttc gccatgaaca catcacgcgc 19680 ttgtgcccac gcctcagcac caccagctag ggtgtgtact tcattaacag tgcgttcagt 19740 atttgaagca gcgttagcga ggtacgtctc agctaaggtc tttgcgtagc cagcgtagtt 19800 gccgaggcgt tcttggagga acttctcgtc aatcaaagag gcatcaccat aacgcaatgc 19860 attacccata gctcgctcta cgtcggcatc ggtacagccc gatacttgag caagcatctg 19920 taagccagca tcaactgcgg cgtcacctgt ctgtagtgta atctgacctg cggctgggtc 19980 tgtttgctct ggttcagcag gaacttgagg ttctggctgt ggagcagcag caggttgtgg 20040 ttgattaccc agtacagggt cgtcggctgg tgcaggttgt ggtgcagggt tctgcgctgg 20100 ttgtggttgg ttcggaacgt cagcaggttt acccgctgct ggtacgcccg gtggaagatt 20160 tgtatcttgc attattgagt caatccttgc tgttcagaca acagtgcagc agagccagca 20220 tcaagtgcca taccggaggc aggggcttga gcagcagctt cctcagccgc ttgtagctgt 20280 tgcagctctt gttctgtgta gaaaatctca gttgggtcaa caccatgacc tgcgaggatt 20340 aggtcaatca ctcggtcagg gttagtacgt tttgtagctt gtgcaagtac agggagaatc 20400 aagttaatgt cgttcgccac tagacctaga cgttgcacca caatactacg gttgatagcc 20460 gcagtaccta cttgaatgtc taggctaacc gcattagctt gcaacagtgc cagtagctca 20520 ggtcgagcct caactgtcag gatgtgcgct aatgggatat gcagaatctc agcaagagta 20580 gcgtagatgc cacccatgtt ttcttctgct tcacgaacgt tgttcttgat ttcttctgct 20640 gttacacgtt cagcatcacg gacgttacca gtgtacatga acgctcgtgc cagctccgca 20700 agaaccatct gtaggtcatt aacgacctca gcaatcttac gtgcttcacc agcttcatgc 20760 gccttgatac cctcaccatc acctgcaacg tactcaccag tctcagcatc attcaatgca 20820 tcaacgtcag cagtactctt aggcgacacg aggttaacga ctcgcattgc ttcaatctca 20880 tacagggtca aggcttgagt tagctctgac acacgtgcga acgcacctgc atggtcttcc 20940 acatggccac gaccataatg ttcaccagaa accaagttcc acaccgcagg gatgtaagga 21000 catagcttct cagggtaaac actaagtgtg ccaacaggct taccatcaat ctgttgagta 21060 atcacaaaaa cgtcaccagc atcacgcgtc tcgcgttgaa tcttggtgta caaccaaaca 21120 tcagtggtag ggtctgtgta tccggttgga cggaattctt ctggcacatc ctgaattggg 21180 atacgctcac gtagaactgt gcacaacact cgaccagcac cgtcacgacg cacaacataa 21240 tcgcgtacac tgtacacggt catacgtccg gttaacgggt cgcggtacag caacgcatta 21300 ccagtgatga taagtaactt cattagatgg gttaactggg caaagccagc gtgagtagtg 21360 agcaacgtca ttgcttcacg ctcgatacca gtaaacacag cagaaagttg cgcgttctca 21420 ctacccagtt gagcaatgat ttcacgcatc ttatctgtgt ctgagatacg gaagaagctc 21480 atcccagttg ggaaaagcgt ccgtgtcagt ttactagcaa ggttgttaac caacatagca 21540 ccgtgagatt ggaagtcgcg ttgaagtgat acgcgctcac cgtctttgtt ctcacgtgta 21600 aaaacggtag gcactgtcca gaaagcataa cgctctgaac tcagagtaac ctcagagtcc 21660 tcatacttag cgaacagagc ctcgtgagtc tcttgagtat gcatacagta ccttataatc 21720 ctaagcttga actaacgtca gccatcttgc gtttacgagt tgtaagacca agagcgtcgt 21780 cggctgctag atttgcagaa ccgcctgtct caaccttaac tacgtcgtca agagctttgt 21840 tagcttggag tgtgttagct tcacgcattc gagtctgttc agcttcgaac tgtttcttct 21900 gcgcatcagc ttggcggcgt gcttcttttg ccgcatcttt accactatcg aatcccatgt 21960 ttcaccttgt agtacttgag tctgtactcg tactcgccct gcttgtgagg gatgagcacc 22020 cagtccagac gttgtgcctt ggcatagcgc ataatcgcac ggtacatgtg atgtccgata 22080 ttgtggttcg ggagcacgta gcttaataca acgcccacgc ctttaccact gatatgcggt 22140 gtcgtgtacg gtgctgttat cgcgcagccc cctattaagt tgcctttgtg atgaatgccc 22200 atacacatga acccatcaac tgcatcggca actgagtctc taaagtcttt agaggtaata 22260 ccctgagcga actcagggtt gttgtggtac gctaggtttg cgacaactcg cgcacctagc 22320 acagcctttc tacctgtgta gaattctaga gtataattgg acacttcgtg tctctcctgt 22380 tcgtgggact ttaaattagc agaagaagaa tggcgaatct ttaaccagtt ccagattcaa 22440 tgtacccatt ggaggcatct cgataccctc aaggtcagca cctgcttcta acgctgcgtc 22500 acggataacc tcaagtgggt tctggtgctc agtgtataga atcacgaact gctcacgcaa 22560 cacacggtgc aggtcgtcaa catcacaagc gtgagtcgca actgaatcgt ggattggaat 22620 gatgtcaccg tcaaagttca agatggtcat cataaggtgt gagctatctc cttggtggat 22680 gaagttcggt gagataccgg atacagcttt ctggcggtgg ttacgctcat agtcgaagtt 22740 gtacagcacg acttggttga tacccatgct gcgcactttg acacgagtct catggcggtt 22800 acagtagcgg ttaatcacca gaccaccaag cactgtcttc cactgcaagt gatgttccat 22860 cggtactcgt tgacatacag ccttgaggta atccatacca cgtgcagcgg ctggcatagc 22920 ctcattaatg ctgtcacgca tcagcggtga tacgaacgaa cacagcttga atagtgtgta 22980 ctcgtctgta ccctcgtagc cctcatcagc agcaccaagg aaaatgtaat ccgtgcagct 23040 acgtaatgtg gcagagtaga agtacgtcat cgttgggcgc ttggtcattg aacgtgtgat 23100 agggttcagt gtccagtaat gcgcttgcac gattgtctca gggtcacgta gagcaacctt 23160 gattttcgag tcagtacgct gcttaacatc catgtacagg tcggccttgg ttaagttgcc 23220 gtcccagaat aggttcgtta gcttaccgcc gattgggtca cgtagcattg cagagaagtg 23280 ttgcccacca gagttagtgg catccaatgc aaccgcaatg tttgactcgt acttctcagg 23340 gtcaggtgag tctagtgctt cgattagttg gatactagcc gcaagcatac accagaactc 23400 gtctgcgtct ttgaagtctt cgttgttcaa tgggtctttc acccactcac gaatcgcatc 23460 aatgcgttcc tcagtccact tggcacgttc ttcgaacagt ggcttatcga aaccaaaaca 23520 tgttgccacg tgtgatttca gccagaacag accacgctta cctagcggct tcttacgtcc 23580 aagtagtagc aacgcctttt ggatgtcaga gccttgtggg ttcaagtgtg acttgaagta 23640 caagcggtaa cgccagtcaa cacaggtcgg gaagtacaac gcatcctcgt ccatgaaacg 23700 gcggcacagg tcaagcgtga tagccagttg gcgaatctgt gatacacgtt tacgttcctt 23760 tgtgtaccac tgacgcatct gggtcttcca aacctcaaac gcctcttggt catcctcaga 23820 ccagtcagcc ttgtcagtgc cgtgtaggtg gaacttaggc tgtggtttct ggtggtggct 23880 tgggatacca acgtcaaagc ctgtagcctt ggcttgcagt actaagtcgt aaatactttt 23940 gttcacgacg taaggcactt cctgtgcttt gttcaacgct tgacggatac cagtcgcctg 24000 tttaaacgcc tcgttcactt cacgtaaacg ggaacgcttg atgtgtcggt tcgagtacgt 24060 atggcgacga ctcaggtcag ttaagtaacc gccgttgtaa atgtcctcat gacgtactgg 24120 aggaacaatc atcggtgggt agtgcaccat agtgtcagcg tgctcaagta catccgagaa 24180 cacaccttgt agttcttcgg ctggctcagc gtacagcaaa cctttcgggc cttgtaccca 24240 ttcaaagata ccagtctcat gcacacactg gagcaacaag cggcctacgt tgaagcaggt 24300 tgtgttgtcc caagggtcag tttcgagttt cacgttcgca ctagcggcac ggaacgtacg 24360 caagatatga gattgggaac gcgtcttgtg ctctttcaag tactcgtaca ctcggttcat 24420 gtacgcaggt gcaaccacct cagcattctg agctatgatt tctgattgca ctgagcgacc 24480 tagtgatgta cctagagact ggatagacga catccccaac tcagttgagg caatgttgtt 24540 cagtagagtg tttaatgtca tcaccgtcag aacatccgac gacactaaac ggatatagcc 24600 acgatactta gcaccaacgc cacgagtctt agaagctttc agctcatcaa ttcgggcgct 24660 aacaccatca tacgccgcat tcagcagacg ctgcacaggt gttaactcag tagcacggcc 24720 attctccaag gcatccagta cagctttacg gctacgctcg attgcctcag tcttaaacgt 24780 ttcttcaatg cgctcttgac gagcggcttt ttcttcgaat gtctcacgat tcttggtgac 24840 agacataaac actccaatac attacaacgt tgttttcaaa ttgcgctgaa tcataagacg 24900 cagcaacttg cgtttctctt tgtccgtaat gccgaggtac agcttccgtg ctgtaccgct 24960 tctacctgat tgaatgtgtt gggcaataac tgcgtactcc ctgtccacat tcacaggtgc 25020 ttcatctacg aagattaatt ccgcagacgc tccacgatgg ttcgacatgc ggcctccggt 25080 ttaccctcaa tcatcactac gtcgaggtat tcaatcgcag gatgtttcac agttaagtag 25140 ctgcggctgt cgttagcgaa cgtacaacca cgaccataca tacggacaac cagtacctca 25200 ccacccatag ggctgtgctc aatgaagtca gcgacgctaa ccgcttcctc tgggaaacca 25260 cagtcagtga ataccgaaat acgcccatat tctagcacct cagcagcacg agcaccagcc 25320 attttaccgt agtaatccgc accaaactga ggcttgatga tgtcctcact cacatgaata 25380 agcacttcac gtggtgactt accacctaac tcaatttgac ctacgtcttt cgtatggcgg 25440 tcagacgcta acgcaatgaa cttaggcaac gccataccca catgcttcgc agcatcacgg 25500 tacagggcat ctttcactgc taggttaaag gcattcatca ggtaatggtc acgtagcatt 25560 tcagctagtg tatccttacc aatacccggt gggccattca gaattaaaca tttaatcatt 25620 gacctttgct ctcccaatgt cgttgcattt gccagtaatg ataagccgtg tcaccgtcct 25680 tagcttcacg actcatacgg gcacaccact ctgagggttt ctcaccactc gccattattc 25740 ttgctcctta gaacgtgcac gcattttacg agcggctgca cgacgggcac gttctttctt 25800 cgcacgagca gcacgcttgt cttcctcagt cttatgaccg ggatacatta agccagttcc 25860 ctctttctcc gtgctttcga ggtagtctgc taggcgacgc agagcaggga tgatttgctg 25920 gtaatccatt ttaccaacaa tccaacgccc tacagcgttt gcaaccttgc cctctcctgc 25980 gttacaggaa cggcaaagcg caccacgaat ctctcctgtc tcgtggttgt ggtctacaac 26040 tacagcacct ttagtactga actgtaattc attcaagcac aggggacaca tgcccccttg 26100 ctcagcccca agtttgcgcc caatggttcg taactgggcg cgagtaacct ttcgtaaagt 26160 cttagatgaa ctcattagct agtgcctcca tgtccatatc ttccttaatc tggttgaacg 26220 ctgactgcca catatcgcgg tcagtatcgc gcagctcaat catggtggct tctagttcag 26280 cacgtgtctc aggtttgctg tccttgtacc acaactctaa ggcacgctca tacaacttta 26340 ctgcatggtt tttactaggc ttcttcatct ggattatcct gtttatgttt gataaccgcc 26400 atgtgatacg tatgcaactg ctccagccag ttgcgatact cagggacagt tacgacttcc 26460 ataaggtagg cgtatccaga atcatcagca gaccgacgta accagagcat ttctgcctca 26520 gctagtgggt cttggcgagc agcagcatac ttaccaatga tggtgttagc tactgttgtc 26580 tcatcatctg gcgaaccagt gataaggtct gttaagaatt cgtaggcttt agttggccca 26640 cacaacttac cgtctaagcg gtcaatgcca cggatgttat cggctgaatc tcccatgagc 26700 atttgtgccc agaagaacgc cgtgccgtga cctttgagtg gcatgttctc accctcgtac 26760 cacttcacat aaccgaagcg attgtcgatt gtgctggtga tagcgagttt ctgctcccag 26820 tacggggcac gagtcatgcg caaatctttg tcaccggaac tgataagccc gttcgggtac 26880 agttcctcac cacgctgtat catcaggtca tcggcttctt cctcgaacgc ccaaatcact 26940 gagataccat caggatgctc atgcagtttc atgttctgca cagcttgctt caacatatca 27000 agcagcggtg gcttctgctt acctttgcgg ttagcctgat acggtttgaa tgttggatac 27060 aatggacgca gacacttagg tgagttgctg tctgtgcaat acacctcagc ggttttacag 27120 ttcgagtaga acatttcctg tagaatcagg gagtggaagc gtcgaatcgc agtcggcaat 27180 gtcttaacag ttgcggctgc ttgatacaag taaaagtccc catcgaggat gaggacttga 27240 cctgagtcaa cgatagcgaa ctggtcgtcc aaggcatcca agtttaaccc atgaatgcct 27300 gtcatgatta tctatcccca gttttaatat aattccacat atcacttgct aagcccttaa 27360 agtctcggaa tgtttccggt acgaactcaa caacgaacca tgtgagggca acgaggatta 27420 gaagtgctgg cagtgctaag atgaacaaga gtgccagtag ggttcgacaa agcttgtact 27480 tgttcaatag ttcaatatca aacaccgtct taccttgccc gttcagccat tcgtaccacc 27540 aagctacaag caccaagtcg atactaagcg ccatgaagtg tccaacgcta ggagcagcag 27600 ccataacatg aatatcatac ccaatgacgg caagtagtgg tactgccaga aaccgcttaa 27660 aagccacccc cacattaaca ctcttgaatt tatcaacgac ttcattgcac ttcttcttta 27720 accatttaaa cattattttt ccttagtacc caaagttagg gtatttcttt tcgaactctt 27780 ggcgagtaag acgctcggcc ttacccacat gaatacgctc acctgttaag acgatggtgg 27840 cttcctctcc atctaacttg agattccagc caccgttgat aacccacagt tccccgttgg 27900 tatcgtctgt gcacagtagc ggtgacgcag catcagggaa gtaggtcaac attataggtc 27960 agcttccttg acgaacacac cgtcacgcat ttcgcctttg cggtctttga tttcaccgta 28020 ggcgaagttt acacactcgt acaacgttag gtcgtagtgg tgcgcaatga tgtacagtgt 28080 acagtacata cgacgtaggt gctcaatagc aatcacagcg tctgctagtg actcgtcatt 28140 gtcgatgtcg tagttataca acgaaccgat aatgtccgac gtctgagcat tcagcgtacc 28200 aataatacca gcacgattct tctcagggtg gagtgcagct actgcttcaa gctgtgccag 28260 catctgagtg tagtcagagg tgttcactac gtcttcgaag tcagcaccaa gctgtgccgc 28320 aatgatgtca agtactacaa cggagtcacc gataccgtct ttaagcaaat ctagtttgcc 28380 tttcacgata gcggtaatca cttcgttcat ttcttctgac agcttacaag cctgagccag 28440 tggggtacag cctttcagaa tgtttcgagc accagcccac tcacggattt taccgaatag 28500 ctcacgtaag ccctcaccgc agttgtatgc tactacttga acttcctgct cggctacgtt 28560 ctgtgcttgt ttctcatcca tttttcagtt ccttatcgtg ggttgatgat ttgatggttg 28620 gtcattgtct tgaagtcttc aagcataacc acgttgtcgt cgttcatgat gccagagtac 28680 tgtgccatgt gagcgccagc atcaccgccg attactagca tgtagttcgt catagcccat 28740 gagcggtact tctgtttaag accaccggac ttgttgatga tgctcaagcg gtacacacga 28800 ggcaacactt taccacggcg gaacacctta acggtttgct ccttgtagtc cacggtaata 28860 cgctcgatgt cggtctggaa cgaatacgct ttcttaatgc gctgtgcaaa ttgagtgatg 28920 ttcattctgt atcctcagtt aaggggcggc ttgcaccgcc cggttagtga ttagactgct 28980 ggtgcgtctg gcattgttgg cattgcaggt gcatcaggca ttgctgcttg agctgctgct 29040 gcgtggtctg cctgtgttgg tgctgactca gctttaggct cgtctgctgg tggcattggt 29100 agaccaccct caagtagtgc ttgaagtggt gaaccctcgt agtccacagc tttcagaatg 29160 tcttcttgaa tgaagttctt agacttacct ttgtcatcag taccatcaat gtacaacgag 29220 tcccaagtct ctttagtcgg gttagaccat aggaacactt ggattttcga agtgtcaaac 29280 tcaggtagtg tgataggctc acctgtttct gggtcgaact taggaagcgg tgacagagtt 29340 tcgaatttca tcgtgttgta ctcacgacca tctttctctt gcacatccag ttccatacgg 29400 aaacacttgt tcagaccttg agcaaggtgc ttcaatgtac cagtgtagtt catgcggtca 29460 aacaacttct tagcgtttgc tttctcgtgg ttgctgactg gcatcttcaa cgagcgaatg 29520 taaacttctt caccattcgg gccgtagaaa cagaaaccaa cacgagcgtt cagtgcagca 29580 gggcgaccag ttggtttacc ttggtgcgtt ggtacaactt taccgtactc gatgtactca 29640 atcatgcggc agttgtatgg gcctttcggc aggatgattt tctcaccacc accagtgcca 29700 gtctcggtca tatcaactgc ttgagtttca acagcagcat ctacaagtga gttcaattgg 29760 tctagtgggt taaatggagt agtcattaat attacctcag tactgttagg tttgttgtct 29820 ttctccctgt acgtgactct ttaaagaggg gagactcaca gggagaacgg tttgtcaagg 29880 gtttcttttg aaaggctgac ttagccctgt aactttttaa ttattatttg acgagtggac 29940 aaatccatgc taggctcggt tcacccgccg cgaggttcca caccctatta ctctctcaac 30000 tcgtctaatt tagacaggtc gtacataaca ggaatgcaat catacttcgt acacaattca 30060 attatcttat ccacacgtcc agtacgcgct gtaccaagca ctagaggata accctgtacg 30120 aatgtatcaa ttgtgtatag atgcccctta acgaagcgca tatcgtctgt ccagacacga 30180 tttaattcgc aatcaaagtc agctaccgct tcgttcatgt taggcttacg actgaacaag 30240 acatcaatct caatgaccgg atgcttaatc ttgataccac cgtacagctt ctcatcgaag 30300 tcactgtcac agttgtcagg gtcatcagta taagcaagga acacttcgct ggctatgtcg 30360 agctggctca gcgtaacgct catacctacc atccagttga acacctcagc ggtacatgag 30420 tcagggccga ctgccacaac gatgtcaagg tcttttggtt tgaagcctcg tgccatatct 30480 cgcaccgccc caccagcaac cgccacttca tatgggcggt tgatagtaga gtgaataagc 30540 tgagtgatgt cacgtgctgc aatatctagt acatgcttat tcatacgtgt tcctatggta 30600 gttgagggac gacagtcaca gcggttgact gcacttcgaa ttcttccggt aggtacttct 30660 cgataccgtc ccaaccgtca gttaactgtt taagcgtctt gaactgctca agggcttgtt 30720 tgtacacacg ctcttgggtg cggcgttctt cgtgccactg ttcaacatca atctcgtact 30780 tgtcatcagc actcttatct ttcaagtact gtaggtacag tgggtcgtcg ttgtaaacta 30840 ctgttgtggc gtttggtaca cggagttgaa cagtaccatt ctcaccatca cggaagtact 30900 cccaaaccgt acccagagct ttacctgtgc ggtcttggat gtacaggtgc agtgatgagc 30960 tagtgtgcag tgccgttggt gtgaggccgt tctgtaccgc tttctcaata cctgagcgtt 31020 tgtaccacag ctcttggtag gctttaacga aagcagctcg tgcattcatt ttaggagcgg 31080 ttggcgcagg atgtttagct ttccactcag tcatgatgtt gttcagaatt gcacgacgtt 31140 tgtcagcgtt tagtcggata gtagccatag tagaatctct cttagtgaat gtgtgttttg 31200 ttgtacattg atgaacccat ttccgcagca gcagggaatg gtacgtcttt caagtgacca 31260 tagttaggcc acagttgcag gatgcggtct ttagcacgct ccatgatgtc tttcacagct 31320 agtgcagcct cacgccccac ttcttcgttc gcagcgtcta ggtacgcagc atcgtgaacg 31380 ttcgtgatta gacatacttg gttgtcgtac cagttcttag caatcatgtg gcgcagaatc 31440 ataccgaacg caaccgccat gaggaagaac gcctcgcctt gacaccagta gttagcaagc 31500 tgtgtatcct tgtagtccat gactttttcc ataccacgac catcggcacg cttaacccaa 31560 cgctcaacct gacggaagct gtaacgcata ccagcaggtg atgtccagta accacggcgg 31620 tacagacgga cagaaccatc gtctgtcatt tcacgcttca agttacccgg taagttacca 31680 gtgcgctcaa cttcgtcacg gattacctga cggaacttag ctgtatcagg gaatagtgcc 31740 gcctcgttgt ctaggaacgc ttgagcgaac tcgacagtac aaccagtagc gaatgcgata 31800 ccagcggctg acgcgccgta ctgggccgca aaactaggtg ctttgattgc agttcgcata 31860 gacttccaca gagcgtggaa ctcgaagccc tcgtcatgac agcgacgata cacttcttcg 31920 tagtctaggt tctcttggaa cgcgagacgg taacagtgca tatcagtacc agctttaagt 31980 agctcaagta gcttcatatc accagtatgt acaacgccca ttacaacttc tagcgcagag 32040 tagtccacct caatgatgcg accatcttcg ccaaagcgtg aggtaaacat ttctttaaca 32100 cgagacttcg cgttgccgtc cttgtcttcc tcagcacgtg gtaggttctg caagttcggg 32160 ttcgatgcag acaagcgacc tgtcactgtt gcacaagtgt tcagtctgtg gtggatgata 32220 ccagaaccgt caggcttctc agggataacg tactgtaaca tcccactaga tttcgtgatg 32280 ttaccgtcat cgtcgtattc atggcgcaga tagtaagtac ctgtgtcttt ctcaagcgtt 32340 gccaatcgag aaatatcact agcgaactca aagccctgct tagctagacc agccatacag 32400 tcaccacttg agctgaacac tggtgaacca tcacacagtg tgcggttctg gcggaactca 32460 ccacgctcac cgaacttgtc ttggaagtgc tcaggtagct ccgagatttt acacaagccc 32520 ggacactgat acacagcttc gccccacttc atcttaagtt cgttcgtgtt ggcacggaat 32580 accttacgta agcccttgtt tttaccggac ttatagatta caacgtcggc tggacacagg 32640 ctgtacttgt cgtacgtctc aggtgtgatg taacgcaacc cgtcaacagg gtcagtccag 32700 acctctttga ctcgcagtac gtggtcatgt agttggtacg catcgacctt gacatacgtt 32760 ggcttgtcat agtgtacttt cttcttgtac ttaatcgggc caccgtaaag caacgccgac 32820 atgtggaagt cagaacccca gttgaagtca atttcttcgg gcatgtcgtc cggtaggtac 32880 gttctgaggc gagtggttag ctccgcaatc tcagcttcct gcaccgcttg gttcttctga 32940 gccactggca tgttcacgta cagaccgaac cactcacaga atgcaaagct tagtaaggca 33000 tccatacgtt cccaaacgag ggggaactta ccttgttcgg caagtgtcgc acactgacca 33060 tagaagcaca gggcggtgtt tgcaacgtca ccgttcggcc ctgcgaggta ctcatgcaag 33120 aggatggggt caatctcgga ggtaaggact ccttgttccc aaagcagttt aataccatcc 33180 actttatgag ttccgccgta tttaggggct gtttcatcga gactcggata cagggaagtt 33240 tggtcagttg caagatactc tccgtgcatg gtacagaaaa ctctaccccc acgtcgaaga 33300 aagccctcaa atgaatctcg tgcgaattgt aagaaccatg aaacttcata agcggcgtta 33360 tgtgcaacga tgagccaaca gtcttcggga atgggtagcc actcagttgt tggagcgcca 33420 acgaaagcct ctcgtgagtt gtatcggaca ctgccgacgc tgccaatttc tgtagttccg 33480 tctggctttt tgcggtcaac acggaacccc gactcaacaa tatagttgtc tggacagtac 33540 ggactggctt tactaccgtg atattcatgg ttctcagtct ccaagtctat aaataatacg 33600 ctactcatgt tctactccat cccaagccgc ccagaacatt ccgttcgact gtacttgttc 33660 gccacgtaga cactttctaa tgcccttgcg acaccaacca tccagctcag cctgtccgat 33720 actgtgatag tagtgctcgt caccagtacg aacacattta cctttgattc gaatagtgtg 33780 gggactacga ttagcccccg ctgctggatg aaacacacgt ttcacctttt ccattaagcc 33840 tccatagccg attctgcgaa gccacaattg aattgacacc gttgagcgtc gaacaggact 33900 tcgaactggt tcacgctcgg tctacctgat accggacact tgttcttagg tgtgctaagc 33960 cctcgcaagc cctcagtatc gttgttaagt ttacccatga acagtgccac atccagagca 34020 ccttgcacac caatcttaga ttgcttcaat gctgtcagcg gtgggtacaa catatcgtaa 34080 ccctcagcac tgaactgcat tgtaccaacc atcaggaagt catgctcaca agccagcgaa 34140 cgcagctcct gccacttcgc ttctaggttc tggtgctctg tctcagcagt accaccacgg 34200 atgtttgcca ccatgtcgat gattaccaca gcagggttca tttcttccac cagcgtagca 34260 atctgaggca tggtgattga gtgtgcatcc ttgacacgga tacgatgcca gtcgcccacc 34320 tcagctaagt acagcggctc gaactgtccc tcgcggtgca gctcacgaat ctctgccagt 34380 gttttaccag ttgctgcttg gtagatacgt ggcacagttc gcacagccat tgattcgttc 34440 actagccaca ggataggtct gtcaccataa cgctcagggg cacacttcat ttggttagca 34500 aagtcagcag cgattgcagc cagcaatgat gtcttacctg catcaacagg tgcggcaact 34560 gccactgtat caccgccacg cagagcacgt atattatgac gaagctgacc aaagcggcgc 34620 aatttgagac cagcgccctc ttcattcgct gcaagtacgt cgtcgagtcc acggtcttcc 34680 catgaaaaca actcgttctg agtcttgaca ttatcaccga actcccgttg aagtttcttc 34740 atttcataca tcaggtcaat ttcttcacct gcttgatact tcgtgaggat gcttgccgct 34800 cgtccagaat acgccagctc atgcatcata gttacaaccc catcgacact ggaatcgtca 34860 accgcttgca cttcatgagt cagtgctttc atggttgcca gttcttcggg tgtgctctgc 34920 gtggctcgta acgaaaccat actattgaat gccgcaaagt ctacttcgtc gtgctctgga 34980 taggtcttcc agtaaagacc aacccaacta agcagtgcgg aagtatccgg tgctaacatg 35040 tctttaggaa cgtaaggcaa taagttgctg tagcgtgtct tgtcacacaa tgcctttacg 35100 ataactgcgt ccataatacc tctttaatct gtgtcggttc acaatctttc gggtctttgc 35160 cgtcgattgt caggggatgt acatttgcga atggtcgtag tactttctga gctttccgtg 35220 ccccgtcaac tcctgcctta tctccgtcgt accaacagag aacgttcggc ttactgccca 35280 tctgctccat aagggaggct aaaagggcgc gggcggttag agagattgta gtgccgagcg 35340 ttgatacgac cagtacgcgt cccttgaaga acaaattcgc cacgtactga acctttatcg 35400 cactcaacgt gtcctcggta agtaccacta gcgatggctg cgtacagtcg ggtttcaagt 35460 tcactaatgt ctggaagttg ctcccatatt gaacccactt cggcgttacg ttcggtgata 35520 cactccgccc aagaaccacg ttctcgaagc ggaagattat ccgccctttg ctctctgacc 35580 actctaagcg acatacgtcc tccaacatgt tcggtgatat acctttgctt gtaaggtatg 35640 aaaacacaaa gtgctgtatt tccgcgctcg tctgactaat gcataaagca tctgcgggta 35700 taggctgcat gcctcgctca tgtacagtgg catcaaggcg tacatatttc ttatactcct 35760 tggtggtctg cttacagcgg ttacaccaca tttcccacga ctgcggattg ttatacaaga 35820 tagccgctgc ggtagcacca cagcagcgaa agcgtctagt ctgaccaatg gctaaacgct 35880 tacaagctct taaccacggc gcgtccatca ctgtcttacc ttttcgtcac tagacttaac 35940 acacagtgtg tactgaacat acgcagtacg gtcaaggcgc ttactctctg agatacaagt 36000 cgcctcgtct tgaataggca cagcaaccgg agcactacca ccccaagtta aagcaatgat 36060 taaccaatac ataagcacct cctaagtgca gtttaagtta ctgaatagtt ttgcgcatca 36120 tacgcactag gtcatcaaga gtacgcccct cgtcatccac agcgcggcga atgtctttac 36180 cgatacaacc acgcgctgtc tgattacctg caaggtacag gatactagcc gccagtagtg 36240 gatgcgtctg cacccactca agcatttcta actcgccctt agcaccgaac tcaagcagca 36300 cctcatcaat gtctgtggcg ttgtcaaggc tattcaagta atcgtcaagg tcaaagtcat 36360 catcaaccca aggcacaggc ttaggtacag gacgctcacc cactggtagc tcagcccaag 36420 tctcaggctt caagcagtgc tcaagcgtat cgtagaactc attcaagttc aacgtctcgc 36480 ggtcagagtg ttcactgtag taacccacac cgatgttggt acactctggg attgtgtcaa 36540 cgaacgttgc tgagtcagtg tagatacctg ttgtgatacg agacttcgga cggtcaaagc 36600 gttcagccag agcttccacg aattcatctg aggcacagcg tccaccaatc tgagtatgga 36660 tgatgtcagt gccacggcgg tcaaagctga tacaccactt cactttctcg aacgactcag 36720 tatcatcttg gtacgctccg gttgaaccga ctcgccccac ttcttcttcg gcaaagaacc 36780 agtactgccc ttgaatgccc tgcttaatca tcataatcat aagcacaaca ccagcaccat 36840 cgtcagcacc aagacaatcc cagataccat cggcatccag tgctaagtgc ccattaagga 36900 cacatagctt tttcttttta ggggtatgcg gttcatcact gtcggggata ccccccatcc 36960 ataacgggat ttgaccatca aagtcaaccg tatctgtgtg gcttgtgaac atgatgtcat 37020 cacgctcacc cacttgtaca aacatgttcc catgcctgtc cgtcgagtac tcaataccct 37080 cagcatcaag cacttcggct accacttctt gcccacgtaa accgggacgt tcttcgctta 37140 ggattgctag tagtagttcg aaagcttttg tgttgtttgg gttagtcatc acgactcctt 37200 aattgtagta gttattattc ttcgtcttcg cggtggtact ctaaccactc gtcaatgact 37260 ccaagttgtt ccgcatcagt tgcagtcaaa cacccacctt gcaggcggtc ttcgaacaca 37320 cgcttaccag tcgccgcaca ctcatccatg cggtcttcgt gcatcgggcc gaccagtgga 37380 cacacttcca cttcatcgtc aaggatgtac tcaccagtgt actcagagta ggtcacatca 37440 ctttggtggt ggtagtcgtg gtcatggtca acccacacgt aatggcggtc acgacagtgc 37500 tcacacactt gacgttcata accaccctca tacacagtgt acatatcatc ttcacgagtg 37560 cggttgtcgc attcgtaaca cgcttctagg ttgatttcaa tgcagccaga tgcgctctgt 37620 gcttcgtatg agctagagta accgtcacac actctccagt actgaccttt cgcaccgtca 37680 cacagctcta cgtgttcttc ataaccgtca aggtaaggca taagtacagt atcagtacta 37740 tttgggtacg gaatgtaacg cagctcttga ccatcaagac agcttgagtt gtggttgtaa 37800 ccagcgttca tcaagtgtcc ctcagtgttg caaccgtatg agcgaacgta ctgcttgtcg 37860 tcaatgttac acacagcgcg accagtgata ttaatgaata catccttagc agagaactcc 37920 ggtgtgtcca gctcaccctc actagccagc acgttgaatt cttctacggt cagctcacgt 37980 ggtgaacggt agcacacggc tagaccaagg ctattgtctt catgacagta cacacgcacc 38040 ggacaacgtt cgtaagggta gtcggtcata cacgaacccg gcccgttctc gtacacaagg 38100 tgccagatat ggtcatactc aggttgagcc ggaatgattg ttgtatggat tggagagttc 38160 agtgctttca ttgcttgcag tgcgtcacgg taatcagaac catcagcgta caagcgtggg 38220 attggtaggt cattgatgat aagacggatt tgcacacgcc cctcagcagt tactacagcg 38280 gtaggcttag cagtgacgtg cttcacttcg ctctcaccgt cccagattag tccgataggg 38340 gtcacacctt ggttctggaa gaagcagttc caagtacgta aaaagcgaac aactaagtca 38400 tcaaggtgtc tgtgcggatt gccgaagtgt aagtcgtaat ctctgtacca acaatcgtgc 38460 agcacttggg ctaggcggat acctttgcag atactatcag cacctaagaa gacaaggttt 38520 gcacagtatt cttcggcaaa agctaataac ttagggtgcg tgaaccctaa aagcttagcg 38580 acttctgata cgtaggtgct taattgatgc acttgcattt catatgacat ctcagcatcg 38640 aagtccacac ccttgattac attaccctta gcgtgaacta acttattcac agcctcgtga 38700 cgggcttcca tgaatattat gtcaggggct ttagtaccac taacttgcgg ttggatactg 38760 tcgtaaccgc catcagtcat gtttaagtag ttcattgcat agtgtactcg tgcaatacgc 38820 tgacatgctt tgaataccca accacgatgg ttgccctcgt acatttcgaa cttctcacgt 38880 agcagtgtag tacgtggtga gatagccttg ctgccacgat tccagccgaa gtttaaccaa 38940 ccagcaagtg agtcaaacat gcctcgttgg tggtgcgctg taagtacttg agtgctgtac 39000 cagtcagggc atagaccaat catcaaggct tctaccttgc ggtcatcagt gccgcgttgg 39060 ttcgagataa agtgacgtaa actaattgtc tctttaccaa ttgccgttgt cacttcaata 39120 tccccacctt gtggtgaggt cttaccgtag tgtggcaagt gctcgtacac cttaccgctt 39180 gcaccaacga agtccacgtt ctctgaggct acacggctgt agaaagctgg catcacagtg 39240 aagcgttcaa ccacattatc agattgctca gctagtaact tgtaaccatc agggtcgtta 39300 acgccgaaca gttggattgc ttcgaatggt tggaaaaact tagacatttg gaactcctta 39360 catgagtagt atcagattag tcaagcatgt cagattgcca tgcatttgct atacgttggg 39420 ttgcctcgaa cattgcactg aatgcctgtt cttgaccgtt ggttaactca gccagtaaat 39480 cgtactgacc atgctctttg agtagcttga gagtacgctt agcatacgtc tcagagtaga 39540 aacgcaccgt gtctagtact tgagtaccat cagttaaggt gcggtgacac agcttgcgct 39600 caagtggtaa cagcttgcgc agtttagggt gtaccagttg tagctcttgg tacgataagc 39660 ggaaagactt ttgttcactc agcataaacc aaatgagcgc agttgaatca cgcagggtta 39720 acttctcacc ttgctcaaga ttctttcgga taatacggtg gatgtgtcca ctgttgaagt 39780 tacgcgctac catttgatag attcgcatct aattctcctt tgaacagaac accacggata 39840 ttgaaacgct tgcagtaagt gataagagac aagcccacta cacgagcacg ctctgagtaa 39900 atatcacgca gcgtttcttg acgtttacgt gtgtggttgt ggtacttgtg gaaagcatct 39960 tgcagtttca agtgctcatt ctttaggtac tcagcgtgtt cctgagcatt catgtatccc 40020 ttggttaaag ttgccatgtt aacatctcct agtacgggat atgattgtgt gcagcataca 40080 cacgattcac gtgttgcggt gtaatgtcta gcatatcggc aatgacttga ctagacacac 40140 ccctttgttt catcttaacg agtttgcgct tactctcggc attaagtttt gaaccggagc 40200 gttttgcatt ccccttgatg ccgttgtaac ggatagcgtg attaatctcg ttgtacgtca 40260 cgccaagata atcacacaat tctttcttag agagacgccc gtaattgtta cggacgtact 40320 caagttgttt agagtttagc gccattactt tgcctttggt agaatatctt taccgtcacg 40380 agcacgcttg agcatggttt caacttcacc gtatgacaag ccagcctcag cccactgtcc 40440 caacaggcgc gccatttgac ctttcaactt gtcttgtttc tggtctagtg atagttcctc 40500 agactttttc ttctctgagt tctcaagcca gtactgctca aggattgggt agaactctgg 40560 gtacgtcatt tcacggttta cacctagttt agccatgatt ttatcggctt tctcgatgtc 40620 acgcttgtaa tgacctttga tgcccttgct aggctctggt ttaacccact tagtagggca 40680 taggacacgt agagttgcgt caagcatagc agcgtcggta cgtgaccagt tatccatcca 40740 agcaacgtta gacttcacta gcgcatggta cagtgcagag attgtcacat tgcggcgcat 40800 agtttcagct ttaacaagtt gagtacggat ttgttttaca gctgttagtt ttggtgcttt 40860 gtttgttgtt acagttgaca tagtttagcc ctcgtttggc gattaaaggt tagtacagat 40920 tagtacgcag attgcattga ttgaccaccg tcgaacagtg atacattagt gagagatagc 40980 ttgtaaccac ctcgcacatt attgatatag ctaacgtgat gcgctagacc gtaagcctcg 41040 ctacgagttc tcgcacatac ttggaagtgt ttcttttgga tacgacgcag accgtcacga 41100 taaaccactt caaagtggta aacacggcgc ggtctaatga aagtcaagta gaaactcttt 41160 gatagttgct ttaaacgttt gaacatagtt cactccaaat aaaataagcg gtggaacaag 41220 ctaccctact agcacattgc acacaaacac ccatagactt tgaccaatgg ttaaaagatt 41280 gcgcttggta ggactgttac accgcttcgt gctagattgc acaagattac atttgatagg 41340 actcagcgaa catatagtgt cagcaaatat gaatgaatac actaagtcct atcaaatgac 41400 caatgctatc gggattggtt gccgctggct cagttgttgc cagtacattg taagcggaaa 41460 actgtatacg gatttcaagc ctgactactt tacacttaca attcaccgga tgcagtggtt 41520 atcagccaca ttacccgtcg tctaactctt acttttaact ctactcgctt cgcttctcgc 41580 tgacccctta cagcagatac aagtgtctcc gtatcgcttg cagatggtgg caataactgc 41640 tctcgcttga gtgttagacc gttgacctaa ttgttaaaga acgatgccct attgggcgga 41700 gactcgctgg ctagtgccgt ttgctctgca atacctcgca aggtttcaga gtgtcccgca 41760 ggactggcta gtgccttgag tcgttagctc tcgctaacat gctttccaat gtatcactaa 41820 gttgttcaac ctgtcaagcg ttttgtgatt cagtgttgag tcagtcggtc acgctgtaag 41880 cggtaattcc aacactttac aactaagcca gcaaacgccg ctttgcattg ttaaacctta 41940 gcatcaaagg cacttagttg tgactcactg agtacgctat ttaagctaca ctacctagtg 42000 attcattgtc aagcatattt ttaaagagcc aagctctata cagtctagct ggctaacacc 42060 ctttgactct cagtatacaa gcaatgtcac cactacttga ttcaccttct actcgtctgg 42120 tggagtcacc cgctgctagt cttattgcca ttgctagtac gttgtgctca gatgttaagc 42180 tggactatat ttttaaagag caaactctat tcgattgtga gtgttgttgg ctgtctactt 42240 ggtttctctg agtgtgcagt cattacccgc cctctgattc attaacatta gcttagctat 42300 cctatctcag cggctacccc tttcaaccgt tcacccctcg cattaccaag agtttacgcg 42360 gtgtttattg tgagtagtca ccgttacaat cgaattttta aagagcgttg tcagcggcta 42420 gtgcctctcg acgggattaa ctatcccata gctagacaga gaacgcaagc ggtatgacta 42480 gaaatgtgag acgtatcaca ataaaagcgg tatatacagt ggttatacag tggtctaaca 42540 atgagttatc cacaagcaat gacctattcg catgatgttc tatccggtat aatgacctta 42600 gcccactgac agtaggcgac cgacacggag cgtagcgaag tgtcccctgt cgccgtatgt 42660 ccactgacta acactagcaa gcattcccta ttgtagcgac ggagcgaagc gaagcgagag 42720 cgacaaagcg caaatcaata ggatagcaac tagatagcat atggatagca gcacgatagc 42780 aagtgttaac aagtggatag caagtgagta gcgcatcaca tcgcagatgt catatgctaa 42840 caagtggata gccagtcgat agccacacgc tgacaggcga caagccagtg agttaacagg 42900 tgagcaaaaa aggcaagcgc agagcacagc cgcgccgccc acgcagccgc aaggctttcc 42960 agtcgctttc ccctgctttc ccacggatag ccacacgcac gcacgcgcac acgctggata 43020 gcccatgcat gcacacgcga gcgcacgcac gccagcgcac acacacgca 43069 <110> Optipharm.CO., LTD <120> Novel Vibrio specific bacteriophage VP2 and antibacterial          composition comprising the same <130> 1.59P <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 43069 <212> DNA <213> Unknown <220> <223> Bacteriophage VP2 <400> 1 cccctacggg ggaaacgcgc gcgtggatgg gggagaaccc cctcgcctga gtataacatt 60 tttcttgtgc agggttttac acgctttccg ctaggaggcc aatggctttc acaggctttc 120 cccatgtcac cactggcttt ccattagctt tccactctct acacatggac acttcgtgtc 180 agaggcttaa ccctcgtaca ccgcgttgtg ttcagcttgc ttcgtttcac ttcgctgcgc 240 ttcaccacga actgccattg ctaagaatgt acgctctttc tctagctcag cctcatacag 300 cttctgcttc atcaggtacg cttcaagagg ccacactttc ttcatggcat ccaccttagc 360 ccaatgctct ccaaccgcct tgtcgaagtt ctctgggtca acacaagctg attcacctgt 420 gacagtgaaa ccattctcca gtgttaacac acagaacgtc agagtagtac ccggcacaac 480 cacgtactgc tcagacttaa cctttgactc tacgagttcc tgtgttacct tatgttcaga 540 catccatctt ccccattact tctttgtatt tatcagagcg gtaatcccat cgggctttct 600 caccgcgtgt atcaacgtgt acaccccagt cgtacagacc gacgccaccg ttagggttca 660 gagctaaagc cgtgtcatac agcttctgga gcttctggaa gttccctcgg tctttgttca 720 gcaagtgcaa gtccactgca tcaccatgta agtgacgaga gttctccgca ccacctacgg 780 ctttattgtg tactgggcag cgataaccgg attcaacctt tagcggctct ccgaagtgct 840 cccgtagcat ttgtaccagc ataagtaggg ctaggtctag tttcaaagcc ccacagtgct 900 gacacgcgaa cttcacaatc ttgaaattcg cagtgtcttt aattagcata attactccta 960 ggctttattg ccaattaggt tgaactcaac aggttgacca atgttgtcct ctagtagcgc 1020 atacgcagca gcagtgaaag cctcaccctc tgcgttctta ggggtttccc aacggttagc 1080 acctgtcttg ataagtgtga tttcctgtgt acgaccaata gtgaatgtgg tgaacaggtc 1140 taagttatca gccagcgtga ggcgtaggaa cttagataca ctgttaacct caagggcgaa 1200 cacattagcg tttgtagtgc cgtctgtgac gtaactaggc gtaagactac cgttgctagg 1260 gttcatacct agaacacctg ttacaatctg tgcagggacg attgtgtggg tgtacgtaac 1320 accagcatcg actgtcaatg cggtactctc agtaacagag ccacctagtg tttggatagt 1380 acactcaatg ttagctgtac ctgctgccac accagtaacc agtccggtac ttgacacagt 1440 aacaaccgca gggttagtgc tattccatgt cttagtgcca actacggctc caaccggaga 1500 tatatccgta atcgtagctt gagcggttgc accaacctcg ataggtgtgt ctagcacgac 1560 ggagaagctc actagcggta ctaaagggtc aacgcctagg ataagctcta cctgcccatc 1620 acggagcact gggtaagcct cacgacattc ttggataagc tgtgtgagga cgtcttgtgg 1680 gattgggtct gggaagtccc aacgcagcaa ctcgttagcg cgggtacgca cagcagcacg 1740 cacccacgga atcactggta atcccggtac tcgatacgga attacaattt gcatacaatc 1800 tcctacattg cagctagctt agtagcgacg ttaacggcgg tctctaagag cttaccgtct 1860 ccacccacta ggtacactgc acccacacct gctaaaaagt acagggtctt tttaacaact 1920 ggggcaactt ccttaaaggt cttcaacatt tcagcggaat cttcacgcat ctgcttcaaa 1980 ctgtccgcaa gttctttctg tgtctgttcc atagcgaata accgtcgttc gtggtcgagc 2040 aggatttctc gggtagtttt atcctcaaag tcctgcatga agacagcctc tttatctgcc 2100 atcacctact ccttacattt tatttttcct tgcgacgaag cacaaagtcg aagattgtac 2160 gtagaagacc gaaccagtct tcgcgtttaa atgagtctaa tcgtttgttc atcatagtct 2220 acctcgtttg cggtgtcggg agttcaagcg tgatagtgtg cctgtaccta ctccgatacc 2280 agttcgttta tgaagtggct tgccagctac cgataggcgg cttgcagtgc ctttgttgcg 2340 tgggcgagtg tctacgcctt ggtaagccat tgggttcaga atgaactcgg ctacacgcat 2400 ttgtagtgct ttctcagcgg ctgaacgttc gtcctcaact aagtgggcat tcagttcttg 2460 aaccagcata gcgatagcat ctgcgcggtc atccttagcc agacagcctt tgtcgtacgt 2520 gatgtcctgt aattgtagga acgcactgta ctgccaacgg cggtcacgtg gatacgacat 2580 gcagcactcg atgtccatgt ctaatgcaga cgtatggaca actaagcggt gacgacgagt 2640 aaccggagag atagtgtcga tgatacgacg ctctttctgc gtactgttac gtaagtcgcg 2700 cacgccaata tgtgcaatgt cacgttgcgc tagggcgttc tggaacagca ttgtaactgt 2760 accgtggccc atgttgctct cgataacgat gtccttaacc tcgaagtcct cggctaagtc 2820 aatcaagcgg ttcatgtttt cctcagacac gccaccttgg aaaccacctg tgccgaacaa 2880 gtgcacgtac gcactagcag caccaccaat tgcaaaggag atttcatcac caccacaacc 2940 agcagggtca attaccatga ttttgtgctt gtacgggata atctcagtcc caatgctcaa 3000 aggagtgtac aagcgtgcgc caaggacacc atcatgcacc tctgggtaaa gggcacgttt 3060 atccgctgtc caactgaaca tgtccggtgc agagttcgaa tcaccagcat ggataatcat 3120 atcagatagt ttgatacgcg tacgcatcgc gtctgatagg gttgtatcca gcatgtactg 3180 taatgcgaaa ccctctgggc cgaagtccag ctctttctct atcaacgcgt cctcgtcgta 3240 acgctgaggg tcagtagttt ctccaagcgt accatcaacg ccgaagccag tgcgtcgtgc 3300 gccctgctca atcagcatct tgatgtacgg ggctaatgtg tcgccgtact tttcctcttg 3360 ctctacagac gggatacgtc cacaccacac tcgtacttca aaaccacgcg agggtagtgt 3420 cttatagata ctgtctttag tctgaggtgt acccagataa agcgtatcac cgtgcgtgtt 3480 gattgccgca aagtctttcg agattgtcaa caagtgctca cgctgtgttt gagtcagacc 3540 gttcttggtc gtctcaatat catcggggat tagcaagtca gcacgcttac cctgtagctg 3600 agcggtgata cccacacaag ccacggatgg tgctttctct agcggtttca agtcacagtg 3660 aacatcgtaa ccctcgtatg aggtacggtc gccacgagca gggtcggcac gtagccaaca 3720 cagcaagtcc catgtctcaa tcaaacggat aacgagggtt gcaacctcgg atgcctgttt 3780 ctcaccaccg gatacaatta gtacacgtgt gctctggtct tggataagac gccacacgct 3840 gtacagagca gcaagcgtac tctttgcctc accacgctgt gcacacacca tcttcttacg 3900 agggccgtgc tgcatgtact gggcaatgtc tgcttgcatg tctgttaagt cgaacccaag 3960 gaatcgcata ccaacgtacg cgaagtcctc gaagttcttg aatgtgattg cgaacatcaa 4020 cgccagctcc tgacgtgtgt ccactgggat gttacgtgga ttttcagcaa gctttttaca 4080 gcgttgtgct aacaacttta atcgtgtaat cacctgtaca gataaaccca cgataacctc 4140 cttagataat acctgctaat tcatcatcgt ccccatcttg gagacggcct agcagttctt 4200 gtttacgtgc ctctcggcgt tgcgcgagtt catcctcgaa ctcgtcggct agttcaccca 4260 tagcctcatc atctgcgtcg gcagaaatgt tgttatcttt gaggaactta gcaatcactg 4320 acttatcggc ggctggaagt ggaatttctt cctcacgcga ttgcttcaat tcctcaatca 4380 aagcgtcagt gaatgcagcg tgcaacgccg ctagacgcga acgactagca gcttttgcca 4440 tgtttactcc ttaaatacgc tctaaccgga tgtcagtcag accctcgtat ttattgtttg 4500 ttgtgttacg acgcaaggat acccgttgtt ggctctgcgc aatgaaagtc aactcatagg 4560 tgtcaccctc gaacattggg atgtcgatta catcattgcc cggctcgaac gtcaccaact 4620 gcaaaccagc ggcttgcgtt gtagtacgtg ccaccttaat tgtcatcttg taacgagcgc 4680 cttgcactaa gttcaagaac tggaattcac cgtttcctga gccttgcaat ttgcggtcat 4740 tacgccactc caatcggtac ggtgtgatgt actgtggtgg gttcttacca gcggctagat 4800 acgacgttgg caaccacgtt ccatcggcat ctttcggcat gtcatcgtca tatggcttag 4860 gtagctcata tggcggtggc agctcgttat agaaaccgga gaagaagttc tcccacttcc 4920 agtctcgcca tacgtcagca gggtatgttt cacctaactg gcgcactggt tcatctggga 4980 acgggatgta gtggaagtag tcaacgtcaa tcgagtcata aatgtagtcg gcactatgtc 5040 cagcccaacg gttgcccggt gcgcgtggat accaaagggc aatccataga cgacccggaa 5100 tgtcagggat gtgtgtggtg ttagtccatt tcaatacacc gtcaacgtag aagtccacac 5160 gtttcgcagg tttccctgct ggtgggttct gttctccagt atgccagtcg aaacgaatct 5220 cgtgccactt accgtcactc aacgcctcat ccacccactt ctcgaagaag tcagtgtatt 5280 cagagaagta gttcgggtca ctctctggta aatcccaagt acgtagctca ccctcccaag 5340 tgttcagacg cgctttatca gccgatacat cctccatatc cacagcatct ttcaatgcag 5400 tcgggtactc gatgtcaatc tcgtggttac gcacaatgta gtaaccatcg tcctcattac 5460 cttgagcgtg taagccctcg tccaagaagc tctgccaacg tgggtcgttc tcgtagattt 5520 cctcgtagtg gaatgtccac aaggctgtcg ctgcaccttt ccggtacgga gagcgcaact 5580 tacagcggaa actgcccggc cctaagtact cacgtgtaac cagtgctgaa cctacctcag 5640 tcttacgacc atcaggggca gggtgtccga tacggtcaac accgatgaca tcaccatcgt 5700 agtacgtacc atgaccacgt agacgtaaca cacctttcac agcgtttgct acatcagagt 5760 agtcagggta aacctcaacg ttctcacgca ccacaccacc gttcagtagc gcggtttggt 5820 ttcccacaac gccagtacca ccccattgct tatgtgcaac gagccagttc tcacgcattt 5880 caatcaagtc agagaagtcg gaatacaact cagtcttgcg gctaatgtca cctttgatgc 5940 gcactggcag cgtgttactc acaccgtagt tggtcttgta ccggataaag tactgaccgt 6000 caggcacatc aggcagcgtg taaatacaac ggcggttatc ctgctcatgc ggccagttca 6060 ccagagaact cgatatatcc tgccacgtgc tgctgccctc agcttgaatc tcaatgttca 6120 agtaaggctg ctcgtagtag cgcggaatca cgttctcgtt gaacgtacca tgcgtgtcga 6180 tgtagaaacg acggatgtca atgttaccaa tccacgactt gtcgtaaaca tcagtaatag 6240 ggacgccagt cgtgcccaca cactcagggc cacagttgaa taatacactg tcaaactgtt 6300 taccccacag cttcaatgag cacgaagctg actgttcatt cggcgtacct ttatcaactg 6360 taatcgtgca gttgtggttc gcgtcctgcg ttagttgcac ttgatgccaa ccactcagac 6420 ttgctaattg cacacgtgcg tacacgtaag tactgttgaa tcgacggtag tacatcaact 6480 caccagagcg tgatacatgc gcaccaaagc taatctgagc actcttttgt tgaccagtgc 6540 cttgttctgt ccaagttaca ttagagccac catacagcag ccacatgcgg tcttgtccat 6600 caacgaagtc acccaatcgg aattgtagtg accaaatgtt cgggtagatt gcagtcggga 6660 ttgtacccac gaatgcaccc acaccccagt tataaccctc ttgaccgggg cgagtaactg 6720 tgtaaagata ctgagcattt gcttgtcgca gcttacacat acggtaaatg ttatcaatac 6780 cgtaagggtt cgctgtaccc tcgtaaccga agttaacata gttctcaagc gtgtcagcac 6840 cattactgta aagcttgtat tgtgagcctg tgaagaatgg gagtgggtct ggtagttctt 6900 tcagaccatc accaaacaag gctacgtcgc cacgataagg cgcacctgtg aagtaacgcg 6960 cgttatcaag gctcactgtg cctacagagt caggattatt gctataggct aagtaccagc 7020 ggttgttgtt accaatccag cggtaaagca cttcgcggta ctggtacact gggtcgtcgt 7080 cttcaccgac attcactgca atgtactgtt tgccattttc gtaccaaacc tgattctgac 7140 cgtggtaatg cttaggggct aaaccctcta gtacatcagc catagatacc tctactcttg 7200 gtctttatcg ttagatgagt taacaaacgc accattgtcc acatcaccaa tcatcaccca 7260 ttcctcgtac tgaaccggaa cggtgtcagg ggatttagtg gagatagtga ttgggcctcg 7320 gcggttcgta cccttaatgt tctcaattgc ggttacacgg tctgcaactt ctttcaattg 7380 gaggaacgtt acagcgtctc ggttctccgt accttgtgcc atgttacgca ctcggtagcg 7440 gtgcatatcc aagtcaccat agaaatcgct aatgccagaa ccctcagtca tttcttgact 7500 taggtacagc aattgcgtgt gcgcgtcgtc taagttagct cgggtgaact cagcaccctc 7560 agagaagata tgacgcagcg cagtgcggtt ggtacgccgt gtaagcgtaa cctctacccc 7620 atcagctaac ggggcgttgg taaagttaat ttgtgttcga ccctgccaag tgtaatctgt 7680 accctcgatt gcaggtgtct catcaagcat aacgaaaatg tcacgcgtgt caaagaattc 7740 aatgctgagg acgatgttgg tcatagtacc gtcgctcgta gcacgttgta cacttaatgc 7800 cataaagcct ctttaaaggg ggattactcc cccatgttgt tgataactgc acgtagcggc 7860 atgaactctt gcgcgaacgg aagaatcttg ctcatacgct gtgtgtccat acgaccatcg 7920 ctgaccatat cctgcaccgc cccgatgctc tgcgtcacgt aacctagtga ggtcagtgag 7980 tgtcgcggag attcaccaag gaatagcggt gacagcaaag tgataccacc aatagctgac 8040 atgttcatcg cagtgttagt gattagttca cgtgggctct gtgcctcttt accgtccaac 8100 gcaagcttag catgggtagc cataagcatg agagggaact ggtacatcat aagcatcgca 8160 agaccagcgt aatcaccatt ctgcgtgtat cggcgcagta gtttgttagt agccgcaatc 8220 gcaaaggatt ggtagccaat cacaatctta cctaccggac tgaattgtgc aaagtgtgac 8280 gtctcaccag tacggatgtt ctgcatcact gagtccatag cacgtgtcga tacaatctcc 8340 aactcacgtt gaatctcgta cgggaacacg ccagtcgggt tctcagcata cgcagtacgt 8400 agcttctgtg tcaactcagg tgtcagacca tgctcagtta gagcgcggct tgtaccacct 8460 ttcaacacgc gttcgacttc atcacaaatg ataccagcgt tcaagttcgc cattaggcgg 8520 tgaaccatgc tgaaaccgtt tgctagtcga gctgcttgcg acaggttctg agttgtctgg 8580 aatgcagcac ttgtgtttgt caagtcaagg ttatcgtcag catatgtgtg gatgtgacgg 8640 tatcgtgctt cgttctggta cgcaccattc aagatgctgt acagacgctc acgagagcca 8700 gtatccgcaa caactgcgtc agccttacgt aacagaccag tgcttgctaa accacgcagt 8760 acacgcgcta aaccgaactc tttcattgcg atactaatgt cagtcatctg gtataaaccg 8820 gagttcttta gcatcacagt gttagcaagg ttgcttgcac cacgcatcag ctcagggatg 8880 tcagtaccga ctgggtagcc tagcaggtgg tcgattgtgt cgtccactgc tttgccccat 8940 gattctgcac cctcacccat acctagtcga gtctcagtga ttgaacggtc aagtgcacgt 9000 aaatcagtga taccacgatt cgcaaatgcc acacgacctg acatacggtt tgtgtaacct 9060 tgcatcaagc caagtgtgtt agtctctaga atgtcagata aacgtagcac atcaccatca 9120 atgtcgaact ctttgttcaa gttcaagtca agacggttac gcaagttctt cactggcgca 9180 gtgctgcctg tcttcggtac gttcacacgt aggaactcat tgattttgtc gtcttcaacg 9240 ccagcatcac gcagagccat aaacaactca tcgttatcca taccttgcat caagttcttc 9300 cattgtgggc caccgttacc ggatacgcca ttcacaatgc ggttgtacca agagtcggct 9360 acacgtgcag ctagcttacc gtccatgtta gtgtaagtat cctgcaacgc agagcgcagc 9420 agtttcttaa ctttaggagg ttcgatgtta cgcactttct cataactata ttgacgaggg 9480 atgtagtact tcgagcgtgg gaactcgtct gcactaatag taccggattt cacaaggtgc 9540 tcatgccaag actcagccca accggaatcg gtgtaggctt tcaccaaacg ctcaatgtcg 9600 gctggtggtg ttgggatttc gcgccctgct ttctcagcac tgtatgcttc atgtaggtac 9660 ttagtcagct tgtcttcttc cacactacga gctgtacgag ccttagcacg gttaaagaac 9720 atatcaaagg tgttaacgcc gtgcttctca cgcatagcag ccacaatcgc atcttcgacc 9780 gcagcagcac gtaggttctg ctcaagagtc aagttacgtt tatggtctac cacactggta 9840 gcacgttcac ccatacgggt agcatcagat acaagtaggt tcgctaggtc ttcactacca 9900 cgagcaatgt tgtcccagag tgcgaacgac tgcttcaaag atttacggac actctgctca 9960 agacgtttag cacgctcaac cgcagtagca ccacggttcg catcatctgc aaggtcagcc 10020 atgctatcag agaagaaagc attcacttta tctaggttat cagcttggta gttagcaacc 10080 ttaacaaagg catcaaccgc ttcatcaaga gctgagtctg tacctttgaa gcctaacatt 10140 ttcagaatct tctcagccac agcttgcaaa gcattctgac ctgcacggac acccggtaac 10200 ttcgctaggt ctttcaccat ctccggctta gacgccaact ctgctaggaa ctcacgattg 10260 ttagtcatag cgtagctgta cttacgagag aactcacggt tgtttttcag tgaggcatgt 10320 aaagcatcaa ggtcacggat agcctctgca atctcaggag caacattacc tttacttgct 10380 tggaataaag tcttagatgt cgcagcatgg ataagctcat gcacagcaac gtctgcatct 10440 acgtggttca acatatcgcc cacggtctta aacactgtac cgtctttcgc tgtagagcgt 10500 agtttcagaa actcagtgtt agcaccaaag gtgtaagatg agcgacgacc agtagcattc 10560 attaaacgca cttccatacc agagatagta tctggtagtg catccataag agcacgagct 10620 gagtctgaca aactatccat agtacggagg tgtgtcacta agtcttgtgc tttaatttga 10680 actgtcttac gtccttgtcg agaggcaaca attcgaccct caacaggtgt gtcctgtaat 10740 gt; acaggtgcgt cagggtcagg cattcgctgt gcagcgacct ctggttttac cacatcaggc 10860 acgatttctg gagcatcact taggacaccg ccagtctgtg ctttacgtgc taaacctgtc 10920 tcagcggtga acacgtgctc agttgcagaa cccaaagcgc ggctctcaac ctctgctgca 10980 ttcgctgctg cacggacacc gcgagttgca cgcgatacgt cgtacagttg gtcaacaccg 11040 attagacctg cggcaaccgc tgtaacccac tcagcctgac ctagttggtc ttgggcgtag 11100 taagtagttg ctaactcacc agcacgcaat gcagcacgca cagccatacc agtacgacca 11160 accacacctg ctgcacccat aggagcaagt aggaacggcg cgtcacctgc taagttacca 11220 gcgataccag cgaacatgtt ctctgcttgt acaccatcac gcttgcgttg gtcttccaca 11280 gcctctttgc ggtactggta ctcatcaagg ctacgtgagc caccgatgaa gttcaattct 11340 tcttctgagt accctagaat gcgcagagag gtatccgctg tcatagtagt accaaggtca 11400 aaggatgtgt cctctgggaa gtccggcatc gttgctttac gcacagccgc acctacgata 11460 ctgttaccaa cacctgctgc gaatgattcg aggccagttg ctttcggtgt ttcaaggcgt 11520 ttaaggttcg cctcacgctc cgcagcgttt aaacctaaat cggtagtgtc gtcccatgct 11580 agtgcaccag cgtccatatc tacgccctta gcattagact ctaagccagc accctgtgaa 11640 gatgggcgac ggttctcaaa aatagccata gtaattcctc aaagtataat gataagggag 11700 gggcaaatgc cccaccactt agtaataggt tgggtacgga ttcttagcag gttgattctg 11760 ttgaagccaa cttgtcaagt cagtacgtaa ttgctcttgt cgagtctttc ctgattgctt 11820 gtacgcagga gtttgcatca tagcgttcat cactagcata cggttagtga ggttgtcgaa 11880 ttggctcaca cccttagcgt aatcagccat agccttagcc acagggttcg cgctaccacc 11940 accaccatga tacgcaaggt caaccagcac agggtacgca gcatcactca actcaagacc 12000 agcagactta gctgctgact taacaattgg gatgtacgta tcttctaggt cgcccttgag 12060 cataccgata gcttgcggga gtgttacctc atcagcaatg gccttaccgg agttagtgtg 12120 acgtccgaag cctacagtct cgccaccacc tgctttcttc ttcttgccaa ggaagccctc 12180 agattccatc agattagcgg ttagcttgcc gaacacaaac gggtcgatac cagacttgtt 12240 cacaccagtc acgttcattg tcacagcgcg gttattcgtt gcgtcataga acgtagcagc 12300 ggttaggcca ctttgttgtg aaccaagacc tgcggcagtc ttaacctcgt catcgtacat 12360 cgtagttgca gcctgttgca gagtgctgcc tgagatactg taacgagtca caataccgtc 12420 ggagtcttcc acagctacaa ccatacccat agcgtcacca cctgctgcac cgatgtcaac 12480 agttacaccg ctcacatcag tctgaatgct tgacagcaca gaagccactt ggttgttcat 12540 agctttaacg aaagtctcct gagagccttt gtatgagcca tagaagtcgt ctagcttctt 12600 acccacagga acgtacgtgt ggatgttgtc accgcccacc tgcaactcca cctgacgcgt 12660 acgggcatct gcataagcag ctttctccat agcgtccaag tcgttagtca gctctgggcg 12720 tttctgtgca atgtacttcc agctctgtgc cacagcagta gcaccagcag tcacctgacg 12780 gtcactttct gtgccgaagt caagccagtt gttcagttta gtaccatcaa tcaaagcgtt 12840 aggcattgtc gcgttctggg cgttgtactt accagaagca atagcctgag cattgcgtcg 12900 caggtcgtcg aacaggatgt tactcgcgtt ctcagggttc tgtaatgcag cattacccat 12960 cgggccacgt aggttctccg gtaggctctg gagcatcgtc tgcttaccaa tggcagtact 13020 tttgctgtac atcatcatcc atgtggcaac ctgcgcctgt tgttcctcac tgacgacatt 13080 atccttgcct gtgaagtcga ttgtcgctaa gacctgacca gtgttcgtac tcatacgttt 13140 agccgctagg gacgccatgt atgagtcgtt agcttgtgaa gcatatgcca tcattgccaa 13200 gttaccttga gcagtgtctg ggaagttctt ctctagctct ttacgagtct tgtctaaatc 13260 cataccgagc tgaccagcta ggacagttga cgggatagca ttgctcagtg cgagttggaa 13320 agtgttcgct tgacgcgctt tcttctccgc ctcgttcaga cgcttcacat acgaaccagc 13380 agtaccagcc tcgataatac cctgctgcgc cttgtcgtcc acgaagttac gtagtcgagt 13440 caactcagcc gcttgttcat ccggtgatag tgctgctaga ccgttgatgt ccgtttcgaa 13500 actcaccata gcctgtgcag cttgctggtt tccggcacgg ttgtactcag tgcgtaacga 13560 cttaataacc gtagggctta gcacgcccat ttccttatca accacgcctt gcagcatgtt 13620 gataatactt ggctcatcag tgcgctgtgc aactgtctgt acgtacgctg tggcacgttg 13680 gattttgtca tcctttgaca agtgtgtaga caggttaatg ctgcgaagac ctttcgcaat 13740 actagcctgt gccatttctg gtgcacccga gtccacaaag ctgtagaact ccgctgctgt 13800 gctgtctagg ttgctgtcta acgcacggtc agcagctacg ttagccattg cagcggagtc 13860 tttctggaac ttgttacgag ccgctacaga cgtttcctgt aggtcttgca acaatgccat 13920 tgccgattgc ttagggatgt gtgggaggta cttactcatc tgactggcga acgcagcgtt 13980 gcgctcttgg ctcagcttgt cgaagtactc taagtcctta ccctgagata cggcattatt 14040 gatttggtcg atttgctcct gacggaactt gcccatttcc tgattcactg ctgcactcaa 14100 gtagcccgat tggtaggact ccttgaatag catgttcacc ttgctgagct tttcatcttg 14160 cttgctcatg gcgtcagtcg tggcagtggc gtctagcgca ccttgaattg cggcatctgc 14220 tgcggcttga ttgtacgcac gttccactgc tggcacggct gtttgcagga atccagcaat 14280 cagcgcacca cttgcgtcag gctcagcctg tacgttctcc atacgtggag aggccgcacc 14340 aatcaacagg ttgccagttg cagctccgag ttgtagcgga ttgctttgga ttggcatatt 14400 atctccttac caacttgtag ttattggttt atccgaataa ctgaaattaa ggtcaatttg 14460 gtcaaaggca ctacgtcccc accaatcgta atcagcgtcg ttctttccta cacttgggac 14520 ttcttgtggc tgtgtctgac tttgtggggt taactggctg aaaccgtagt tcgcagcagc 14580 gcccaccatg ataccagcag cctgcccgta tgcagcgttc atgatttgtt caccggatga 14640 tgccacacca ctttgtagag cattactgcc ctcgtcggta gtcttacgca acatcatgtt 14700 taaaccctcg ttcgtgatta catgttggtt ctcaactgta ccttgagcac ggtcagcgtt 14760 gaccatgact gattgcgcag cgtcacgtgc actaacgccc acagtgtccg tggcagcagc 14820 catagcacgt gtttggttag cagcagcgtt cttctgcgca ttgatattaa atagcgcact 14880 agcggtcttc tggttctcaa tagagcgttg caagttaatc tgcgacacag actgagctgt 14940 gctgagcatg actttcttgt tgtactccat gatagcctga tttcgagcta cggtggcttt 15000 gttctccgcc ttagctgctt gaccaccagt gaacagtgct gacgccgcac ctactacagc 15060 accgatagca gcaccccaag ggccacctac cgatgcaccc atagcagcac cagacgtagc 15120 accacctggcg acaccgccac cacttgcacc tgccatatta actccttata cgcgacgtcg 15180 gcgttggttg taacgaatga tatattcaat ttccagaatg ttcatatcct gtgtaccatc 15240 agtactcaag tacatttcag tagactgcgc attgcttcga cacggcacag ggacgcgtgc 15300 taggtcagca agtaacgcca tatctggctg aacgctatca gtaccaaaca caaggctagt 15360 tgtgctatct gagtaatcca atccgatact tgggtcaacg atgcgtacat cgaactcacc 15420 agtatttcga gttgttaact cataacgcaa cagacgaacc ggagcagtac tgattagagt 15480 atcgttctgg tctttcagca ttggaggtgt tggtgcgaac atagactcgt accggaaacc 15540 tagaaagatt tcttcatgtg ttgagcctga ttcaacagtg aactcccaag tgatttcgtc 15600 tacgtcaata gccacttcct cagaggcacg tgcaccagtc gctacagcac cagtgagctt 15660 gccctccgct acccaaggac gcatatagat tggcacgata cctttaccct cagtcgtaat 15720 aggtacacgg ctcattgcgt ctagatgtgg taatcggtca acatcaaacg tcacaccgcc 15780 ctgtttaggg tcgatagttg tgatagtgat tttgtcggta ctaccgtcgt catctgcggc 15840 aaacaacaca acacggtcac gagcaaagtg cagactagcg acacggtacg gaaagagcca 15900 tttatgccat gcttggtgta ctttaccttg gttcgtgaag tgatactcgt gtgcaatcac 15960 ttgacggttg tcaccagtag ttgccattag cacgatgtta gcagcacttg cactctgcat 16020 gaaacgagcc tcgccctcaa tgtaacgggg gatatgagtc gtaacgtctt gagatacgta 16080 ctgagacgat gtgtattgtg atggtacaag ctccagaact gcactgaatg cttcgcttcg 16140 aggtgcaggg tacatcaggg tctgtgaggt tgtaacaggt gctacaaacg cattacaggc 16200 caaatctgag gtcaacacca cactagcgtt atcaggcgca agtaactgtt gcagggatgg 16260 tactaccgcc tgtgtactgt caccaagcag aatcaagtcc ttgttgaact gcaacgcttg 16320 gcggaacact gagttctgag ccgaaccgga tgcaatatca atacggtctg ttgggtcaag 16380 ggaactcacg gtagagcgga agaaacggtc tggttcgcca gtggcactca tacagacgta 16440 tgcaccagat agcagcacta agcggccttg gaacgtaccg ataccagtga tgcggcgttc 16500 ttcaaggaat gtcggtgcag ggttagttaa gtcgtcacct gcgagacgcc cctccatgat 16560 atgttgctct atgttgtcat cgacaatctt atagggcaca tccacgctaa tagcagtcac 16620 ggagttgtag tcaccagatt ccaaccacac acccttttcg gcgctgtaac gataccacac 16680 aagtgcacgc tctgattgac ccacagcaca caacgcacca tcagcagacg ggtgcagacg 16740 cgcagggagg tcagtctcta ggtttacctg tgactggttc gagtagccaa cgtaaggtga 16800 acctgacgta ctggtgattt taaggtcagt acctgtaatt aattcaaagt agatgtacgg 16860 gccgacacgc acagcgaagt ccacacccac agcaattaac ttctcgacca gcttacgtgc 16920 aattgcttct ggcacagatt ggtctgcgtc acctgctgtt gtaccgtcag gtgtagtata 16980 tgtcacggtt tggctaccga gcgaccacac tacggacagg tcgtactctt tactgaacgc 17040 accactcttg acgaagtaga aacctgttgt cttcgggtct ttcttgtcag agttgtcagt 17100 cgttgcagac ggacgtttct cagtgttcag aatgtacgtg ataccaccca tactggtagt 17160 ctggatacta cgacggtcag ccgctaggag gtatgacaag ttaccctcac tcacgatgtt 17220 cttagcctcg cggtctaaga gccaccagcc gcctgtgttg gtgttgatta ctaggtgtcg 17280 tccatcagca cctcgttcga gatactgtgt gtacagcgca tcaccctcac caatcgggcc 17340 gaaatcgtgc acactcacga acaaactacc gggacgacgg cggatgcttg ttactgggtc 17400 ggacagcata ttaacttgtg cagagcattg tccagccaca cgcgcacgtg ggacttgttg 17460 cgacacgcct tgcagcaggt cattcaatgc accctcaaat ggtcgagcca tgattcacct 17520 ctatgtatat agtgcagagc ggatacgacg gtaacgccct gtcttgcttg tactgtagcg 17580 gcggttacgt aggttctgct tgtgcagcac tcggaacgcc tcttgcgact gacggtctag 17640 gttctggtac tggttgtcca cacctaaatc gtttgcataa acttgcatag ccgcttcata 17700 cgcgacgatg tgttgggcca tttctggcat gtgttcaaat ggtgtatcaa acacgatacg 17760 caatagcttg gactcatcga atacgatgtt attggtggcg taatccaaca gcaatccgtc 17820 ttcattctcg ccgtacatca catcactagc cgcatcgtaa atgtcgatgg ttgcgctagg 17880 taggtagata tgcccctcaa catcagggta cagcgtcatt tcacgctcgt tgaaccacca 17940 accatcgctc agcaatagtt tgcgtttagt ttcaagagtg tccttgacga taccaatggt 18000 agggttgcga acagtgtcgg atgtgatgcg gtactcacca atcgcagtca acgagatatt 18060 gattgcatct agtaaggtca tattttaagc actccaaaca aaaaagggga gcagccgaag 18120 ccactcccct tgggtattat tgctgattag ctctcagtct ctgttttctc tggtgcacca 18180 gt; gtccacttag cgcgtaggtc gattacttta gccatttatt tgctccttat actgctgctg 18300 gttcttcgaa cttagtaacg attgcagtgt ctggacgacg ctgaccaact gtgtacatag 18360 cgtaacagtc aagtacgtta cagaactctt tctcgtcatc ccagatacga gttgtgaatg 18420 ggtgtgcttc cacagtcaca agtgtacgtg atttagagaa agtaaccatt tgacatttag 18480 catcttccgc agatacgttg aacgaagtgc caagtgggtg agcagcgatt gcaccagttg 18540 ggaactcggt acctcaact actggtacac cgttcatacg aacgacacga cgaccgccgt 18600 agtcagacat atcaacgttg aactcttggt tcagcagttt tggatgctct actagcgcag 18660 agtaaatgtc tggtgataca agggtaattt ggtcagtcag aggaacttta cgtttcacta 18720 gctcagtcac accagccttg tgagcttcgt tgatagagat agcgttcgct tcaagttctt 18780 cttgagaaac cgcagcagcc ttgtaagttg ccagtacttc tacaccgtca ctgaaagcag 18840 gtttcaagtg gtcaggcgcg tcccaagtac gacccttgat gatttggata aggtgcgctt 18900 ggtcgaagac ctcagcgaac tcagagccgt tgttctgacc catttcggtt aggaagtcag 18960 gtgcagtcca gtcgtcttgg tagtcgattg ggttacggat gtacagcact gtatccacga 19020 cgataatcat tttatcgtta cgtacaggcg tgttatccag agtttcaccg gaacgacgac 19080 ctttcactgt tgaagtgttc aagcggtcaa tacggtaagt gttagagcgt tcagcagttg 19140 aacgttgtgc agacaggcca cggaacatag ctgcgtactg gaaacgagtg tcaacttcgt 19200 tttggtacac ttctaggtgg atgtcttggt cggaagcagc accaccccaa tgtgggcgag 19260 ttagacctgc tttgtatgaa gtatcagcca ttttaaatcc ttatggttga aaagtttaaa 19320 tctggagttt cagacgccag aaacgcaaaa aggggcttat cgccccagtt gtttgccacg 19380 tgcgcggcgt gcaagcagat tctgatacgc ttgaccagca gggccacttt caaaagaagt 19440 gttaccaaac ttggtacgta aatccgcaag ttctttcgag aactcagcag ccgacaacgc 19500 agtgttgcca gcttgaccac ccattccatg aatgtgttgg ccttgagttg gtaaagcacc 19560 agactgacgg gcgtagtcta gtaccatctg agctgcgtcc ttggctagac cgctgtttgc 19620 catagtcttg accgcagtct gaatatgagc aggagcgttc gccatgaaca catcacgcgc 19680 ttgtgcccac gcctcagcac caccagctag ggtgtgtact tcattaacag tgcgttcagt 19740 atttgaagca gcgttagcga ggtacgtctc agctaaggtc tttgcgtagc cagcgtagtt 19800 gccgaggcgt tcttggagga acttctcgtc aatcaaagag gcatcaccat aacgcaatgc 19860 attacccata gctcgctcta cgtcggcatc ggtacagccc gatacttgag caagcatctg 19920 taagccagca tcaactgcgg cgtcacctgt ctgtagtgta atctgacctg cggctgggtc 19980 tgtttgctct ggttcagcag gaacttgagg ttctggctgt ggagcagcag caggttgtgg 20040 ttgattaccc agtacagggt cgtcggctgg tgcaggttgt ggtgcagggt tctgcgctgg 20100 ttgtggttgg ttcggaacgt cagcaggttt acccgctgct ggtacgcccg gtggaagatt 20160 tgtatcttgc attattgagt caatccttgc tgttcagaca acagtgcagc agagccagca 20220 tcaagtgcca taccggaggc aggggcttga gcagcagctt cctcagccgc ttgtagctgt 20280 tgcagctctt gttctgtgta gaaaatctca gttgggtcaa caccatgacc tgcgaggatt 20340 aggtcaatca ctcggtcagg gttagtacgt tttgtagctt gtgcaagtac agggagaatc 20400 aagttaatgt cgttcgccac tagacctaga cgttgcacca caatactacg gttgatagcc 20460 gcagtaccta cttgaatgtc taggctaacc gcattagctt gcaacagtgc cagtagctca 20520 ggtcgagcct caactgtcag gatgtgcgct aatgggatat gcagaatctc agcaagagta 20580 gcgtagatgc cacccatgtt ttcttctgct tcacgaacgt tgttcttgat ttcttctgct 20640 gttacacgtt cagcatcacg gacgttacca gtgtacatga acgctcgtgc cagctccgca 20700 agaaccatct gtaggtcatt aacgacctca gcaatcttac gtgcttcacc agcttcatgc 20760 gccttgatac cctcaccatc acctgcaacg tactcaccag tctcagcatc attcaatgca 20820 tcaacgtcag cagtactctt aggcgacacg aggttaacga ctcgcattgc ttcaatctca 20880 tacagggtca aggcttgagt tagctctgac acacgtgcga acgcacctgc atggtcttcc 20940 acatggccac gaccataatg ttcaccagaa accaagttcc acaccgcagg gatgtaagga 21000 catagcttct cagggtaaac actaagtgtg ccaacaggct taccatcaat ctgttgagta 21060 atcacaaaaa cgtcaccagc atcacgcgtc tcgcgttgaa tcttggtgta caaccaaaca 21120 tcagtggtag ggtctgtgta tccggttgga cggaattctt ctggcacatc ctgaattggg 21180 atacgctcac gtagaactgt gcacaacact cgaccagcac cgtcacgacg cacaacataa 21240 tcgcgtacac tgtacacggt catacgtccg gttaacgggt cgcggtacag caacgcatta 21300 ccagtgatga taagtaactt cattagatgg gttaactggg caaagccagc gtgagtagtg 21360 agcaacgtca ttgcttcacg ctcgatacca gtaaacacag cagaaagttg cgcgttctca 21420 ctacccagtt gagcaatgat ttcacgcatc ttatctgtgt ctgagatacg gaagaagctc 21480 atcccagttg ggaaaagcgt ccgtgtcagt ttactagcaa ggttgttaac caacatagca 21540 ccgtgagatt ggaagtcgcg ttgaagtgat acgcgctcac cgtctttgtt ctcacgtgta 21600 aaaacggtag gcactgtcca gaaagcataa cgctctgaac tcagagtaac ctcagagtcc 21660 tcatacttag cgaacagagc ctcgtgagtc tcttgagtat gcatacagta ccttataatc 21720 ctaagcttga actaacgtca gccatcttgc gtttacgagt tgtaagacca agagcgtcgt 21780 cggctgctag atttgcagaa ccgcctgtct caaccttaac tacgtcgtca agagctttgt 21840 tagcttggag tgtgttagct tcacgcattc gagtctgttc agcttcgaac tgtttcttct 21900 gcgcatcagc ttggcggcgt gcttcttttg ccgcatcttt accactatcg aatcccatgt 21960 ttcaccttgt agtacttgag tctgtactcg tactcgccct gcttgtgagg gatgagcacc 22020 cagtccagac gttgtgcctt ggcatagcgc ataatcgcac ggtacatgtg atgtccgata 22080 ttgtggttcg ggagcacgta gcttaataca acgcccacgc ctttaccact gatatgcggt 22140 gtcgtgtacg gtgctgttat cgcgcagccc cctattaagt tgcctttgtg atgaatgccc 22200 atacacatga acccatcaac tgcatcggca actgagtctc taaagtcttt agaggtaata 22260 ccctgagcga actcagggtt gttgtggtac gctaggtttg cgacaactcg cgcacctagc 22320 acagcctttc tacctgtgta gaattctaga gtataattgg acacttcgtg tctctcctgt 22380 tcgtgggact ttaaattagc agaagaagaa tggcgaatct ttaaccagtt ccagattcaa 22440 tgtacccatt ggaggcatct cgataccctc aaggtcagca cctgcttcta acgctgcgtc 22500 acggataacc tcaagtgggt tctggtgctc agtgtataga atcacgaact gctcacgcaa 22560 cacacggtgc aggtcgtcaa catcacaagc gtgagtcgca actgaatcgt ggattggaat 22620 gatgtcaccg tcaaagttca agatggtcat cataaggtgt gagctatctc cttggtggat 22680 gaagttcggt gagataccgg atacagcttt ctggcggtgg ttacgctcat agtcgaagtt 22740 gtacagcacg acttggttga tacccatgct gcgcactttg acacgagtct catggcggtt 22800 acagtagcgg ttaatcacca gaccaccaag cactgtcttc cactgcaagt gatgttccat 22860 cggtactcgt tgacatacag ccttgaggta atccatacca cgtgcagcgg ctggcatagc 22920 ctcattaatg ctgtcacgca tcagcggtga tacgaacgaa cacagcttga atagtgtgta 22980 ctcgtctgta ccctcgtagc cctcatcagc agcaccaagg aaaatgtaat ccgtgcagct 23040 acgtaatgtg gcagagtaga agtacgtcat cgttgggcgc ttggtcattg aacgtgtgat 23100 agggttcagt gtccagtaat gcgcttgcac gattgtctca gggtcacgta gagcaacctt 23160 gattttcgag tcagtacgct gcttaacatc catgtacagg tcggccttgg ttaagttgcc 23220 gtcccagaat aggttcgtta gcttaccgcc gattgggtca cgtagcattg cagagaagtg 23280 ttgcccacca gagttagtgg catccaatgc aaccgcaatg tttgactcgt acttctcagg 23340 gtcaggtgag tctagtgctt cgattagttg gatactagcc gcaagcatac accagaactc 23400 gtctgcgtct ttgaagtctt cgttgttcaa tgggtctttc acccactcac gaatcgcatc 23460 aatgcgttcc tcagtccact tggcacgttc ttcgaacagt ggcttatcga aaccaaaaca 23520 tgttgccacg tgtgatttca gccagaacag accacgctta cctagcggct tcttacgtcc 23580 aagtagtagc aacgcctttt ggatgtcaga gccttgtggg ttcaagtgtg acttgaagta 23640 caagcggtaa cgccagtcaa cacaggtcgg gaagtacaac gcatcctcgt ccatgaaacg 23700 gcggcacagg tcaagcgtga tagccagttg gcgaatctgt gatacacgtt tacgttcctt 23760 tgtgtaccac tgacgcatct gggtcttcca aacctcaaac gcctcttggt catcctcaga 23820 ccagtcagcc ttgtcagtgc cgtgtaggtg gaacttaggc tgtggtttct ggtggtggct 23880 tgggatacca acgtcaaagc ctgtagcctt ggcttgcagt actaagtcgt aaatactttt 23940 gttcacgacg taaggcactt cctgtgcttt gttcaacgct tgacggatac cagtcgcctg 24000 tttaaacgcc tcgttcactt cacgtaaacg ggaacgcttg atgtgtcggt tcgagtacgt 24060 atggcgacga ctcaggtcag ttaagtaacc gccgttgtaa atgtcctcat gacgtactgg 24120 aggaacaatc atcggtgggt agtgcaccat agtgtcagcg tgctcaagta catccgagaa 24180 cacaccttgt agttcttcgg ctggctcagc gtacagcaaa cctttcgggc cttgtaccca 24240 ttcaaagata ccagtctcat gcacacactg gagcaacaag cggcctacgt tgaagcaggt 24300 tgtgttgtcc caagggtcag tttcgagttt cacgttcgca ctagcggcac ggaacgtacg 24360 caagatatga gattgggaac gcgtcttgtg ctctttcaag tactcgtaca ctcggttcat 24420 gtacgcaggt gcaaccacct cagcattctg agctatgatt tctgattgca ctgagcgacc 24480 tagtgatgta cctagagact ggatagacga catccccaac tcagttgagg caatgttgtt 24540 cagtagagtg tttaatgtca tcaccgtcag aacatccgac gacactaaac ggatatagcc 24600 acgatactta gcaccaacgc cacgagtctt agaagctttc agctcatcaa ttcgggcgct 24660 aacaccatca tacgccgcat tcagcagacg ctgcacaggt gttaactcag tagcacggcc 24720 attctccaag gcatccagta cagctttacg gctacgctcg attgcctcag tcttaaacgt 24780 ttcttcaatg cgctcttgac gagcggcttt ttcttcgaat gtctcacgat tcttggtgac 24840 agacataaac actccaatac attacaacgt tgttttcaaa ttgcgctgaa tcataagacg 24900 cagcaacttg cgtttctctt tgtccgtaat gccgaggtac agcttccgtg ctgtaccgct 24960 tctacctgat tgaatgtgtt gggcaataac tgcgtactcc ctgtccacat tcacaggtgc 25020 ttcatctacg aagattaatt ccgcagacgc tccacgatgg ttcgacatgc ggcctccggt 25080 ttaccctcaa tcatcactac gtcgaggtat tcaatcgcag gatgtttcac agttaagtag 25140 ctgcggctgt cgttagcgaa cgtacaacca cgaccataca tacggacaac cagtacctca 25200 ccacccatag ggctgtgctc aatgaagtca gcgacgctaa ccgcttcctc tgggaaacca 25260 cagtcagtga ataccgaaat acgcccatat tctagcacct cagcagcacg agcaccagcc 25320 attttaccgt agtaatccgc accaaactga ggcttgatga tgtcctcact cacatgaata 25380 agcacttcac gtggtgactt accacctaac tcaatttgac ctacgtcttt cgtatggcgg 25440 tcagacgcta acgcaatgaa cttaggcaac gccataccca catgcttcgc agcatcacgg 25500 tacagggcat ctttcactgc taggttaaag gcattcatca ggtaatggtc acgtagcatt 25560 tcagctagtg tatccttacc aatacccggt gggccattca gaattaaaca tttaatcatt 25620 gacctttgct ctcccaatgt cgttgcattt gccagtaatg ataagccgtg tcaccgtcct 25680 tagcttcacg actcatacgg gcacaccact ctgagggttt ctcaccactc gccattattc 25740 ttgctcctta gaacgtgcac gcattttacg agcggctgca cgacgggcac gttctttctt 25800 cgcacgagca gcacgcttgt cttcctcagt cttatgaccg ggatacatta agccagttcc 25860 ctctttctcc gtgctttcga ggtagtctgc taggcgacgc agagcaggga tgatttgctg 25920 gtaatccatt ttaccaacaa tccaacgccc tacagcgttt gcaaccttgc cctctcctgc 25980 gttacaggaa cggcaaagcg caccacgaat ctctcctgtc tcgtggttgt ggtctacaac 26040 tacagcacct ttagtactga actgtaattc attcaagcac aggggacaca tgcccccttg 26100 ctcagcccca agtttgcgcc caatggttcg taactgggcg cgagtaacct ttcgtaaagt 26160 cttagatgaa ctcattagct agtgcctcca tgtccatatc ttccttaatc tggttgaacg 26220 ctgactgcca catatcgcgg tcagtatcgc gcagctcaat catggtggct tctagttcag 26280 cacgtgtctc aggtttgctg tccttgtacc acaactctaa ggcacgctca tacaacttta 26340 ctgcatggtt tttactaggc ttcttcatct ggattatcct gtttatgttt gataaccgcc 26400 atgtgatacg tatgcaactg ctccagccag ttgcgatact cagggacagt tacgacttcc 26460 ataaggtagg cgtatccaga atcatcagca gaccgacgta accagagcat ttctgcctca 26520 gctagtgggt cttggcgagc agcagcatac ttaccaatga tggtgttagc tactgttgtc 26580 tcatcatctg gcgaaccagt gataaggtct gttaagaatt cgtaggcttt agttggccca 26640 cacaacttac cgtctaagcg gtcaatgcca cggatgttat cggctgaatc tcccatgagc 26700 atttgtgccc agaagaacgc cgtgccgtga cctttgagtg gcatgttctc accctcgtac 26760 cacttcacat aaccgaagcg attgtcgatt gtgctggtga tagcgagttt ctgctcccag 26820 tacggggcac gagtcatgcg caaatctttg tcaccggaac tgataagccc gttcgggtac 26880 agttcctcac cacgctgtat catcaggtca tcggcttctt cctcgaacgc ccaaatcact 26940 gagataccat caggatgctc atgcagtttc atgttctgca cagcttgctt caacatatca 27000 agcagcggtg gcttctgctt acctttgcgg ttagcctgat acggtttgaa tgttggatac 27060 aatggacgca gacacttagg tgagttgctg tctgtgcaat acacctcagc ggttttacag 27120 ttcgagtaga acatttcctg tagaatcagg gagtggaagc gtcgaatcgc agtcggcaat 27180 gtcttaacag ttgcggctgc ttgatacaag taaaagtccc catcgaggat gaggacttga 27240 cctgagtcaa cgatagcgaa ctggtcgtcc aaggcatcca agtttaaccc atgaatgcct 27300 gtcatgatta tctatcccca gttttaatat aattccacat atcacttgct aagcccttaa 27360 agtctcggaa tgtttccggt acgaactcaa caacgaacca tgtgagggca acgaggatta 27420 gaagtgctgg cagtgctaag atgaacaaga gtgccagtag ggttcgacaa agcttgtact 27480 tgttcaatag ttcaatatca aacaccgtct taccttgccc gttcagccat tcgtaccacc 27540 aagctacaag caccaagtcg atactaagcg ccatgaagtg tccaacgcta ggagcagcag 27600 ccataacatg aatatcatac ccaatgacgg caagtagtgg tactgccaga aaccgcttaa 27660 aagccacccc cacattaaca ctcttgaatt tatcaacgac ttcattgcac ttcttcttta 27720 accatttaaa cattattttt ccttagtacc caaagttagg gtatttcttt tcgaactctt 27780 ggcgagtaag acgctcggcc ttacccacat gaatacgctc acctgttaag acgatggtgg 27840 cttcctctcc atctaacttg agattccagc caccgttgat aacccacagt tccccgttgg 27900 tatcgtctgt gcacagtagc ggtgacgcag catcagggaa gtaggtcaac attataggtc 27960 agcttccttg acgaacacac cgtcacgcat ttcgcctttg cggtctttga tttcaccgta 28020 ggcgaagttt acacactcgt acaacgttag gtcgtagtgg tgcgcaatga tgtacagtgt 28080 acagtacata cgacgtaggt gctcaatagc aatcacagcg tctgctagtg actcgtcatt 28140 gtcgatgtcg tagttataca acgaaccgat aatgtccgac gtctgagcat tcagcgtacc 28200 aataatacca gcacgattct tctcagggtg gagtgcagct actgcttcaa gctgtgccag 28260 catctgagtg tagtcagagg tgttcactac gtcttcgaag tcagcaccaa gctgtgccgc 28320 aatgatgtca agtactacaa cggagtcacc gataccgtct ttaagcaaat ctagtttgcc 28380 tttcacgata gcggtaatca cttcgttcat ttcttctgac agcttacaag cctgagccag 28440 tggggtacag cctttcagaa tgtttcgagc accagcccac tcacggattt taccgaatag 28500 ctcacgtaag ccctcaccgc agttgtatgc tactacttga acttcctgct cggctacgtt 28560 ctgtgcttgt ttctcatcca tttttcagtt ccttatcgtg ggttgatgat ttgatggttg 28620 gtcattgtct tgaagtcttc aagcataacc acgttgtcgt cgttcatgat gccagagtac 28680 tgtgccatgt gagcgccagc atcaccgccg attactagca tgtagttcgt catagcccat 28740 gagcggtact tctgtttaag accaccggac ttgttgatga tgctcaagcg gtacacacga 28800 ggcaacactt taccacggcg gaacacctta acggtttgct ccttgtagtc cacggtaata 28860 cgctcgatgt cggtctggaa cgaatacgct ttcttaatgc gctgtgcaaa ttgagtgatg 28920 ttcattctgt atcctcagtt aaggggcggc ttgcaccgcc cggttagtga ttagactgct 28980 ggtgcgtctg gcattgttgg cattgcaggt gcatcaggca ttgctgcttg agctgctgct 29040 gcgtggtctg cctgtgttgg tgctgactca gctttaggct cgtctgctgg tggcattggt 29100 agaccaccct caagtagtgc ttgaagtggt gaaccctcgt agtccacagc tttcagaatg 29160 tcttcttgaa tgaagttctt agacttacct ttgtcatcag taccatcaat gtacaacgag 29220 tcccaagtct ctttagtcgg gttagaccat aggaacactt ggattttcga agtgtcaaac 29280 tcaggtagtg tgataggctc acctgtttct gggtcgaact taggaagcgg tgacagagtt 29340 tcgaatttca tcgtgttgta ctcacgacca tctttctctt gcacatccag ttccatacgg 29400 aaacacttgt tcagaccttg agcaaggtgc ttcaatgtac cagtgtagtt catgcggtca 29460 aacaacttct tagcgtttgc tttctcgtgg ttgctgactg gcatcttcaa cgagcgaatg 29520 taaacttctt caccattcgg gccgtagaaa cagaaaccaa cacgagcgtt cagtgcagca 29580 gggcgaccag ttggtttacc ttggtgcgtt ggtacaactt taccgtactc gatgtactca 29640 atcatgcggc agttgtatgg gcctttcggc aggatgattt tctcaccacc accagtgcca 29700 gtctcggtca tatcaactgc ttgagtttca acagcagcat ctacaagtga gttcaattgg 29760 tctagtgggt taaatggagt agtcattaat attacctcag tactgttagg tttgttgtct 29820 ttctccctgt acgtgactct ttaaagaggg gagactcaca gggagaacgg tttgtcaagg 29880 gtttcttttg aaaggctgac ttagccctgt aactttttaa ttattatttg acgagtggac 29940 aaatccatgc taggctcggt tcacccgccg cgaggttcca caccctatta ctctctcaac 30000 tcgtctaatt tagacaggtc gtacataaca ggaatgcaat catacttcgt acacaattca 30060 attatcttat ccacacgtcc agtacgcgct gtaccaagca ctagaggata accctgtacg 30120 aatgtatcaa ttgtgtatag atgcccctta acgaagcgca tatcgtctgt ccagacacga 30180 tttaattcgc aatcaaagtc agctaccgct tcgttcatgt taggcttacg actgaacaag 30240 acatcaatct caatgaccgg atgcttaatc ttgataccac cgtacagctt ctcatcgaag 30300 tcactgtcac agttgtcagg gtcatcagta taagcaagga acacttcgct ggctatgtcg 30360 agctggctca gcgtaacgct catacctacc atccagttga acacctcagc ggtacatgag 30420 tcagggccga ctgccacaac gatgtcaagg tcttttggtt tgaagcctcg tgccatatct 30480 cgcaccgccc caccagcaac cgccacttca tatgggcggt tgatagtaga gtgaataagc 30540 tgagtgatgt cacgtgctgc aatatctagt acatgcttat tcatacgtgt tcctatggta 30600 gttgagggac gacagtcaca gcggttgact gcacttcgaa ttcttccggt aggtacttct 30660 cgataccgtc ccaaccgtca gttaactgtt taagcgtctt gaactgctca agggcttgtt 30720 tgtacacacg ctcttgggtg cggcgttctt cgtgccactg ttcaacatca atctcgtact 30780 tgtcatcagc actcttatct ttcaagtact gtaggtacag tgggtcgtcg ttgtaaacta 30840 ctgttgtggc gtttggtaca cggagttgaa cagtaccatt ctcaccatca cggaagtact 30900 cccaaaccgt acccagagct ttacctgtgc ggtcttggat gtacaggtgc agtgatgagc 30960 tagtgtgcag tgccgttggt gtgaggccgt tctgtaccgc tttctcaata cctgagcgtt 31020 tgtaccacag ctcttggtag gctttaacga aagcagctcg tgcattcatt ttaggagcgg 31080 ttggcgcagg atgtttagct ttccactcag tcatgatgtt gttcagaatt gcacgacgtt 31140 tgtcagcgtt tagtcggata gtagccatag tagaatctct cttagtgaat gtgtgttttg 31200 ttgtacattg atgaacccat ttccgcagca gcagggaatg gtacgtcttt caagtgacca 31260 tagttaggcc acagttgcag gatgcggtct ttagcacgct ccatgatgtc tttcacagct 31320 agtgcagcct cacgccccac ttcttcgttc gcagcgtcta ggtacgcagc atcgtgaacg 31380 ttcgtgatta gacatacttg gttgtcgtac cagttcttag caatcatgtg gcgcagaatc 31440 ataccgaacg caaccgccat gaggaagaac gcctcgcctt gacaccagta gttagcaagc 31500 tgtgtatcct tgtagtccat gactttttcc ataccacgac catcggcacg cttaacccaa 31560 cgctcaacct gacggaagct gtaacgcata ccagcaggtg atgtccagta accacggcgg 31620 tacagacgga cagaaccatc gtctgtcatt tcacgcttca agttacccgg taagttacca 31680 gtgcgctcaa cttcgtcacg gattacctga cggaacttag ctgtatcagg gaatagtgcc 31740 gcctcgttgt ctaggaacgc ttgagcgaac tcgacagtac aaccagtagc gaatgcgata 31800 ccagcggctg acgcgccgta ctgggccgca aaactaggtg ctttgattgc agttcgcata 31860 gacttccaca gagcgtggaa ctcgaagccc tcgtcatgac agcgacgata cacttcttcg 31920 tagtctaggt tctcttggaa cgcgagacgg taacagtgca tatcagtacc agctttaagt 31980 agctcaagta gcttcatatc accagtatgt acaacgccca ttacaacttc tagcgcagag 32040 tagtccacct caatgatgcg accatcttcg ccaaagcgtg aggtaaacat ttctttaaca 32100 cgagacttcg cgttgccgtc cttgtcttcc tcagcacgtg gtaggttctg caagttcggg 32160 ttcgatgcag acaagcgacc tgtcactgtt gcacaagtgt tcagtctgtg gtggatgata 32220 ccagaaccgt caggcttctc agggataacg tactgtaaca tcccactaga tttcgtgatg 32280 ttaccgtcat cgtcgtattc atggcgcaga tagtaagtac ctgtgtcttt ctcaagcgtt 32340 gccaatcgag aaatatcact agcgaactca aagccctgct tagctagacc agccatacag 32400 tcaccacttg agctgaacac tggtgaacca tcacacagtg tgcggttctg gcggaactca 32460 ccacgctcac cgaacttgtc ttggaagtgc tcaggtagct ccgagatttt acacaagccc 32520 ggacactgat acacagcttc gccccacttc atcttaagtt cgttcgtgtt ggcacggaat 32580 accttacgta agcccttgtt tttaccggac ttatagatta caacgtcggc tggacacagg 32640 ctgtacttgt cgtacgtctc aggtgtgatg taacgcaacc cgtcaacagg gtcagtccag 32700 acctctttga ctcgcagtac gtggtcatgt agttggtacg catcgacctt gacatacgtt 32760 ggcttgtcat agtgtacttt cttcttgtac ttaatcgggc caccgtaaag caacgccgac 32820 atgtggaagt cagaacccca gttgaagtca atttcttcgg gcatgtcgtc cggtaggtac 32880 gttctgaggc gagtggttag ctccgcaatc tcagcttcct gcaccgcttg gttcttctga 32940 gccactggca tgttcacgta cagaccgaac cactcacaga atgcaaagct tagtaaggca 33000 tccatacgtt cccaaacgag ggggaactta ccttgttcgg caagtgtcgc acactgacca 33060 tagaagcaca gggcggtgtt tgcaacgtca ccgttcggcc ctgcgaggta ctcatgcaag 33120 aggatggggt caatctcgga ggtaaggact ccttgttccc aaagcagttt aataccatcc 33180 actttatgag ttccgccgta tttaggggct gtttcatcga gactcggata cagggaagtt 33240 tggtcagttg caagatactc tccgtgcatg gtacagaaaa ctctaccccc acgtcgaaga 33300 aagccctcaa atgaatctcg tgcgaattgt aagaaccatg aaacttcata agcggcgtta 33360 tgtgcaacga tgagccaaca gtcttcggga atgggtagcc actcagttgt tggagcgcca 33420 acgaaagcct ctcgtgagtt gtatcggaca ctgccgacgc tgccaatttc tgtagttccg 33480 tctggctttt tgcggtcaac acggaacccc gactcaacaa tatagttgtc tggacagtac 33540 ggactggctt tactaccgtg atattcatgg ttctcagtct ccaagtctat aaataatacg 33600 ctactcatgt tctactccat cccaagccgc ccagaacatt ccgttcgact gtacttgttc 33660 gccacgtaga cactttctaa tgcccttgcg acaccaacca tccagctcag cctgtccgat 33720 actgtgatag tagtgctcgt caccagtacg aacacattta cctttgattc gaatagtgtg 33780 gggactacga ttagcccccg ctgctggatg aaacacacgt ttcacctttt ccattaagcc 33840 tccatagccg attctgcgaa gccacaattg aattgacacc gttgagcgtc gaacaggact 33900 tcgaactggt tcacgctcgg tctacctgat accggacact tgttcttagg tgtgctaagc 33960 cctcgcaagc cctcagtatc gttgttaagt ttacccatga acagtgccac atccagagca 34020 ccttgcacac caatcttaga ttgcttcaat gctgtcagcg gtgggtacaa catatcgtaa 34080 ccctcagcac tgaactgcat tgtaccaacc atcaggaagt catgctcaca agccagcgaa 34140 cgcagctcct gccacttcgc ttctaggttc tggtgctctg tctcagcagt accaccacgg 34200 atgtttgcca ccatgtcgat gattaccaca gcagggttca tttcttccac cagcgtagca 34260 atctgaggca tggtgattga gtgtgcatcc ttgacacgga tacgatgcca gtcgcccacc 34320 tcagctaagt acagcggctc gaactgtccc tcgcggtgca gctcacgaat ctctgccagt 34380 gttttaccag ttgctgcttg gtagatacgt ggcacagttc gcacagccat tgattcgttc 34440 actagccaca ggataggtct gtcaccataa cgctcagggg cacacttcat ttggttagca 34500 aagtcagcag cgattgcagc cagcaatgat gtcttacctg catcaacagg tgcggcaact 34560 gccactgtat caccgccacg cagagcacgt atattatgac gaagctgacc aaagcggcgc 34620 aatttgagac cagcgccctc ttcattcgct gcaagtacgt cgtcgagtcc acggtcttcc 34680 catgaaaaca actcgttctg agtcttgaca ttatcaccga actcccgttg aagtttcttc 34740 atttcataca tcaggtcaat ttcttcacct gcttgatact tcgtgaggat gcttgccgct 34800 cgtccagaat acgccagctc atgcatcata gttacaaccc catcgacact ggaatcgtca 34860 accgcttgca cttcatgagt cagtgctttc atggttgcca gttcttcggg tgtgctctgc 34920 gtggctcgta acgaaaccat actattgaat gccgcaaagt ctacttcgtc gtgctctgga 34980 taggtcttcc agtaaagacc aacccaacta agcagtgcgg aagtatccgg tgctaacatg 35040 tctttaggaa cgtaaggcaa taagttgctg tagcgtgtct tgtcacacaa tgcctttacg 35100 ataactgcgt ccataatacc tctttaatct gtgtcggttc acaatctttc gggtctttgc 35160 cgtcgattgt caggggatgt acatttgcga atggtcgtag tactttctga gctttccgtg 35220 ccccgtcaac tcctgcctta tctccgtcgt accaacagag aacgttcggc ttactgccca 35280 tctgctccat aagggaggct aaaagggcgc gggcggttag agagattgta gtgccgagcg 35340 ttgatacgac cagtacgcgt cccttgaaga acaaattcgc cacgtactga acctttatcg 35400 cactcaacgt gtcctcggta agtaccacta gcgatggctg cgtacagtcg ggtttcaagt 35460 tcactaatgt ctggaagttg ctcccatatt gaacccactt cggcgttacg ttcggtgata 35520 cactccgccc aagaaccacg ttctcgaagc ggaagattat ccgccctttg ctctctgacc 35580 actctaagcg acatacgtcc tccaacatgt tcggtgatat acctttgctt gtaaggtatg 35640 aaaacacaaa gtgctgtatt tccgcgctcg tctgactaat gcataaagca tctgcgggta 35700 taggctgcat gcctcgctca tgtacagtgg catcaaggcg tacatatttc ttatactcct 35760 tggtggtctg cttacagcgg ttacaccaca tttcccacga ctgcggattg ttatacaaga 35820 tagccgctgc ggtagcacca cagcagcgaa agcgtctagt ctgaccaatg gctaaacgct 35880 tacaagctct taaccacggc gcgtccatca ctgtcttacc ttttcgtcac tagacttaac 35940 acacagtgtg tactgaacat acgcagtacg gtcaaggcgc ttactctctg agatacaagt 36000 cgcctcgtct tgaataggca cagcaaccgg agcactacca ccccaagtta aagcaatgat 36060 taaccaatac ataagcacct cctaagtgca gtttaagtta ctgaatagtt ttgcgcatca 36120 tacgcactag gtcatcaaga gtacgcccct cgtcatccac agcgcggcga atgtctttac 36180 cgatacaacc acgcgctgtc tgattacctg caaggtacag gatactagcc gccagtagtg 36240 gatgcgtctg cacccactca agcatttcta actcgccctt agcaccgaac tcaagcagca 36300 cctcatcaat gtctgtggcg ttgtcaaggc tattcaagta atcgtcaagg tcaaagtcat 36360 catcaaccca aggcacaggc ttaggtacag gacgctcacc cactggtagc tcagcccaag 36420 tctcaggctt caagcagtgc tcaagcgtat cgtagaactc attcaagttc aacgtctcgc 36480 ggtcagagtg ttcactgtag taacccacac cgatgttggt acactctggg attgtgtcaa 36540 cgaacgttgc tgagtcagtg tagatacctg ttgtgatacg agacttcgga cggtcaaagc 36600 gttcagccag agcttccacg aattcatctg aggcacagcg tccaccaatc tgagtatgga 36660 tgatgtcagt gccacggcgg tcaaagctga tacaccactt cactttctcg aacgactcag 36720 tatcatcttg gtacgctccg gttgaaccga ctcgccccac ttcttcttcg gcaaagaacc 36780 agtactgccc ttgaatgccc tgcttaatca tcataatcat aagcacaaca ccagcaccat 36840 cgtcagcacc aagacaatcc cagataccat cggcatccag tgctaagtgc ccattaagga 36900 cacatagctt tttcttttta ggggtatgcg gttcatcact gtcggggata ccccccatcc 36960 ataacgggat ttgaccatca aagtcaaccg tatctgtgtg gcttgtgaac atgatgtcat 37020 cacgctcacc cacttgtaca aacatgttcc catgcctgtc cgtcgagtac tcaataccct 37080 cagcatcaag cacttcggct accacttctt gcccacgtaa accgggacgt tcttcgctta 37140 ggattgctag tagtagttcg aaagcttttg tgttgtttgg gttagtcatc acgactcctt 37200 gt; ccaagttgtt ccgcatcagt tgcagtcaaa cacccacctt gcaggcggtc ttcgaacaca 37320 cgcttaccag tcgccgcaca ctcatccatg cggtcttcgt gcatcgggcc gaccagtgga 37380 cacacttcca cttcatcgtc aaggatgtac tcaccagtgt actcagagta ggtcacatca 37440 ctttggtggt ggtagtcgtg gtcatggtca acccacacgt aatggcggtc acgacagtgc 37500 tcacacactt gacgttcata accaccctca tacacagtgt acatatcatc ttcacgagtg 37560 cggttgtcgc attcgtaaca cgcttctagg ttgatttcaa tgcagccaga tgcgctctgt 37620 gcttcgtatg agctagagta accgtcacac actctccagt actgaccttt cgcaccgtca 37680 cacagctcta cgtgttcttc ataaccgtca aggtaaggca taagtacagt atcagtacta 37740 tttgggtacg gaatgtaacg cagctcttga ccatcaagac agcttgagtt gtggttgtaa 37800 ccagcgttca tcaagtgtcc ctcagtgttg caaccgtatg agcgaacgta ctgcttgtcg 37860 tcaatgttac acacagcgcg accagtgata ttaatgaata catccttagc agagaactcc 37920 ggtgtgtcca gctcaccctc actagccagc acgttgaatt cttctacggt cagctcacgt 37980 ggtgaacggt agcacacggc tagaccaagg ctattgtctt catgacagta cacacgcacc 38040 ggacaacgtt cgtaagggta gtcggtcata cacgaacccg gcccgttctc gtacacaagg 38100 tgccagatat ggtcatactc aggttgagcc ggaatgattg ttgtatggat tggagagttc 38160 agtgctttca ttgcttgcag tgcgtcacgg taatcagaac catcagcgta caagcgtggg 38220 cattgatgat gtaggcttag cagtgacgtg cttcacttcg ctctcaccgt cccagattag tccgataggg 38340 gtcacacctt ggttctggaa gaagcagttc caagtacgta aaaagcgaac aactaagtca 38400 tcaaggtgtc tgtgcggatt gccgaagtgt aagtcgtaat ctctgtacca acaatcgtgc 38460 agcacttggg ctaggcggat acctttgcag atactatcag cacctaagaa gacaaggttt 38520 gcacagtatt cttcggcaaa agctaataac ttagggtgcg tgaaccctaa aagcttagcg 38580 acttctgata cgtaggtgct taattgatgc acttgcattt catatgacat ctcagcatcg 38640 aagtccacac ccttgattac attaccctta gcgtgaacta acttattcac agcctcgtga 38700 cgggcttcca tgaatattat gtcaggggct ttagtaccac taacttgcgg ttggatactg 38760 tcgtaaccgc catcagtcat gtttaagtag ttcattgcat agtgtactcg tgcaatacgc 38820 tgacatgctt tgaataccca accacgatgg ttgccctcgt acatttcgaa cttctcacgt 38880 agcagtgtag tacgtggtga gatagccttg ctgccacgat tccagccgaa gtttaaccaa 38940 ccagcaagtg agtcaaacat gcctcgttgg tggtgcgctg taagtacttg agtgctgtac 39000 cagtcagggc atagaccaat catcaaggct tctaccttgc ggtcatcagt gccgcgttgg 39060 ttcgagataa agtgacgtaa actaattgtc tctttaccaa ttgccgttgt cacttcaata 39120 tccccacctt gtggtgaggt cttaccgtag tgtggcaagt gctcgtacac cttaccgctt 39180 gcaccaacga agtccacgtt ctctgaggct acacggctgt agaaagctgg catcacagtg 39240 aagcgttcaa ccacattatc agattgctca gctagtaact tgtaaccatc agggtcgtta 39300 acgccgaaca gttggattgc ttcgaatggt tggaaaaact tagacatttg gaactcctta 39360 catgagtagt atcagattag tcaagcatgt cagattgcca tgcatttgct atacgttggg 39420 ttgcctcgaa cattgcactg aatgcctgtt cttgaccgtt ggttaactca gccagtaaat 39480 cgtactgacc atgctctttg agtagcttga gagtacgctt agcatacgtc tcagagtaga 39540 aacgcaccgt gtctagtact tgagtaccat cagttaaggt gcggtgacac agcttgcgct 39600 caagtggtaa cagcttgcgc agtttagggt gtaccagttg tagctcttgg tacgataagc 39660 ggaaagactt ttgttcactc agcataaacc aaatgagcgc agttgaatca cgcagggtta 39720 acttctcacc ttgctcaaga ttctttcgga taatacggtg gatgtgtcca ctgttgaagt 39780 tacgcgctac catttgatag attcgcatct aattctcctt tgaacagaac accacggata 39840 ttgaaacgct tgcagtaagt gataagagac aagcccacta cacgagcacg ctctgagtaa 39900 atatcacgca gcgtttcttg acgtttacgt gtgtggttgt ggtacttgtg gaaagcatct 39960 tgcagtttca agtgctcatt ctttaggtac tcagcgtgtt cctgagcatt catgtatccc 40020 ttggttaaag ttgccatgtt aacatctcct agtacgggat atgattgtgt gcagcataca 40080 cacgattcac gtgttgcggt gtaatgtcta gcatatcggc aatgacttga ctagacacac 40140 ccctttgttt catcttaacg agtttgcgct tactctcggc attaagtttt gaaccggagc 40200 gttttgcatt ccccttgatg ccgttgtaac ggatagcgtg attaatctcg ttgtacgtca 40260 cgccaagata atcacacaat tctttcttag agagacgccc gtaattgtta cggacgtact 40320 caagttgttt agagtttagc gccattactt tgcctttggt agaatatctt taccgtcacg 40380 agcacgcttg agcatggttt caacttcacc gtatgacaag ccagcctcag cccactgtcc 40440 caacaggcgc gccatttgac ctttcaactt gtcttgtttc tggtctagtg atagttcctc 40500 agactttttc ttctctgagt tctcaagcca gtactgctca aggattgggt agaactctgg 40560 gtacgtcatt tcacggttta cacctagttt agccatgatt ttatcggctt tctcgatgtc 40620 acgcttgtaa tgacctttga tgcccttgct aggctctggt ttaacccact tagtagggca 40680 taggacacgt agagttgcgt caagcatagc agcgtcggta cgtgaccagt tatccatcca 40740 agcaacgtta gacttcacta gcgcatggta cagtgcagag attgtcacat tgcggcgcat 40800 agtttcagct ttaacaagtt gagtacggat ttgttttaca gctgttagtt ttggtgcttt 40860 gtttgttgtt acagttgaca tagtttagag ctcgtttggc gattaaaggt tagtacagat 40920 tagtacgcag attgcattga ttgaccaccg tcgaacagtg atacattagt gagagatagc 40980 ttgtaaccac ctcgcacatt attgatatag ctaacgtgat gcgctagacc gtaagcctcg 41040 ctacgagttc tcgcacatac ttggaagtgt ttcttttgga tacgacgcag accgtcacga 41100 taaaccactt caaagtggta aacacggcgc ggtctaatga aagtcaagta gaaactcttt 41160 gatagttgct ttaaacgttt gaacatagtt cactccaaat aaaataagcg gtggaacaag 41220 ctaccctact agcacattgc acacaaacac ccatagactt tgaccaatgg ttaaaagatt 41280 gcgcttggta ggactgttac accgcttcgt gctagattgc acaagattac atttgatagg 41340 actcagcgaa catatagtgt cagcaaatat gaatgaatac actaagtcct atcaaatgac 41400 caatgctatc gggattggtt gccgctggct cagttgttgc cagtacattg taagcggaaa 41460 actgtatacg gatttcaagc ctgactactt tacacttaca attcaccgga tgcagtggtt 41520 atcagccaca ttacccgtcg tctaactctt acttttaact ctactcgctt cgcttctcgc 41580 tgacccctta cagcagatac aagtgtctcc gtatcgcttg cagatggtgg caataactgc 41640 tctcgcttga gtgttagacc gttgacctaa ttgttaaaga acgatgccct attgggcgga 41700 gactcgctgg ctagtgccgt ttgctctgca atacctcgca aggtttcaga gtgtcccgca 41760 ggactggcta gtgccttgag tcgttagctc tcgctaacat gctttccaat gtatcactaa 41820 gttgttcaac ctgtcaagcg ttttgtgatt cagtgttgag tcagtcggtc acgctgtaag 41880 cggtaattcc aacactttac aactaagcca gcaaacgccg ctttgcattg ttaaacctta 41940 gcatcaaagg cacttagttg tgactcactg agtacgctat ttaagctaca ctacctagtg 42000 attcattgtc aagcatattt ttaaagagcc aagctctata cagtctagct ggctaacacc 42060 ctttgactct cagtatacaa gcaatgtcac cactacttga ttcaccttct actcgtctgg 42120 tggagtcacc cgctgctagt cttattgcca ttgctagtac gttgtgctca gatgttaagc 42180 tggactatat ttttaaagag caaactctat tcgattgtga gtgttgttgg ctgtctactt 42240 ggtttctctg agtgtgcagt cattacccgc cctctgattc attaacatta gcttagctat 42300 cctatctcag cggctacccc tttcaaccgt tcacccctcg cattaccaag agtttacgcg 42360 gtgtttattg tgagtagtca ccgttacaat cgaattttta aagagcgttg tcagcggcta 42420 gtgcctctcg acgggattaa ctatcccata gctagacaga gaacgcaagc ggtatgacta 42480 gaaatgtgag acgtatcaca ataaaagcgg tatatacagt ggttatacaga tggtctaaca 42540 atgagttatc cacaagcaat gacctattcg catgatgttc tatccggtat aatgacctta 42600 gcccactgac agtaggcgac cgacacggag cgtagcgaag tgtcccctgt cgccgtatgt 42660 ccactgacta acactagcaa gcattcccta ttgtagcgac ggagcgaagc gaagcgagag 42720 cgacaaagcg caaatcaata ggatagcaac tagatagcat atggatagca gcacgatagc 42780 aagtgttaac aagtggatag caagtgagta gcgcatcaca tcgcagatgt catatgctaa 42840 caagtggata gccagtcgat agccacacgc tgacaggcga caagccagtg agttaacagg 42900 tgagcaaaaa aggcaagcgc agagcacagc cgcgccgccc acgcagccgc aaggctttcc 42960 agtcgctttc ccctgctttc ccacggatag ccacacgcac gcacgcgcac acgctggata 43020 gcccatgcat gcacacgcga gcgcacgcac gccagcgcac acacacgca 43069

Claims (11)

비브리오균(Vibrio)에 특이적인 사멸능을 갖고, 서열번호 1로 표시되는 염기서열로 이루어진 박테리오파지 VP2.
A bacteriophage VP2 having the nucleotide sequence shown in SEQ ID NO: 1, which has a killing activity specific to Vibrio .
제1항에 있어서,
상기 박테리오파지는 수탁번호 KFCC11727P인, 박테리오파지 VP2.
The method according to claim 1,
The bacteriophage is the bacteriophage VP2, which is Accession No. KFCC11727P.
제1항에 있어서,
상기 비브리오균은 장염비브리오균(Vibrio parahaemolyticus)인, 박테리오파지 VP2.
The method according to claim 1,
The Vibrio fungus may be selected from the group consisting of Vibrio parahaemolyticus ), bacteriophage VP2.
제1항의 박테리오파지 VP2를 포함하는, 항생용 조성물.
An antibiotic composition comprising the bacteriophage VP2 of claim 1.
제1항의 박테리오파지 VP2를 포함하는, 사료 첨가용 조성물.
A composition for feed addition comprising the bacteriophage VP2 of claim 1.
제5항의 박테리오파지 VP2를 포함하는, 사료.
A feed comprising bacteriophage VP2 of claim 5.
제1항의 박테리오파지 VP2를 포함하는, 소독제.
A disinfectant comprising bacteriophage VP2 of claim 1.
제1항의 박테리오파지 VP2를 포함하는, 세척제.
A cleansing agent comprising the bacteriophage VP2 of claim 1.
제1항의 박테리오파지 VP2를 인간을 제외한 개체에 투여하는 단계를 포함하는, 비브리오균에 의한 감염성 질병의 예방 또는 치료 방법.
A method for preventing or treating an infectious disease caused by Vibrio bacteria, comprising administering the bacteriophage VP2 of claim 1 to an individual other than a human.
제9항에 있어서,
상기 비브리오균은 장염비브리오균(Vibrio parahaemolyticus)인, 비브리오균에 의한 감염성 질병의 예방 또는 치료 방법.
10. The method of claim 9,
The Vibrio fungus may be selected from the group consisting of Vibrio parahaemolyticus ) for preventing or treating infectious diseases caused by Vibrio bacteria.
제9항에 있어서,
상기 비브리오균에 의한 감염성 질병은 장염비브리오성 식중독, 콜레라, 비브리오 패혈증 및 어류 비브리오 감염증으로 이루어진 군에서 선택되는 1종 이상인, 비브리오균에 의한 감염성 질병의 예방 또는 치료 방법.
10. The method of claim 9,
Wherein the infectious disease caused by the Vibrio sp. Is at least one selected from the group consisting of Vibrio parahaemolyticus, cholera, Vibrio septicemia, and fish Vibrio infections.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210157183A (en) * 2020-06-19 2021-12-28 이화여자대학교 산학협력단 Novel bacteriophage VPT02 infecting vibrio and use thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210157183A (en) * 2020-06-19 2021-12-28 이화여자대학교 산학협력단 Novel bacteriophage VPT02 infecting vibrio and use thereof

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