KR101511712B1 - Novel infectious bursal disease virus IBD K7 and infectious bursal disease vaccine using the same - Google Patents

Novel infectious bursal disease virus IBD K7 and infectious bursal disease vaccine using the same Download PDF

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KR101511712B1
KR101511712B1 KR1020130036421A KR20130036421A KR101511712B1 KR 101511712 B1 KR101511712 B1 KR 101511712B1 KR 1020130036421 A KR1020130036421 A KR 1020130036421A KR 20130036421 A KR20130036421 A KR 20130036421A KR 101511712 B1 KR101511712 B1 KR 101511712B1
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송창선
최강석
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Abstract

본 발명은 신규 전염성 F낭병 바이러스 IBD K7주 및 이를 이용한 전염성 F낭병 백신에 관한 것이다.The present invention relates to a novel infectious F pancreatic virus IBD K7 strain and a communicable F pancreatic vaccine using the same.

Description

신규 전염성 Ｆ낭병 바이러스 ＩＢＤ Ｋ7주 및 이를 이용한 전염성 Ｆ낭병 백신{Novel infectious bursal disease virus IBD K7 and infectious bursal disease vaccine using the same} [0001] The present invention relates to a new infectious F cystic virus IBD K7 strain and a contagious F cystic vaccine using the same, and more particularly, to a novel infectious bursal disease virus IBD K7 and an infectious bursal disease vaccine using the same.

본 발명은 신규 전염성 F낭병 바이러스 IBD K7주 및 이를 이용한 전염성 F낭병 백신에 관한 것이다.The present invention relates to a novel infectious F pancreatic virus IBD K7 strain and a communicable F pancreatic vaccine using the same.

닭의 전염성 F낭병(infectious bursal disease, IBD) 또는 감보로병(Gumboro disease)로 불리는 이 병은, IBD 바이러스(IBDV)에 의하여 발병되는 전염병이다. IBDV는 Family Birnaviridae에 속하며 genome은 두 가닥(double-stranded)의 A와 B 2개의 RNA 분절로 이루어져 있으며 분절 A에는 VP5, VP2, VP4, VP3 유전자가 있고, 분절 B에는 VP1 유전자가 있다. IBDV는 1형과 2형 두 개의 혈청형이 존재하는데 이들 두 가지 혈청형은 바이러스 중화(VN) 테스트에 의해 분류될 수 있다. 혈청형 1형 바이러스는 닭에 대해 병원성을 나타내는 것으로 확인된 반면, 혈청형 2형 IBDV는 칠면조에서만 아급성 질병을 유발한다. 전염성 F낭병은 주요 표적으로서 F낭(Bursa of Fabricius) 세포에 감염이 일어나 면역억제를 일으키는 병이다. 3~6주령의 닭에 감염되면 높은 이환율과 폐사율을 보이는 임상형 전염성 F낭병이 유발되며, 3주령 이전에 감염될 경우 항체를 생산하는 F낭 내 림프구를 급격히 파괴시켜 면역억제가 심하게 나타나며, 이로 인하여 각종 백신에 대한 면역효과가 저하되고 전염성 질병에 대한 저항성도 약화된다. 따라서 정상계군에서는 불현성 또는 잠복감염으로 증상이 관찰되지 않던 질병들이 전염성 F낭병에 감염된 계군에서는 임상증상을 나타내거나 더욱 심한 임상증상을 일으켜 양계산업에 막대한 경제적 손실을 가져오고 있다.This disease, called infectious bursal disease (IBD) or gumboro disease, is an infectious disease caused by the IBD virus (IBDV). IBDV belongs to the family Birnaviridae. The genome consists of double-stranded A and B RNA segments, segment A has VP5, VP2, VP4 and VP3 genes, and segment B has the VP1 gene. There are two serotypes of IBDV, type 1 and type 2, and these two serotypes can be classified by the virus neutralization (VN) test. Serotype 1 viruses have been shown to be pathogenic to chickens, while serotype 2 IBDV only causes subacute disease in turkeys. Infectious F pancreatitis is a disease that causes immunosuppression by infecting F cells (Bursa of Fabricius) cells as a main target. Infections with 3-6 week old chickens lead to clinical infectious F pancytia with high morbidity and mortality. In case of infection before 3 weeks of age, the immunocompromise is markedly destroyed by rapidly destroying the lymphocytes in the F cyst producing antibody. Immune effects against various vaccines are reduced and resistance to infectious diseases is weakened. Therefore, diseases in which the symptoms are not observed due to incoherent or latent infections in the normal system are manifesting clinical symptoms or more severe clinical symptoms in the infectious system infected with infectious fungal disease, resulting in a great economic loss to the poultry industry.

이 병은 1962년 Cosgrove에 의해 처음 보고된 이후 전 세계에서 이 병이 존재하고 있음이 확인되었고, 우리나라에서는 1979년에 가축위생연구소의 연구팀에 의해 50% 이상의 닭이 감염되어 있음이 확인되었다. 그 후 유럽에서 1987년 처음으로 강독형 전염성 F낭병(very virulent IBD)이 출현하여 심각한 병증을 일으켰으며, 우리나라에서는 1992년 충남 성환 지역 9주령 산란계에서 처음으로 보고되었다. 그 당시 발생농장은 심한 임상증상과 함께 30% 이상의 높은 폐사율을 보이는 것으로 보고되었다. 강독형 전염성 F낭병은 기존 고전형과 달리 3주령 이하나 6주령 이후 주령에서도 높은 폐사를 유발하고 면역장기인 F낭에서 부종이나 충혈 등의 염증반응 없이 곧바로 위축됨으로써 병증이 빠르게 진행되는 특징을 나타냈다.This disease was first reported by Cosgrove in 1962 and has been confirmed to be present worldwide. In Korea, in 1979, more than 50% of chickens were infected by the research team of the Livestock Hygiene Research Institute. In 1987, a very virulent IBD was developed in Europe for the first time in 1987 and it was first reported in Korea in 1992 in a 9-week-old laying hens in Chungnam Seonghwan area. The resulting farms were reported to have high mortality rates of over 30% with severe clinical symptoms. Indocyanine fungicide (FCP), which is different from the existing type, induces high mortality in the age of 3 weeks or less after 6 weeks of age, and rapidly progresses in the disease progression without any inflammatory reaction such as edema or redness in the immunocompetent F cyst .

현재 발생하는 야외 전염성 F낭병은 고전형이나 변이형 전염성 F낭병이 거의 없고 사실상 모두 강독형 전염성 F낭병(79%)이다. 병증이 심각하게 나타나는 강독형 감보로 병을 예방하기 위하여 국내외에서 사용되고 있는 전염성 F낭병 생독백신은 독력과 모체이행항체 수준을 극복하는 능력에 따라 약독(M; mild), 중간독(I; intermediate), 중간독 플러스(I+; intermediate plus)로 구별되는데, 조기 감염이 되는 강독형 전염성 F낭병을 예방하기 위해 모체이행항체를 극복할 수 있는 독력이 강한 중간독 및 중간독 플러스 백신이 현재 주로 사용되고 있다.Current outbreaks of outbreaks of F-type pneumonia are almost all cases of persistent F-type pneumonia (79%). Infectious F pancreatic venom, which is used domestically and abroad to prevent intractable schizophrenia with severe pathologic symptoms, is classified as weak (M; mild), intermediate (I, intermediate) according to its ability to cope with toxicity and maternal antibody level, , And intermediate plus (I +). In order to prevent indocyanous fungal infections, which are early infections, potent mid-dose and mid-dose plus vaccines that are able to overcome maternal antibodies are currently in use .

동물약품협회의 자료에 근거하여 연도별 전염성 F낭병 백신 스트레인별 판매비율을 살펴보면 중간독 플러스형 백신(전체의 74~79%), 중간독형 백신(16~22%), 약독형(4~9%) 순서로 나타났다. 이는 강독형 전염성 F낭병의 조기감염을 예방하기 위해 보다 빨리 모체이행항체를 극복할 수 있는 중간독 플러스 백신 사용을 선호하고 있는 것을 나타낸다고 할 수 있다. (그림 2) 중간독 플러스 백신이 강한 면역반응을 일으켜 효과적인 전염성 F낭병 예방을 가능하게 하지만 강한 백신 반응으로 강독형 전염성 F낭병에 감염된 것과 유사한 증상을 일으키고, 면역장기인 F낭에 심각한 손상을 일으키는 것으로 알려져 있다. (Rautenschlein S et al., Protective efficacy of intermediate and intermediate plus infectious bursal disease virus (IBDV) vaccines against very virulent IBDV in commercial broilers, Avian Dis. 2005 Jun;49(2):231-7)Based on data from the Animal Health Association, annual sales of infectious F pseudotyped vaccine strains were as follows: middle-dose plus-dose vaccine (74-79% of total), intermediate-dose vaccine (16-22% %) In order. This suggests that they prefer to use a mid-dose plus vaccine that can overcome maternal transit antibody to prevent early infection of read-only infectious F pancreatitis. (Fig. 2) The mid-dose plus vaccine induces a strong immune response to prevent effective communicable F pancreatitis. However, the strong vaccine reaction causes symptoms similar to those infected with the infectious fungal pancreatitis, causing severe damage to the immune organs F cyst . (Rautenschlein S et al., Protective efficacy of intermediate and intermediate plus infectious bursal disease virus (IBDV) vaccines against very virulent IBDV in commercial broilers, Avian Dis 2005 Jun; 49 (2): 231-7)

현재 국내에서 주로 사용되고 있는 중간독 플러스 백신주들은 전염성 F낭병에 대한 강한 면역력을 형성하지만 면역장기인 F낭에 심각한 손상을 일으켜 다른 질병에 대한 저항성을 저하시킬 수 있기 때문에 기존의 중간독 플러스형 백신의 독력에 비해 낮은 독력을 나타내며, 중간독형 전염성 F낭병 백신보다 높은 면역원성을 지닌 백신 개발이 절실히 요구되고 있는 실정이다.The mid-dose plus vaccine strains currently used domestically constitute strong immunity against infectious F pancreatitis, but they can cause serious damage to the immune organs F sac, which may reduce the resistance to other diseases. Therefore, It is necessary to develop a vaccine having immunogenicity lower than that of poisoning and higher immunogenicity than the medium-toxic infectious F pancreatitis vaccine.

관련 특허로 대한민국 특허공개번호 제1020110116315호는 전염성 F낭병에 대하여 백신 효능이 우수한 신규한 비병원성 전염성 F낭병 바이러스(BP-IBDVac), 이를 포함하는 전염성 F낭병 백신, 및 이의 제조 방법이 기재되어 있다.
Korean Patent Publication No. 1020110116315 discloses a novel non-pathogenic infectious F pandemic virus (BP-IBDVac) having excellent vaccine efficacy against infectious F pancytopia, a contagious F pancreatic vaccine containing the same, and a method for producing the same.

본 발명은 상기의 문제점을 해결하고 상기의 필요성에 의하여 안출된 것으로서 본 발명의 목적은 신규 전염성 F낭병 바이러스를 제공하는 것이다.Disclosure of Invention Technical Problem [8] The present invention has been made in view of the above problems, and it is an object of the present invention to provide a novel infectious F. psoriasis virus.

본 발명의 다른 목적은 신규한 전염성 F낭병 백신을 제공하는 것이다.Another object of the present invention is to provide a novel infective F pandemic vaccine.

상기의 목적을 달성하기 위하여 본 발명은 기탁번호 KCTC 12376BP로 기탁된 전염성 F낭병 바이러스 IBD K7주를 제공한다. In order to achieve the above object, the present invention provides an infectious F pancreatic virus IBD K7 strain deposited with Accession No. KCTC 12376BP.

상기 균주는 2013년 3월 11일자로 국제기탁기관인 한국생명공학연구원 유전자은행(대한민국 대전시 유성구 소재)에 수탁번호 KCTC 12376BP로 기탁되었다. On March 11, 2013, the strain was deposited with KCTC 12376BP under accession number KCTC 12376BP at the Korea Biotechnology Research Institute Gene Bank (located in Yuseong-gu, Daejeon, Republic of Korea).

일 구현예에 상기 바이러스는 국내 육계로부터 분리한 것이 바람직하고, 상기 바이러스의 시그먼트 A 및 B는 각각 서열번호 1 및 2에 기재된 염기서열을 가지는 것이 바람직하나 이에 한정되지 아니한다.In one embodiment, the virus is preferably isolated from the domestic broiler, and the viruses of the viruses A and B preferably have the nucleotide sequences shown in SEQ ID NOS: 1 and 2, respectively, but are not limited thereto.

일 구현예에 상기 바이러스의 시그먼트 A는 서열번호 3(VP 5) 및 4(VP2-VP4-VP3)에 기재된 아미노산 서열을 가지며 시그먼트 B는 서열번호 5(VP 1)에 기재된 아미노산 서열을 가지며, 서열번호 3에서는 45번, 74번, 133번, 서열번호 4에서는 222번, 242번, 256번, 279번, 284번, 294번, 299번, 451번, 680번, 715번, 751번, 981번, 1005번째 잔기, 그리고 서열번호 5에서는 4번, 13번, 61번, 145번, 146번, 147번, 242번, 287번, 390번, 393번, 508번, 511번, 562번, 687번, 695번째 잔기가 다른 균주들과 차이가 있는 것이 바람직하나 이에 한정되지 아니한다.In one embodiment, the virus A has an amino acid sequence as set forth in SEQ ID NO: 3 (VP 5) and 4 (VP2-VP4-VP3), and the sequence B has an amino acid sequence as set forth in SEQ ID NO: 5 , SEQ ID NO: 3, 45, 74, and 133, and SEQ ID NO: 4, 222, 242, 256, 279, 284, 294, 299, 451, 680, 715, 751 , 981 and 1005 residues and in SEQ ID NO: 5, 4, 13, 61, 145, 146, 147, 242, 287, 390, 393, 508, 511, 562 687, and 695 residues are different from other strains, but are not limited thereto.

또한 본 발명은 상기 본 발명의 바이러스 및 약학적 허용 담체 또는 희석제를 포함하는, 전염성 F낭병 바이러스 감염으로부터 유발된 질병에 대한 가금 보호용 백신을 제공한다.The present invention also provides a vaccine for poultry protection against a disease caused by an infectious F pandemic virus infection comprising the virus of the present invention and a pharmaceutically acceptable carrier or diluent.

일 구현예에 있어서, 상기 바이러스가 살아있는 형태임이 바람직하고, 상기 백신이 보조제를 추가로 포함하는 것을 바람직하며, 상기 백신이 가금에 감염성이 있는 다른 병원균의 백신성분을 하나 이상 추가로 포함하는 것이 바람직하나 이에 한정되지 아니한다.In one embodiment, it is preferred that the virus is in a live form, and that the vaccine further comprises an adjuvant, and it is preferred that the vaccine further comprises one or more vaccine components of other pathogens infectious to the poultry But not limited to this.

또 본 발명은 a) 감염되기 쉬운 기질에 제1항 내지 제3항의 어느 한 항에 정의된 전염성 F낭병 바이러스를 접종하는 단계; b) 상기 전염성 F낭병 바이러스를 증식시키는 단계; 및 c) 상기 전염성 F낭병 바이러스 함유 물질을 수거하는 단계를 포함하는 전염성 F낭병 바이러스의 제조 방법을 제공한다.The invention also relates to a method for the treatment of infectious disease comprising the steps of: a) inoculating a susceptible substrate with an infective F pandemic virus as defined in any one of claims 1 to 3; b) propagating said infectious F pancreatic virus; And c) collecting the infectious F parasitic virus-containing substance.

일 구현예에 있어서, 상기의 기질은 닭 간의 배세포(CEL), 닭의 배 섬유아세포(CEF), 닭의 신장세포(CK), 및 베로(Vero) 세포주로 구성된 군으로부터 선택된 하나의 세포인 것이 바람직하나 이에 한정되지 아니한다.In one embodiment, the substrate is a cell selected from the group consisting of chicken intestine cell (CEL), chicken embryonic fibroblast cell (CEF), chicken kidney cell (CK), and Vero cell line But is not limited thereto.

또 본 발명은 상기 본 발명의 백신을 조류에 투여하는 것을 포함하는 가금에서 전염성 F낭병 감염으로부터 유발된 질병을 제어하는 방법을 제공한다.The present invention also provides a method for controlling a disease caused by an infectious F pancreatic infection in a poultice comprising administering the vaccine of the present invention to an alga.

살아있는 바이러스를 함유하는 본 발명의 백신은 동결건조된 형태나 (동결된) 현탁액의 형태로 제조되어 시판된다. 백신은 상기 조성물에 통상적으로 사용되는 희석제 또는 약학적 허용 담체를 추가로 포함한다. 담체는 안정화제, 보존제 및 완충액을 포함한다. 적당한 안정화제는 예를 들어 SPGA, 탄화수소(예로, 소비톨, 만니톨, 전분, 자당, 덱스트란, 글루타메이트, 또는 글루코스), 단백질(예로, 건조된 우유혈청, 알부민 또는 카세인)또는 그것의 분해 산물이다. 적당한 완충액은 예를 들어 알칼리 금속 포스페이트이다. 적당한 보존제는 티메로살, 메티올레이트, 겐타마이신이다. 희석액은 물, 수성 완충액(예로 완충 염수), 알코올 및 폴리올(예로 글리세롤)이다.The vaccine of the present invention containing live virus is manufactured and sold in the form of a lyophilized form or a (frozen) suspension. The vaccine further comprises a diluent or pharmaceutically acceptable carrier conventionally used in the composition. Carriers include stabilizers, preservatives and buffers. Suitable stabilizers are, for example, SPGA, hydrocarbons (e.g., sorbitol, mannitol, starch, sucrose, dextran, glutamate, or glucose), proteins (such as dried milk serum, albumin or casein) . Suitable buffers are, for example, alkali metal phosphates. Suitable preservatives are thimerosal, methiolate, gentamycin. Diluents are water, aqueous buffers (such as buffered saline), alcohols and polyols (such as glycerol).

필요한 경우, 본 발명의 살아있는 백신은 보조제를 함유할 수 있다. 보조적 활성을 가진 적당한 화합물과 조성물의 예는 불활성화된 백신의 제조를 위해 아래에 언급된 것과 동일하다.If desired, the live vaccine of the present invention may contain adjuvants. Examples of suitable compounds and compositions with adjuvant activity are the same as those mentioned below for the preparation of inactivated vaccines.

본 발명의 살아있는 백신은 예를 들어 근육내, 피하내 주사에 의한 투여가 가능하지만, 살아있는 백신은 전염성F낭병 바이러스 백신접종에 통상적으로 사용되는 저가의 대량 투여 기법에 의해 투여되는 것이 바람직하다. 이 기법은 음료수 및 분무 백신접종을 포함한다.Although the live vaccine of the present invention can be administered, for example, by intramuscular or subcutaneous injection, it is preferred that the live vaccine is administered by a low-dose, high-dose technique commonly used in the infectious F pandemic virus vaccination. This technique involves beverage and spray vaccination.

살아있는 백신의 투여를 위한 또 다른 방법은 난내(in ovo) 투여, 점안 투여 및 부리를 담그는 투여(beak dipping administration)를 포함한다.Other methods for administration of live vaccines include in ovo administration, topical administration, and beak dipping administration.

본 발명의 백신은 활성 성분으로서 전염성F낭병 바이러스를 유효량, 즉 독성 바이러스에 의한 공격(항원 투여)에 대항하여 백신 접종된 조류 또는 그들의 자손에서 면역을 유도할 전염성F낭병 바이러스 물질을 면역화시키는 양으로 포함한다. 본원에서 면역은 백신접종되지 않은 군에 비해 백신접종 후 조류 집단에서 상당히 높은 레벨의 보호를 유도하는 것으로 정의된다.The vaccine of the present invention comprises an effective amount of an infectious F pandemic virus as an active ingredient, that is, an amount that immunizes a vaccinated bird or an infectious F pandemic virus substance that will induce immunity in their offspring against an attack by a toxic virus (antigen administration) . Immunity is defined herein as inducing a significantly higher level of protection in a population of birds following vaccination compared to the non-vaccinated population.

통상적으로, 본 발명의 살아있는 백신은 조류 1 마리 당 102-109TCID50 , 바람직하게는 102-106 TCID50의 용량으로 투여될 수 있다. Typically, the live vaccine of the present invention is the bird 1 10 2 -10 9 TCID50, preferably per grain can be administered at a dose of 10 2 -10 6 TCID50.

본 발명의 전염성F낭병 바이러스 백신은 닭에 효과적으로 사용될 수 있으나, 칠면조, 기니아 파울(guinea fowl) 및 메추라기와 같은 기타의 가금도 효과적으로 백신으로 접종될 수 있다. 닭은 식용 닭, 복제 가축, 및 알을 낳는 가축을 포함한다.The infective F pandemic virus vaccine of the present invention can be effectively used for chickens, but other poultry such as turkey, guinea fowl and quail can also be effectively vaccinated. Chickens include edible chickens, replica livestock, and livestock.

본 발명에 따른 살아있는 백신을 투여받은 동물의 나이는 현재 시판되는 살아있는 전염성F낭병 바이러스 백신을 투여받은 동물의 나이와 동일하다. 예를 들면, 식용 닭은 생후 1일부터, 본 발명의 살아있는 백신으로 직접 백신접종된다. 식용 닭의 종축과 같은 부모 가축의 백신접종은 본 발명의 살아있는 백신으로 수행될 수 있다. 이런 유형의 면역주사 프로그램의 장점은 수직적으로 새끼에게 전달되는 모계 유래의 항체에 의해 제공된 생후 1일된 자손의 직접적인 보호를 포함한다. 전형적인 종축의 백신접종 프로그램은 생후 2~3주의 종축에게 살아있는 약화된 백신을 접종한 후 생후 14-18주에 불활성화된 백신을 접종하는 것을 포함한다.
The age of the animal receiving the live vaccine according to the present invention is the same as the age of the animal on which the currently live live infective F pandemic virus vaccine is administered. For example, edible chickens are vaccinated directly from the first day of life with the live vaccine of the present invention. Vaccination of the parent animal, such as the breeder of the edible chicken, can be performed with the live vaccine of the present invention. The advantages of this type of vaccination program include the direct protection of one day old offspring provided by maternally derived antibodies transmitted vertically to the offspring. A typical longitudinal vaccination program involves inoculating an inactivated vaccine at 14 to 18 weeks of age after inoculation with a live attenuated vaccine for two to three weeks after breeding.

본 발명에서는 새로운 전염성 F낭병 바이러스, 이를 포함하는 백신 및 이들의 제조 방법을 제공함으로써 전염성 F낭병으로부터 유효하게 가금을 보호할 수 있다.In the present invention, poultry can be effectively protected from infectious F pancreatitis by providing a new infectious F pancreatic virus, a vaccine containing the same, and a method for producing the same.

도 1은 국내 분리 전염성 F낭병 바이러스의 독력 유형별 분류 <출처: 농림수산검역검사본부>
도 2는 국내 전염성 F낭병 생독백신 종류별 판매비율을 나타낸 그래프,
도 3은 타 병원체 오염여부 시험 사진,
도 4는 VP2 초가변 부위의 계통학적 분석Figure 1 shows the classification of domestic isolates of F-virulent virulence viruses according to their virulence types. Source: Ministry of Agriculture, Forestry and Fisheries Quarantine Inspection Headquarters
FIG. 2 is a graph showing sales ratios of the domestic infectious F pancreatic virulence type vaccine,
FIG. 3 is a photograph showing the contamination test of other pathogens,
Figure 4 shows phylogenetic analysis of the VP2 hypervariable region

이하 비한정적인 실시예를 통하여 본 발명을 더욱 상세하게 설명한다. 단 하기 실시예는 본 발명을 예시하기 위한 의도로 기재한 것으로서 본 발명의 범위는 하기 실시예에 의하여 제한되는 것으로 해석되지 아니한다.The present invention will now be described in more detail by way of non-limiting examples. The following examples are intended to illustrate the present invention and the scope of the present invention is not to be construed as being limited by the following examples.

실시예Example 1:국내 야외농장에서의  1: At a domestic outdoor farm infectiousinfectious bursalbursal diseasedisease virusvirus IBDIBD K7K7 주 분리, 동정 기타병원체 부정 시험Primary isolation, identification, and other pathogen maltreatment

2007년 충남소재 육계 농장으로부터 의뢰된 육계로부터 F낭을 채취하여 멸균 인산완충용액(pH7.2)을 넣어 10중량%의 유제액을 만들었다. 이를 3,000g에서 15분간 원심분리하여 세포와 조직을 침전시키고, 상층액을 0.45㎛의 주사기 여과기로 여과하였다. 11일령 SPF 발육란의 장뇨막(chorioallantoic membrane: CAM) 접종법으로 분리하여 역전사 중합효소연쇄반응(RT-PCR)을 통해 IBD K7 스트레인을 최종 동정하였다.In 2007, fungi were taken from broiler chickens from broiler farms in Chungnam Province and sterilized phosphate buffer solution (pH 7.2) was added to make a 10% by weight emulsion. This was centrifuged at 3,000 g for 15 minutes to precipitate cells and tissues, and the supernatant was filtered with a 0.45 μm syringe filter. The IBD K7 strain was finally identified by reverse transcription-polymerase chain reaction (RT-PCR) using a chorioallantoic membrane (CAM) inoculation method at 11 days of SPF development.

실시예Example 2: 2: 신규한New IBDVIBDV 야외분리주Outdoor separator 내 기타병원체 부정 시험  My other pathogen fraud

계태아란의 장요막강액 및 장뇨막에서 증식이 가능한 기타 병원체의 존재 유무를 확인하기 위하여 전염성 기관지염 바이러스 (Infectious bronchitis virus, IBV), 뉴캣슬병 바이러스 (Newcastle disease, NDV), 조류아데노바이러스(Fowl adeno virus, FAdV), 닭전염성빈혈증 바이러스(Chicken infectious anemia virus, CIAV), 마이코플라스마(Mycoplasma)에 대한 역전사 중합효소연쇄반응 (RT-PCR) 및 중합효소연쇄반응(PCR)을 실시하여 전염성 F낭병 바이러스 분리주(IBD K7) 외에 기타 병원체가 없음을 확인하였다.Infectious bronchitis virus (IBV), Newcastle disease (NDV), Fowl adenovirus (Fowl adenovirus), and fowl adenovirus , FAdV), Chicken infectious anemia virus (CIAV), Mycoplasma (RT-PCR), and Polymerase chain reaction (PCR) (IBD K7) and other pathogens.

실시예Example 3:게놈 염기서열 분석 3: Genome sequencing

11일령 SPF 발육란에 바이러스를 접종하고 5일 후 수확한 장뇨막액 150 ㎕를 활용하여 RNA를 추출하였다. 상기에서 추출한 RNA 1 ㎍, 랜덤 헥사머(random hexamer) 10 pmol, deoxynucleotide triphosphates (dNTPs) 20pmol, Superscript II reverse transcriptase(Invitrogen) 200 units 의 농도로 섞어서 42℃에서 60분간 반응시키고, 역전사 효소는 70℃에서 15분 동안 처리하여 불활성화시켰다.RNA was extracted using 150 μl of the nasal membranes harvested 5 days after inoculation with the virus in the 11-day-old SPF developmental field. 1 μg of the above extracted RNA, 10 pmol of a random hexamer, 20 pmol of deoxynucleotide triphosphates (dNTPs) and 200 units of Superscript II reverse transcriptase (Invitrogen) were mixed and reacted at 42 ° C. for 60 minutes. The reverse transcriptase was reacted at 70 ° C. Lt; / RTI &gt; for 15 minutes.

이와 같이 얻어진 cDNA를 4배 희석한 cDNA 용액 1 ㎕, 10x PCR 버퍼 1 ㎕, 2.5 mM dNTPs 0.2 ㎕, 각각의 프라이머 2 ㎕, Taq 중합효소 1 ㎕ 및 DW 7.2 ㎕를 혼합하였다. (I. Lojkic et al., Sequence Analysis of Both Genome Segments of Three Croatian Infectious Bursal Disease Field Viruses, AVIAN DISEASES 52:513-519, 2008)1 μl of the cDNA solution obtained by diluting the obtained cDNA 4 times, 1 μl of 10 × PCR buffer, 0.2 μl of 2.5 mM dNTPs, 2 μl of each primer, 1 μl of Taq polymerase and 7.2 μl of DW were mixed. (I. Lojkic et al., Sequence Analysis of Both Genome Segments of Three Croatian Infectious Bursal Disease Field Viruses, AVIAN DISEASES 52: 513-519, 2008)

상기 반응액을 활용하여 얻은 PCR 산물을 정제하여 ABI3100 자동염기서열분석기를 사용하여 염기서열을 결정하였다. 그 결과 분절 A(Segment A)와 분절 B(segment B)로 표시되는 본 발명의 IBD K7의 게놈 염기 서열(IBD K7 바이러스의 Segment A 염기서열과 IBD K7 바이러스의 Segment B 염기서열은 각각 서열번호 1 및 2)을 얻었으며 VP1, VP5-VP2-VP4-VP3의 단백질의 아미노산 서열을 얻었다.The PCR product was purified using the above reaction solution, and the base sequence was determined using an ABI3100 automatic base sequence analyzer. As a result, the genome sequence of IBD K7 of the present invention (Segment A base sequence of IBD K7 virus and Segment B nucleotide sequence of IBD K7 virus of the present invention, represented by segment A and segment B, And 2), and the amino acid sequence of the protein of VP1, VP5-VP2-VP4-VP3 was obtained.

Figure 112013029113217-pat00001
Figure 112013029113217-pat00001

표 1은 Segment A 염기 분석에 사용된 primer set의 염기 서열 및 위치를 나타낸 표Table 1 shows the nucleotide sequences and positions of the primer sets used in Segment A base analysis

Figure 112013029113217-pat00002
Figure 112013029113217-pat00002

표 2는 Segment B 염기 분석에 사용된 primer set의 염기 서열 및 위치를 나타낸 표Table 2 shows the nucleotide sequences and positions of the primer sets used in Segment B base analysis

실시예Example 4: 4: IBDIBD K7K7 바이러스의 분자생물학적 특성 Molecular biological characteristics of virus

IBD K7 바이러스 전체 유전자를 GenBank 상에 등록된 국내외 전염성 F낭병 바이러스들과 BLAST 검색(www.ncbi.nlm.nih.gov/blast)과 Bioedit 프로그램을 사용하여 비교 분석하였다. 그 결과 IBD K7는 강독형 바이러스보다 약독형 바이러스에 상동성이 상대적으로 높은 것으로 나타났다.(표 3)The entire IBD K7 virus gene was compared and analyzed with the BLAST search (www.ncbi.nlm.nih.gov/blast) and the Bioedit program with the domestic and foreign communicable F pandemic viruses registered on GenBank. As a result, IBD K7 was found to be more homologous to the attenuated virus than the depressed virus (Table 3).

또한 계통 분석에 적용하는 VP2 초가변 부위 분석결과 기존의 어느 타입에도 속하지 않는 새로운 타입임이 밝혀졌으며 방어단백질 VP2의 병원성관련 모티브의 경우 IBD K7은 강독형과 약독형 바이러스 모티브를 동시에 가지고 있는 매우 독특한 항원구조를 가지고 있는 것으로 나타났다.(도 4 및 표 4) 그러나 VP2단백질 이외의 다른 구조단백질의 경우 IBD K7은 전형적인 약독형 바이러스 모티브를 가지고 있었다.(표 4 및 5)In addition, the analysis of VP2 hypervariable region applied to the systematic analysis revealed that it is a new type that does not belong to any existing type. In the case of the pathogenic motif of the protective protein VP2, IBD K7 is a very unique antigen (Figure 4 and Table 4). However, in the case of other structural proteins other than the VP2 protein, IBD K7 had a typical attenuated viral motif (Tables 4 and 5).

결론적으로 IBD K7은 강독형 바이러스 항원모티브를 일부 가진 기존의 바이러스와 비교하여 신규한 재조합 IBD 바이러스임을 나타내고 있다.In conclusion, IBD K7 is a novel recombinant IBD virus compared to existing viruses that have some of the depressed virus antigen motif.

Figure 112013029113217-pat00003
Figure 112013029113217-pat00003

표 3은 IBD K7와 기존 IBDV 스트레인과 전체 유전자 상동성 비교Table 3 compares the overall gene homology with IBD K7 and conventional IBDV strain

Figure 112013029113217-pat00004
Figure 112013029113217-pat00004

표 4는 IBD K7와 백신주 및 야외주의 VP5, VP2, VP4, VP3 비교분석Table 4 compares IBD K7 and VP5, VP2, VP4, VP3 for vaccine and open field

Figure 112013029113217-pat00005
Figure 112013029113217-pat00005

표 5는 IBD K7와 백신주 및 야외주의 VP1 비교분석Table 5 compares IBD K7 with the VP1 of vaccine and open field

실시예Example 5: 5: IBDIBD K7K7 스트레인의 병원성 실험 Pathogenic experiment of strain

백신 스트레인으로서의 가능성을 조사하고자 SPF 닭 3주령에 IBD K7 스트레인과 기존 IBDV 백신주인 Winterfield 스트레인, D78 스티레인을 각 그룹당 8수의 닭에 103ELD50/dose로 경구접종을 실시한 후 4일째 4수, 7일째 나머지를 안락사시켜 체중 및 BB-율[(F-낭 중량(g)/체중(g))x1000] 및 항체반응 검사를 실시하였다. 그 결과 Winterfield 스트레인 접종군과 IBD K7 스트레인 접종군에서 4일이내 각각 1수씩 폐사하였고 나머지 그룹은 모두 7일간 생존하였다. 체중과 BB-율을 비교분석한 결과 중간독 플러스와 중간독 중간의 병원성을 가진 것으로 조사되었으며 백신 접종 후 4일과 7일에 채혈하여 분리한 혈청의 ELISA 역가 확인 결과 면역반응은 중간독 플러스보다는 다소 낮지만 중간독 보다는 강한 것으로 확인되었다.(표 6 및 7)To investigate the possibility of vaccine strain, we injected IBD K7 strain at 3 weeks of age with SPF chicken, Winterfield strain and D78 styrenes, which were the original IBDV vaccine hosts, at 8 × 103 chickens per group at 103 ELD50 / The remaining body was euthanized, and body weight and BB-ratio [(F-sac weight (g) / body weight (g)) x1000] and antibody reaction test were performed. As a result, the mice were killed by the Winterfield strain inoculation group and the IBD K7 strain inoculation group within 4 days, and the remaining groups survived for 7 days. As a result of comparing the body weight and BB-ratio, it was found that the vaccine had a pathogenicity intermediate between the middle dose and the middle dose. The ELISA of the serum separated by blood sampling on the 4th and 7th day after the vaccination showed that the immune response was somewhat (Table 6 and 7). &Lt; tb &gt; &lt; TABLE &gt;

Figure 112013029113217-pat00006
Figure 112013029113217-pat00006

표 6은 IBD K7 스트레인, 중간독 플러스, 중간독 백신 접종 4일 및 7일 후 체중 및 BB율Table 6 shows the results of IBD K7 strain, mid-dose plus, mid-venous vaccination 4 days and 7 days post-

Figure 112013029113217-pat00007
Figure 112013029113217-pat00007

표 7은 IBD K7 스트레인, 중간독 플러스, 중간독 백신 접종 4일 및 7일 후 ELISA 역가
Table 7 shows the results of the IBD K7 strain, mid-dose plus mid-dose vaccination 4 and 7 days after ELISA

실시예Example 6: 6: SPFSPF 닭에서의Chicken 면역원성 및  Immunogenicity and 방어능Defense 실험 Experiment

IBD K7의 생독백신으로써의 효능을 알아보기 위하여, 3주령 SPF 닭에 IBD K7 스트레인과 기존 백신 스트레인 Winterfield 스트레인 및 D78을 1수당 103ELD50씩 경구로 백신접종하고, 백신접종 2주 후 최근 유행 강독형 전염성 F낭병 바이러스를 1수당 105ELD50씩 점안으로 공격접종하였다. 대조군도 동일한 바이러스를 동일한 방법으로 공격접종하였다. 접종 후 5일간 침울이나 깃털 역립, 폐사등의 임상증상을 관찰하였으며 체중 변화를 측정하였다. To determine the efficacy of IBD K7 as a live vaccine, 3-week-old SPF chickens were orally vaccinated with IBD K7 strain, and the original vaccine strain Winterfield strain and D78 were given at a rate of 10 3 ELD 50 per dose. Two weeks after vaccination, Inducible infectious F pandemic virus was inoculated in 10 5 ELD 50 doses per dose. Controls were also inoculated with the same virus in the same manner. After 5 days of inoculation, clinical symptoms such as drowsiness, feathering, and mortality were observed and weight change was measured.

백신 2주 후 공격접종 전 ELISA 항체가 및 임상증상을 볼 때 중간독 백신인 D78에 비하여 우수한 면역원성을 나타냈으며 중간독 플러스 백신인 Winterfield 스트레인 접종군과 유의적인 차이를 나타내지 않는 것을 확인할 수 있었다.(표 8)The ELISA antibody and the clinical symptoms of the vaccine two weeks after the vaccination showed excellent immunogenicity compared with the middle dose vaccine, D78, and the vaccine did not show any significant difference from the Winterfield strain inoculation group. (Table 8)

Figure 112013029113217-pat00008
Figure 112013029113217-pat00008

표 8은 IBD K7 스트레인 백신 닭에서의 강독형 IBDV 공격접종에 대한 방어능 및 면역원성을 나타낸 표이다.Table 8 is a table showing the protective and immunogenicity against impregnated IBDV challenge in IBD K7 strain vaccinated chickens.

한국생명공학연구원Korea Biotechnology Research Institute KCTC12376BPKCTC12376BP 2013031120130311

<110> Konkuk University Industrial Cooperation Corp. <120> Novel infectious bursal disease virus IBD K7 and infectious bursal disease vaccine using the same <160> 5 <170> KopatentIn 1.71 <210> 1 <211> 3131 <212> DNA <213> infectious bursal disease virus IBD K7 <400> 1 gggacaggcc gtcaaggcct tgttccagga tggaactcct ccttctacaa cgctatcatt 60 gatggttagt agagatcaga caaacgatcg cagcgatgac aaacctgcaa gatcaaaccc 120 aacagattgt tccgttcata cggagccttc tgatgccaac aaccggaccg gcgtccattc 180 cggacgacac cctggagaag cacactctca ggtcagagac ctcgacctac aatttgactg 240 tgggggacac agggtcaggg ctaattgtct ttttccctgg attccctggc tcaactgtgg 300 gtgctcacta cacactgcag agcaatggga actacaagtt cgatcagatg ctcctgactg 360 cccagaacct accggccagt tacaactact gcaggctagt gagtcggagt ctcacagtga 420 ggtcaagcac acttcctggt ggcgtttatg cactaaacgg caccataaac gccgtgacct 480 tccaaggaag cctgagtgaa ctgacagatg ttagctacaa tgggttgatg tctgcaacag 540 ccaacatcaa cgacaaaatt gggaacgtcc tggtagggga aggggtcacc gtcctcagct 600 tacccacatc atatgatctt gggtatgtga ggcttggtga ccccattccc gcaatagggc 660 ttgacccaaa aatggtagcc acatgtgaca gcagcgacag acccagagtc tacaccataa 720 ctgcagccga tgattaccaa ttctcatcac agtaccaatc aggtggggta acaatcacac 780 tgttctcagc caacattgat gccatcacca gcctcagcgt tgggggggag ctcgtgtttc 840 aaacaagcgt ccaaggcctt atactgggcg ccaccatcta ccttataggc tttgatggga 900 caacggtaat caccagagct gtggccgcaa acaatgggct gacggccggc accgacaatc 960 ttatgccatt caatcttgtg attccaacca acgagataac ccagccaatc acatccatca 1020 aactggagat agtgacctcc aaaagtggtg gtcaggcagg ggatcagatg tcatggtcgg 1080 taagtgggag cctagcagtg acgatccatg gtggcaacta cccaggggcc ctccgtcccg 1140 tcacgctagt agcctacgaa agagtggcaa caggatccgt cgttacggtc gctggggtga 1200 gcaacttcga gctgatccca aatcctgaac tagcaaagaa cctggttaca gaatacggcc 1260 gatttgaccc aggagccatg aactacacaa aattgatact gagtgagagg gaccgtcttg 1320 gcatcaggac cgtctggcca acaagggagt acactgactt tcgtgaatac ttcatggagg 1380 tggccgacct caactctccc ctgaagattg caggagcatt cggcttcaaa gacataatcc 1440 gggccataag gaggatagct gtgccggtgg tctccacatt gttcccacct gccgctcccc 1500 tagcccatgc aattggggaa ggtgtagact acctgctggg cgatgaggca caggctgctt 1560 caggaactgc tcgagccgcg tcaggaaaag caagagctgc ctcaggccgc ataaggcagc 1620 tgactctcgc cgccgacaag gggtacgagg tagtcgcgaa tctattccag gtgccccaga 1680 atcccgtagt cgacgggatt cttgcttcac ctggggtact ccgcggtgcg cacaacctcg 1740 actgcgtgtt aagagagggt gccacgctat tccccgtggt cattacgaca gtggaagacg 1800 ccatgacacc caaagcattg aacagcaaaa tgtttgctgt cattgaaggc gtgcgagaag 1860 acctccaacc tccatctcaa agaggatcct tcatacgaac tctctctgga cacagagtct 1920 atggatatgc tccagatggg gtacttccac tggagactgg gagagactac accgttgtcc 1980 caatagatga tgtctgggac gacagcatta tgctgtccaa agaccccata cctcctattg 2040 tgggaaacag tggaaaccta gccatagctt acatggatgt gtttcgaccc aaagtcccta 2100 tccatgtggc tatgacggga gccctcaatg cttgtggcga gattgagaaa gtaagcttta 2160 gaagcaccaa gctcgccact gcacaccgac ttggcctcaa gttggctggt cccggagcat 2220 tcgatgtaaa caccgggccc aactgggcaa cgttcatcaa acgtttccct cacaatccac 2280 gcgactggga caggctcccc tacctcaacc taccatacct tccacccaat gcaggacgcc 2340 agtaccacct tgccatggct gcatcagagt tcaaagagac ccccgaactc gagagcgccg 2400 tcagagcaat ggaagcagca gccaacgtgg acccactatt ccaatctgca ctcagtgtgt 2460 tcatgtggct ggaagagaat gggattgtga ctgacatggc caacttcgca ctcagcgacc 2520 caaacgccca tcggatgcga aattttcttg caaacgcacc acaagcaggt agcaagtcgc 2580 aaagggccaa gtacgggaca gcaggctacg gagtggaggc ccggggcccc acaccagagg 2640 aagcacagag ggaaaaagac acacggatct caaagaagat ggagaccatg ggcatctact 2700 ttgcaacacc agaatgggta gcactcaatg ggcaccgagg gccaagcccc ggccagctaa 2760 agtactggca gaacacacga gaaataccgg acccaaacga ggactatcta gactacgtgc 2820 atgcagagaa gagccggttg gcatcagaag aacaaatcca acgggcagct acgtcgatct 2880 acggggctcc aggacaggca gagccacccc aagctttcat agacgaagtt gccaaagtct 2940 atgaaatcaa ccatggacgt ggcccaaacc aagaacagat gaaagatctg ctcttgactg 3000 cgatggagat gaagcatcgc aatcccaggc gggctccacc aaagcccaag ccaaaaccca 3060 atgctccaac acagagaccc cctggtcggc tgggccgctg gatcaggact gtctctgatg 3120 aggaccttga g 3131 <210> 2 <211> 2687 <212> DNA <213> infectious bursal disease virus IBD K7 <400> 2 tcctcttctt gatgattctg ccaccatgag tgacattttc aacagtccac aggcgcgaag 60 cacgatctca gcagcgttcg gcataaagcc tactgctgga caagacgtgg aagaactctt 120 gatccctaaa gtttgggtgc cacctgagga tccgcttgcc agccctagtc gactggcaaa 180 gttcctcaga gagaacggct acaaagtttt gcagccacgg tctctgcccg agaatgagga 240 ttatgagacc gaccaaatac tcccagactt agcatggatg cgacaggtag aaggggctgt 300 tttaaaacct actctatctc tccctattgg agatcaggag tacttcccaa agtactaccc 360 aacacatcgc cctagcaagg agaagcccaa tgcgtacccg ccagacattg cactactcaa 420 gcagatgatt tacctgtttc tccaggttcc agaggccaac gagggcctaa aggatgaagt 480 aaccctcttg acccaaaaca taagggacaa ggcctatgga agtgggacct acatgggaca 540 agcaactcga cttgtggcca tgaaggaggt cgccactggg agaaacccaa acaaggatcc 600 tctaaagctt gggtacactt ttgagagcat cgcgcagcta cttgacatca cactaccggt 660 aggcccaccc ggtgaggatg acaagccctg ggtgccactc acaagagtgc cgtcacggat 720 gttggtgctg acgggagacg tagatggcga ctttgaggtt gaagactacc ttcccaaaat 780 caacctcaag tcatcaagtg gactaccata tgtaggtcgc accaaaggag agacaattgg 840 cgagatgata gctatctcaa accagtttct cagagagcta tcaacactgt tgaagcaagg 900 tgcagggaca aaggggtcaa acaagaagaa gctactcagc atgttaagtg accattggta 960 cttatcatgc gggcttttgt ttccaaaggc tgaaaggtac gacaaaagta catggctcac 1020 caagacccgg aacatatggt cagctccatc ccctacacac ctcatgatct ccatgatcac 1080 ctggcccgtg atgtccaaca gcccaaataa cgtgttgaac attgaagggt gtccatcact 1140 ctacaaattc aacccgttca gaggagggtt gaacaggatc gtcgagtgga tattggcccc 1200 ggaagaaccc aaggctcttg tatatgcgga caacatatac attgtccact caaacacgtg 1260 gtactcaatt gacctagaga agggcgaggc aaactgcact cgccaacaca tgcaagctgc 1320 aatgtactac atactcacca gagggtggtc agacaacggc gacccaatgt tcaatcaaac 1380 atgggccacc tttgccatga acattgcccc tgctctagtg gtagactcat cgtgcctgat 1440 aatgaacctg caaattaaga cctatggtca aggcagcggg aatgcagcca cgttcatcaa 1500 caaccacctc ttgagcacgc tagtgcttga ccagtggaac ctgatgagac agcccagacc 1560 agacagcgag gagttcaaat caattgagga caagctaggt atcaacttta agattgagag 1620 gtccattgat gacatcaggg gcaagctgag acagcttgtc ctccttgcac aaccagggta 1680 cctgagtggg ggggttgaac cagaacaatc cagcccaact gttgagcttg acctactagg 1740 gtggtcagct acatacagca aagatctcgg gatctatgtg ccggtgcttg acaaggaacg 1800 cctattttgt tctgctgcgt atcccaaggg agtagagaac aagagtctca agtccaaagt 1860 cgggatcgag caggcataca aggtagtcag gtatgaggcg ttgaggttgg taggtggttg 1920 gaactaccca ctcctgaaca aagcctgcaa gaataacgca ggcgccgctc ggcggcatct 1980 ggaggccaag gggttcccac tcgacgagtt cctagccgag tggtctgagc tgtcagagtt 2040 cggtgaggcc ttcgaaggct tcaatatcaa gctgaccgta acatctgaga gcctagccga 2100 actgaacaag ccagtacccc ccaagccccc aaatgtcaac agaccagtca acactggggg 2160 actcaaggca gtcagcaacg ccctcaagac cggtcggtac aggaacgaag ccggactgag 2220 tggtctcgtc cttctagcca cagcaagaag ccgtctgcaa gatgcagtta aggccaaggc 2280 agaagccgag aaactccaca agtccaagcc tgacgacccc gatgcagact ggttcgaaag 2340 atcagaaact ctgtcagacc ttctggagaa agccgacatc gccagcaagg tcgcccactc 2400 agcactcgtg gaaacaagcg acgcccttga agcagttcag tcgacttccg tgtacacccc 2460 caagtaccca gaagtcaaga acccacagac cgcctccaac cccgttgttg ggctccacct 2520 gcccgccaag agagccaccg gtgtccaggc cgctcttctc ggagcaggaa cgagcagacc 2580 aatggggatg gaggccccaa tacggtccaa gaacgccgtg aaaatggcca aacggcggca 2640 acgccaaaag gagagccgct aacagccatg atgggaacca ctcaaga 2687 <210> 3 <211> 145 <212> PRT <213> infectious bursal disease virus IBD K7 <400> 3 Met Val Ser Arg Asp Gln Thr Asn Asp Arg Ser Asp Asp Lys Pro Ala 1 5 10 15 Arg Ser Asn Pro Thr Asp Cys Ser Val His Thr Glu Pro Ser Asp Ala 20 25 30 Asn Asn Arg Thr Gly Val His Ser Gly Arg His Pro Gly Glu Ala His 35 40 45 Ser Gln Val Arg Asp Leu Asp Leu Gln Phe Asp Cys Gly Gly His Arg 50 55 60 Val Arg Ala Asn Cys Leu Phe Pro Trp Ile Pro Trp Leu Asn Cys Gly 65 70 75 80 Cys Ser Leu His Thr Ala Glu Gln Trp Glu Leu Gln Val Arg Ser Asp 85 90 95 Ala Pro Asp Cys Pro Glu Pro Thr Gly Gln Leu Gln Leu Leu Gln Ala 100 105 110 Ser Glu Ser Glu Ser His Ser Glu Val Lys His Thr Ser Trp Trp Arg 115 120 125 Leu Cys Thr Lys Arg His His Lys Arg Arg Asp Leu Pro Arg Lys Pro 130 135 140 Glu 145 <210> 4 <211> 1012 <212> PRT <213> infectious bursal disease virus IBD K7 <400> 4 Met Thr Asn Leu Gln Asp Gln Thr Gln Gln Ile Val Pro Phe Ile Arg 1 5 10 15 Ser Leu Leu Met Pro Thr Thr Gly Pro Ala Ser Ile Pro Asp Asp Thr 20 25 30 Leu Glu Lys His Thr Leu Arg Ser Glu Thr Ser Thr Tyr Asn Leu Thr 35 40 45 Val Gly Asp Thr Gly Ser Gly Leu Ile Val Phe Phe Pro Gly Phe Pro 50 55 60 Gly Ser Thr Val Gly Ala His Tyr Thr Leu Gln Ser Asn Gly Asn Tyr 65 70 75 80 Lys Phe Asp Gln Met Leu Leu Thr Ala Gln Asn Leu Pro Ala Ser Tyr 85 90 95 Asn Tyr Cys Arg Leu Val Ser Arg Ser Leu Thr Val Arg Ser Ser Thr 100 105 110 Leu Pro Gly Gly Val Tyr Ala Leu Asn Gly Thr Ile Asn Ala Val Thr 115 120 125 Phe Gln Gly Ser Leu Ser Glu Leu Thr Asp Val Ser Tyr Asn Gly Leu 130 135 140 Met Ser Ala Thr Ala Asn Ile Asn Asp Lys Ile Gly Asn Val Leu Val 145 150 155 160 Gly Glu Gly Val Thr Val Leu Ser Leu Pro Thr Ser Tyr Asp Leu Gly 165 170 175 Tyr Val Arg Leu Gly Asp Pro Ile Pro Ala Ile Gly Leu Asp Pro Lys 180 185 190 Met Val Ala Thr Cys Asp Ser Ser Asp Arg Pro Arg Val Tyr Thr Ile 195 200 205 Thr Ala Ala Asp Asp Tyr Gln Phe Ser Ser Gln Tyr Gln Ser Gly Gly 210 215 220 Val Thr Ile Thr Leu Phe Ser Ala Asn Ile Asp Ala Ile Thr Ser Leu 225 230 235 240 Ser Val Gly Gly Glu Leu Val Phe Gln Thr Ser Val Gln Gly Leu Ile 245 250 255 Leu Gly Ala Thr Ile Tyr Leu Ile Gly Phe Asp Gly Thr Thr Val Ile 260 265 270 Thr Arg Ala Val Ala Ala Asn Asn Gly Leu Thr Ala Gly Thr Asp Asn 275 280 285 Leu Met Pro Phe Asn Leu Val Ile Pro Thr Asn Glu Ile Thr Gln Pro 290 295 300 Ile Thr Ser Ile Lys Leu Glu Ile Val Thr Ser Lys Ser Gly Gly Gln 305 310 315 320 Ala Gly Asp Gln Met Ser Trp Ser Val Ser Gly Ser Leu Ala Val Thr 325 330 335 Ile His Gly Gly Asn Tyr Pro Gly Ala Leu Arg Pro Val Thr Leu Val 340 345 350 Ala Tyr Glu Arg Val Ala Thr Gly Ser Val Val Thr Val Ala Gly Val 355 360 365 Ser Asn Phe Glu Leu Ile Pro Asn Pro Glu Leu Ala Lys Asn Leu Val 370 375 380 Thr Glu Tyr Gly Arg Phe Asp Pro Gly Ala Met Asn Tyr Thr Lys Leu 385 390 395 400 Ile Leu Ser Glu Arg Asp Arg Leu Gly Ile Arg Thr Val Trp Pro Thr 405 410 415 Arg Glu Tyr Thr Asp Phe Arg Glu Tyr Phe Met Glu Val Ala Asp Leu 420 425 430 Asn Ser Pro Leu Lys Ile Ala Gly Ala Phe Gly Phe Lys Asp Ile Ile 435 440 445 Arg Ala Ile Arg Arg Ile Ala Val Pro Val Val Ser Thr Leu Phe Pro 450 455 460 Pro Ala Ala Pro Leu Ala His Ala Ile Gly Glu Gly Val Asp Tyr Leu 465 470 475 480 Leu Gly Asp Glu Ala Gln Ala Ala Ser Gly Thr Ala Arg Ala Ala Ser 485 490 495 Gly Lys Ala Arg Ala Ala Ser Gly Arg Ile Arg Gln Leu Thr Leu Ala 500 505 510 Ala Asp Lys Gly Tyr Glu Val Val Ala Asn Leu Phe Gln Val Pro Gln 515 520 525 Asn Pro Val Val Asp Gly Ile Leu Ala Ser Pro Gly Val Leu Arg Gly 530 535 540 Ala His Asn Leu Asp Cys Val Leu Arg Glu Gly Ala Thr Leu Phe Pro 545 550 555 560 Val Val Ile Thr Thr Val Glu Asp Ala Met Thr Pro Lys Ala Leu Asn 565 570 575 Ser Lys Met Phe Ala Val Ile Glu Gly Val Arg Glu Asp Leu Gln Pro 580 585 590 Pro Ser Gln Arg Gly Ser Phe Ile Arg Thr Leu Ser Gly His Arg Val 595 600 605 Tyr Gly Tyr Ala Pro Asp Gly Val Leu Pro Leu Glu Thr Gly Arg Asp 610 615 620 Tyr Thr Val Val Pro Ile Asp Asp Val Trp Asp Asp Ser Ile Met Leu 625 630 635 640 Ser Lys Asp Pro Ile Pro Pro Ile Val Gly Asn Ser Gly Asn Leu Ala 645 650 655 Ile Ala Tyr Met Asp Val Phe Arg Pro Lys Val Pro Ile His Val Ala 660 665 670 Met Thr Gly Ala Leu Asn Ala Cys Gly Glu Ile Glu Lys Val Ser Phe 675 680 685 Arg Ser Thr Lys Leu Ala Thr Ala His Arg Leu Gly Leu Lys Leu Ala 690 695 700 Gly Pro Gly Ala Phe Asp Val Asn Thr Gly Pro Asn Trp Ala Thr Phe 705 710 715 720 Ile Lys Arg Phe Pro His Asn Pro Arg Asp Trp Asp Arg Leu Pro Tyr 725 730 735 Leu Asn Leu Pro Tyr Leu Pro Pro Asn Ala Gly Arg Gln Tyr His Leu 740 745 750 Ala Met Ala Ala Ser Glu Phe Lys Glu Thr Pro Glu Leu Glu Ser Ala 755 760 765 Val Arg Ala Met Glu Ala Ala Ala Asn Val Asp Pro Leu Phe Gln Ser 770 775 780 Ala Leu Ser Val Phe Met Trp Leu Glu Glu Asn Gly Ile Val Thr Asp 785 790 795 800 Met Ala Asn Phe Ala Leu Ser Asp Pro Asn Ala His Arg Met Arg Asn 805 810 815 Phe Leu Ala Asn Ala Pro Gln Ala Gly Ser Lys Ser Gln Arg Ala Lys 820 825 830 Tyr Gly Thr Ala Gly Tyr Gly Val Glu Ala Arg Gly Pro Thr Pro Glu 835 840 845 Glu Ala Gln Arg Glu Lys Asp Thr Arg Ile Ser Lys Lys Met Glu Thr 850 855 860 Met Gly Ile Tyr Phe Ala Thr Pro Glu Trp Val Ala Leu Asn Gly His 865 870 875 880 Arg Gly Pro Ser Pro Gly Gln Leu Lys Tyr Trp Gln Asn Thr Arg Glu 885 890 895 Ile Pro Asp Pro Asn Glu Asp Tyr Leu Asp Tyr Val His Ala Glu Lys 900 905 910 Ser Arg Leu Ala Ser Glu Glu Gln Ile Gln Arg Ala Ala Thr Ser Ile 915 920 925 Tyr Gly Ala Pro Gly Gln Ala Glu Pro Pro Gln Ala Phe Ile Asp Glu 930 935 940 Val Ala Lys Val Tyr Glu Ile Asn His Gly Arg Gly Pro Asn Gln Glu 945 950 955 960 Gln Met Lys Asp Leu Leu Leu Thr Ala Met Glu Met Lys His Arg Asn 965 970 975 Pro Arg Arg Ala Pro Pro Lys Pro Lys Pro Lys Pro Asn Ala Pro Thr 980 985 990 Gln Arg Pro Pro Gly Arg Leu Gly Arg Trp Ile Arg Thr Val Ser Asp 995 1000 1005 Glu Asp Leu Glu 1010 <210> 5 <211> 830 <212> PRT <213> Infectious bursal disease virus IBD K7 <400> 5 Met Ser Asp Ile Phe Asn Ser Pro Gln Ala Arg Ser Thr Ile Ser Ala 1 5 10 15 Ala Phe Gly Ile Lys Pro Thr Ala Gly Gln Asp Val Glu Glu Leu Leu 20 25 30 Ile Pro Lys Val Trp Val Pro Pro Glu Asp Pro Leu Ala Ser Pro Ser 35 40 45 Arg Leu Ala Lys Phe Leu Arg Glu Asn Gly Tyr Lys Val Leu Gln Pro 50 55 60 Arg Ser Leu Pro Glu Asn Glu Asp Tyr Glu Thr Asp Gln Ile Leu Pro 65 70 75 80 Asp Leu Ala Trp Met Arg Gln Val Glu Gly Ala Val Leu Lys Pro Thr 85 90 95 Leu Ser Leu Pro Ile Gly Asp Gln Glu Tyr Phe Pro Lys Tyr Tyr Pro 100 105 110 Thr His Arg Pro Ser Lys Glu Lys Pro Asn Ala Tyr Pro Pro Asp Ile 115 120 125 Ala Leu Leu Lys Gln Met Ile Tyr Leu Phe Leu Gln Val Pro Glu Ala 130 135 140 Asn Glu Gly Leu Lys Asp Glu Val Thr Leu Leu Thr Gln Asn Ile Arg 145 150 155 160 Asp Lys Ala Tyr Gly Ser Gly Thr Tyr Met Gly Gln Ala Thr Arg Leu 165 170 175 Val Ala Met Lys Glu Val Ala Thr Gly Arg Asn Pro Asn Lys Asp Pro 180 185 190 Leu Lys Leu Gly Tyr Thr Phe Glu Ser Ile Ala Gln Leu Leu Asp Ile 195 200 205 Thr Leu Pro Val Gly Pro Pro Gly Glu Asp Asp Lys Pro Trp Val Pro 210 215 220 Leu Thr Arg Val Pro Ser Arg Met Leu Val Leu Thr Gly Asp Val Asp 225 230 235 240 Gly Asp Phe Glu Val Glu Asp Tyr Leu Pro Lys Ile Asn Leu Lys Ser 245 250 255 Ser Ser Gly Leu Pro Tyr Val Gly Arg Thr Lys Gly Glu Thr Ile Gly 260 265 270 Glu Met Ile Ala Ile Ser Asn Gln Phe Leu Arg Glu Leu Ser Thr Leu 275 280 285 Leu Lys Gln Gly Ala Gly Thr Lys Gly Ser Asn Lys Lys Lys Leu Leu 290 295 300 Ser Met Leu Ser Asp His Trp Tyr Leu Ser Cys Gly Leu Leu Phe Pro 305 310 315 320 Lys Ala Glu Arg Tyr Asp Lys Ser Thr Trp Leu Thr Lys Thr Arg Asn 325 330 335 Ile Trp Ser Ala Pro Ser Pro Thr His Leu Met Ile Ser Met Ile Thr 340 345 350 Trp Pro Val Met Ser Asn Ser Pro Asn Asn Val Leu Asn Ile Glu Gly 355 360 365 Cys Pro Ser Leu Tyr Lys Phe Asn Pro Phe Arg Gly Gly Leu Asn Arg 370 375 380 Ile Val Glu Trp Ile Leu Ala Pro Glu Glu Pro Lys Ala Leu Val Tyr 385 390 395 400 Ala Asp Asn Ile Tyr Ile Val His Ser Asn Thr Trp Tyr Ser Ile Asp 405 410 415 Leu Glu Lys Gly Glu Ala Asn Cys Thr Arg Gln His Met Gln Ala Ala 420 425 430 Met Tyr Tyr Ile Leu Thr Arg Gly Trp Ser Asp Asn Gly Asp Pro Met 435 440 445 Phe Asn Gln Thr Trp Ala Thr Phe Ala Met Asn Ile Ala Pro Ala Leu 450 455 460 Val Val Asp Ser Ser Cys Leu Ile Met Asn Leu Gln Ile Lys Thr Tyr 465 470 475 480 Gly Gln Gly Ser Gly Asn Ala Ala Thr Phe Ile Asn Asn His Leu Leu 485 490 495 Ser Thr Leu Val Leu Asp Gln Trp Asn Leu Met Arg Gln Pro Arg Pro 500 505 510 Asp Ser Glu Glu Phe Lys Ser Ile Glu Asp Lys Leu Gly Ile Asn Phe 515 520 525 Lys Ile Glu Arg Ser Ile Asp Asp Ile Arg Gly Lys Leu Arg Gln Leu 530 535 540 Val Leu Leu Ala Gln Pro Gly Tyr Leu Ser Gly Gly Val Glu Pro Glu 545 550 555 560 Gln Ser Ser Pro Thr Val Glu Leu Asp Leu Leu Gly Trp Ser Ala Thr 565 570 575 Tyr Ser Lys Asp Leu Gly Ile Tyr Val Pro Val Leu Asp Lys Glu Arg 580 585 590 Leu Phe Cys Ser Ala Ala Tyr Pro Lys Gly Val Glu Asn Lys Ser Leu 595 600 605 Lys Ser Lys Val Gly Ile Glu Gln Ala Tyr Lys Val Val Arg Tyr Glu 610 615 620 Ala Leu Arg Leu Val Gly Gly Trp Asn Tyr Pro Leu Leu Asn Lys Ala 625 630 635 640 Cys Lys Asn Asn Ala Gly Ala Ala Arg Arg His Leu Glu Ala Lys Gly 645 650 655 Phe Pro Leu Asp Glu Phe Leu Ala Glu Trp Ser Glu Leu Ser Glu Phe 660 665 670 Gly Glu Ala Phe Glu Gly Phe Asn Ile Lys Leu Thr Val Thr Ser Glu 675 680 685 Ser Leu Ala Glu Leu Asn Lys Pro Val Pro Pro Lys Pro Pro Asn Val 690 695 700 Asn Arg Pro Val Asn Thr Gly Gly Leu Lys Ala Val Ser Asn Ala Leu 705 710 715 720 Lys Thr Gly Arg Tyr Arg Asn Glu Ala Gly Leu Ser Gly Leu Val Leu 725 730 735 Leu Ala Thr Ala Arg Ser Arg Leu Gln Asp Ala Val Lys Ala Lys Ala 740 745 750 Glu Ala Glu Lys Leu His Lys Ser Lys Pro Asp Asp Pro Asp Ala Asp 755 760 765 Trp Phe Glu Arg Ser Glu Thr Leu Ser Asp Leu Leu Glu Lys Ala Asp 770 775 780 Ile Ala Ser Lys Val Ala His Ser Ala Leu Val Glu Thr Ser Asp Ala 785 790 795 800 Leu Glu Ala Val Gln Ser Thr Ser Val Tyr Thr Pro Lys Tyr Pro Glu 805 810 815 Val Lys Asn Pro Gln Thr Ala Ser Asn Pro Val Val Gly Leu 820 825 830 <110> Konkuk University Industrial Cooperation Corp. <120> Novel infectious bursal disease virus IBD K7 and infectious          bursal disease <160> 5 <170> Kopatentin 1.71 <210> 1 <211> 3131 <212> DNA <213> infectious bursal disease virus IBD K7 <400> 1 gggacaggcc gtcaaggcct tgttccagga tggaactcct ccttctacaa cgctatcatt 60 gatggttagt agagatcaga caaacgatcg cagcgatgac aaacctgcaa gatcaaaccc 120 aacagattgt tccgttcata cggagccttc tgatgccaac aaccggaccg gcgtccattc 180 cggacgacac cctggagaag cacactctca ggtcagagac ctcgacctac aatttgactg 240 tgggggacac agggtcaggg ctaattgtct ttttccctgg attccctggc tcaactgtgg 300 gtgctcacta cacactgcag agcaatggga actacaagtt cgatcagatg ctcctgactg 360 cccagaacct accggccagt tacaactact gcaggctagt gagtcggagt ctcacagtga 420 ggtcaagcac acttcctggt ggcgtttatg cactaaacgg caccataaac gccgtgacct 480 tccaaggaag cctgagtgaa ctgacagatg ttagctacaa tgggttgatg tctgcaacag 540 ccaacatcaa cgacaaaatt gggaacgtcc tggtagggga aggggtcacc gtcctcagct 600 tacccacatc atatgatctt gggtatgtga ggcttggtga ccccattccc gcaatagggc 660 ttgacccaaa aatggtagcc acatgtgaca gcagcgacag acccagagtc tacaccataa 720 ctgcagccga tgattaccaa ttctcatcac agtaccaatc aggtggggta acaatcacac 780 tgttctcagc caacattgat gccatcacca gcctcagcgt tgggggggag ctcgtgtttc 840 aaacaagcgt ccaaggcctt atactgggcg ccaccatcta ccttataggc tttgatggga 900 caacggtaat caccagagct gtggccgcaa acaatgggct gacggccggc accgacaatc 960 ttatgccatt caatcttgtg attccaacca acgagataac ccagccaatc acatccatca 1020 aactggagat agtgacctcc aaaagtggtg gtcaggcagg ggatcagatg tcatggtcgg 1080 taagtgggag cctagcagtg acgatccatg gtggcaacta cccaggggcc ctccgtcccg 1140 tcacgctagt agcctacgaa agagtggcaa caggatccgt cgttacggtc gctggggtga 1200 gcaacttcga gctgatccca aatcctgaac tagcaaagaa cctggttaca gaatacggcc 1260 gatttgaccc aggagccatg aactacacaa aattgatact gagtgagagg gaccgtcttg 1320 gcatcaggac cgtctggcca acaagggagt acactgactt tcgtgaatac ttcatggagg 1380 tggccgacct caactctccc ctgaagattg caggagcatt cggcttcaaa gacataatcc 1440 gggccataag gaggatagct gtgccggtgg tctccacatt gttcccacct gccgctcccc 1500 tagcccatgc aattggggaa ggtgtagact acctgctggg cgatgaggca caggctgctt 1560 caggaactgc tcgagccgcg tcaggaaaag caagagctgc ctcaggccgc ataaggcagc 1620 tgactctcgc cgccgacaag gggtacgagg tagtcgcgaa tctattccag gtgccccaga 1680 atcccgtagt cgacgggatt cttgcttcac ctggggtact ccgcggtgcg cacaacctcg 1740 actgcgtgtt aagagagggt gccacgctat tccccgtggt cattacgaca gtggaagacg 1800 ccatgacacc caaagcattg aacagcaaaa tgtttgctgt cattgaaggc gtgcgagaag 1860 acctccaacc tccatctcaa agaggatcct tcatacgaac tctctctgga cacagagtct 1920 atggatatgc tccagatggg gtacttccac tggagactgg gagagactac accgttgtcc 1980 caatagatga tgtctgggac gacagcatta tgctgtccaa agaccccata cctcctattg 2040 tgggaaacag tggaaaccta gccatagctt acatggatgt gtttcgaccc aaagtcccta 2100 tccatgtggc tatgacggga gccctcaatg cttgtggcga gattgagaaa gtaagcttta 2160 gaagcaccaa gctcgccact gcacaccgac ttggcctcaa gttggctggt cccggagcat 2220 tcgatgtaaa caccgggccc aactgggcaa cgttcatcaa acgtttccct cacaatccac 2280 gcgactggga caggctcccc tacctcaacc taccatacct tccacccaat gcaggacgcc 2340 agtaccacct tgccatggct gcatcagagt tcaaagagac ccccgaactc gagagcgccg 2400 tcagagcaat ggaagcagca gccaacgtgg acccactatt ccaatctgca ctcagtgtgt 2460 tcatgtggct ggaagagaat gggattgtga ctgacatggc caacttcgca ctcagcgacc 2520 caaacgccca tcggatgcga aattttcttg caaacgcacc acaagcaggt agcaagtcgc 2580 aaagggccaa gtacgggaca gcaggctacg gagtggaggc ccggggcccc acaccagagg 2640 aagcacagag ggaaaaagac acacggatct caaagaagat ggagaccatg ggcatctact 2700 ttgcaacacc agaatgggta gcactcaatg ggcaccgagg gccaagcccc ggccagctaa 2760 agtactggca gaacacacga gaaataccgg acccaaacga ggactatcta gactacgtgc 2820 atgcagagaa gagccggttg gcatcagaag aacaaatcca acgggcagct acgtcgatct 2880 acggggctcc aggacaggca gagccacccc aagctttcat agacgaagtt gccaaagtct 2940 atgaaatcaa ccatggacgt ggcccaaacc aagaacagat gaaagatctg ctcttgactg 3000 cgatggagat gaagcatcgc aatcccaggc gggctccacc aaagcccaag ccaaaaccca 3060 atgctccaac acagagaccc cctggtcggc tgggccgctg gatcaggact gtctctgatg 3120 aggaccttga g 3131 <210> 2 <211> 2687 <212> DNA <213> infectious bursal disease virus IBD K7 <400> 2 tcctcttctt gatgattctg ccaccatgag tgacattttc aacagtccac aggcgcgaag 60 cacgatctca gcagcgttcg gcataaagcc tactgctgga caagacgtgg aagaactctt 120 gatccctaaa gtttgggtgc cacctgagga tccgcttgcc agccctagtc gactggcaaa 180 gttcctcaga gagaacggct acaaagtttt gcagccacgg tctctgcccg agaatgagga 240 ttatgagacc gaccaaatac tcccagactt agcatggatg cgacaggtag aaggggctgt 300 tttaaaacct actctatctc tccctattgg agatcaggag tacttcccaa agtactaccc 360 aacacatcgc cctagcaagg agaagcccaa tgcgtacccg ccagacattg cactactcaa 420 gcagatgatt tacctgtttc tccaggttcc agaggccaac gagggcctaa aggatgaagt 480 aaccctcttg acccaaaaca taagggacaa ggcctatgga agtgggacct acatgggaca 540 agcaactcga cttgtggcca tgaaggaggt cgccactggg agaaacccaa acaaggatcc 600 tctaaagctt gggtacactt ttgagagcat cgcgcagcta cttgacatca cactaccggt 660 aggcccaccc ggtgaggatg acaagccctg ggtgccactc acaagagtgc cgtcacggat 720 gttggtgctg acgggagacg tagatggcga ctttgaggtt gaagactacc ttcccaaaat 780 caacctcaag tcatcaagtg gactaccata tgtaggtcgc accaaaggag agacaattgg 840 cgagatgata gctatctcaa accagtttct cagagagcta tcaacactgt tgaagcaagg 900 tgcagggaca aaggggtcaa acaagaagaa gctactcagc atgttaagtg accattggta 960 cttatcatgc gggcttttgt ttccaaaggc tgaaaggtac gacaaaagta catggctcac 1020 caagacccgg aacatatggt cagctccatc ccctacacac ctcatgatct ccatgatcac 1080 ctggcccgtg atgtccaaca gcccaaataa cgtgttgaac attgaagggt gtccatcact 1140 ctacaaattc aacccgttca gaggagggtt gaacaggatc gtcgagtgga tattggcccc 1200 ggaagaaccc aaggctcttg tatatgcgga caacatatac attgtccact caaacacgtg 1260 gtactcaatt gacctagaga agggcgaggc aaactgcact cgccaacaca tgcaagctgc 1320 aatgtactac atactcacca gagggtggtc agacaacggc gacccaatgt tcaatcaaac 1380 atgggccacc tttgccatga acattgcccc tgctctagtg gtagactcat cgtgcctgat 1440 aatgaacctg caaattaaga cctatggtca aggcagcggg aatgcagcca cgttcatcaa 1500 caaccacctc ttgagcacgc tagtgcttga ccagtggaac ctgatgagac agcccagacc 1560 agacagcgag gagttcaaat caattgagga caagctaggt atcaacttta agattgagag 1620 gtccattgat gacatcaggg gcaagctgag acagcttgtc ctccttgcac aaccagggta 1680 cctgagtggg ggggttgaac cagaacaatc cagcccaact gttgagcttg acctactagg 1740 gtggtcagct acatacagca aagatctcgg gatctatgtg ccggtgcttg acaaggaacg 1800 cctattttgt tctgctgcgt atcccaaggg agtagagaac aagagtctca agtccaaagt 1860 cgggatcgag caggcataca aggtagtcag gtatgaggcg ttgaggttgg taggtggttg 1920 gaactaccca ctcctgaaca aagcctgcaa gaataacgca ggcgccgctc ggcggcatct 1980 ggaggccaag gggttcccac tcgacgagtt cctagccgag tggtctgagc tgtcagagtt 2040 cggtgaggcc ttcgaaggct tcaatatcaa gctgaccgta acatctgaga gcctagccga 2100 actgaacaag ccagtacccc ccaagccccc aaatgtcaac agaccagtca acactggggg 2160 actcaaggca gtcagcaacg ccctcaagac cggtcggtac aggaacgaag ccggactgag 2220 tggtctcgtc cttctagcca cagcaagaag ccgtctgcaa gatgcagtta aggccaaggc 2280 agaagccgag aaactccaca agtccaagcc tgacgacccc gatgcagact ggttcgaaag 2340 atcagaaact ctgtcagacc ttctggagaa agccgacatc gccagcaagg tcgcccactc 2400 agcactcgtg gaaacaagcg acgcccttga agcagttcag tcgacttccg tgtacacccc 2460 caagtaccca gaagtcaaga acccacagac cgcctccaac cccgttgttg ggctccacct 2520 gcccgccaag agagccaccg gtgtccaggc cgctcttctc ggagcaggaa cgagcagacc 2580 aatggggatg gaggccccaa tacggtccaa gaacgccgtg aaaatggcca aacggcggca 2640 acgccaaaag gagagccgct aacagccatg atgggaacca ctcaaga 2687 <210> 3 <211> 145 <212> PRT <213> infectious bursal disease virus IBD K7 <400> 3 Met Val Ser Arg Asp Gln Thr Asn Asp Arg Ser Asp Asp Lys Pro Ala   1 5 10 15 Arg Ser Asn Pro Thr Asp Cys Ser Val His Thr Glu Pro Ser Asp Ala              20 25 30 Asn Asn Arg Thr Gly Val His Ser Gly Arg His Pro Gly Glu Ala His          35 40 45 Ser Gln Val Arg Asp Leu Asp Leu Gln Phe Asp Cys Gly Gly His Arg      50 55 60 Val Arg Ala Asn Cys Leu Phe Pro Trp Ile Pro Trp Leu Asn Cys Gly  65 70 75 80 Cys Ser Leu His Thr Ala Glu Gln Trp Glu Leu Gln Val Arg Ser Serp                  85 90 95 Ala Pro Asp Cys Pro Glu Pro Thr Gly Gln Leu Gln Leu Leu Gln Ala             100 105 110 Ser Glu Ser Glu Ser His Ser Glu Val Lys His Thr Ser Trp Trp Arg         115 120 125 Leu Cys Thr Lys Arg His His Lys Arg Arg Asp Leu Pro Arg Lys Pro     130 135 140 Glu 145 <210> 4 <211> 1012 <212> PRT <213> infectious bursal disease virus IBD K7 <400> 4 Met Thr Asn Leu Gln Asp Gln Thr Gln Gln Ile Val Pro Phe Ile Arg   1 5 10 15 Ser Leu Leu Met Pro Thr Thr Gly Pro Ala Ser Ile Pro Asp Asp Thr              20 25 30 Leu Glu Lys His Thr Leu Arg Ser Glu Thr Ser Thr Tyr Asn Leu Thr          35 40 45 Val Gly Asp Thr Gly Ser Gly Leu Ile Val Phe Phe Pro Gly Phe Pro      50 55 60 Gly Ser Thr Val Gly Ala His Tyr Thr Leu Gln Ser Asn Gly Asn Tyr  65 70 75 80 Lys Phe Asp Gln Met Leu Leu Thr Ala Gln Asn Leu Pro Ala Ser Tyr                  85 90 95 Asn Tyr Cys Arg Leu Val Ser Ser Ser Leu Thr Val Arg Ser Ser Thr             100 105 110 Leu Pro Gly Gly Val Tyr Ala Leu Asn Gly Thr Ile Asn Ala Val Thr         115 120 125 Phe Gln Gly Ser Leu Ser Glu Leu Thr Asp Val Ser Tyr Asn Gly Leu     130 135 140 Met Ser Ala Thr Ala Asn Ile Asn Asp Lys Ile Gly Asn Val Leu Val 145 150 155 160 Gly Glu Gly Val Thr Val Leu Ser Leu Pro Thr Ser Tyr Asp Leu Gly                 165 170 175 Tyr Val Arg Leu Gly Asp Pro Ile Pro Ala Ile Gly Leu Asp Pro Lys             180 185 190 Met Val Ala Thr Cys Asp Ser Ser Asp Arg Pro Arg Val Tyr Thr Ile         195 200 205 Thr Ala Asp Asp Tyr Gln Phe Ser Ser Gln Tyr Gln Ser Gly Gly     210 215 220 Val Thr Ile Thr Leu Phe Ser Ala Asn Ile Asp Ala Ile Thr Ser Leu 225 230 235 240 Ser Val Gly Gly Glu Leu Val Phe Gln Thr Ser Val Gln Gly Leu Ile                 245 250 255 Leu Gly Ala Thr Ile Tyr Leu Ile Gly Phe Asp Gly Thr Thr Val Ile             260 265 270 Thr Arg Ala Val Ala Asn Asn Gly Leu Thr Ala Gly Thr Asp Asn         275 280 285 Leu Met Pro Phe Asn Leu Val Ile Pro Thr Asn Glu Ile Thr Gln Pro     290 295 300 Ile Thr Ser Ile Lys Leu Glu Ile Val Thr Ser Lys Ser Gly Gly Gln 305 310 315 320 Ala Gly Asp Gln Met Ser Trp Ser Val Ser Gly Ser Leu Ala Val Thr                 325 330 335 Ile His Gly Gly Asn Tyr Pro Gly Ala Leu Arg Pro Val Thr Leu Val             340 345 350 Ala Tyr Glu Arg Val Ala Thr Gly Ser Val Val Thr Val Ala Gly Val         355 360 365 Ser Asn Phe Glu Leu Ile Pro Asn Pro Glu Leu Ala Lys Asn Leu Val     370 375 380 Thr Gly Tyr Gly Arg Phe Asp Pro Gly Ala Met Asn Tyr Thr Lys Leu 385 390 395 400 Ile Leu Ser Glu Arg Asp Arg Leu Gly Ile Arg Thr Val Trp Pro Thr                 405 410 415 Arg Glu Tyr Thr Asp Phe Arg Glu Tyr Phe Met Glu Val Ala Asp Leu             420 425 430 Asn Ser Pro Leu Lys Ile Ala Gly Ala Phe Gly Phe Lys Asp Ile Ile         435 440 445 Arg Ala Ile Arg Arg Ile Ala Val Pro Val Val Ser Thr Leu Phe Pro     450 455 460 Pro Ala Ala Pro Leu Ala His Ale Ile Gly Glu Gly Val Asp Tyr Leu 465 470 475 480 Leu Gly Asp Glu Ala Gln Ala Ala Ser Gly Thr Ala Arg Ala Ala Ser                 485 490 495 Gly Lys Ala Arg Ala Ala Ser Gly Arg Ile Arg Gln Leu Thr Leu Ala             500 505 510 Ala Asp Lys Gly Tyr Glu Val Val Ala Asn Leu Phe Gln Val Pro Gln         515 520 525 Asn Pro Val Val Asp Gly Ile Leu Ala Ser Pro Gly Val Leu Arg Gly     530 535 540 Ala His Asn Leu Asp Cys Val Leu Arg Glu Gly Ala Thr Leu Phe Pro 545 550 555 560 Val Val Ile Thr Thr Val Glu Asp Ala Met Thr Pro Lys Ala Leu Asn                 565 570 575 Ser Lys Met Phe Ala Val Ile Glu Gly Val Arg Glu Asp Leu Gln Pro             580 585 590 Pro Ser Gln Arg Gly Ser Phe Ile Arg Thr Leu Ser Gly His Arg Val         595 600 605 Tyr Gly Tyr Ala Pro Asp Gly Val Leu Pro Leu Glu Thr Gly Arg Asp     610 615 620 Tyr Thr Val Val Pro Ile Asp Asp Val Trp Asp Asp Ser Ile Met Leu 625 630 635 640 Ser Lys Asp Pro Ile Pro Pro Ile Val Gly Asn Ser Gly Asn Leu Ala                 645 650 655 Ile Ala Tyr Met Asp Val Phe Arg Pro Lys Val Pro Ile His Val Ala             660 665 670 Met Thr Gly Ala Leu Asn Ala Cys Gly Glu Ile Glu Lys Val Ser Phe         675 680 685 Arg Ser Thr Lys Leu Ala Thr Ala His Arg Leu Gly Leu Lys Leu Ala     690 695 700 Gly Pro Gly Ala Phe Asp Val Asn Thr Gly Pro Asn Trp Ala Thr Phe 705 710 715 720 Ile Lys Arg Phe Pro His Asn Pro Arg Asp Trp Asp Arg Leu Pro Tyr                 725 730 735 Leu Asn Leu Pro Tyr Leu Pro Pro Asn Ala Gly Arg Gln Tyr His Leu             740 745 750 Ala Met Ala Ala Ser Glu Phe Lys Glu Thr Pro Glu Leu Glu Ser Ala         755 760 765 Val Arg Ala Met Glu Ala Ala Ala Asn Val Asp Pro Leu Phe Gln Ser     770 775 780 Ala Leu Ser Val Phe Met Trp Leu Glu Glu Asn Gly Ile Val Thr Asp 785 790 795 800 Met Ala Asn Phe Ala Leu Ser Asp Pro Asn Ala His Arg Met Arg Asn                 805 810 815 Phe Leu Ala Asn Ala Pro Gln Ala Gly Ser Lys Ser Gln Arg Ala Lys             820 825 830 Tyr Gly Thr Ala Gly Tyr Gly Val Glu Ala Arg Gly Pro Thr Pro Glu         835 840 845 Glu Ala Gln Arg Glu Lys Asp Thr Arg Ile Ser Lys Lys Met Glu Thr     850 855 860 Met Gly Ile Tyr Phe Ala Thr Pro Glu Trp Val Ala Leu Asn Gly His 865 870 875 880 Arg Gly Pro Ser Pro Gly Gln Leu Lys Tyr Trp Gln Asn Thr Arg Glu                 885 890 895 Ile Pro Asp Pro Asn Glu Asp Tyr Leu Asp Tyr Val His Ala Glu Lys             900 905 910 Ser Arg Leu Ala Ser Glu Glu Gln Ile Gln Arg Ala Ala Thr Ser Ile         915 920 925 Tyr Gly Ala Pro Gly Gln Ala Glu Pro Pro Gln Ala Phe Ile Asp Glu     930 935 940 Val Ala Lys Val Tyr Glu Ile Asn His Gly Arg Gly Pro Asn Gln Glu 945 950 955 960 Gln Met Lys Asp Leu Leu Leu Thr Ala Met Glu Met Lys His Arg Asn                 965 970 975 Pro Arg Arg Ala Pro Pro Lys Pro Lys Pro Lys Pro Asn Ala Pro Thr             980 985 990 Gln Arg Pro Pro Gly Arg Leu Gly Arg Trp Ile Arg Thr Val Ser Asp         995 1000 1005 Glu Asp Leu Glu    1010 <210> 5 <211> 830 <212> PRT <213> Infectious bursal disease virus IBD K7 <400> 5 Met Ser Asp Ile Phe Asn Ser Pro Gln Ala Arg Ser Thr Ile Ser Ala   1 5 10 15 Ala Phe Gly Ile Lys Pro Thr Ala Gly Gln Asp Val Glu Glu Leu Leu              20 25 30 Ile Pro Lys Val Trp Val Pro Pro Glu Asp Pro Leu Ala Ser Pro Ser          35 40 45 Arg Leu Ala Lys Phe Leu Arg Glu Asn Gly Tyr Lys Val Leu Gln Pro      50 55 60 Arg Ser Leu Pro Glu Asn Glu Asp Tyr Glu Thr Asp Gln Ile Leu Pro  65 70 75 80 Asp Leu Ala Trp Met Arg Gln Val Glu Gly Ala Val Leu Lys Pro Thr                  85 90 95 Leu Ser Leu Pro Ile Gly Asp Gln Glu Tyr Phe Pro Lys Tyr Tyr Pro             100 105 110 Thr His Arg Pro Ser Lys Glu Lys Pro Asn Ala Tyr Pro Pro Asp Ile         115 120 125 Ala Leu Leu Lys Gln Met Ile Tyr Leu Phe Leu Gln Val Pro Glu Ala     130 135 140 Asn Glu Gly Leu Lys Asp Glu Val Thr Leu Leu Thr Gln Asn Ile Arg 145 150 155 160 Asp Lys Ala Tyr Gly Ser Gly Thr Tyr Met Gly Gln Ala Thr Arg Leu                 165 170 175 Val Ala Met Lys Glu Val Ala Thr Gly Arg Asn Pro Asn Lys Asp Pro             180 185 190 Leu Lys Leu Gly Tyr Thr Phe Glu Ser Ile Ala Gln Leu Leu Asp Ile         195 200 205 Thr Leu Pro Val Gly Pro Pro Gly Glu Asp Asp Lys Pro Trp Val Pro     210 215 220 Leu Thr Arg Val Ser Ser Met Met Leu Val Leu Thr Gly Asp Val Asp 225 230 235 240 Gly Asp Phe Glu Val Glu Asp Tyr Leu Pro Lys Ile Asn Leu Lys Ser                 245 250 255 Ser Ser Gly Leu Pro Tyr Val Gly Arg Thr Lys Gly Glu Thr Ile Gly             260 265 270 Glu Met Ile Ala Ile Ser Asn Gln Phe Leu Arg Glu Leu Ser Thr Leu         275 280 285 Leu Lys Gln Gly Aly Gly Thr Lys Gly Ser Asn Lys Lys Lys Leu Leu     290 295 300 Ser Met Leu Ser Asp His Trp Tyr Leu Ser Cys Gly Leu Leu Phe Pro 305 310 315 320 Lys Ala Glu Arg Tyr Asp Lys Ser Thr Trp Leu Thr Lys Thr Arg Asn                 325 330 335 Ile Trp Ser Ala Pro Ser Pro Thr His Leu Met Ile Ser Met Ile Thr             340 345 350 Trp Pro Val Met Ser Asn Ser Pro Asn Asn Val Leu Asn Ile Glu Gly         355 360 365 Cys Pro Ser Leu Tyr Lys Phe Asn Pro Phe Arg Gly Gly Leu Asn Arg     370 375 380 Ile Val Glu Trp Ile Leu Ala Pro Glu Glu Pro Lys Ala Leu Val Tyr 385 390 395 400 Ala Asp Asn Ile Tyr Ile Val His Ser Asn Thr Trp Tyr Ser Ile Asp                 405 410 415 Leu Glu Lys Gly Glu Ala Asn Cys Thr Arg Gln His Met Gln Ala Ala             420 425 430 Met Tyr Tyr Ile Leu Thr Arg Gly Trp Ser Asp Asn Gly Asp Pro Met         435 440 445 Phe Asn Gln Thr Trp Ala Thr Phe Ala Met Asn Ile Ala Pro Ala Leu     450 455 460 Val Val Asp Ser Ser Cys Leu Ile Met Asn Leu Gln Ile Lys Thr Tyr 465 470 475 480 Gly Gln Gly Ser Gly Asn Ala Ala Thr Phe Ile Asn Asn His Leu Leu                 485 490 495 Ser Thr Leu Val Leu Asp Gln Trp Asn Leu Met Arg Gln Pro Arg Pro             500 505 510 Asp Ser Glu Glu Phe Lys Ser Ile Glu Asp Lys Leu Gly Ile Asn Phe         515 520 525 Lys Ile Glu Arg Ser Ile Asp Asp Ile Arg Gly Lys Leu Arg Gln Leu     530 535 540 Val Leu Leu Ala Gln Pro Gly Tyr Leu Ser Gly Gly Val Glu Pro Glu 545 550 555 560 Gln Ser Ser Pro Thr Val Glu Leu Asp Leu Leu Gly Trp Ser Ala Thr                 565 570 575 Tyr Ser Lys Asp Leu Gly Ile Tyr Val Pro Val Leu Asp Lys Glu Arg             580 585 590 Leu Phe Cys Ser Ala Ala Tyr Pro Lys Gly Val Glu Asn Lys Ser Leu         595 600 605 Lys Ser Lys Val Gly Ile Glu Gln Ala Tyr Lys Val Val Arg Tyr Glu     610 615 620 Ala Leu Arg Leu Val Gly Gly Trp Asn Tyr Pro Leu Leu Asn Lys Ala 625 630 635 640 Cys Lys Asn Asn Ala Gly Ala Ala Arg Arg His Leu Glu Ala Lys Gly                 645 650 655 Phe Pro Leu Asp Glu Phe Leu Ala Glu Trp Ser Glu Leu Ser Glu Phe             660 665 670 Gly Glu Ala Phe Glu Gly Phe Asn Ile Lys Leu Thr Val Thr Ser Glu         675 680 685 Ser Leu Ala Glu Leu Asn Lys Pro Val Pro Pro Lys Pro Pro Asn Val     690 695 700 Asn Arg Pro Val Asn Thr Gly Gly Leu Lys Ala Val Ser Asn Ala Leu 705 710 715 720 Lys Thr Gly Arg Tyr Arg Asn Glu Ala Gly Leu Ser Gly Leu Val Leu                 725 730 735 Leu Ala Thr Ala Arg Ser Ser Leu Gln Asp Ala Val Lys Ala Lys Ala             740 745 750 Glu Ala Glu Lys Leu His Lys Ser Lys Pro Asp Asp Pro Asp Ala Asp         755 760 765 Trp Phe Glu Arg Ser Glu Thr Leu Ser Asp Leu Leu Glu Lys Ala Asp     770 775 780 Ile Ala Ser Lys Val Ala His Ser Ala Leu Val Glu Thr Ser Asp Ala 785 790 795 800 Leu Glu Ala Val Gln Ser Thr Ser Val Tyr Thr Pro Lys Tyr Pro Glu                 805 810 815 Val Lys Asn Pro Gln Thr Ala Ser Asn Pro Val Val Gly Leu             820 825 830

Claims (11)

시그먼트 A 및 B로 구성되고, 상기 시그먼트 A는 서열번호 3 및 4에 기재된 아미노산 서열로 이루어지고, 상기 시그먼트 B는 서열번호 5에 기재된 아미노산 서열로 이루어진, 기탁번호 KCTC 12376BP로 기탁된 전염성 F낭병 바이러스 IBD K7주. Wherein Segment A comprises the amino acid sequence of SEQ ID NOS: 3 and 4, and Segment B comprises the amino acid sequence of SEQ ID NO: 5, F pandemic virus IBD K7 strain. 제 1항에 있어서, 상기 바이러스는 국내 육계로부터 분리한 전염성 F낭병 바이러스 IBD K7주.The virus according to claim 1, wherein the virus is an infectious F pancreatic virus IBD K7 strain isolated from domestic broiler chickens. 제 1항에 있어서, 상기 바이러스의 시그먼트 A 및 B는 각각 서열번호 1 및 2에 기재된 염기서열을 가지는 것을 특징으로 하는 전염성 F낭병 바이러스 IBD K7주.The Infectious Fungal disease IBD K7 strain according to claim 1, wherein the viruses have the nucleotide sequences of SEQ ID NOS: 1 and 2, respectively. 삭제delete 제1항의 바이러스 및 약학적 허용 담체 또는 희석제를 포함하는, 전염성 F낭병 바이러스 감염으로부터 유발된 질병에 대한 가금 보호용 백신. A vaccine for the protection of poultry against diseases caused by infectious F pandemic virus infection, comprising the virus of claim 1 and a pharmaceutically acceptable carrier or diluent. 제5항에 있어서, 상기 전염성 F낭병 바이러스가 살아있는 형태임을 특징으로 하는 백신.6. The vaccine according to claim 5, wherein the infectious F pancreatic virus is in a live form. 제5항에 있어서, 상기 백신이 보조제를 추가로 포함하는 것을 특징으로 하는 백신.6. The vaccine of claim 5, wherein the vaccine further comprises an adjuvant. 제5항에 있어서, 상기 백신이 가금에 감염성이 있는 다른 병원균의 백신성분을 하나 이상 추가로 포함하는 것을 특징으로 하는 백신.6. A vaccine according to claim 5, wherein the vaccine further comprises one or more vaccine components of other pathogens infectious to the poultry. 삭제delete 삭제delete 제5항의 백신을 조류에 투여하는 것을 포함하는 가금에서 전염성 F낭병 감염으로부터 유발된 질병을 제어하는 방법.A method for controlling a disease caused by an infectious F pancreatic infection in a poultice comprising administering the vaccine of claim 5 to the algae.
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