KR20010033398A - 오스트리니아 누빌라리스에 활성인 독소 - Google Patents
오스트리니아 누빌라리스에 활성인 독소 Download PDFInfo
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- KR20010033398A KR20010033398A KR1020007006867A KR20007006867A KR20010033398A KR 20010033398 A KR20010033398 A KR 20010033398A KR 1020007006867 A KR1020007006867 A KR 1020007006867A KR 20007006867 A KR20007006867 A KR 20007006867A KR 20010033398 A KR20010033398 A KR 20010033398A
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- asn
- gly
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Abstract
본 발명은 비포유 동물 해충(non-mamalian pests), 구체적으로 식물 해충 박멸에 유용한 물질 및 방법에 관한 것이다. 특정 구체예에서, 본 발명은 인시류(Lepidopterans) 박멸에 유용한 신규의 바실러스 츄린겐시스(Bacillus thuringensis) 독소를 제공한다. 바람직한 구체예에서, 본 발명의 독소는 유럽 옥수수 좀벌레인 오스트리니아 누빌라리스(Ostrinia nubilalis) 박멸에 사용된다. 본 발명은 또한 본 발명의 독소를 코드화하는 뉴클레오티드 서열을 추가로 제공한다. 본 발명의 뉴클레오티드 서열은 식물과 같은 숙주를 형질 전환시켜 본 발명의 살충 독소를 발현시킬 수 있다. 뿐만 아니라, 본 발명은 해충에 활성인 독소를 코드화하는 유전자 동정용 신규의 뉴클레오티드 프라이머에 관한 것이다. 상기 프라이머들은 PCR 기술에 사용되어 상기 독소들을 코드화하는 유전자의 특징을 이루는 유전자 절편을 생성하는데에 유용하다. 상기 프라이머들은 또한 상기 독소-코드화 유전자를 검출하는 프로브로서도 유용하다.
Description
토양 미생물인 바실러스 츄린겐시스(Bacillus thuringensis)(B.t)는 그램-양성, 포자-형성 세균이다. 대부분의 비.티.(B.t) 균주는 살충 활성을 나타내지 않는다. 몇몇 비.티. 균주들은 파라스포랄 결정성 단백질 함유물(parasporal crystalline protein inclusion)을 생산하는 것으로 특징지워진다. "δ-내독소(δ-endotoxin)"는 외독소와는 상이하여, 비특이적 숙주 범위를 갖는다. 상기 함유물들은 종종 현미경적으로 결정의 형태로서 관찰된다. 이들 단백질은 해충에 매우 독성이 강하며 이들의 독소 활성에 대해서도 특이적이다. 어떤 비.티. 독소 유전자는 분리후 시퀀싱한 결과, 재조합 DNA-기초 비.티. 산물을 생산하여 사용할 수 있게 되었다. 뿐만 아니라, 해충 저항성에 대한 비.티. 독소 유전자에 의하여 유전학적으로 조작된 식물 및 비.티. 독소 전달 비히클로서 원형 그대로의 안정화된 미생물 세포를 사용하는 것을 포함하는 유전 공학 기술을 사용하여, 비.티. 독소를 농업적 환경에 전달하는 신규의 방법을 개발중에 있다[Gaertner, F.H., L.Kim(1988) TIBTECH 6:S4-S7]. 따라서, 분리된 비.티. 내독소 유전자는 상업적으로 시판되고 있다.
지난 15년 동안, 시판되고 있던 비.티. 살충제는 좁은 범위의 인시류(lepidoptera)(모충) 해충에만 사용이 국한되어 왔다. 수년 동안 인시류 해충의 상업적 해충제로서 비.츄린겐시스 쿠르스타키 아종(B. thuringiensis subsp. kurstaki)의 포자 및 결정체를 제조하여 사용하였다. 예를 들어, 비.츄린겐시스 쿠르스타키 변종 HD-1(B. thuringiensis var. kurstaki HD-1)은 다수의 인시류 해충 애벌레에 독성이 있는 결정성 δ-내독소를 생산한다.
현재 연구자들은 이전보다 더욱 광범위한 해충에 특이성이 있는 비.티. 살충제를 발견하였는데, 이들은 예를 들어, 비.티.의 다른 종들 즉, 이스라엘렌시스(israelensis) 및 모리소니(morrisoni)[ 테네브리오니스(tenebrionis), 비.티.M-7(B.t. M-7), 비.티. 샌 디에고(B.t. san diego)라고도 알려져 있음]는 쌍시류(Diptera) 목 및 초시류(Coleoptera) 목 곤충들을 박멸하는데에 상업적으로 사용되고 있다[Gaertner, F.H.(1989)"Cellular Delivery Systems for Insecticidal Proteins:Living and Non-Living Microorganisms", in Controlled Delivery of Crop Protection Agents, R.M.Wilkins, ed.,Taylor and Francis, New York and London,1990, pp.245-255]. 또한 Couch, T.L.(1980)저, "Mosquito Pathogenicity of Bacillus thuringiensis var. israelensis", Developments in Industrial Microbiology 22:61-76 ; 및 Beegle, C.C.(1978)저 "Use of Entomogenenous Bacteria in Agroecosystems", Developments in Industrial Microbiology 20:97-104를 참조. Krieg, A., A.M. Huger, G.A.Langenbrush,W.Schnetter 저(1983) Z.ang.Ent. 95:500-508 에는 초시목에 속하는 2종류의 딱정 벌레에 대하여 활성이 있다고 하는 바실루스 츄린겐시스 테네브리오니스 변종(Bacillus thuringiensis var tenebrionis)에 대하여 기술되어 있다.
이들은 콜로라도 감자 딱정벌레, 렙티노타르사 디셈리네아타(Leptinotarsa decemlineata) 및 아젤라스티카 알니(Agelastica alni)이다.
더욱 최근에, 비.티.의 신규한 아종들이 동정되었는데, 이들로부터 활성 δ-내독소 단백질의 기능을 담당하는 유전자가 분리되었다[Hofte, H.,H.R.Whitely(1989) Microbiological Reviews 52(2):242-255]. Hofte와 Whitely는 비.티. 결정성 단백질 유전자를 4개의 주된 군으로 구분하였다. 즉, CryI(인시류-특이적), CryⅡ(인시류-특이적 및 쌍시류-특이적), CryⅢ(초시류-특이적), 및 CryⅣ(쌍시류-특이적)이었다.
다른 해충들에 대해서 특이적으로 독성을 나타내는 균주들의 발견에 관하여도 보고되었다[Feitelson, J.S., J.Payne, L.Kim(1992) Bio/Technology 10:271-275]. CryⅤ는 선충류에 대해 특이적인 독소군으로 제안되었다. 램버트외 다수[Lambert, B.,L. Buysse, C.Decock, S.Jansens, C.Piens, B.Saey, J.Seurinck, K. van Audenhove, J.Van Rie, A.Van Vliet, M.Peferoen(1996) Appl. Environ. Microbiol 62(1):80-86 및 Shevelev et al.(1993) FEBS Lett. 336:79-82]는 인시류에 독성을 갖는 Cry9 독소의 특성에 대하여 기술하고 있다. 국제공개 WO 94/05771 및 WO 94/24264에서도 인시류 곤충에 대한 활성을 갖는 비.티. 분리주에 관하여 기술되어 있다. Gleave외 다수[(1991) JGM 138:55-62] 및 Smulevitch외 다수[(1991)FEBS Lett. 293:25-26]도 또한 비.티.독소에 관하여 기술하고 있다. 다수의 기타 비.티. 유전자 군이 현재 동정되어 있다.
에스케리챠 콜라이 (Escherichia coli)내에서 비.티. 결정성 단백질 유전자를 클로닝 및 발현시키는 것에 대하여는 발간된 논문에 기술되어 있다[Schnepf, H.E., H.R. Whitely(1981) Proc. Natl. Acad. Sci. USA 78:2893-2897]. 미국 특허 제 4,448,885호 및 미국 특허 제 4,467,036호 모두에는 이.콜라이(E.Coli)내에서 비.티. 결정성 단백질을 발현시키는 것에 대하여 기술되어 있다. 미국 특허 제4,990,332호; 제 5,039,523호; 제 5,126,133호; 제 5,164,180호; 및 제 5,169,629호에는 인시류에 대한 활성을 갖는 비.티. 독소에 대하여 기술되어 있다. PCT 출원 WO96/05314에는 인시류 해충에 활성인 PS86W1, PS86V1 및 기타 비.티. 분리주에 관하여 개시되어 있다. WO94/24264 및 WO94/05771로 발행된 PCT 특허 출원 공보에는 인시류 해충에 활성인 비.티. 분리주 및 독소에 관하여 기술되어 있다. 밤나방과(Noctuidae)에 대하여 활성이 있는 비.티. 단백질은 Lambert et al.(상동)에 기술되어 있으며, 미국 특허 제 4,797,276호 및 제 4,853,331호에는 다양한 환경에서 초시류(coleopteran) 해충을 박멸하는데에 사용될 수 있는 비.츄린겐시스 균주 테네브리오니스에 관하여 개시되어 있다. 미국 특허 제 4,918,006호는 쌍시류에 활성인 비.티. 독소에 관하여 기술되어 있다. 미국 특허 제 5,151,363호 및 미국 특허 제 4,948,734호에는 선충류에 활성인 임의의 비.티. 분리주에 관하여 기술되어 있다. 선충류에 활성인 것으로 기술된 다른 미국 특허로서는 미국 특허 제 5,093,120호; 제 5,236,843호; 제 5,262,399호; 제 5,270,448호; 제 5,281,530호; 제 5,322,932호; 제 5,350,577호; 제 5,426,049호; 및 5,439,881호를 포함한다. 연구 및 투자 자원을 확대시킨 결과, 다른 특허에서도 비.티. 분리주 및 상기 비.티. 분리주의 신규 용도에 관하여 기술하고 있다. Feitelson et al., (상동) 참조. 그러나, 신규의 비.티. 분리주 및 공지의 비.티.분리주의 용도를 발견한 것은 경험에 의한 것으로서, 예기치 못했던 기술이다.
유력한 독소 유전자를 분리하는 것은 경험칙에 의한 방법에 의하여 서서히 이루어 진 것이다. Carozzi외 다수 공저(Carozzi, N.B., V.C.Kramer, G.W.Warren, S.Evola, G.Koziel(1991) Appl. Env.Microbiol. 57(11):3057-3061)에서는 신규의 비.티. 분리주 동정 방법에 관하여 기술하고 있다. 상기 보고서에는 인시류-활성 독소 유전자에 대한 본 발명의 특이적인 프라이머, 프로브, 독소 및 유전자에 관하여 기술되어 있지 않다. 미국 특허 제 5,204,237호에는 비.티. 독소 유전자 분리용 특이적 프로브 및 보편적 프로브에 관하여 기술되어 있다. 그러나, 상기 특허에는 본 발명의 프로브, 프라이머, 독소 및 유전자에 관해서는 기술되어 있지 않다.
WO94/21795 및 Estruch, J.J.외 다수 공저[(1996) PNAS 93:5389-5394]에는 바실러스 미생물로부터 수득된 독소에 관하여 기술되어 있다. 상기 독소들은 무성 세포 생장기(vegetative cell growth)동안에 생산되기 때문에 영양 살충 단백질(vegetative insecticidal protein)(VIP)로 불리워졌던 것으로 보고된다. 뿐만 아니라, 상기 독소들은 결정 형성 δ-내독소와는 구별되는 것으로 보고되었다. 인시류 해충에 활성인 독소들의 활성에 관하여도 보고된 바 있다.
흑색 컷웜(black cutworm)[아그로티스 입실론(Agrotis ipsilon)(Hufnagel); 인시류(Lepidoptera): 밤나방(Noctuidae)]은 콘, 면, 평지속 곡물(cole crops)(브라시카(Brassica), 브로커리, 양배추, 중국 양배추와 같은) 및 잔디를 포함하는 다수의 곡물에 심각한 해충이다. 2차적 숙주 식물로서는 비트 뿌리, 캡시큠(Capsicum), 이집트콩, 파바 콩(faba bean), 양상추, 자주개자리, 양파, 감자, 무우, 평지(카놀라), 쌀, 간장콩, 딸기, 사탕 무우, 담배, 토마토 및 산림을 포함한다. 북미의 아글로티스(Aglotis)속 해충은 클로버, 콘, 담배, 대마, 양파, 딸기, 검은 딸기, 나무 딸기, 알팔파, 보리, 콩, 양배추, 귀리, 완두, 감자, 고구마, 토마토, 화훼, 잔디, 자주개자리, 옥수수, 아스파라거스, 포도, 거의 모든 종류의 식물 잎, 잎담배 및 다수의 기타 곡물 및 정원용 식물등을 먹고 산다. 기타 아그로니티 족(Tribe Agroniti) 컷웜들로서는 해충, 구체적으로 펠티아(Feltia) 속의 해충[예를 들어, 에프.쟈큘리페라(F.jaculifera(Guenee); 듀센스 서브고티카(ducens subgothica)와 동등] 및 유조아(Euxoa)[예를 들어, 이.메소리아(E. messoria), 이.스캔든스(E.scandens)(Riley), 이.옥실리아리스 스미스(E.auxiliaris Smith), 이.디테르사(E.detersa)(Walker), 이.테셀라타(E.tessellata)(Harris), 이.오크로개스터(E.ochrogaster)(Guenee)]들이 있다. 숙주 식물로서는 평지(rape)를 포함하는 다수의 곡물이 있다.
컷웜은 또란 북미 대륙 이외의 지역에도 서식하는데, 이들은 경제적으로 중요한 영향을 미치는 해충으로서, 이집트콩, 밀, 야채, 사탕 무우, 자주개자리, 옥수수, 감자, 순무, 평지, 상추, 딸기, 로건베리, 아마, 면, 간장콩, 담배, 비트 뿌리, 중국 양배추, 토마토, 가지, 사탕수수, 목초, 양배추, 땅콩, 호리병박, 순무, 해바라기, 브라시카(Brassica), 양파, 부추, 셀러리, 참깨, 아스파라거스, 장군풀, 치커리, 온실 재배 곡물 및 시금치등을 공격한다. 흑색 컷웜인 에이.입실론은 중앙 아메리카, 유럽, 아시아, 오스트레일리아, 아프리카, 인도, 타이완, 멕시코, 이집트 및 뉴질랜드를 포함하는 북미 대륙 이외의 지역에 서식한다.
컷웜은 몇몇 영(instar)을 거쳐서 유충으로서 진화된다. 비록 후기 영의 유충에 의해 실생 식물(seedling)이 절단되면 뚜렷한 손상 및 경제적 손실을 초래한다 할지라도, 보통 옥수수와 같은 곡물에 있어서는 잎을 갉아 먹히면 수확량에 손실이 발생하게 된다. 제 4단계의 유충 영에 도달할 경우, 유충은 특히 실생 식물 및 식물의 일부분을 갉아서 절단하기 시작한다. 섭생 활동에 변화가 생기기 때문에, 경제적으로 손실을 입은 곡물들은 초기 경고 신호 조차도 없이 예기치 않게 증진될 수 있다. 뿐만 아니라 이들의 야행성 및 땅속에 잠복하는 습성은 이들을 검출하는데에 애로사항이 되고 있다. 거대한 컷웜들은 매일 약간씩의 실생 식물들을 파괴시킬 수 있으며, 다량 출몰할 경우 곡물 전체의 줄기를 갉아먹어 없앨 수 있다.
궐련 잎맥(peripheral weed) 대조구와 같은 에이.입실론에 대한 대조구 배양물은 이들 컷웜의 다량 출몰 현상을 방지하는데에 도움이 될 수 있다; 그러나, 이러한 방법들이 항상 편리하고 효과적인 것만은 아니다. 출몰 현상(infestation)은 매우 산발적이며, 살충제 처리후 식재하거나 또는 식재함과 동시에 살충제를 처리하는 것과 같은 과거의 방법은 그리 효과적이지 못했다. 어떤 미끼(bait)는 곡물에 있어서 컷웜을 박멸시키는데에 유용하다. 덩쿨 잔디와 같은 잔디 풀을 보호하기 위해서 화학 살충제가 사용되어 오고 있다. 사람들과 처리된 지역(예를 들어, 골프 잔디, 운동장, 공원 및 기타 오락 장소, 전문적인 조망을 제공하는 장소, 가정용 잔디)이 가까이 접하기 때문에 화학 살충제는 특히 잔디에 사용되고 있다. 일반적으로 천연 산물(예를 들어, 선충류의 애저디라크틴(azadirachtin))은 효과가 덜어진다. 지금까지, 상기 컷웜을 효과적으로 박멸시킬 수 있는 독소가 시판되고 있지 않기 때문에, 바실러스 츄린겐시스 산물이 흑색 컷웜을 박멸하는데에 널리 사용되어 오고 있다.
[발명의 요약]
본 발명은 비포유 동물 해충(non-mamalian pests), 구체적으로 식물 해충 박멸에 유용한 물질 및 방법에 관한 것이다. 특정 구체예에서, 본 발명은 인시류(Lepidopterans) 박멸에 유용한 신규의 독소를 제공한다. 특히 바람직한 구체예에서, 본 발명의 독소는 흑색 컷웜(black cutworm)을 박멸하는데에 사용된다. 본 발명은 본 발명의 인시류-활성 독소를 코드화하는 뉴클레오티드 서열을 추가로 제공한다. 본 발명은 살충 독소를 코드화하는 유전자의 동정 및 특징화에 유용한 뉴클레오티드 서열 및 방법을 추가로 제공한다. 본 발명은 살충 활성을 갖는 신규의 바실러스 츄린겐시스 분리주를 추가로 제공한다.
하나의 구체예에서, 본 발명은 PCR 기술에 있어서 프라이머로서 유용한 특이한 뉴클레오티드 서열에 관한다. 상기 프라이머는 특이 독소 유전자의 동정 및 분리에 사용될 수 있는 특징적인 유전자 절편을 합성한다. 본 발명의 뉴클레오티드 서열은 앞서 기술된 δ-내독소와 구별되는 독소를 코드화한다.
본 발명의 하나의 구체예에서, 비.티. 분리주는 미생물이 다량 증식할 수 있는 조건하에서 배양될 수 있다. 상기 미생물을 처리하여 단일 사슬 게놈성 핵산을 수득한후, 상기 DNA를 본 발명의 프라이머와 접촉시켜 PCR 증폭시킬 수 있다. 독소 코드화 유전자의 특징적인 절편은 상기 방법으로 증폭되어 독소-코드화 유전자의 존재를 동정하는데에 사용될 것이다.
본 발명의 다른 양상은 본원에 개시된 뉴클레오티드 서열을 프로브로서 사용하여 인시류에 활성인 비.티. 독소를 코드화하는 유전자를 검출, 동정 및 특징화하는데에 사용하는 것에 관한다. 따라서 동정된 유전자는 인시류에 활성인 독소를 코드화한다. 이와 유사하게, 상기 분리주는 이들 해충에 대하여 활성을 가질 것이다.
본 발명의 신규 살충 비.티. 분리주에는 PS31G1, PS185U2, PS11B, PS218G2, PS213E5, PS28C, PS86BB1, PS89J3, PS94R1, PS27J2, PS101DD 및 PS202S를 포함한다.
본원에 기술된 바와 같이, 본 발명에 유용한 독소는 다수의 독소들의 일부분들을 결합시켜 제조된 키메라 독소일 수 있다.
바람직한 구체예에서, 본 발명은 표적 해충에 의하여 섭취되는 형질 전환된 식물 세포가 조직내에서 살충 독소를 발현시키도록 본 발명의 하나 이상의 폴리뉴클레오티드 서열로 형질 전환된 식물 세포에 관한 것이다. 이러한 식물의 형질 전환은 당 업계의 숙련자들에게 널리 공지된 기술을 사용하여 수행될 수 있으며 통상적으로 유전자를 변형시켜 식물내에서 독소의 발현을 최적화하는 것을 포함한다.
이와는 달리, 본 발명의 비.티.분리 균주, 또는 본원에 기술된 독소를 발현시키는 재조합 미생물은 해충 박멸에 사용될 수 있다. 이러한 관점에서, 본 발명은 실질적으로 원형 그대로인 비.티. 세포 및/또는 본 발명의 발현 독소를 포함하는 재조합 세포를 처리하여 실질적으로 원형 그대로인 세포들이 표적 해충의 환경에 적용될때 살충 활성을 연장시키는 것을 포함한다. 처리된 세포는 살충 독소에 대한 보호막으로서 작용한다. 상기 독소는 표적 해충에 의하여 섭취됨에 따라서 활성화된다.
[서열의 간단한 설명]
서열 1은 본 발명에 유용한 정방향 프라이머(forward primer)이다.
서열 2는 본 발명에 유용한 역방향 프라이머(reverse primer)이다.
서열 3은 본 발명에 유용한 정방향 프라이머이다.
서열 4는 본 발명에 유용한 역방향 프라이머이다.
서열 5는 본 발명에 유용한 정방향 프라이머이다.
서열 6은 본 발명에 유용한 역방향 프라이머이다.
서열 7은 11B1AR 독소의 아미노산 서열이다.
서열 8은 독소 11B1AR의 아미노산 서열(서열 7)을 코드화하는 뉴클레오티드 서열이다.
서열 9는 11B1BR 독소의 아미노산 서열이다.
서열 10은 독소 11B1BR의 아미노산 서열(서열 9)을 코드화하는 뉴클레오티드 서열이다.
서열 11은 1291A 독소의 아미노산 서열이다.
서열 12는 독소 1291A의 아미노산 서열(서열 11)을 코드화하는 뉴클레오티드 서열이다.
서열 13은 1292A 독소의 아미노산 서열이다.
서열 14는 독소 1292A의 아미노산 서열(서열 13)을 코드화하는 뉴클레오티드 서열이다.
서열 15는 1292B 독소의 아미노산 서열이다.
서열 16은 독소 1292B의 아미노산 서열(서열 15)을 코드화하는 뉴클레오티드 서열이다.
서열 17은 31GA 독소의 아미노산 서열이다.
서열 18은 독소 31GA의 아미노산 서열(서열 17)을 코드화하는 뉴클레오티드 서열이다.
서열 19는 31GBR 독소의 아미노산 서열이다.
서열 20은 독소 31GBR의 아미노산 서열(서열 19)을 코드화하는 뉴클레오티드 서열이다.
서열 21은 본 발명의 방법에 의하여 동정된 85N1R 독소의 아미노산 서열이다.
서열 22는 독소 85N1R의 아미노산 서열(서열 21)을 코드화하는 뉴클레오티드 서열이다.
서열 23은 85N2 독소의 아미노산 서열이다.
서열 24는 독소 85N2의 아미노산 서열(서열 21)을 코드화하는 뉴클레오티드 서열이다.
서열 25는 85N3 독소의 아미노산 서열이다.
서열 26은 독소 85N3의 아미노산 서열(서열 25)을 코드화하는 뉴클레오티드 서열이다.
서열 27은 86V1C1 독소의 아미노산 서열이다.
서열 28은 독소 86V1C1의 아미노산 서열(서열 27)을 코드화하는 뉴클레오티드 서열이다.
서열 29는 86V1C2 독소의 아미노산 서열이다.
서열 30은 독소 86V1C2의 아미노산 서열(서열 29)을 코드화하는 뉴클레오티드 서열이다.
서열 31은 86V1C3R 독소의 아미노산 서열이다.
서열 32는 독소 86V1C3R의 아미노산 서열(서열 31)을 코드화하는 뉴클레오티드 서열이다.
서열 33은 F525A 독소의 아미노산 서열이다.
서열 34는 독소 F525A의 아미노산 서열(서열 33)을 코드화하는 뉴클레오티드 서열이다.
서열 35는 F525B 독소의 아미노산 서열이다.
서열 36은 독소 F525B의 아미노산 서열(서열 35)을 코드화하는 뉴클레오티드 서열이다.
서열 37은 F525C 독소의 아미노산 서열이다.
서열 38은 독소 F525C의 아미노산 서열(서열 37)을 코드화하는 뉴클레오티드 서열이다.
서열 39는 F573A 독소의 아미노산 서열이다.
서열 40은 독소 F573A의 아미노산 서열(서열 39)을 코드화하는 뉴클레오티드 서열이다.
서열 41은 F573B 독소의 아미노산 서열이다.
서열 42는 독소 F573B의 아미노산 서열(서열 41)을 코드화하는 뉴클레오티드 서열이다.
서열 43은 F573C 독소의 아미노산 서열이다.
서열 44는 독소 F573C의 아미노산 서열(서열 43)을 코드화하는 뉴클레오티드 서열이다.
서열 45는 FBB1A 독소의 아미노산 서열이다.
서열 46은 독소 FBB1A의 아미노산 서열(서열 45)을 코드화하는 뉴클레오티드 서열이다.
서열 47은 FBB1BR 독소의 아미노산 서열이다.
서열 48은 독소 FBB1BR의 아미노산 서열(서열 47)을 코드화하는 뉴클레오티드 서열이다.
서열 49는 FBB1C 독소의 아미노산 서열이다.
서열 50은 독소 FBB1C의 아미노산 서열(서열 49)을 코드화하는 뉴클레오티드 서열이다.
서열 51은 FBB1D 독소의 아미노산 서열이다.
서열 52는 독소 FBB1D의 아미노산 서열(서열 51)을 코드화하는 뉴클레오티드 서열이다.
서열 53은 J31AR 독소의 아미노산 서열이다.
서열 54는 독소 J31AR의 아미노산 서열(서열 53)을 코드화하는 뉴클레오티드 서열이다.
서열 55는 J32AR 독소의 아미노산 서열이다.
서열 56은 독소 J32AR의 아미노산 서열(서열 55)을 코드화하는 뉴클레오티드 서열이다.
서열 57은 W1FAR 독소의 아미노산 서열이다.
서열 58은 독소 W1FAR의 아미노산 서열(서열 57)을 코드화하는 뉴클레오티드 서열이다.
서열 59는 W1FBR 독소의 아미노산 서열이다.
서열 60은 독소 W1FBR의 아미노산 서열(서열 59)을 코드화하는 뉴클레오티드 서열이다.
서열 61은 W1FC 독소의 아미노산 서열이다.
서열 62는 독소 W1FC(서열 61)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 63은 본 발명에 의한 PCR 프라이머 또는 혼성화 프로브로서 유용한 올리고뉴클레오티드이다.
서열 64는 본 발명에 의한 PCR 프라이머 또는 혼성화 프로브로서 유용한 올리고뉴클레오티드이다.
서열 65는 본 발명에 의한 PCR 프라이머 또는 혼성화 프로브로서 유용한 올리고뉴클레오티드이다.
서열 66은 본 발명에 의한 PCR 프라이머 또는 혼성화 프로브로서 유용한 올리고뉴클레오티드이다.
서열 67은 본 발명에 의한 PCR 프라이머 또는 혼성화 프로브로서 유용한 올리고뉴클레오티드이다.
서열 68은 본 발명에 의한 PCR 프라이머 또는 혼성화 프로브로서 유용한 올리고뉴클레오티드이다.
서열 69는 본 발명에 의한 PCR 프라이머 또는 혼성화 프로브로서 유용한 올리고뉴클레오티드이다.
서열 70은 86BB1(a) 독소의 아미노산 서열이다.
서열 71은 독소 86BB1(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 72는 86BB1(b) 독소의 아미노산 서열이다.
서열 73은 독소 86BB1(b)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 74는 31G1(a) 독소의 아미노산 서열이다.
서열 75는 독소 31G1(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 76은 129HD 키메라 독소의 아미노산 서열이다.
서열 77은 독소 129HD 키메라 독소의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 78은 11B(a) 독소의 아미노산 서열이다.
서열 79는 독소 11B(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 80은 31G1(b) 독소의 아미노산 서열이다.
서열 81은 독소 31G1(b)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 82는 86BB1(c) 독소의 아미노산 서열이다.
서열 83은 독소 86BB1(c)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 84는 86V1(a) 독소의 아미노산 서열이다.
서열 85는 독소 86V1(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 86은 86W1(a) 독소의 아미노산 서열이다.
서열 87은 독소 86W1(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 88은 94R1(a) 독소의 일부 아미노산 서열이다.
서열 89는 독소 94R1(a)의 아미노산 서열을 코드화하는 일부 뉴클레오티드 서열이다.
서열 90은 185U2(a) 독소의 아미노산 서열이다.
서열 91은 독소 185U2(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 92는 202S(a) 독소의 아미노산 서열이다.
서열 93은 독소 202S(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 94는 213E5(a) 독소의 아미노산 서열이다.
서열 95는 독소 213E5(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 96은 218G2(a) 독소의 아미노산 서열이다.
서열 97은 독소 218G2(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 98은 29HD(a) 독소의 아미노산 서열이다.
서열 99는 독소 29HD(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 100은 110HD(a) 독소의 아미노산 서열이다.
서열 101은 독소 110HD(a)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 102는 129HD(b) 독소의 아미노산 서열이다.
서열 103은 독소 129HD(b)의 아미노산 서열을 코드화하는 뉴클레오티드 서열이다.
서열 104는 573HD(a) 독소의 일부 아미노산 서열이다.
서열 105는 독소 573HD(a)의 아미노산 서열을 코드화하는 일부 뉴클레오티드 서열이다.
본 발명은 비포유 동물 해충 박멸 물질 및 방법에 관한 것이다. 특정 구체예에서, 본 발명은 인시류에 대하여 활성을 갖는 신규의 바실러스 츄린겐시스 분리 균주 및 독소에 관한 것이다. 특히 바람직한 구체예에서, 본원에 기술된 독소 및 방법은 혹색 컷웜을 박멸하는데에 사용될 수 있다. 본 발명은 살충 독소를 코드화하는 신규의 유전자 및 유용한 특성을 갖는 독소를 코드화하는 비.티. 유전자를 동정 및 특징화하는 신규의 방법에 관한다. 본 발명은 이들 독소를 코드화하는 폴리뉴클레오티드 서열 및 이들 폴리뉴클레오티드 서열들을 사용하여 상기 독소를 발현하는 재조합 숙주를 생산하는 것에 관한다.
본 발명의 임의의 단백질들은 이미 바실러스 츄린겐시스로부터 분리된 결정 즉 "Cry" 단백질과는 구별되는 것이다.
본 발명의 다른 양상은 신규의 분리 균주 및 이들 분리 균주로부터 수득 가능한 독소 및 유전자에 관한다. 본 발명에 의한 신규의 비.티. 분리주들은 PS31G1, PS185U2, PS11B, PS218G2, PS213E5, PS28C, PS86BB1, PS89J3, PS94R1, PS27J2, PS101DD 및 PS202S로 명명되었다.
본원에 제공된 신규의 독소 및 폴리펩타이드 서열은 몇가지의 파라미터에 따라서 정의된다. 본원에 기술된 독소들중 가장 특징적인 것은 살충 활성이다. 특정 구체예에서, 상기 독소들은 인시류 해충에 대하여 활성을 갖는다. 본 발명의 독소 및 유전자는 그들의 아미노산 및 뉴클레오티드 서열에 의하여 추가로 정의될 수 있다. 상기 분자들의 서열들은 임의의 구체화된 서열에 대한 상동성 및 혼성화되는 능력의 관점에서 정의될 수 있으며, 또는 임의의 구체화된 프로브 및 프라이머에 의해서 증폭될 수 있다. 뿐만 아니라, 본원에 제공된 독소는 임의의 항체와의 면역 침강성을 기초로 하여 동정될 수 있다.
비.티. 결정성 단백질의 일부분을 결합시킴으로써 유용한 키메라 독소를 제조하는 방법들이 개발되어 오고 있다. 결합된 부분들은 이들 부분들이 결합되어 살충 활성을 갖는 키메라 단백질을 합성하는한, 상기 부분들 자체는 살충 활성일 필요는 없다. 상기 방법은 예를 들어, 유럽 특허 제 0 228 838호; Ge, A.Z., N.L.Shivarova, D.II. Dean(1989) Proc. Natl. Acad. Sci. USA 86:4037-4041; Ge, A.Z., D.Rivers, R.Milne, D.II.Dean(1991) J.Biol.Chem.266:17954-17958; Schnepf, H.E., K.Tomezak, J.P.Ortega, H.R.Whiteley(1990) J.Biol.Chem.265:20923-10930; Honee, G.D.Convents, J.Van Rie, S.Jansens, M.Perferoen, B.Visser(1991) Mol.Microbiol. 5:2799-2806 에 기술된 바와 같이 제한 효소를 사용하여 수행될 수 있다. 이와는 달리, 세포성 재조합 기작을 사용하는 재조합 방법을 사용하여 유사한 결과를 얻어내는데에 사용될 수 있다. 예를 들어, Caramori, T.,A.M.Albertini, A.Galizzi(1991)Gene 98:37-44; Widner, W.R.,H.R.Whiteley(1990) J.Bacteriol.172:2826-2832; Bosch, D.,B.Schipper, H.van der Kliej, R.A. de Maagd, W.J.Stickema(1994)Biotechnology 12:915-918 을 참조하시오. 이러한 키메라 DNA를 제조할 수 있는 기타 다수의 방법들이 당 업계에 공지되어 있다. 본 발명은 본원에서 동정된 신규의 서열을 이용하는 키메라 단백질을 포함하는 것을 의미한다.
본원의 교시에 따라서, 당 업계의 숙련자는 본원에 기술된 다양한 독소 및 폴리뉴클레오티드 서열을 용이하게 제조 및 사용할 수 있다.
본 발명에 유용한 비.티. 분리 균주는 1815 North University Street, Peoria, Illinois 61604, USA 소재, Agricultural Research Service Patent Culture Collection(NRRL), Northern Regional Research Center에 기탁하였다. 비.티. 균주의 기탁 번호는 다음과 같다:
본 발명의 배양균은 본원의 계류중에 37 CFR 1.14 및 35 U.S.C. 122에 의거 미합중국 특허상표청장이 분양 받을 권리를 부여한 자에 대한 상기 배양균의 분양을 보장할 것을 조건으로 기탁되었다. 상기 기탁물은 본원의 대응출원 또는 그의 자출원이 출원된 나라에서 외국 특허법에 의해 요구될 때 입수될 수 있다. 그러나, 이러한 기탁물의 입수 가능성이 정부의 행위에 의해 수여된 특허권을 훼손하는 본원 발명의 실시허락을 구성하는 것은 아니라는 사실을 이해하여야만 한다.
더욱이, 본 발명의 배양균 기탁물은 미생물 기탁에 관한 부다페스트 조약의 규정에 따라 보관되고 공중에게 분양될 수 있는 상태에 놓일 것이다. 즉, 이들 상기 균주들은 미생물 시료의 분양에 대한 최근의 요청후 최소한 5년간, 어떠한 경우라도, 기탁일로부터 최소한 30년간 또는 그 미생물을 개시한 특허의 존속 기간 동안 생존하고 오염되지 않도록 유지하는데 필요한 모든 주의를 기울여 보관될 것이다. 기탁자는 기탁물의 상태때문에 기탁 기관이 분양 요청시 분양할 수 없는 경우에 기탁물(들)을 대체할 의무를 지닌다는 것을 인식한다. 본원의 배양균에 대한 공중의 입수에 대한 모든 제한은 그들을 개시한 특허의 허여시에 결정적으로 철회될 것이다.
다음은 본 발명에 유용한 임의의 분리 균주의 특징을 제시하는 표이다.
ND = 측정 안됨
하나의 구체예에서, 본 발명은 인시류 해충에 활성인 단백질 독소를 코드화하는 바실러스 츄린겐시스(비.티.) 유전자의 분리 및 동정용 뉴클레오티드 프라이머 및 프로브를 포함하는 물질 및 방법에 관한 것이다. 본원에 기술된 뉴클레오티드 서열은 또한 신규의 살충 비.티. 분리주를 동정하는데에 사용될 수 있다. 뿐만 아니라, 본 발명은 본원에 개시된 방법 및 물질을 사용하여 동정된 유전자, 분리 균주 및 독소에 관한 것이다.
유전자 및 독소
본 발명에 유용한 유전자 및 독소는 본원에 특징적으로 구체화된 독소의 특징적인 살충 활성을 갖는 전체 길이의 서열 뿐만 아니라 이들 서열의 절편, 변이체(variant), 돌연 변이체 및 융합 단백질을 포함한다. 또한 본 발명의 교시에 따라서 하나 이상의 비.티. 독소 또는 유전자의 일부분들을 결합하여 제조된 키메라 유전자 및 독소가 이용될 수도 있다. 본원에서 사용된 바와 같이, 유전자의 "변이체(variants)" 또는 "변이(variation)"란 용어는 동일한 독소를 코드화하거나 또는 살균 활성을 갖는 독소와 동등한 독소를 코드화하는 뉴클레오티드 서열을 의미한다. 본원에서 사용된 바와 같이, "동등한 독소(equivalent toxin)"란 용어는 구체화된 독소와 같이 표적 해충에 대하여 동일한 또는 본질적으로 동일한 독소를 의미한다.
활성 독소를 코드화하는 유전자가 몇가지 방법을 통하여 동정되어 수득가능하다는 사실은 본 업계의 숙련자들에게는 자명한 일일 것이다. 본원에 구체화된 특정 유전자는 상술된 배양액 기탁소에 기탁된 분리 균주로부터 수득될 수 있다.
상기 유전자, 또는 이의 일부 또는 변이체는 또한 예를 들어, 유전자 합성기를 사용하여 합성되어 구성될 수 있다. 유전자의 변이는 점 돌연 변이를 유발시키는 표준적인 기술을 사용하여 용이하게 구성될 수 있다. 뿐만 아니라, 상기 유전자의 절편들도 시판되고 있는 표준적 방법에 의한 엑소뉴클레아제 또는 엔도뉴클레아제를 사용하여 제조될 수 있다. 예를 들어, Bal31과 같은 효소 또는 부위-특이적 돌연변이에 의해서 상기 유전자의 말단으로부터 뉴클레오티드들을 체계적으로 절단할 수 있는 것이다. 뿐만 아니라, 활성 절편을 코드화하는 유전자는 다양한 종류의 제한 효소를 사용하여 수득될 수 있다. 단백질 분해 효소는 이들 독소의 활성 절편을 직접적으로 수득하는데에 사용될 수 있다.
동등한 독소 및/또는 이들 동등한 독소를 코드화하는 유전자는 본원에 제공된 교시를 사용하여 비.티. 분리 균주 및/또는 DNA 라이브러리로부터 수득할 수 있다. 본 발명의 살충 독소를 수득하는 다수의 방법들이 존재한다. 예를 들어, 본원에 개시되어 청구된 살충 독소에 대한 항체는 단백질 혼합물로부터 기타 독소들을 동정하여 분리하는데에 사용될 수 있다. 구체적으로, 항체는 다른 비.티. 독소들과 가장 눈에 띄게 특징적이고 가장 항상적인(constant) 독소의 일부분에 대해서 증가할 수 있다. 이후 상기 항체는 특징적인 활성을 갖는 동등한 독소를 면역 침강법(immunoprecipitation), 효소 연결 면역 흡수 검정법(enzyme linked immunosorbent assay;ELISA) 또는 웨스턴 블럿팅(western blotting)에 의해서 특이적으로 동정하는데에 사용될 수 있다. 본원에 개시된 독소, 또는 동등한 독소 ,또는 상기 독소의 절편에 대한 항체는 본 업계의 표준적인 방법을 사용하여 용이하게 제조될 수 있다. 이후 상기 독소를 코드화하는 유전자는 미생물로부터 수득될 수 있다.
구체화된 독소의 살충 활성을 갖는 절편들 및 동등체는 본 발명의 범위내에 포함된다. 뿐만 아니라, 유전자 코드가 풍부하기 때문에, 다수의 상이한 DNA 서열이 본원에 기술된 아미노산 서열을 코드화할 수 있다. 이와 같은 동일한 독소 또는 본질적으로 동일한 독소를 코드화하는 선택적 DNA 서열(alternative DNA sequences)을 제조하는 것은 당 업계의 숙련자들에게는 널리 공지된 것이다. 이들 변이체 DNA 서열은 본 발명의 범위내에 포함된다. 본원에 사용된 바와 같이, "본질적으로 동일한(essentially the same)" 서열은 실질적으로 살충 활성에 영향을 주지 않는 아미노산의 치환 , 결실, 부가 또는 삽입이 발생한 서열에 관한 것이다. 뿐만 아니라, 살충 활성을 갖는 절편들은 본 발명의 범위내에 포함된다.
본 발명의 독소 및 유전자를 동정하는 추가의 방법은 올리고뉴클레오티드 프로브를 사용하는 것이다. 상기 프로브들은 검출 가능한 뉴클레오티드 서열이다. 프로브들은 본 발명의 독소-코드화 유전자를 동정하는 신속한 방법을 제공한다. 본 발명에서 프로브로서 사용된 뉴클레오티드 서열은 DNA 합성기 및 표준적 방법에 따라 합성될 수 있다.
본 발명의 임의의 독소는 본원에 특징적으로 구체화되어 있다. 상기 독소는 본 발명의 독소에 대한 단순한 일례에 불과하기 때문에, 본 발명이 본원에 구체화된 독소와 동일하거나 또는 유사한 살충 활성을 갖는 변이체 또는 동등 독소(및 동등 독소를 코드화하는 뉴클레오티드 서열)를 포함한다는 것은 자명한 일이다. 동등 독소는 구체화된 독소와 상동성인 아미노산을 가질 것이다. 이러한 아미노산 동일성은 통상적으로 60% 이상, 바람직하게는 75% 이상, 더욱 바람직하게는 80% 이상, 더더욱 바람직하게는 90% 이상이고 95% 이상일 수도 있다. 아미노산 상동성은 생물학적 활성에 관여하거나 또는 생물학적 활성에 중대한 영향을 미치는 3차원 구조를 결정하는데에 관여하는 독소의 중요한 지역에서 가장 높을 것이다. 이러한 관점에서, 만일 상기 치환이 활성에 중요하지 않은 지역에 발생되었거나 또는 상기 분자의 3차원적 구조에 영향을 미치지 않는 보존적 아미노산 치환일 경우, 임의의 아미노산 치환은 허용 가능한 것이며 또한 예상할 수 있는 것이다. 예를 들어, 아미노산은 다음의 군에 속할 수 있다 : 비극성, 비하전 극성, 염기성 및 산성. 하나의 군에 속하는 아미노산이 동일한 타입의 다른 아미노산으로 치환된 보존적 치환은 상기 치환이 실질적으로 상기 화합물의 생물학적 활성을 실질적으로 변경시키지 않는한 본 발명의 범위내에 포함된다. 표 2는 각각의 군에 속하는 아미노산의 리스트를 제공한다.
| 아미노산 군 | 아미노산 예 |
| 비극성(nonpolar) | Ala, Val, Leu, Ile, Pro, Met, Phe, Trp |
| 비하전 극성(uncharged polar) | Gly, Ser, Thr, Cys, Tyr, Asn, Gln |
| 산성(acidic) | Asp, Glu |
| 염기성(basic) | Lys, Arg, His |
몇몇의 경우에서, 비보존적 치환이 발생할 수도 있다. 중요한 인자는 이들 치환이 독소의 생물학적 활성을 상당한 정도로 감소시켜서는 안된다는 것이다.
뿐만 아니라 본 발명의 독소는 상술된 바와 같이 독소 함유물의 모양 및 위치의 관점에서도 특징지워질 수 있다.
본원에서 사용된 바와 같이, "분리된(isolated)" 폴리뉴클레오티드 및/또는 "정제된(purified)" 독소란 상기 분자들이 자연상태에서 결합하는 다른 분자들과 결합하지 않은 상태에 있는 분자들을 의미한다. 그러므로, "정제된" 독소는 예를 들어, 식물체내에서 발현되는 독소를 포함하는 것이다. "분리되어 정제된(isolated and purified)"이란 의미는 본원에 기술된 바와 같이 "인위적으로 조작된 것(hand of man)"을 의미한다. 키메라 독소 및 유전자 또한 "인위적으로 조작된 것(hand of man)"을 포함한다.
재조합 숙주
상기 분리 균주들에 보유되어 있는 본 발명의 독소-코드화 유전자는 광범위한 미생물 또는 식물 숙주에 도입될 수 있다. 독소 유전자를 발현시키면 직간접적으로 살충제를 생성 및 보유시킬 수 있다. 미생물 숙주로서는 예를 들어, 슈도모나스(Pseudomonas)와 같은 미생물 이 해충이 있는 지역에 사용될 수 있는데, 상기 지역에서 미생물이 증식 및 섭취될 것이다. 그 결과 해충이 박멸된다. 이와는 달리, 독소 활성을 연장시키고 세포를 안정화시키는 조건하에서 독소 유전자를 보유하는 미생물을 사멸 및 처리시킬 수 있다. 상기 처리된 세포, 즉 독소 활성을 보유하는 세포는 이후 표적 해충의 환경에 적용될 수 있다.
비.티. 독소 유전자가 적합한 벡터를 통하여 미생물 숙주에 도입된후 상기 숙주가 살아있는 상태로 환경에 적용될 경우, 특정의 숙주 미생물 사용이 필수적이다. 미생물 숙주는 하나 이상의 관심 있는 농작물의 "피토스피어(phytosphere)" [필로플레인(phylloplane), 필로스피어(phyllosphere), 리조스피어(rhizosphere) 및/또는 리조플레인(rhizoplane)]에 서식하는 것으로 알려진 숙주들 중에서 선택된다. 폴리펩타이드 살충제를 발현시키는 유전자를 유지 및 발현시키기 위하여, 바람직하게는 상기 살충제가 환경적으로 파괴 및 불활성화되는 것으로부터 보호하기 위하여 이와 같은 미생물들은 특정 환경(농작물 및 다른 곤충의 서식지)에서 야생형 미생물과의 생존 경쟁에서 성공적으로 승리할 수 있는 것으로 선택한다.
다수의 미생물들이 각종의 중요한 작물의 엽층(식물 잎의 표면) 및/또는 구근권(식물 뿌리 주위의 토양)에 서식하는 것으로 알려져 있다. 이들 미생물들은 세균, 조류 및 진균류를 포함한다. 특히 중요한 것은, 예컨데 속 슈도모나스 (Pseudomonas), 에르위니아 (Erwinia), 세라티아 (Serratia), 클렙시엘라 (Klebsiella), 크산토모나스 (Xanthomonas), 스트렙토마이세스 (Streptomyces), 리조비움 (Rhizobium), 로도슈도모나스 (Rhodopseudomonas), 메틸로필리어스 (Methylophilius), 아그로박테리움 (Agrobacterium), 아세토박터 (Acetobacter), 락토바실러스 (Lactobacillus), 아르스로박터 (Arthrobacter), 아조토박터 (Azotobacter), 류코노스톡 (Leuconostoc) 및 알칼리게네스 (Alcaligenes)들과 같은 일례의 세균; 속 사카로마이세스 (Saccharomyces), 크립토코커스 (Cryptococcus), 클뤼베로마이세스 (Kluyveromyces), 스포로볼로마이세스 (Sporobolomyces), 로도토룰라 (Rhodotorula) 및 아우레오바시디움 (Aureobasidium)과 같은 일례의 진균류(fungi)를 포함한다. 특히 중요한 것은 슈도모나스 시린가에 (Pseudomonas syringae), 슈도모나스 플루오레스센스 (Pseudomonas fluorescens), 세라티아 마르세스센스 (Serratia marcescens), 아세토박터 크실리눔 (Acetobacter xylinum), 아그로박테리움 투메파시엔스 (Agrobacterium tumefaciens), 로도슈도모나스 스페로이데스 (Rhodopseudomonas spheroides), 크산토모나스 캄페스트리스 (Xanthomonas campestris), 리조비움 멜리오티 (Rhizobium melioti), 알칼리게네스 엔트로퍼스 (Alcaligenes entrophus), 및 아조토박터 빈란디 (Azotobacter vinlandii)와 같은 피토스피어 세균종 ; 및 로도토룰라 루브라 (Rhodotorula rubra), 알. 글루티니스 (R. glutinis), 알. 마리나 (R. marina), 알. 아우란티아카 (R. aurantiaca), 크립토코커스 알비두스 (Cryptococcus albidus), 시. 디플루엔스 (C. diffluens), 시. 라우렌티 (C. laurentii), 사카로마이세스 로세이 (Saccharomyces rosei), 에스. 프레토리엔시스 (S. pretoriensis), 에스. 세레비시아에 (S. cerevisiae), 스포로볼로마이세스 로세우스 (Sporobolomyces roseus), 에스. 오도루스 (S. odorus), 클뤼베로마이세스 베로나에 (Kluyveromyces veronae), 및 아우레오바시디움 폴루란스 (Aureobasidium pollulans)와 같은 피토스피어 효모 종들을 포함한다. 특히 중요한 것은 색소화된 미생물(pigmented microorganism)이다.
독소를 코드화하는 비.티. 유전자들을 유전자의 안정한 유지 및 발현을 허용하는 조건하에서 미생물 숙주내로 도입시키는데는 여러 가지 방법을 이용할 수 있다. 이들 방법들은 당업자들에게 잘 알려져 있고, 예컨대 본원에 참고자료로 첨부된 미국특허 제5,135,867호에 기술되어 있다.
본 발명의 분리주, 독소 및 유전자를 사용하여 흑색 컷웜을 포함하는 동시류(Homopteran)를 박멸하는 방법은 당 업계의 숙련자들에게 공지된 여러 가지 방법으로 수행될 수 있다. 이와 같은 방법에는 예를 들어, 비.티. 분리주를 해충(또는 해충이 서식하는 지역)에 적용시키는 방법, 재조합 미생물을 해충(또는 해충이 서식하는 지역)에 적용시키는 방법 및 식물을 본 발명의 살충 독소를 코드화하는 유전자로 형질전환시키는 방법을 포함한다. 재조합 미생물로서는, 예를 들어 비.티., 이. 콜라이 또는 슈도모나스일 수 있다. 형질 전환은 표준적인 기술을 사용하여 당 업계의 숙련자들에 의해 수행될 수 있다. 상기 형질 전환에 필요한 재료들에 관하여는 본원에 기술되어 있거나 그렇지 않으면 업계의 숙련자에 의하여 용이하게 입수할 수 있다.
숙주 세포를 형질 전환시키기 위하여 본 발명의 신규의 독소와 기능적으로 동등한 합성 유전자도 또한 사용될 수 있다. 상기 합성 유전자 제조 방법에 관하여는 예를 들어, 미국 특허 제 5,380,831호에 기술되어 있다.
세포 처리
상술한 바와 같이, 독소 활성을 연장시키고 세포를 안정화시키기 위하여 비.티. 또는 비.티. 독소를 발현시키는 재조합 세포가 처리될 수 있다. 제조된 살충제 마이크로캡슐은 안정화되어 상기 마이크로캡슐이 표적 해충 환경에 적용될 경우 독소를 보호할 수 있는 세포 구조체(cellular structure)내에 비.티. 독소를 포함한다. 적당한 숙주 세포로서는 원핵 세포 또는 진핵 세포를 포함할 수 있는데, 보통은 포유류와 같은 고등 생물체에서 독성을 갖는 물질을 생성시키지 않는 세포들에 한정된다. 그러나, 상기 독성 물질이 불안정하거나 또는 그 독성이 포유류 숙주에서 발생될 수 있는 독소로 인한 영향을 피하기에 충분히 낮은 경우, 고등 생물체에 독성을 갖는 물질을 생성시키는 생물체가 사용될 수도 있다. 특히 관심 있는 숙주는 원핵 생물 및 곰팡이와 같은 하등 진핵 생물일 것이다.
처리시의 세포는 -어떤 경우에는 포자가 사용될 수도 있지만- 포자형이기 보다는 일반적으로 원형 그대로(intact)의 형태 및 실질적으로는 증식형일 것이다.
예를 들어, 비.티. 독소 유전자를 포함하는 미생물 세포의 처리 방법은 그 기술이 독소의 성질에 유해한 영향을 미치거나 또는 상기 독소를 보호하는 세포의 능력을 감소시키지 않는한, 화학적 또는 물리적 방법에 의하거나 또는 화학적 및/또는 물리적 방법을 조합하여 사용할 수 있다. 화학 시약의 예로서는 할로겐화제를 들 수 있으며, 구체적으로 원자 번호 17-80의 할로겐이 바람직하다. 더욱 바람직하게는, 온화한 조건 및 원하는 결과를 얻기에 충분한 시간 동안 요오드가 사용될 수 있다. 다른 적당한 기술로서는 글루타르알데히드와 같은 알데히드; 제피란 클로라이드 및 세틸로피리디늄 클로라이드와 같은 항감염제; 이소프로판올 및 에탄올과 같은 알코올; Lugol 요오드, Bouin 고정제, 다양한 산 및 Helly 고정제와 같은 다양한 조직학적 고정제[Humanson, Gretchen L., Animal Tissue Techniques, W.H.Freeman and Company, 1967 참조]; 또는 세포가 숙주 환경에 투입되었을때 세포내에서 생산되는 독소의 활성을 보존 또는 연장시키는 물리적(열) 및 화학적 제제를 조합하여 처리하는 방법을 포함한다. 물리적 방법의 예에는 예를 들어 감마선 및 X선 처리법, 동결법, UV 광선 처리법, 동결 건조법등이 있다. 미생물 세포의 처리 방법에 관하여는 미합중국 특허 제 4,695,455호 및 제 4,695,462호에 기술되어 있으며 본원에 참고 문헌으로서 첨부되어 있다.
일반적으로 세포는 환경 조건에 대한 저항성이 강화되도록 구조적 안정성을 강화시킬 것이다. 살충제가 원형(proform)으로 존재하는 경우, 세포 처리 방법은 표적 해충 병원균에 의하여 살충제가 원형에서 성숙한 형태로 변화하는 과정을 방해하지 않는 방법으로 선택되어야 한다. 예를 들어, 포름알데히드는 단백질을 가교 결합시킬 것이며 이로써 원형 폴리펩타이드 살충제의 변화 과정을 방해하게 될 것이다. 따라서 처리 방법은 적어도 독소의 생체 이용률 또는 독소의 생체 활성에 대한 실질적인 부분들을 보유해야 한다.
독소 생성을 위하여 숙주 세포를 선택하는데에 있어서 특히 관심 있는 특징에는 비.티. 유전자를 숙주 세포내에 도입 시키는 것이 용이하다는 점, 발현 시스템의 유용성, 발현 효율, 숙주내 살충제의 안정성 및 기타 부수적인 유전적 능력등을 포함한다. 살충제 마이크로 캡슐을 사용하는데에 있어서 관심 있는 특징에는 두꺼운 세포벽, 색소 침착, 및 세포내 패키징(intracelluar packaging) 또는 봉입체(inclusion body) 형성과 같은 살충제 보호 특성; 수성 환경에서의 생존성; 포유 류 독성의 흠결; 해충에 대한 섭취 선호성; 독소에 해를 끼치지 않고 용이하게 살충 및 고정시키는 능력등을 포함한다. 다른 특징으로는 제조 및 조작의 용이성, 경제성, 저장 안정성등을 포함한다.
세포의 성장
선택적 배지내의 실질적으로 모든 세포가 비.티. 유전자를 보유하면, 비.티. 살충 유전자를 포함하는 숙주 세포는 DNA 구조체(DNA construct)가 선택적인 이점을 제공하는 특정의 영양 배지내에서 성장할 수 있다. 이후 상기 세포들은종래의 방법에 따라서 회수될 수 있다. 이와는 달리, 상기 세포들을 처리후 회수할 수도 있다.
본 발명의 비.티. 세포는 표준적인 배지 및 발효 기술을 사용하여 배양될 수 있다. 발효 싸이클을 마치면 당 업계에 널리 공지된 방법을 사용하여 발효액으로부터 비.티. 포자 및 결정체를 1차적으로 분리하여 회수할 수 있다. 특정의 비.티. 포자 및 결정체는 널리 공지된 기술을 사용하여 수득된후 종래의 σ-내독소 비.티. 제재로서 사용될 수 있다. 예를 들어서, 상기 포자 및 결정체는 계면 활성제, 분산제, 비활성 담체 및 특정의 표적 해충으로의 처리 및 적용을 촉진시키는 다른 성분을 부가하여 습윤성 분말, 농축액, 입자 및 기타의 제재로 조제될 수 있다. 이와 같은 조제 방법 및 적용 방법은 당 업계에 널리 공지되어 있다. 이와는 달리, 본 발명에 의한 독소를 수득하기 위하여 발효 공정으로부터 수득된 상등액이 사용될 수 있다. 이후 널리 공지된 기술을 사용하여 가용성, 분비성 독소를 분리 및 정제시킨다.
해충 박멸 방법 및 제재
본 발명에 따른 분리주, 독소 및 유전자를 사용하여 동시류를 박멸시키는 방법은 당 업계의 숙련자들에게 알려진 다양한 방법으로써 수행될 수 있다. 상기 방법은 예를 들어, 해충(또는 해충 서식지)에 비.티. 분리주를 처리하는 방법, 해충(또는 해충 서식지)에 재조합 미생물을 처리하는 방법 및 본 발명의 살충 독소를 코드화하는 유전자로 식물을 형질 전환시키는 방법을 포함한다. 재조합 미생물은 예를 들어, 비.티., 이.콜라이, 또는 슈도모나스일 수 있다. 형질 전환은 표준적인 기술을 사용하여 당 업계의 숙련자들에 의해서 수행될 수 있다. 이와 같은 형질 전환에 필요한 재료들에 관하여는 본원에 기술되어 있거나 그렇지 않으면 업계의 숙련자에 의하여 용이하게 입수할 수 있다.
비.티. 분리주 또는 재조합 미생물의 독소 및 유인물을 함유하는 조제 미끼 입자는 본원에 기술된 비.티. 분리주로부터 수득할 수 있는 유전자를 포함하며 토양에 사용될 수 있다. 조제 산물은 또한 윤작(crop cycle) 후반기에 종자-피복방식, 뿌리에 처리하는 방식 또는 식물 전체에 처리하는 방식등으로 적용될 수 있다. 식물 및 토양 처리시 비.티. 세포는 무기물(필로실리케이트, 카보네이트, 설페이트, 포스페이트등)과 같은 다양한 비활성 재료들과 혼합되거나 또는 식물성 재료들(옥수수대 분말, 쌀겨, 호두 껍질등)과 혼합되어 습윤성 분말, 입자 또는 분진으로 사용될 수 있다. 상기 제재에는 스프레더 스티커 에쥬번트(spreader sticker adjuvant), 안정화제, 기타 살충 부가제 또는 계면 활성제를 포함할 수 있다. 조제액은 수성-기초 또는 비수성일 수 있으며 거품, 겔, 현탁액, 유화 농축액등의 형태로 사용될 수 있다. 상기 성분들은 유동화제(rheological agent), 계면 활성제, 유화제, 분산제 또는 중합체를 포함할 수 있다.
당 업계의 숙련자에 의하여 이해될 수 있는 바와 같이, 살충제의 농도는 특정 제재의 성질, 구체적으로 농축되어 사용되는지 아니면 직접 사용되는지에 따라서 다양할 것이다. 살충제는 1-100중량%일 수 있다. 건조 제재의 함량은 살충제의 약 1-95중량%인 반면에, 액상 제재 함량은 일반적으로 액상내 고체의 약 1-60중량%이다. 일반적으로 제재은 약 102-104cell/mg 일것이다. 세포 함유 제재들은 1헥타르당 약 50㎎(액상 또는 건조) - 1㎏ 이상의 양으로 투여될 것이다.
상기 제재는 해충의 환경, 예를 들어 토양 및 잎에 분무(spraying), 살포(dust) 및 살수(sprinkling)등의 방법으로 적용시킬 수 있다.
돌연 변이
본 발명에 의하여 수득할 수 있는 신규 분리주를 당 업계에 널리 공지된 기술에 의하여 돌연 변이시킬 수 있다. 예를 들어, 에틸메탄 설포네이트(EMS) 돌연 변이법을 통하여 분리주를 무아포성(asporogenic) 돌연 변이시킬 수 있다. 당 업계에 널리 공지된 방법으로 자외선 및 니트로소구아니딘 처리하여 돌연 변이 시킬 수 있다.
발효 기간이 연장되면 더욱 소량의 무아포성 돌연 변이체가 원형 그대로인 용균되지 않은 상태로 잔류하게 될 것이다 : 이와 같은 균종을 용균 음성(-)으로 나타낸다. 용균 음성 균종은 발효 공정의 최종 단계에서 교반 플라스크 배지내의 무아포성 돌연 변이체를 스크리닝시킨후 여전히 원형 그대로의 상태이며 독소 결정체를 함유하는 돌연 변이체를 선별함으로써 동정될 수 있다. 용균 음성 균종은 보호 및 캡슐화된 독소 단백질이 수득될 수 있는 세포 처리 공정에 적합하다.
상기 무아포성 돌연 변이체의 파지 저항성 변이체를 제조하기 위하여, 소량의 파지 용균물을 영양 아가상에 도포후 건조시킨다. 이후 상기 소량의 파지 감수성 박테리아 균종을 상기의 건조 용균물에 직접 도말시킨후 건조시킨다. 상기 배양 접시를 30℃에서 인큐베이팅시킨다. 상기 배양 접시를 2일 동안 인큐베이팅시키는데, 이때 아가상에 다수의 콜로니가 자라난 것을 볼 수 있다. 상기 콜로니들중 몇개를 찍어서 영양 아가 접시상에서 재배양시킨다. 이와 같은 내성 배양균을 파지 용균물과 교차 스트리킹(cross streaking)시켜 내성 평가를 수행한다. 파지 용균물로 이루어진 선을 상기 접시상에서 스트리킹시킨후 건조시킨다. 이어서 내성 배양균으로 추측되는 배양균들을 상기 파지의 선(phage line)을 가로질러 스트리킹시킨다. 30℃에서 밤새 인큐베이팅시킨후 관찰한 결과, 내성 박테리아 배양균은 어디에서도 용균되지 않았다. 이후 영양 아가 접시상에 내성 균주 론(lawn)을 도말시켜 파지에 대한 내성을 재확인하였다. 감수성 균종이 양성 대조구로서 사용되는 것과 같은 동일한 방식으로 도말되었다. 건조시킨후, 파지 용균물 수적을 상기 접시의 중앙 부분에 떨어뜨린후 건조시켰다. 30℃에서 24시간 동안 인큐베이팅시킨후, 파지 용균물이 존재하는 지역에서는 내성 배양균이 용균되지 않았음을 알 수 있었다.
폴리뉴클레오타이드 프로브
DNA가 염기 상보성이라고 불리우는 기본 특성을 갖는다는 사실은 널리 공지된 바이다. 본질적으로, DNA는 한 쌍의 역 평행 사슬의 형태로 존재하는데, 각각의 사슬에 존재하는 염기는 각 사슬로부터 반대쪽 사슬을 향하여 뻗어져 있다. 한쪽 사슬에 존재하는 아데닌(A) 염기는 항상 반대쪽 사슬상의 티민(T) 염기와 쌍을 이루며, 구아닌(G) 염기는 시토신(C) 염기와 쌍을 이룬다. 염기들은 이들의 수소 결합력에 의해서 특별한 방식으로 병치된다. 각각의 결합이 상대적으로 약함에도 불구하고, 다수의 인접한 수소 결합 염기에 의한 전체 영향력 및 염기 차단(base-stacking) 효과들에 의해서 2개의 상보적 사슬이 안정하게 결합된다. 상기 결합은 고 pH 처리 또는 고온 처리와 같은 처리에 의하여 파괴될 수 있으며, 이와 같은 조건들은 2개의 사슬의 분해, 즉 "변성(denaturation)"을 일으킨다. 이후 DNA를 염기의 수소 결합이 열역학적으로 선호될 수 있는 조건하에 방치시키면, 상기 DNA 사슬은 어닐링(annealing), 즉 "혼성화(hybridization)"되어 본래의 DNA 이중 사슬 구조를 형성하게 될 것이다. 상기 혼성화 반응이 적당한 조건하에서 수행되면, 상기 반응의 특이성은 매우 높아질 수 있다. 즉, 염기의 상보성이 높은 사슬만이 안정한 이중 사슬 구조를 형성할 수 있을 것이다. 반응 조건과 혼성화 특이성과의 관계는 널리 공지되어 있다. 그러므로, 혼성화 반응은 2 개의 DNA 단편이 염기 배열에 있어서 상보적인지 여부를 평가하는데에 사용될 수 있다. 프로브를 사용하여 관심 있는 DNA 서열을 용이하게 검출하고 특징화시키는 것의 원리는 혼성화 기작이다.
상기 프로브는 DNA 또는 RNA일 수 있다. 통상적으로 프로브의 길이는 약 10base 이상, 더욱 통상적으로는 약 18base 이상인데, 이는 약 50base 이상까지 제조될 수 있으며, 만일 상기 프로브가 합성하여 제조되는 경우에는 약 200base 미만인 것이 보통이다. 그러나, 더욱 장쇄인 프로브가 용이하게 사용될 수 있는데, 이러한 프로브의 길이는 예를 들어, 수 킬로베이스에까지 달한다. 상기 프로브 서열은 관심있는 독소를 코드화하는 유전자의 일부와 실질적으로 상보적이도록 디자인된다. 상기 프로브는 그것과 혼성화되는 서열과 완벽하게 상보적일 필요는 없다, 상기 프로브는 당 업계의 숙련자들에게 널리 공지된 기술을 사용하여 라벨링될 수 있다.
본 발명을 프로브로서 사용하는 하나의 연구 방법에는 우선 모든 DNA가 본원에 개시된 뉴클레오티드 서열과 상동인 비.티. 분리주의 유전자 뱅크를 서던 블럿 분석법으로 동정하는 것을 포함한다. 그러므로, 생물학적 분석법의 도움 없이, 다수의 신규 비.티. 분리 균주 및 상기 비.티. 분리 균주에 의해 발현된 각각의 내독소 유전자 산물의 존재 가능한 활성을 미리 파악할 수 있다. 이러한 프로브 분석법은 다양한 비.티. 아종내에 존재하며 시판되고 있을지도 모르는 살충 내독소 유전자를 동정하는 신속한 방법을 제공한다.
통상적으로 본 발명에 따른 유용한 혼성화 방법중의 하나는 관심있는 DNA 샘플을 화학적으로 분리시킨후 정제하는 초기 과정을 포함한다. 용균된 세균 또는 세균으로부터 분리된 분획 핵산 전부가 사용될 수 있다. 세포들은 공지된 기술을 사용하여 처리될 수 있으며 그 결과 DNA (및/또는 RNA)를 유리시킬 수 있다. 적당한 제한 효소를 사용하여 DNA 샘플을 절단할 수 있다. 그 결과 생성된 절편들은 일반적으로 아가로즈 또는 아크릴아미드와 같은 겔내에서 수행되는 전기 영동에 의해서 크기별로 분리될 수 있다. 이후 관심 있는 단편들을 그 형태를 유지시키면서 고정화 막으로 이동시킬 수 있다. 이후 상기 막을 건조시킨후 미리 혼성화(prehybridization)시켜 이후의 혼성화 용액내 침수시를 대비하여 평형화시킨다. 핵산을 고형 지지체에 고정시키는 방법은 다양할 수 있다. 이후 과정에서의 DNA 고정화(fixing)는 실험실에서 사용하는 것을 떠나서, 이 기술이 속한 분야의 연구에 사용되기에 고부가 가치를 갖는다.
본 발명에서는 특정의 혼성화 기술이 필수적인 것은 아니다. 혼성화 기술이 개선됨에 따라 이들을 용이하게 적용시킬 수 있다.
당 업계에 널리 공지된 바와 같이, 프로브 분자 및 핵산 샘플이 그들 사이에 강한 비공유 결합을 형성하여 혼성화되면, 상기 프로브 및 샘플이 근본적으로 동일하다는 것을 추측할 수 있다. 상기 프로브의 검출성(detectable) 라벨은 혼성화가 발생되었는지 여부를 알 수 있는 수단을 제공한다.
본 발명의 폴리뉴클레오타이드 및 이들로부터 수득된 프로브는 표준 방법에 따라 DNA 합성기를 사용하여 합성될 수 있다. 뉴클레오타이드 단편을 프로브로서 사용하는 경우, 특정 프로브는 방사성 라벨 또는 비방사성 라벨을 포함하는 당 업계의 숙련자들에게 공지된 특정의 적당한 라벨로 라벨링된다. 일반적인 방사성 라벨에는32P,35S등을 포함한다. 방사성 동위 원소로 라벨링된 프로브는 DNase 및 DNA 중합효소를 사용하는 종래의 닉 트랜스레이션(nick-translation) 반응에 의하여 DNA 샘플과 상보적인 뉴클레오타이드 서열로부터 제조될 수 있다. 이후 상기 프로브 및 샘플을 혼성화 완충 용액내에서 화합시킨후 어닐링(annealing)될 때까지 적당한 온도에 방치시킨다. 이후 상기 막을 세척하여 통상적으로 방사선 사진 촬영 및/또는 액상 섬광 계수 측정법에 의하여 검출 및 정량되는 샘플 및 이에 결합된 프로브 분자에 남아있는 외재성 잔류 물질(extraneous materials)을 제거시킨다. 합성 프로브에 있어서, 프로브로서 사용되는 DNA의 말단을 라벨링시키는데에 폴리뉴클레오타이드 키나제 또는 말단 전이 효소와 같은 효소를 사용하는 것이 가장 바람직할 수 있다.
비방사성 라벨로서는 예를 들어, 바이오틴 또는 티록신과 같은 리간드 및 하이드롤라제 또는 퍼옥시다제와 같은 효소 또는 루시페린과 같은 다양한 화학 발광 물질 또는 플루오세인 및 이의 유도체와 같은 형광성 화합물을 포함한다. 상기 프로브는 본래부터 형광성을 갖도록 제조될 수 있는데, 이에 관하여는 국제 특허 출원 제 WO93/16094에 기술되어 있다. 상기 프로브는 또한 양쪽 말단에 분리가 용이한 다른 형태의 라벨로 라벨링될 수 있는데, 예를 들어서 한쪽 말단에는 상술된 바와 같은 동위 원소를 사용할 수 있으며 다른 쪽 말단에는 바이오틴 라벨을 사용하여 라벨링시킬 수 있다.
혼성화 용액내 존재하는 라벨링된 프로브의 양은 매우 다양한데, 이는 라벨의 특성, 필터에 결합될 수 있는 라벨링된 프로브 및 혼성화의 엄격성(stringency)에 따라서 다르다. 일반적으로 프로브가 과량으로 존재할 경우 고정된 DNA에 프로브가 결합되는 속도가 증가될 것이다.
다양한 정도의 혼성화 엄격성(stringency)이 적용될 수 있다. 상기 조건이 극할수록 이중 사슬 구조 형성에 필요한 상보성은 더욱 증가하게 될 것이다. 상기 조건의 극성(severity)은 온도, 프로브 농도, 프로브 길이, 이온 강도, 시간등에 의해서 조절될 수 있다. 바람직하게, 혼성화는 예를 들어 Keller, G.H.,M.M.Manak(1987) DNA Probes, Stockton Press, New York, NY.,pp.169-170에 기술된 바와 같이 당 업계에 널리 공지된 기술에 의하여 극한 조건하에서 수행될 수 있다.
본원에 사용된 혼성화에 "엄격한(stringent)" 조건이란, 본원 발명의 출원인이 사용하는 조건과 혼성화의 특이성(specificity) 정도가 동일하거나 또는 거의 동일한 정도의 조건을 의미한다. 특히, 고정화된 DNA의 혼성화는 P32-라벨링된 유전자 특이적 프로브를 사용하는 서던 블럿(Southern blot)시의 표준적 방법에 의하여 수행된다[Maniatis,T.,E.F.Fritsch, J.Sambrook(1982)Molecular Cloning:A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY]. 일반적으로, 혼성화 및 그 후속 조치로서 행하여 지는 세척은 본 발명의 독소 유전자와 상동성을 갖는 표적 서열을 검출할 수 있는 엄격한 조건하에서 수행된다. 이중 DNA 유전자 프로브에 있어서 혼성화는 6X SSPE, 5X Denhartdt 용액, 0.1% SDS, 0.1㎎/㎖의 변성 DNA 존재하, DNA 혼성체의 용융 온도(melting temperature; 이하 Tm이라 칭함) 이하인 20-25℃에서 밤새 수행된다. 상기 용융 온도는 하기 공식에 의하여 설명될 수 있다[Beltz, G.A., K.A.Jacobs, T.H.Eickbush, P.T. Cherbas, 및 F.C.Kafatos(1983) Methods of Enzymology, R.Wu, L. Grossman 및 K.Moldave(eds.) Academic Press, New York 100:266-285 참조].
Tm = 81.5℃+16.6 Log[Na+]+0.41(% G+C)-0.61(% 포름알데히드)-600/이중 염기쌍의 길이
통상적으로 세척은 하기와 같이 수행된다.
(1) 1X SSPE, 0.1% SDS 내, 실온에서 15분 동안 2회 세척(저 엄격성 세척;low stringency wash)
(2) 0.2X SSPE, 0.1% SDS 내, Tm 20℃에서 15분 동안 1회 세척(중간 엄격성 세척;moderate stringency wash)
올리고뉴클레오타이드 프로브에 있어서 혼성화는 6X SSPE, 5X Denhartdt 용액, 0.1% SDS, 0.1㎎/㎖의 변성 DNA 존재하, DNA 혼성체의 용융 온도(melting temperature; 이하 Tm이라 칭함) 이하인 10-20℃에서 밤새 수행된다. 상기 용융 온도는 하기 공식에 의하여 설명될 수 있다
Tm(℃) = 2(T/A 염기쌍 수) + 4(G/C 염기쌍 수)
[Suggs, S.V.,T.Miyake, E.H.Kawashime, M.J.Johnson, K.Itakura 및 R.B.Wallace(1981) ICN-UCLA Symp. Dev. Biol. Using Purified Genes, D.D.Brown(ed.), Academic Press, New York, 23:683-693 참조]
통상적으로 세척은 하기와 같이 수행된다.
(1) 1X SSPE, 0.1% SDS내, 실온에서 15분 동안 2회 세척(저 엄격성 세척;low stringency wash)
(2) 1X SSPE, 0.1% SDS내, 15분 동안 1회 세척(중간 엄격성 세척;moderate stringency wash)
이중 구조체 형성 및 안정성은 혼성체의 2 개의 사슬 사이의 실질적 상보성에 따라서 다르며, 상기된 바와 같이, 어느 정도의 미스매취(mismatch)는 크게 문제되지 않는다. 따라서 본 발명의 뉴클레오타이드 서열은 돌연 변이(단일 부위 또는 다수 부위), 결실, 삽입 및 이들을 조합한 경우를 포함하는데, 이때 상기 돌연 변이, 삽입 및 결실의 경우는 관심 있는 표적 폴리뉴클레오타이드 서열과 안정적으로 혼성화될 수 있다. 돌연 변이, 삽입 및 결실은 주어진 폴리뉴클레오타이드 서열내에서 다양한 방식으로 발생될 수 있으며, 이와 같은 방법들은 보통의 숙련자에게 공지된 방법이다. 다른 방법들도 장래 공지될 수 있을 것이다.
공지의 방법은
(1) 서열을 화학적으로 합성시키거나 또는 공지의 서열에 인위적으로 돌연 변이, 삽입, 결실시키는 단계;
(2) 본 발명의 뉴클레오타이드 서열을 프로브로서 사용하여 혼성화시켜 신규의 서열을 수득하거나 또는 프로브 서열을 돌연 변이, 삽입 또는 결실시키는 단계;
(3) 시험용 서열을 시험관내 또는 생체내에서 돌연 변이, 삽입 또는 결실시키는 단계
를 포함하나, 이에 한정되는 것은 아니다.
주어진 프로브로부터 생성된 돌연 변이성, 삽입성 및 결실성 변이체가 원래의 프로브보다 다소 효율적이라는 것을 알 수 있는 것은 매우 중요하다. 이와 같은 효율상의 차이점에도 불구하고, 상기 변이체들은 본 발명의 범주내에 포함된다.
그러므로 기술된 뉴클레오타이드 서열의 돌연 변이성, 삽입성 및 결실성 변이체는 당 업계의 숙련자들에게 널리 공지된 방법으로 제조될 수 있다. 상기 변이체들은 또한 그들이 원래 서열과 실질적으로 상동성이 있는한 동일한 방식으로 프라이머 서열로서 사용될 수도 있다. 본원에 사용된 '서열의 실질적 상동성(substantial homology)'이란, 상기 변이체가 원래의 프로브와 동일한 기능을 갖기에 충분히 상동적임을 의미한다. 바람직하게, 변이체는 구체화된 서열과 아미노산 또는 뉴클레오타이드에 있어서 동일성(identity)이 50% 이상; 더욱 바람직하게는 75% 이상; 및 가장 바람직하게는 90% 이상일 것이다. 상기 변이체가 의도하는 바와 같은 기능을 갖는데에 필요로 하는 상동성의 정도는 서열의 의도된 용도에 따라 상이할 것이다. 서열의 기능을 개선시키거나 또는 방법론적으로 유리하도록 디자인된 돌연 변이체 즉, 삽입성 및 결실성 돌연 변이체를 제조하는 방법은 당 업계의 숙련자들에게 공지되어 있다.
PCR 기술
중합 효소 연쇄 반응(Polymerase Chain Reaction;PCR)은 핵산 서열의 반복적이고 효소적인, 프라이머에 의한 합성 방법이다. 상기 방법은 당 업계의 숙련자들에 의하여 널리 공지되고 일반적으로 사용되는 방법이다[Mullis, 미국 특허 제 4,683,195 호, 제 4,683,202 호 및 제 4,800,159 호; Saiki, Randall K., Stephen Scharf, Fred Faloona, Kary B. Mullis, Glenn T. Horn, Henry A. Erlich, Norman Arnheim(1985) "Enzymatic Amplification of β-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia", Science 230:1350-1354 참조]. PCR은 표적 서열의 반대편 서열에 혼성화되는 2개의 올리고뉴클레오타이드 프라이머와 접하고 있는, 관심 있는 DNA 단편을 효소적으로 증폭시키는 것에 그 원리를 두고 있다. 상기 프라이머는 3' 말단이 서로를 향하게 방향지워 진다. PCR은 주형의 열변성, 상보적 서열로의 프라이머의 어닐링 및 어닐링된 프라이머를 DNA 중합효소로 연장시켜 PCR 프라이머의 5' 말단으로 한정되는 단편을 증폭시키는 단계를 반복적으로 순환시킴으로써 수행된다. 각각의 프라이머의 연장 생성물이 다른 프라이머의 주형으로 사용될 수 있기 때문에, 근본적으로 각각의 단계는 전 단계에서 생성된 DNA 단편의 양을 2배로 증폭시킨다. 이로써 특정 표적 단편은 단 몇시간 내에 수백만배 이상으로 기하 급수적으로 축적된다. 서머스 아쿠아티쿠스(Thermus Aquaticus)와 같은 호열성 세균으로부터 분리된 Taq 중합 효소와 같은 내열성 DNA 중합 효소를 사용하여 증폭 과정을 완전히 자동적으로 진행시킬 수 있다.
본 발명에 의하여 수득될 수 있는 DNA 서열은 PCR 증폭 반응용 프라이머로서 사용될 수 있다. PCR 증폭 반응 수행시, 프라이머와 주형 사이의 어느 정도의 미스매취는 큰 문제가 되지 않는다. 그러므로 본원의 관점에서 수득된 프라이머를 돌연 변이, 결실 및 삽입(특히 5' 말단에 부가하는 것)시키는 것은 본 발명의 범주내에 속한다. 주어진 프라이머내에서의 돌연 변이, 삽입 및 결실은 통상의 숙련공에게 공지된 방법에 의하여 제조될 수 있다. 주어진 프로브로부터 생성된 돌연 변이성, 삽입성 및 결실성 변이체가 원래의 프로브보다 다소 효율적이라는 것을 알 수 있는 것은 매우 중요하다. 이와 같은 효율상의 차이점에도 불구하고, 상기 변이체들은 본 발명의 범주내에 포함된다.
본원과 관련된 모든 미국 특허는 본원에 참고 문헌으로서 첨부되어 있다.
이하는 본 발명을 실시하는 방법을 나타내는 실시예이다. 하기 실시예는 본원 발명을 한정하는 것으로서 해석되어서는 안된다. 하기의 모든 퍼센트는 중량부이며 용매 혼합 비율은 특별히 언급되지 않은한 부피부이다.
실시예 1 - 본 발명에 유용한 비.티. 분리주의 배양
비.티. 분리 균주 또는 이의 돌연 변이체의 하위 배양액은 다음과 같은 펩톤, 글루코즈, 염의 배지에 접종시키는데에 사용될 수 있다.
박토 펩톤(Bacto Peptone) 7.5g/l
글루코즈 1.0g/l
KH2PO43.4g/l
K2HPO44.35g/l
염 용액 5.0ml/l
CaCl2용액 5.0ml/l
pH 7.2
염 용액(100ml)
MgSO4 7H2O 2.46g
MnSO4 H2O 0.04g
ZnSO4 7H2O 0.28g
FeSO4 7H2O 0.40g
CaCl2용액(100ml)
CaCl2 2H2O 3.66g
상기 염 용액 및 CaCl2용액을 여과지로 살균시킨후 접종시 고압 가열법(autoclave)에 의해서 가열된 배양액에 가하였다. 플라스크를 30℃, 200rpm의 회전 교반기에서 64시간 동안 배양시켰다.
상기 방법은 당 업계에 널리 공지된 방법에 의하여 거대한 발효조로 용이하게 규모를 확대시킬 수 있다. 자주 사용되는 방법은 수득된 발효액에 예를 들어, 원심 분리와 같은 분리 기술을 적용시키는 것이다.
이와는 달리, 비.티. 분리 균주, 또는 이의 돌연 변이체의 하위 배양액은 TB 배양액이라 알려진 이하의 배지를 접종시키는데에 사용될 수 있다.
트립톤 12g/l
효모 추출액 24g/l
글리세롤 4g/l
KH2PO42.1g/l
K2HPO414.7g/l
pH 7.4
냉각후 포타슘 포스페이트를 고압 가열된 배양액에 가하였다. 플라스크를 30℃, 250rpm의 회전 교반기에서 24-36시간 동안 배양시켰다.
상기 방법은 당 업계에 공지된 방법에 따라 거대한 발효조로 용이하게 규모가 확대될 수 있다.
상기 발효를 통하여 수득된 비.티.는 당 업계에 널리 공지된 방법에 의하여 분리될 수 있다. 자주 사용되는 방법은 수득된 발효액에 예를 들어, 원심 분리와 같은 분리 기술을 적용시키는 것이다. 특정 구체예에서, 본 발명에 유용한 비.티. 단백질은 상등액으로부터 수득될 수 있다. 활성 단백질을 포함하는 배양액의 상등액은 이하에 기술된 바와 같이 생물 검정법에 사용될 수 있다.
실시예 2 - 신규의 인시류 활성 바실러스 츄린겐시스 독소를 코드화하는 유전자의 동정
비.티. 독소 유전자의 3' 말단에 인접한 다형성 DNA 절편(polymorphic DNA fragment)을 PCR 증폭시켜 신규의 독소 유전자를 동정 및 분류하는 데에 유용한 2개의 프라이머를 디자인하였다. 이들 올리고뉴클레오티드 프라이머는 Cry7, Cry8 또는 Cry9의 아과(subfamily)의 독소를 코드화하는 유전자들을 보다 보편적인 Cry1 아과의 인시류-활성 독소 유전자와 증폭된 DNA에 대한 크기의 차이를 기초로 하여 구분한다. 이들 프라이머 서열들은 다음과 같다 :
정방향 1 5'CGTGGCTATATCCTTCGTGTYAC3' (서열 1)
역방향 1 5'ACRATRAATGTTCCTTCYGTTTC3' (서열 2)
정방향 2 5'GGATATGTMTTACGTGTAACWGC3' (서열 3)
역방향 2 5'CTACACTTTCTATRTTGAATRYACCTTC3' (서열 4)
본 발명의 프라이머 쌍 1(서열 1 및 2)을 사용하는 표준적인 PCR 증폭 방법 결과, cry1 아과와 관련된 비.티. 독소 유전자로부터 길이가 약 415-440 염기쌍인 DNA 절편을 수득하였다.
본 발명에 의한 프라이머 쌍 2(서열 3 및 4)를 사용하는 PCR 증폭 방법 결과, cry7, cry8 또는 cry9 아과 독소 유전자로부터 길이가 약 230-290 염기쌍인 DNA 절편을 수득하였다.
본 발명에 의한 프라이머들은 신규의 독소를 코드화하는 유전자를 동정하는데에 사용될 수 있다. PCR용 크루드한 DNA 주형을 비.티. 균주로부터 제조하였다. 밤새 배양시킨 바실러스 츄린겐시스 플레이트 배양액으로부터 세포들을 긁어내어 300ml TE 완충액(10mM Tris-Cl, 1mM EDTA, pH 8.0)에 재현탁시켰다. 단백질 분해효소 K(Proteinase K) 0.1mg/ml를 가한후 세포 현탁액을 15분 동안 55℃로 가열시켰다. 이후 상기 현탁액을 15분 동안 가열시켰다. 미세 원심 분리로 세포 조각을 펠렛화시켜 DNA를 포함하는 상등액을 clcan 튜브로 옮겼다.
PCR은 상술된 정방향 2(서열 3) 및 역방향 2(서열 4) 올리고뉴클레오티드로 구성된 프라이머 쌍을 사용하여 수행되었다. 균주들은 길이가 약 230-290bp인 DNA 절편을 증폭시켜 특징화된 유전자들을 함유한다는 사실이 동정되었다. 이들 균주들로부터의 포자-결정체 제조물을 아그로티스 입실론 및 추가의 인시류 표적의 생물 활성에 대하여 시험하였다.
프라이머 1 및 2(서열 1 및 2, 서열 3 및 4)모두를 사용하여 PS185U2를 관찰하였다. 이들 균주에서, 프라이머쌍 1(서열 1 및 2)은 상기 cry1 아과와 관련된 독소 유전자에서 예상되는 크기를 갖는 DNA 밴드를 나타내었다.
실시예 3 - 인시류-활성 균주에 존재하는 바실러스 츄린겐시스 독소 유전자의 제한 절편 길이 다형성(Restriction Fragment Length Polymorphism ; RFLP) 분석법
흡광도가 600nm에서 1.0일때까지 생장시킨 바실러스 츄린겐시스(비.티.) 균주로부터 전체 세포성 DNA(Cellular DNA)를 제조하였다. 세포들을 원심 분리시켜 펠렛화한후 원형질체 완충액(protoplast buffer)(0.3M 설탕, 25mM Tris-Cl(pH 8.0), 25mM EDTA내 20mg/ml 리소자임)에 재현탁시켰다. 37℃에서 1시간 동안 배양시킨후, 원형질을 2회 동결-냉동 순환에 의해서 용균시켰다. 0.1M NaCl, 0.1% SDS, 0.1M Tris-Cl 용액 9 부피를 가하여 용균을 완료하였다. 분리된 용균물을 페놀:클로로포름(1:1)로 2회 추출하였다. 핵산을 2부피의 에탄올로 침전시킨후 원심 분리하여 펠렛화시켰다. 상기 펠렛을 TE 완충액내에 재현탁시킨후 RNase를 가하여 최종 농도가 50g/ml가 되도록 하였다. 37℃, 1시간 동안 배양시킨후, 각각의 용액을 페놀:클로로포름(1:1) 및 TE-포화 클로로포름으로 추출하였다. 3M NaOAc 0.1 부피 및 에탄올 2 부피를 가하여 수성 상으로부터 DNA를 침전시켰다. 이후 DNA를 원심 분리에 의하여 펠렛화시킨후 70% 에탄올로 세정하여 건조시키고 다시 TE 완충액내에 재현탁시켰다.
PCR-증폭되고32P-라벨링된 DNA 프로브의 2가지 형태는 전체의 세포성 비.티. DNA에 대한 표준적인 서던 혼성화 방법을 사용하여 RFLP로 독소 유전자를 특징화할 수 있다. 첫번째 프로브(A)는 이하의 프라이머를 사용하여 증폭된 DNA 절편이다.
정방향 3 : 5' CCAGWTTTAYAGGAGG 3'
역방향 3 : 5' GTAAACAAGCTCGCCACCGC 3'
두번째 프로브(B)는 상기 실시예에 기술된 프라이머로 증폭된 230-290bp 또는 415-440bp인 DNA 절편이다.
상술한32P-라벨링된 DNA 프로브를 사용하는 서던 블럿시 고정화된 DNA의 혼성화는 표준적인 방법으로 수행되었다[Maniatis, T.,E.F.Fritsch, J.Sambrook(1982)Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY]. 일반적으로, 혼성화 및 이후의 세정은 온화한 엄격성(moderate stringency)하에서 수행된다. 이중 사슬 DNA 유전자 프로브에 있어서, 혼성화는 6X SSPE, 5X Denhardt 용액, 0.1% SDS, 0.1mg/ml 변성 DNA내에서, DNA 혼성체의 용융 온도(Tm)이하인 20-25℃에서 밤새 수행된다. 용융 온도는 이하의 식으로 나타내어 진다[Beltz, G.A., K.A., Jacobs, T.H.Eickbush, P.T.Cherbas 및 F.C.Kafatos(1983)In Methods in Enzymology, R.Wu.,L.Grossman 및 K.Moldave(eds.), Academic Press, New York. 100:266-285].
Tm = 81.5℃ + 16.6Log[Na+]+0.41(G+C%)-0.61(포름아미드%)-600/이중체 염기쌍 길이
통상적으로 세정은 다음과 같이 수행된다:
(1)1X SSPE, 0.1% SDS내 15분 동안 실온에서 2회 세정 (낮은 엄격성 세정)
(2)0.2X SSPE, 0.1% SDS내 15분 동안 Tm - 20℃에서 1회 세정(온화한 엄격성 세정)
아그로티스 입실론에서 가장 활성이 높은 10개의 균주에 대한 RFLP 데이터가 얻어졌다(표 3 및 4). 본원에 기술된 DNA 밴드의 혼성화에는 연구된 신규의 독소 유전자의 전부 또는 일부를 포함한다.
실시예 4 - 독소 유전자의 DNA 시퀀싱
아그로티스 입실론에 활성인 비.티. 균주에 존재하는 독소 유전자의 PCR-증폭된 단편을 시퀀싱하였다. 이를 수행하기 위해서, 정방향 프라이머 3(서열 5) 및 역방향 프라이머 3(서열 6)을 사용하여 수득된 증폭 DNA 절편을 공급자(San Diego, CA 소재, Invitrogen)에 의하여 기술된 바와 같이, 맨처음 PCR DNA TA-클로닝 플라스미드 벡터인, pCRII에 클로닝하였다. 시퀀싱용으로 각각의 비.티. 균주로부터 증폭시켜 얻어진 DNA 절편의 혼합물로부터 각각의 pCRII 클론들을 선택하였다. 가열하여 콜로니들을 용균시켜 크루드한 플라스미드 DNA를 방출시켰다. 자동화 시퀀싱용 DNA 주형을 프라이머의 다중 클로닝 부위(multiple cloning site)에 측접하는 벡터-특이적 프라이머를 사용하는 PCR에 의하여 증폭시켰다. 상기 DNA 주형은 자동화된 시퀀싱 방법인 Applied Biosystems(Foster City, CA)을 사용하여 시퀀싱되었다. 이하 기술된(서열 62 이후의 7개의 서열) 독소 유전자 서열 및 이에 상응하는 뉴클레오티드 서열은 상기 방법으로 동정되었다. 이들 서열들을 표 5에 나열하였다. 상기 뉴클레오티드 서열들로부터 추론된 폴리펩타이드 서열들도 나타내었다.
이러한 부분적인 서열로부터, PCR 프라이머로서 유용한 7개의 올리고뉴클레오티드 또는 혼성화 프로브를 디자인하였다. 상기 올리고뉴클레오티드들은 다음과 같다 :
5' GTTCATTGGTATAAGAGTTGGTG 3' (서열 63)
5' CCACTGCAAGTCCGGACCAAATTCG 3' (서열 64)
5' GAATATATTCCCGTCYATCTCTGG 3' (서열 65)
5' GCACGAATTACTGTAGCGATAGG 3' (서열 66)
5' GCTGGTAACTTTGGAGATATGCGTG 3' (서열 67)
5' GATTTCTTTGTAACACGTGGAGG 3' (서열 68)
5' CACTACTAATCAGAGCGATCTG 3' (서열 69)
특정 유전자 독소 서열 및 이들 서열을 혼성화법 또는 상기된 역방향 프라이머 3과 관련된 PCR에 의하여 동정할 수 있는 올리고뉴클레오티드 프로브에 관해서는 표 5에 나열되어 있다.
실시예 5 - 전체 길이 독소 유전자의 분리 및 DNA 시퀀싱
당 업계에 공지된 표준적인 방법을 사용하여 비.티. 균주로부터 전체 세포성 DNA를 추출하였다. 예를 들어, 상기 실시예 3을 참조하시오. 크기별로 분획화된 전체 세포성 DNA의 Sau3A 부분 제한 절편을 박테리오파지 벡터, Lamda-Gem11내에 구성하였다. 재조합 파지를 패키징시킨후 이.콜라이 KW251 세포에 도말하였다. 플라크를 올리고뉴클레오티드 프라이머인 서열 5 및 6으로 PCR-증폭시킨 DNA 절편으로부터 수득된 방사능 라벨링된 유전자-특이적 프로브로 혼성화시켜 스크리닝하였다. 혼성화 파지는 플라크-정제되어 표준적인 방법으로 DNA를 분리시키기 위한 이.콜라이 KW251 세포의 배양액을 감염시키는데에 사용되었다[Maniatis, T.,E.F.Fritsch, J.Sambrook(1982)Molecular Cloning:A Laboratory Manual, Cold Spring Harbor, Cold Spring Harbor, NY]. 이후 독소 유전자는 DNA 서열 분석용인 pBluescript 벡터(Stratagene)에 서브 클로닝되었다.
이하에 나열된 전체 길이 독소 유전자는 자동화된 시퀀싱 방법인 Applied Biosystems(Foster City, CA)를 사용하여 시퀀싱되었다. 독소 유전자 서열 및 각각의 예상 폴리펩타이드 서열들을 이하에 나열하였다.
상기 독소들을 코드화하는 플라스미드를 포함하는 재조합 이.콜라이 NM522 균주는 NRRL에 1997년 6월 27일 기탁하였다.
실시예 6 - 슈도모나스 플레오레스센스(피.에프.)내에서 신규의 비.티. 독소의 이종 발현(heterologous expression)
전체 길이의 독소 유전자를 표준적 DNA 클로닝 방법에 의하여 플라스미드 벡터내에 조작하여 이를 발현시키기 위하여 슈도모나스 플레오레스센스를 형질 전환시켰다. 재조합 세균 균주(표 6)를 발현시키기 위해서 교반 플라스크내에서 생장시켜 독소를 생산하여 다양한 인시류 해충에 대하여 정량 생물 검정법을 수행하였다.
실시예 7 - 트립신을 사용한 내독소의 프로세싱
슈도모나스 플루오레스센스 배양액을 표준 방법에 따라 48시간 동안 생장시켰다. 원심 분리로 세포 펠렛을 수득한후 물로 3회 세정하여 -70℃에 저장하였다. 내독소 함유물을 리소자임 및 DNase로 처리후 차동 원심분리(differential centrifugation)시켜 세포로부터 분리하였다. 이후 이러한 방식으로 분리된 독소를 트립신 분해시켜 펩타이드를 절단한후 인시류 해충에 대한 생물 검정법을 실행하였다.
상세한 방법을 이하에 기술하였다.
용균 완충액(Lysis buffer) 4L(1일 사용량)
gm
Tris 염기 24.22
NaCl 46.75
글리세롤 252
디티오트레이톨 0.62
EDTA 디소듐 염 29.78
Triton X-100 20 ml
pH를 HCl로 7.5로 맞춘후 증류수를 가하여 최종 농도가 4L되도록 하였다.
1. 37℃ 수조에서 냉동 세포 펠렛을 해동시켰다.
2. 500ml들이 폴리카보네이트 원심 분리 용기가 약 400ml정도로 채워질때까지 용균 완충액을 가하였다. 상기 용기를 뒤집거나 또는 낮은 rpm에서 Polytron을 사용하여 분산시켰다.
3. 4℃, 20분 동안 원심 분리(10,000 × g)시켰다.
4. 상등액을 가만히 따른후 버렸다.
5. 낮은 rpm에서 Polytron을 사용하여 펠렛의 단위 그램당 5ml의 용균 완충액에 상기 펠렛을 재현탁시켜 펠렛을 분산시켰다.
6. 25mg/ml 리소자임 용액을 상기 현탁액에 가하여 최종 농도가 0.6mg/ml가 되도록 하였다.
7. 37℃에서 4분 동안 배양시켰으며, 30초마다 용기를 뒤집어주었다.
8. 현탁액을 얼음에 1시간 동안 방치시켰다,
9. 상기 튜브에 2.5M MgCl6H2O를 가하여 최종 농도가 60mM이 되도록 하였다. 40mg/ml의 디옥시리보뉴클레아제 I(Sigma) 용액을 가하여 최종 농도가 0.5mg/ml가 되도록 한다.
10. 4℃에서 밤새 배양하였다.
11. 낮은 rpm의 Polytron을 사용하여 상기 용균액을 균질화시켰다.
12. 4℃, 20분 동안 10,000g에서 원심 분리하였다. 상등액을 따라내어 버렸다.
13. 용균 완충액내에 함유물 팰렛을 재현탁시켰다. 용균이 완전히 이루어졌는지 현미경으로 체크하였다.
14. 용균 완충액으로 상기 함유물 펠렛을 5회 세정하였다.(2 - 5 단계 반복)
15. 10mM Tris-Cl, pH 7.5, 0.1mM PMSF의 현탁액 형태로 -70℃에서 1.5ml들이 에피튜브에 저장하였다.
트립신으로 함유물을 분해하는 방법은 다음과 같이 수행되었다.
분해 용액(digestion solution)
1. 2ml 1M NaCAPS pH 10.5
2. 함유 제조물(100mg 단백질)
3. 절단될 단백질의 양과 트립신의 비율을 1:100으로 맞춘 용액(본 방법의 과정중에 첨가)
4. 최종 부피 10ml가 되도록 하는 H2O
트립신 처리는 다음과 같이 수행되었다.
1. 상기 분해 용액을 37℃에서 15분 동안 트립신 없이 배양시켰다.
2. 1:100(트립신:독소 단백질 중량/중량) 비율의 트립신을 가하였다.
3. 상기 용액을 37℃에서 2시간 동안 가끔씩 뒤집어 주어 혼합시키면서 배양하였다.
4. 상기 분해 용액을 4℃, 15분 동안 15,000g에서 원심 분리시켰다.
5. 상등액을 분리하여 보관하였다.
6. 상등액을 SDS-PAGE 전기 영동으로 분석하여 이하에서 논의되는 바와 같이 생물 검정법에 사용하였다.
실시예 8 - 키메라 구성물(chimeric construct)내에서 비.티. 균주 HD129로부터의 유전자의 발현
유전자를 비.티. HD129 균주로부터 분리하였다. 상기 유전자는 명확한 해독 개시 코돈이 존재하지 않는 가성 유전자(pseudogene)이다. HD129 균주로부터 수득한 유전자를 발현시키기 위하여, 슈도모나스 발현 시스템내 cry1Ac의 처음 28 코돈을 융합시킨 유전자를 디자인하여 구성하였다. 상기 키메라 독소의 뉴클레오티드 및 펩타이드 서열을 서열 76 및 77에 나타내었다. 유도시, 상기 신규의 키메라 독소를 포함하는 재조합 피.플루오레스센스는 예상 크기를 갖는 폴리펩타이드를 발현하였다.
실시예 9 - 독소 유전자의 추가의 시퀀싱
표 7에 나열한 분리 균주로부터 수득한 가용성 독소들의 DNA를 시퀀싱하였다. 이로써 얻어진 서열들의 서열 번호도 표 7에 나타내었다.
실시예 10 - 흑색 컷웜 생물 검정법
비.티. 분리 균주를 포함하는 분말 현탁액을 적당량의 분말과 증류수를 혼합한후 격렬하게 교반시켜 제조하였다. 이후 상기 현탁액을 알팔파 분말 28 그램(Bioserv) 및 최종 먹이의 단위 리터당 1.2ml의 포르말린으로 보강된 흑색 컷웜 인공 먹이(BioServ, Frenchtown, NJ)와 혼합하였다. 이후 상기 현탁액을 현탁액 3ml와 먹이 27ml의 비율이 되도록 최종 인공 먹이와 혼합하였다. 교반후, 상기 혼합물을 3ml의 웰로 구획화된 플라스틱 트레이에 부었다[Nutrend Container Corporation, Jacksonville, FL]. 비.티.를 함유하지 않는 물을 대조구로서 사용하였다. 초기 제 1 영의 아그로티스 입실론 유충(Frnch Agricultural Services, Lamberton, MN)을 상기 먹이 혼합물상에 방치시켰다. 이후 웰을 고정용 철을 사용하여 "MYLAR" 시이트(ClearLam Packaging, IL)로 밀봉한후 공기가 통하도록 각각의 웰에 구멍을 몇개씩 뚫어주었다. 유충을 29℃, 4일 동안 명기:암기의 비율이 14:10인 방치소에 방치하였다. 4일후 사멸율을 측정하였다.
이하의 비.티. 분리 균주들이 흑색 컷웜에 대하여 활성을 갖는 것으로 관찰되었다 : PS185U2, PS11B, PS218G2, PS213E5, PS86W1, PS28C, PS86BB1, PS89J3, PS86V1, PS94R1, HD525, HD573, PS27J2, HD110, HD10, PS202S, HD29, PS202S, HD29, PS101DD, HD129 및 PS31G1. 생물 검정 결과는 표 8에 나타내었다.
실시예 11 - 아그로티스 입실론에 대한 비.티. 분리 균주의 활성
배양액의 상등액으로서의 균주들을 시험하였다. 시료를 흑색 컷웜 인공 먹이(BioServ, Frenchtown, NJ)에 적용하여 공기 건조시킨후 유충을 투여하였다. 비.티.를 함유하지 않는 용액을 대조구로서 사용하였다. 처리된 각각의 웰에 알을 투여한후 이후 웰을 고정용 철을 사용하여 "MYLAR" 시이트(ClearLam Packaging, IL)로 밀봉한후 공기가 통하도록 각각의 웰에 구멍을 몇개씩 뚫어주었다. 상기 알을 29℃, 4일 동안 명기:암기의 비율이 14:10인 방치소에 방치하였다. 4일후 사멸율을 측정하였다. 에이.입실론에 대하여 사멸율을 나타내는(물 대조구 이상의 사멸율) 균주들을 표 9에 나타내었다.
실시예 12 - 헬리오티스 비레스센스(Heliothis virescence)(Fabricius) 및 헬리코베르파 지아(Helicoverpa zea)(Boddie)
동결 슈도모나스 플루오레스센스 클론 또는 비.티. 상등 배양액 시료들의 형태로서의 균주들을 시험하였다. 증류수와 시료들을 각각 혼합한후 격렬하게 교반시켜 클론의 현탁액을 제조하였다. 먹이 혼입 생물 검정법을 수행하기 위하여, 6mL 현탁액 및 54mL 먹이의 비율로 현탁액을 인공 먹이와 혼합시켰다. 교반후, 상기 혼합물을 3ml의 웰로 구획화된 플라스틱 트레이에 부었다[Nutrend Container Corporation, Jacksonville, FL]. 시료의 상등액을 상기 요약한 바와 같이 먹이와 3-6ml의 비율로 혼합하였다. 상부 로드 생물 검정법(top load bioassay)에서, 현탁액 또는 상등액을 상기 인공 먹이의 상부에 적용하여 공기 건조시킨후 유충을 투입시켰다. 물 블랭크를 대조구로서 사용하였다. 제 1 영의 유충(USDA-ARS, Stoneville, MS)을 상기 목이 혼합물상에 방치시켰다. 이후 웰을 고정용 철을 사용하여 "MYLAR" 시이트(ClearLam Packaging, IL)로 밀봉한후 공기가 통하도록 각각의 웰에 구멍을 몇개씩 뚫어주었다. 상기 유충을 25℃, 6일 동안 명기:암기의 비율이 14:10인 방치소에 방치하였다. 6일후 사멸율을 측정하였다.
결과들은 다음과 같았다 :
실시예 13 - 오스트리니아 누빌라리스(Ostrinia nubilalis)(유럽 옥수수 좀 벌레 ; European Corn Borer)
본 발명의 분리 균주 및 독소를 유럽 옥수수 좀벌레(ECB)인 오스트리니아 누빌라리스를 박멸하는데에 사용하였다. ECB에 대한 활성은 예를 들어, 제 1 영의 유충을 사용하는 표준적 인공 먹이 혼입 곤충 생물 검정법에 의해서 확인하였다. 특정 구체예를 통해서, 31G1(a) 유전자를 발현하는 트립신-처리된 클론의 LC50 수치는 0.284(㎍/ml)라는 것이 밝혀졌다.
실시예 14 - 식물에 독소 유전자를 삽입하는 방법
본 발명의 하나의 양상은 본 발명의 살충 독소를 코드화하는 유전자로 식물을 형질 전환시키는 것이다. 형질 전환된 식물은 표적 해충의 공격에 대하여 내성을 갖는다.
본원에 기술된 살충 독소를 코드화하는 유전자는 당 업계에 널리 공지된 다양한 방법으로 식물 세포내 삽입될 수 있다. 예를 들어, 고등 식물에 외부 유전자를 삽입시키기 위한 제조물로서는 이.콜라이내 복제 시스템 및 형질 전환된 세포가 선별될 수 있도록 만들어 주는 마커를 포함하는 다수의 클로닝 벡터가 유용하다. 상기 벡터에는 예를 들어, pBR322, 일련의 pUC들, 일련의 M13mp들, pACYC184등이 있다. 따라서 비.티. 독소를 코드화하는 서열이 벡터내 적당한 제한 부위에 삽입될 수 있다. 결과로 제조된 플라스미드를 이.콜라이 내로의 형질전환시 사용하였다. 상기 이.콜라이 세포를 적당한 영양 배지내에서 배양시킨후 수득하여 용균시켰다. 그 결과, 플라스미드가 수득되었다. 분석 방법으로서는 서열 분석, 제한 분석, 전기 영동 및 기타 생화학-분자 생물학적 방법이 일반적으로 수행된다. 각각의 조작을 수행한후, 사용된 DNA 서열을 절단시켜 다음의 DNA 서열에 연결시켰다. 각각의 플라스미드 서열을 동일한 또는 다른 플라스미드내에 클로닝시켰다. 원하는 유전자를 식물내에 삽입시키는 방법에 따라서, 기타의 DNA 서열이 필요할 수도 있다. 예를 들어, 만일 Ti 또는 Ri 플라스미드가 식물 세포의 형질 전환시 사용되려면, 최소한 Ti 또는 Ri 플라스미드 T-DNA의 우측 경계 부위 - 때때로 Ti 또는 Ri 플라스미드 T-DNA의 좌측 또는 우측 경계 부위 - 가 삽입된 유전자의 측면과 접하여 연결된다.
식물 세포의 형질 변환시 사용되는 T-DNA 의 사용에 관하여는 EP 120 516; Hoekema(1985)저 "The Binary Plant Vector System" Offset-durkkerij Kanters B.V., Alblasserdam, Chapter5; Fraley외 다수, Crit.Rev.Pant Sci. 4:1-46; 및 An외 다수(1985) EMBO J. 4:277-287에 충분히 기술되어 있다.
삽입된 DNA가 일단 게놈내 융합되면, 이 DNA는 게놈내에서 비교적 안정하여 대게는 다시 외부로 방출되지 않는다. 일반적으로 상기 DNA는 예를 들어 카나마이신, G418, 블레오마이신, 하이그로마이신 또는 클로르암페니콜과 같은 항생제에 대한 내성을 식물에게 부여하는 선별 마커를 포함한다. 각각 사용된 마커는 DNA가 삽입되지 않은 세포보다 형질 전환된 세포의 선별을 가능하게 한다.
다수의 기술들이 식물 숙주 세포에 DNA를 삽입시키는데에 유용하다. 이와 같은 기술에는 아그로박테리움 튜미페이션스(Agrobacterium tumefaciens) 또는 아그로박테리움 리조제네스(Agrobacterium rhizogenes)를 형질 전환제로서 사용하는 T-DNA 형질 전환, 융합, 주사, 바이오리스틱스(biolistics)(미세 입자 충격법;microparticle bombardment) 또는 전기 영동 및 다른 가능한 방법을 포함한다. 아그로박테리아가 형질 전환에 사용되는 경우, 삽입될 DNA는 특정 플라스미드, 즉 중간 벡터(intermediate vector) 또는 2원 벡터(binary vector)내에 클로닝되어야 한다. 상기 중간 벡터는 T-DNA내 서열과 상동적인 서열로 인한 상동 재조합(homologous recombination)에 의하여 Ti 플라스미드 또는 Ri 플라스미드내에 삽입될 수 있다. 상기 Ti 플라스미드 또는 Ri 플라스미드는 상기 T-DNA를 전이시키는데에 필요한 vir 영역을 포함한다. 중간 벡터 자체는 아그로박테리아내에서 복제하지 못한다. 상기 중간 벡터는 헬퍼 플라스미드(helper plasmid)를 사용하여 아그로박테리움 튜미페이션스(Agrobacterium tumefaciens)내에 전이될 수 있다(컨쥬게이션). 2원 벡터 자체는 이.콜라이와 아가로박테리아 모두에서 복제하지 못한다. 이들은 우측 및 좌측 T-DNA 경계 부위로 틀이 구성된 선별 마커 유전자 및 링커 또는 폴리링커를 포함한다. 이들은 아가로박테리아내에 직접적으로 형질 전환된다[Holsters외 다수, 1978, Mol.Gen. Genet. 163:181-187]. 숙주 세포로 사용된 아가로박테리아는 vir 영역을 갖는 플라스미드를 포함할 것이다. 상기 vir 영역은 T-DNA를 식물 세포내에 전이시키는데에 필요하다. 추가의 T-DNA가 포함될 수도 있다. 이와 같이 형질 전환된 박테리아는 식물 세포의 형질 전환에 사용된다. 식물 세포내에 DNA를 전이시키기 위하여 아그로박테리움 튜미페이션스(Agrobacterium tumefaciens) 또는 아그로박테리움 리조제네스(Agrobacterium rhizogenes)와 식물 외식편(plant explant)을 함께 배양시키는 것이 바람직하다. 이후 적당한 배지에서 감염시킨 식물 재료(예를 들어 입 조각, 줄기 조각, 분열 조직, 뿌리 및 원형질 또는 현탁 배양 세포)로부터 식물 개체를 재발생시킬 수 있는데, 이때 선별을 위하여 항생제 또는 살생물제를 포함할 수도 있다. 이후 이와 같이 수득된 식물에 대하여 삽입 DNA의 존재 여부를 시험하였다. 주사(injection) 또는 전기 천공법(electroporation) 수행시 특별한 조건이 필요한 것은 아니다. 예를 들어 pUC 유도체와 같은 보통의 플라스미드를 사용할 수도 있다. 바이오리스틱스 형질 전환(biolistics transformation)시, 플라스미드 DNA 또는 선형 DNA가 사용될 수 있다.
상기 형질 전환된 세포는 통상의 방식에 따라 형태학적으로 정상인 식물로 재발생된다. 만일 형질 전환 세포가 생식 세포를 포함하는 경우에는, 삽입된 DNA 및 이에 상응하는 형질은 자손 식물에 전이될 것이다. 이와 같은 식물은 정상적으로 성장할 수 있으며 형질 전환된 유전 인자 또는 기타의 유전 인자를 갖는 식물과 교잡될 수 있다. 결과로 형성된 잡종 개체는 이에 상응하는 표현형을 나타낼 것이다.
본 발명의 바람직한 구체예에서, 식물은 식물에 사용되기에 최적화된 코돈인 유전자로 형질 전환될 것이다. 예를 들어, 본원에 참고 문헌으로서 첨부된 미합중국 특허 제 5,380,831호를 참조. 바람직하게는 분절된 독소를 코드화하는 식물이 사용될 것이다. 상기 분절된 독소는 일반적으로 전체 길이 독소의 약 55-80%를 코드화할 것이다. 식물에 사용되는 비.티. 유전자를 합성하는 방법은 당 업계에 공지되어 있다.
본원에 기술된 실시예 및 구체예는 본 발명을 단지 예시하기 위하여 기술된 것으로서 당 업계의 숙련자들에 의하여 다양하게 변경 및 수정이 가능하며 이는 또한 본원 발명의 사상과 범주 및 첨부된 청구 범위내에 포함된다.
〈110〉 MYCOGEN CORPORATION
〈120〉 Toxins Active Against Ostrinia nubilalis
〈150〉 US 09/002285
〈151〉 1997-12-31
〈160〉 105
〈170〉 KOPATIN 1.5
〈210〉 1
〈211〉 23
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 1
cgtggctata tccttcgtgt yac 23
〈210〉 2
〈211〉 23
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 2
acratraatg ttccttcygt ttc 23
〈210〉 3
〈211〉 23
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 3
ggatatgtmt tacgtgtaac wgc 23
〈210〉 4
〈211〉 28
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 4
ctacactttc tatrttgaat ryaccttc 28
〈210〉 5
〈211〉 16
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 5
ccagwtttay aggagg 16
〈210〉 6
〈211〉 20
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 6
gtaaacaagc tcgccaccgc 20
〈210〉 7
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 7
Pro Gly Phe Xaa Gly Gly Asp Ile Leu Arg Arg Thr Ser Pro Xaa Gln
1 5 10 15
Ile Ser Xaa Leu Arg Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr
20 25 30
Arg Val Arg Ile Xaa Xaa Ala Ser Thr Thr Xaa Xaa Gln Phe His Thr
35 40 45
Ser Ile Xaa Gly Arg Pro Ile Asn Gln Gly Asn Phe Ser Xaa Thr Met
50 55 60
Ser Ser Gly Ser Asn Leu Gln Ser Gly Xaa Phe Arg Thr Val Gly Phe
65 70 75 80
Thr Thr Pro Xaa Asn Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser
85 90 95
Xaa His Val Phe Asn Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu
100 105 110
Phe Val Pro Ala Glu Val Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg
115 120 125
Ala Xaa Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 8
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 8
ccaggattta yaggaggaga tattcttcga agaacttcac ctgkscagat ttcawcctta 60
agagtaaata ttactgcacc attatcacaa agatatcggg taagaattcr cwacgcttct 120
acyacawatt twcaattcca tacatcaatt grcggaagac ctattaatca gggkaatttt 180
tcascaacta tgagtagtgg gagtaattta cagtccggaa kctttaggac tgtaggtttt 240
actactccgt ktaacttttc aaatggatca agtgtattta cgttaagtkc tcatgtcttc 300
aattcaggca atgaagttta tatagatcga attgaatttg ttccggcaga agtaaccttt 360
gaggcagaat atgatttaga aagagcacma aaggcggtgg cgagcttgtt tac 413
〈210〉 9
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 9
Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Asp Gly Gly Xaa
1 5 10 15
Val Gly Thr Ile Arg Ala Asn Val Asn Ala Pro Leu Thr Gln Gln Tyr
20 25 30
Arg Ile Arg Leu Arg Tyr Ala Ser Thr Thr Ser Phe Val Val Asn Leu
35 40 45
Phe Val Asn Asn Ser Ala Ala Gly Phe Thr Leu Pro Ser Thr Met Ala
50 55 60
Gln Asn Gly Ser Leu Thr Xaa Glu Ser Phe Asn Thr Leu Glu Val Thr
65 70 75 80
His Xaa Ile Arg Phe Ser Gln Ser Asp Thr Thr Leu Arg Leu Asn Ile
85 90 95
Phe Pro Ser Ile Ser Gly Gln Xaa Val Tyr Val Asp Lys Xaa Glu Ile
100 105 110
Val Pro Xaa Asn Pro Thr Arg Glu Ala Glu Glu Asp Leu Glu Asp Xaa
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 10
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 10
ccaggwttta caggagggga tatacttcga agaacggacg gtggtrcagt tggaacgatt 60
agagctaatg ttaatgcccc attaacacaa caatatcgta taagattacg ctatgcttcg 120
acaacaagtt ttgttgttaa tttatttgtt aataatagtg cggctggctt tactttaccg 180
agtacaatgg ctcaaaatgg ttctttaaca yrcgagtcgt ttaatacctt agaggtaact 240
catwctatta gattttcaca gtcagatact acacttaggt tgaatatatt cccgtcyatc 300
tctggtcaag ragtgtatgt agataaacwt gaaatcgttc cawttaaccc gacacgagaa 360
gcggaagaag atttagaaga tscaaagaaa gcggtggcga gcttgtttac 410
〈210〉 11
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 11
Pro Gly Phe Xaa Gly Gly Asp Ile Leu Arg Arg Thr Gly Val Gly Thr
1 5 10 15
Phe Gly Thr Ile Arg Val Arg Xaa Thr Ala Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Ile Arg Phe Arg Phe Ala Xaa Thr Thr Asn Leu Phe Ile Gly Ile
35 40 45
Arg Val Gly Asp Arg Gln Val Asn Tyr Phe Asp Phe Gly Arg Thr Met
50 55 60
Asn Arg Gly Asp Glu Leu Arg Tyr Glu Ser Phe Ala Thr Arg Glu Phe
65 70 75 80
Thr Thr Asp Phe Asn Phe Arg Gln Pro Gln Glu Leu Ile Ser Val Phe
85 90 95
Ala Asn Ala Phe Ser Ala Gly Gln Glu Val Tyr Phe Asp Arg Ile Glu
100 105 110
Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Lys Glu Asp Leu Glu Ala
115 120 125
Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 12
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 12
ccaggtttta yaggagggga tatactccga agaacagggg ttggtacatt tggaacaata 60
agggtaagga ytactgcccc cttaacacaa agatatcgca taagattccg tttcgcttyt 120
accacaaatt tgttcattgg tataagagtt ggtgatagac aagtaaatta ttttgacttc 180
ggaagaacaa tgaacagagg agatgaatta aggtacgaat cttttgctac aagggagttt 240
actactgatt ttaattttag acaacctcaa gaattaatct cagtgtttgc aaatgcattt 300
agcgctggtc aagaagttta ttttgataga attgagatta tccccgttaa tcccgcacga 360
gaggcgaaag aggatytaga agcagcaaag aaagcggtgg cgagcttgtt tac 413
〈210〉 13
〈211〉 135
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 13
Gly Phe Ile Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser Leu
1 5 10 15
Gly Val Leu Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr Arg
20 25 30
Ile Xaa Val Arg Tyr Ala Xaa Thr Thr Asn Ile Arg Leu Ser Val Asn
35 40 45
Gly Ser Phe Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg Leu
50 55 60
Gly Glu Asp Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn Thr
65 70 75 80
Ser Ile Arg Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile Glu
85 90 95
Pro Ser Phe Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe Ile
100 105 110
Pro Val Asn Pro Thr Arg Glu Ala Lys Glu Asp Leu Glu Ala Ala Lys
115 120 125
Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 14
〈211〉 407
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 14
ggmtttatag gaggagctct acttcaaagg actgaccatg gttcgcttgg agtattgagg 60
gtccaatttc cacttcactt aagacaacaa tatcgtatta sagtccgtta tgcttytaca 120
acaaatattc gattgagtgt gaatggcagt ttcggtacta tttctcaaaa tctccctagt 180
acaatgagat taggagagga tttaagatac ggatcttttg ctataagaga gtttaatact 240
tctattagac ccactgcaag tccggaccaa attcgattga caatagaacc atcttttatt 300
agacaagagg tctatgtaga tagaattgag ttcattccag ttaatccgac gcgagaggcg 360
aaagaggatc tagaagcagc aaaaaaagcg gtggcgagct tgtttac 407
〈210〉 15
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 15
Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln
1 5 10 15
Ile Ser Thr Leu Arg Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr
20 25 30
Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr
35 40 45
Ser Ile Asp Gly Arg Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met
50 55 60
Ser Ser Gly Ser Asn Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe
65 70 75 80
Thr Thr Pro Phe Asn Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser
85 90 95
Ala His Val Phe Asn Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu
100 105 110
Phe Val Pro Ala Glu Val Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg
115 120 125
Ala Gln Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 16
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 16
ccaggattta caggaggaga tattcttcga agaacttcac ctggccagat ttcaacctta 60
agagtaaata ttactgcacc attatcacaa agatatcggg taagaattcg ctacgcttct 120
accacaaatt tacaattcca tacatcaatt gacggaagac ctattaatca ggggaatttt 180
tcagcaacta tgagtagtgg gagtaattta cagtccggaa gctttaggac tgtaggtttt 240
actactccgt ttaacttttc aaatggatca agtgtattta cgttaagtgc tcatgtcttc 300
aattcaggca atgaagttta tatagatcga attgaatttg ttccggcaga agtaaccttt 360
gaggcagaat atgatttaga aagagcgcaa aaggcggtgg cgagcttgtt tac 413
〈210〉 17
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 17
Pro Gly Phe Xaa Gly Gly Asp Ile Leu Arg Arg Thr Asp Gly Gly Ala
1 5 10 15
Val Gly Thr Ile Arg Ala Asn Val Asn Ala Pro Leu Thr Gln Gln Tyr
20 25 30
Arg Ile Arg Leu Arg Tyr Ala Ser Thr Thr Ser Phe Val Val Asn Leu
35 40 45
Phe Val Asn Asn Ser Ala Ala Gly Phe Thr Leu Pro Ser Thr Met Ala
50 55 60
Gln Asn Gly Ser Leu Thr Tyr Glu Ser Phe Asn Thr Leu Glu Val Thr
65 70 75 80
His Thr Ile Arg Phe Ser Gln Ser Asp Thr Thr Leu Arg Leu Asn Ile
85 90 95
Phe Pro Ser Ile Ser Gly Gln Glu Val Tyr Val Asp Lys Leu Glu Ile
100 105 110
Val Pro Ile Asn Pro Thr Arg Glu Ala Glu Glu Asp Leu Glu Asp Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 18
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 18
ccaggwttta yaggagggga tatacttcga agaacggacg gtggtgcagt tggaacgatt 60
agagctaatg ttaatgcccc attaacacaa caatatcgta taagattacg ctatgcttcg 120
acaacaagtt ttgttgttaa tttatttgtt aataatagtg cggctggctt tactttaccg 180
agtacaatgg ctcaaaatgg ttctttaaca tacgagtcgt ttaatacctt agaggtaact 240
catactatta gattttcaca gtcagatact acacttaggt tgaatatatt cccgtctatc 300
tctggtcaag aagtgtatgt agataaactt gaaatcgttc caattaaccc gacacgagaa 360
gcggaagaag atttagaaga tgcaaagaaa gcggtggcga gcttgtttac 410
〈210〉 19
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 19
Pro Gly Phe Xaa Gly Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln
1 5 10 15
Ile Ser Thr Leu Arg Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr
20 25 30
Arg Val Arg Ile Arg Tyr Ala Xaa Thr Thr Asn Leu Gln Phe His Thr
35 40 45
Ser Ile Asp Gly Arg Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met
50 55 60
Ser Ser Gly Ser Asn Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe
65 70 75 80
Thr Thr Pro Phe Asn Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser
85 90 95
Ala His Val Phe Asn Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu
100 105 110
Phe Val Pro Ala Glu Val Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg
115 120 125
Ala Gln Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 20
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 20
ccaggwttta yaggaggaga tattcttcga agaacttcac ctggccagat ttcaacctta 60
agagtaaata ttactgcacc attatcacaa agatatcggg taagaattcg ctacgcttyt 120
acyacaaatt tacaattcca tacatcaatt gacggaagac ctattaatca gggkaatttt 180
tcagcaacta tgagtagtgg gagtaattta cagtccggaa gctttaggac tgtaggtttt 240
actactccgt ttaacttttc aaatggatca agtgtattta cgttaagtgc tcatgtcttc 300
aattcaggca atgaagttta tatagatcga attgaatttg ttccggcaga agtaaccttt 360
gaggcagaat atgatttaga aagagcacaa aaggcggtgg cgagcttgtt tac 413
〈210〉 21
〈211〉 106
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 21
Phe Thr Gly Gly Asp Ile Leu Arg Arg Asn Thr Ile Gly Glu Phe Val
1 5 10 15
Ser Leu Gln Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu
20 25 30
Arg Phe Arg Tyr Ala Ser Ser Arg Asp Ala Arg Ile Thr Val Ala Ile
35 40 45
Gly Gly Gln Ile Arg Val Asp Met Thr Leu Glu Lys Thr Met Glu Ile
50 55 60
Gly Glu Ser Leu Thr Xaa Arg Thr Phe Ser Tyr Thr Asn Phe Ser Asn
65 70 75 80
Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Arg Ile Ala Glu Glu
85 90 95
Leu Pro Ile Arg Gly Gly Glu Leu Val Tyr
100 105
〈210〉 22
〈211〉 318
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 22
tttacaggag gggatatcct tcgaagaaat accattggtg agtttgtgtc tttacaagtc 60
aatattaact caccaattac ccaaagatac cgtttaagat ttcgttatgc ttccagtagg 120
gatgcacgaa ttactgtagc gataggagga caaattagag tagatatgac ccttgaaaaa 180
accatggaaa ttggggagag cttaacatyt agaacattta gctataccaa ttttagtaat 240
cctttttcat ttagggctaa tccagatata attagaatag ctgaagaact tcctattcgc 300
ggtggcgagc ttgtttac 318
〈210〉 23
〈211〉 96
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 23
Ile Pro Leu Val Ser Leu Cys Leu Tyr Lys Ser Ile Leu Thr His Gln
1 5 10 15
Leu Pro Lys Asp Thr Val Xaa Xaa Phe Val Met Leu Pro Val Gly Met
20 25 30
His Glu Leu Leu Xaa Arg Xaa Glu Asp Lys Leu Glu Xaa Ile Xaa Pro
35 40 45
Leu Lys Lys Pro Trp Lys Leu Gly Arg Ala Xaa His Leu Glu His Leu
50 55 60
Ala Ile Pro Ile Leu Val Ile Leu Phe His Leu Gly Leu Ile Gln Ile
65 70 75 80
Xaa Leu Glu Xaa Leu Lys Asn Phe Leu Phe Ala Val Ala Ser Leu Phe
85 90 95
〈210〉 24
〈211〉 292
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 24
aaataccatt ggtgagtttg tgtctttaca agtcaatatt aactcaccaa ttacccaaag 60
ataccgttta aratttcgtt atgcttccag tagggatgca cgaattactg tagcgatagg 120
aggacaaatt agagtagata tgacccttga aaaaaccatg gaaattgggg agagcttaac 180
atctagaaca tttagctata ccaattttag taatcctttt tcatttaggg ctaatccaga 240
tataattaga atagctgaag aacttcctat tcgcggtggc gagcttgttt ac 292
〈210〉 25
〈211〉 108
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 25
Pro Gly Phe Xaa Gly Gly Asp Ile Leu Arg Arg Asn Thr Ile Gly Glu
1 5 10 15
Phe Val Ser Leu Gln Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr
20 25 30
Arg Leu Arg Phe Arg Tyr Ala Ser Ser Arg Asp Ala Arg Ile Thr Val
35 40 45
Ala Ile Gly Gly Gln Ile Arg Val Xaa Met Thr Leu Glu Lys Thr Met
50 55 60
Glu Ile Gly Glu Ser Leu Thr Ser Arg Thr Phe Ser Tyr Thr Asn Phe
65 70 75 80
Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Arg Ile Ala
85 90 95
Glu Glu Leu Pro Ile Arg Gly Gly Glu Leu Val Tyr
100 105
〈210〉 26
〈211〉 324
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 26
ccaggwttta yaggagggga tatccttcga agaaatacca ttggtgagtt tgtgtcttta 60
caagtcaata ttaactcacc aattacccaa agataccgtt taagatttcg ttatgcttcc 120
agtagggatg cacgaattac tgtagcgata ggaggacaaa ttagagtaka tatgaccctt 180
gaaaaaacca tggaaattgg ggagagctta acatctagaa catttagcta taccaatttt 240
agtaatcctt tttcatttag ggctaatcca gatataatta gaatagctga agaacttcct 300
attcgcggtg gcgagcttgt ttac 324
〈210〉 27
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 27
Gly Phe Xaa Gly Gly Asp Val Ile Arg Arg Thr Asn Thr Gly Gly Phe
1 5 10 15
Gly Ala Ile Arg Val Ser Val Thr Gly Pro Leu Thr Gln Arg Tyr Arg
20 25 30
Ile Arg Phe Arg Tyr Ala Ser Thr Ile Asp Phe Asp Phe Phe Val Thr
35 40 45
Arg Gly Gly Thr Thr Ile Asn Asn Phe Arg Phe Thr Arg Thr Met Asn
50 55 60
Arg Gly Gln Glu Ser Arg Tyr Glu Ser Tyr Arg Thr Val Glu Phe Thr
65 70 75 80
Thr Pro Phe Asn Phe Thr Gln Ser Gln Asp Ile Ile Arg Thr Xaa Ile
85 90 95
Gln Gly Leu Ser Gly Asn Gly Glu Val Tyr Leu Asp Arg Ile Glu Ile
100 105 110
Ile Pro Val Asn Pro Thr Arg Glu Ala Glu Glu Asp Leu Glu Ala Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 28
〈211〉 411
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 28
aggatttaya ggaggagatg taatccgaag aacaaatact ggtggattcg gagcaataag 60
ggtgtcggtc actggaccgc taacacaacg atatcgcata aggttccgtt atgcttcgac 120
aatagatttt gatttctttg taacacgtgg aggaactact ataaataatt ttagatttac 180
acgtacaatg aacaggggac aggaatcaag atatgaatcc tatcgtactg tagagtttac 240
aactcctttt aactttacac aaagtcaaga tataattcga acayctatcc agggacttag 300
tggaaatggg gaagtatacc ttgatagaat tgaaatcatc cctgtaaatc caacacgaga 360
agcggaagar gatttagaag cggcgaagaa agcggtggcg agcttgttta c 411
〈210〉 29
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 29
Pro Gly Phe Ile Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser
1 5 10 15
Leu Gly Val Leu Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr
20 25 30
Arg Ile Arg Val Arg Tyr Ala Ser Thr Thr Asn Ile Arg Leu Ser Val
35 40 45
Asn Gly Ser Phe Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg
50 55 60
Leu Gly Glu Asp Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn
65 70 75 80
Thr Ser Ile Arg Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile
85 90 95
Glu Pro Ser Phe Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe
100 105 110
Ile Pro Val Asn Pro Thr Arg Glu Ala Lys Glu Asp Leu Glu Ala Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 30
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 30
ccaggattta taggaggagc tctacttcaa aggactgacc atggttcgct tggagtattg 60
agggtccaat ttccacttca cttaagacaa caatatcgta ttagagtccg ttatgcttct 120
acaacaaata ttcgattgag tgtgaatggc agtttcggta ctatttctca aaatctccct 180
agtacaatga gattaggaga ggatttaaga tacggatctt ttgctataag agagtttaat 240
acttctatta gacccactgc aagtccggac caaattcgat tgacaataga accatctttt 300
attagacaag aggtctatgt agatagaatt gagttcattc cagttaatcc gacgcgagag 360
gcgaaagagg atctagaagc agcaaaaaaa gcggtggcga gcttgtttac 410
〈210〉 31
〈211〉 142
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 31
Pro Gly Phe Xaa Gly Gly Gly Ile Leu Arg Arg Thr Thr Asn Gly Thr
1 5 10 15
Phe Gly Thr Leu Arg Val Thr Val Asn Ser Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Val Arg Val Arg Phe Ala Ser Ser Gly Asn Phe Ser Ile Arg Ile
35 40 45
Leu Arg Gly Asn Thr Ser Ile Ala Tyr Gln Arg Phe Gly Ser Thr Met
50 55 60
Asn Arg Gly Gln Glu Leu Thr Tyr Glu Ser Phe Val Thr Ser Glu Phe
65 70 75 80
Thr Thr Asn Gln Ser Asp Leu Pro Phe Thr Phe Thr Gln Ala Gln Glu
85 90 95
Asn Leu Thr Ile Leu Ala Glu Gly Val Ser Thr Gly Ser Glu Tyr Phe
100 105 110
Ile Asp Arg Ile Glu Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Glu
115 120 125
Glu Asp Leu Glu Ala Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135 140
〈210〉 32
〈211〉 428
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 32
ccaggwttta yaggaggggg tatactccga agaacaacta atggcacatt tggaacgtta 60
agagtaacag ttaattcacc attaacacaa agatatcgcg taagagttcg ttttgcttca 120
tcaggaaatt tcagcataag gatactgcgt ggaaatacct ctatagctta tcaaagattt 180
gggagtacaa tgaacagagg acaggaacta acttacgaat catttgtcac aagtgagttc 240
actactaatc agagcgatct gccttttaca tttacacaag ctcaagaaaa tttaacaatc 300
cttgcagaag gtgttagcac cggtagtgaa tattttatag atagaattga aatcatccct 360
gtgaacccgg cacgagaagc agaagaggat ttagaagcrg cgaagaaagc ggtggcgagc 420
ttgtttac 428
〈210〉 33
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 33
Pro Gly Phe Ile Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser
1 5 10 15
Leu Gly Val Leu Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr
20 25 30
Arg Ile Arg Val Arg Tyr Ala Ser Thr Thr Asn Ile Arg Leu Ser Val
35 40 45
Asn Gly Ser Phe Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg
50 55 60
Leu Gly Glu Asp Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn
65 70 75 80
Thr Ser Ile Arg Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile
85 90 95
Glu Pro Ser Phe Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe
100 105 110
Ile Pro Val Asn Pro Thr Arg Glu Ala Lys Glu Asp Leu Glu Ala Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 34
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 34
accaggattt ataggaggag ctctacttca aaggactgac catggttcgc ttggagtatt 60
ggggtccaat ttccacttca cttaagacaa caatatcgta ttagagtccg ttatgcttct 120
acaacaaata ttcgattgag tgtgaatggc agtttcggta ctatttctca aaatctccct 180
agtacaatga gattaggaga ggatttaaga tacggatctt ttgctataag agagtttaat 240
acttctatta gacccactgc aagtccggac caaattcgat tgacaataga accatctttt 300
attagacaag aggtctatgt agatagaatt gagttcattc cagttaatcc gacgcgagag 360
gcgaaagagg atctagaagc agcaaaaaaa gcggtggcga gcttgtttac 410
〈210〉 35
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 35
Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Gly Val Gly Thr
1 5 10 15
Phe Gly Thr Ile Arg Val Arg Thr Thr Ala Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Ile Arg Phe Arg Phe Ala Ser Thr Thr Asn Leu Phe Ile Gly Ile
35 40 45
Arg Val Gly Asp Arg Gln Val Asn Tyr Phe Asp Phe Gly Arg Thr Met
50 55 60
Asn Arg Gly Asp Glu Leu Arg Tyr Glu Ser Phe Ala Thr Arg Glu Phe
65 70 75 80
Thr Thr Asp Phe Asn Phe Arg Gln Pro Gln Glu Leu Ile Ser Val Phe
85 90 95
Ala Asn Ala Phe Ser Ala Gly Gln Glu Val Tyr Phe Asp Arg Ile Glu
100 105 110
Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Lys Glu Asp Leu Glu Ala
115 120 125
Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 36
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 36
ccaggtttta caggagggga tatactccga agaacagggg ttggtacatt tggaacaata 60
agggtaagga ctactgcccc cttaacacaa agatatcgca taagattccg tttcgcttct 120
accacaaatt tgttcattgg tataagagtt ggtgatagac aagtaaatta ttttgacttc 180
ggaagaacaa tgaacagagg agatgaatta aggtacgaat cttttgctac aagggagttt 240
actactgatt ttaattttag acaacctcaa gaattaatct cagtgtttgc aaatgcattt 300
agcgctggtc aagaagttta ttttgataga attgagatta tccccgttaa tcccgcacga 360
gaggcgaaag aggatctaga agcagcaaag aaagcggtgg cgagcttgtt tac 413
〈210〉 37
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 37
Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Ser Pro Gly Gln
1 5 10 15
Ile Ser Thr Leu Arg Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr
20 25 30
Arg Val Arg Ile Arg Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr
35 40 45
Ser Ile Asp Gly Arg Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met
50 55 60
Ser Ser Gly Ser Asn Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe
65 70 75 80
Thr Thr Pro Phe Asn Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser
85 90 95
Ala His Val Phe Asn Ser Gly Asn Glu Val Tyr Ile Asp Arg Ile Glu
100 105 110
Phe Val Pro Ala Glu Val Thr Phe Glu Ala Glu Tyr Asp Leu Glu Arg
115 120 125
Ala Gln Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 38
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 38
ccaggwttta caggaggaga tattcttcga agaacttcac ctggccagat ttcaacctta 60
agagtaaata ttactgcacc attatcacaa agatatcggg taagaattcg ctacgcttct 120
accacaaatt tacaattcca tacatcaatt gacggaagac ctattaatca ggggaatttt 180
tcagcaacta tgagtagtgg gagtaattta cagtccggaa gctttaggac tgtaggtttt 240
actactccgt ttaacttttc aaatggatca agtgtattta cgttaagtgc tcatgtcttc 300
aattcaggca atgaagttta tatagatcga attgaatttg ttccggcaga agtaaccttt 360
gaggcagaat atgatttaga aagagcacar aaggcggtgg cgagcttgtt tac 413
〈210〉 39
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 39
Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Gly Val Gly Thr
1 5 10 15
Phe Gly Thr Ile Arg Val Arg Thr Thr Ala Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Ile Arg Phe Arg Phe Ala Ser Thr Thr Asn Leu Phe Ile Gly Ile
35 40 45
Arg Val Gly Asp Arg Gln Val Asn Tyr Phe Asp Phe Gly Arg Thr Met
50 55 60
Asn Arg Gly Asp Glu Leu Arg Tyr Glu Ser Phe Ala Thr Arg Glu Phe
65 70 75 80
Thr Thr Asp Phe Asn Phe Arg Gln Pro Gln Glu Leu Ile Ser Val Phe
85 90 95
Ala Asn Ala Phe Ser Ala Gly Gln Glu Val Tyr Phe Asp Arg Ile Glu
100 105 110
Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Lys Glu Asp Leu Glu Ala
115 120 125
Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 40
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 40
ccaggtttta caggagggga tatactccga agaacagggg ttggtacatt tggaacaata 60
agggtaagga ctactgcccc cttaacacaa agatatcgca taagattccg tttcgcttct 120
accacaaatt tgttcattgg tataagagtt ggtgatagac aagtaaatta ttttgacttc 180
ggaagaacaa tgaacagagg agatgaatta aggtacgaat cttttgctac aagggagttt 240
actactgatt ttaattttag acaacctcaa gaattaatct cagtgtttgc aaatgcattt 300
agcgctggtc aagaagttta ttttgataga attgagatta tccccgttaa tcccgcacga 360
gaggcgaaag aggatctaga agcagcaaag aaagcggtgg cgagcttgtt tac 413
〈210〉 41
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 41
Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Asn Ala Gly Asn
1 5 10 15
Phe Gly Asp Met Arg Val Asn Ile Thr Ala Pro Leu Ser Gln Arg Tyr
20 25 30
Arg Val Arg Ile Arg Tyr Ala Ser Thr Ala Asn Leu Gln Phe His Thr
35 40 45
Ser Ile Asn Gly Arg Ala Ile Asn Gln Ala Asn Phe Pro Ala Thr Met
50 55 60
Asn Ser Gly Glu Asn Leu Gln Ser Gly Ser Phe Arg Val Ala Gly Phe
65 70 75 80
Thr Thr Pro Phe Thr Phe Ser Asp Ala Leu Ser Thr Phe Thr Ile Gly
85 90 95
Ala Phe Ser Phe Ser Ser Asn Asn Glu Val Tyr Ile Asp Arg Ile Glu
100 105 110
Phe Val Pro Ala Glu Val Thr Phe Ala Thr Glu Ser Asp Gln Asp Arg
115 120 125
Ala Gln Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 42
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 42
ccaggwttta caggagggga tatccttcga agaacgaatg ctggtaactt tggagatatg 60
cgtgtaaaca ttactgcacc actatcacaa agatatcgcg taaggattcg ttatgcttct 120
actgcaaatt tacaattcca tacatcaatt aacggaagag ccattaatca ggcgaatttc 180
ccagcaacta tgaacagtgg ggagaattta cagtccggaa gcttcagggt tgcaggtttt 240
actactccat ttaccttttc agatgcacta agcacattca caataggtgc ttttagcttc 300
tcttcaaaca acgaagttta tatagatcga attgaatttg ttccggcaga agtaacattt 360
gcaacagaat ctgatcagga tagagcacaa aaggcggtgg cgagcttgtt tac 413
〈210〉 43
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 43
Pro Gly Phe Ile Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser
1 5 10 15
Leu Gly Val Leu Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr
20 25 30
Arg Ile Arg Val Arg Tyr Ala Ser Thr Thr Asn Ile Arg Leu Ser Val
35 40 45
Asn Gly Ser Phe Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg
50 55 60
Leu Gly Glu Asp Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn
65 70 75 80
Thr Ser Ile Arg Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile
85 90 95
Glu Pro Ser Phe Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe
100 105 110
Ile Pro Val Asn Pro Thr Arg Glu Ala Lys Glu Asp Leu Xaa Ala Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 44
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 44
ccaggattta taggaggagc tctacttcaa aggactgacc atggttcgct tggagtattg 60
agggtccaat ttccacttca cttaagacaa caatatcgta ttagagtccg ttatgcttct 120
acaacaaata ttcgattgag tgtgaatggc agtttcggta ctatttctca aaatctccct 180
agtacaatga gattaggaga ggatttaaga tacggatctt ttgctataag agagtttaat 240
acttctatta gacccactgc aagtccggac caaattcgat tgacaataga accatctttt 300
attagacaag aggtctatgt agatagaatt gagttcattc cagttaatcc gacgcgagag 360
gcgaaagagg atctakaagc agcaaaaaaa gcggtggcga gcttgtttac 410
〈210〉 45
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 45
Gln Xaa Leu Ser Gly Gly Asp Val Ile Arg Arg Thr Asn Thr Gly Gly
1 5 10 15
Phe Gly Ala Ile Arg Val Ser Val Thr Gly Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Ile Arg Phe Arg Tyr Ala Ser Thr Ile Asp Phe Asp Phe Phe Val
35 40 45
Thr Arg Gly Gly Thr Thr Ile Asn Asn Phe Arg Phe Thr Arg Thr Met
50 55 60
Asn Arg Gly Gln Glu Ser Arg Tyr Glu Ser Tyr Arg Thr Val Glu Phe
65 70 75 80
Thr Thr Pro Phe Asn Phe Thr Gln Ser Gln Asp Ile Ile Arg Thr Ser
85 90 95
Ile Gln Gly Leu Ser Gly Asn Gly Glu Val Tyr Leu Asp Arg Ile Glu
100 105 110
Ile Ile Pro Val Asn Pro Thr Arg Glu Ala Glu Glu Asp Leu Glu Ala
115 120 125
Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 46
〈211〉 414
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 46
ccaggwttta tcaggaggag atgtaatccg aagaacaaat actggtggat tcggagcaat 60
aagggtgtcg gtcactggac cgctaacaca acgatatcgc ataaggttcc gttatgcttc 120
gacaatagat tttgatttct ttgtaacacg tggaggaact actataaata attttagatt 180
tacacgtaca atgaacaggg gacaggaatc aagatatgaa tcctatcgta ctgtagagtt 240
tacaactcct tttaacttta cacaaagtca agatataatt cgaacatcta tccagggact 300
tagtggaaat ggggaagtat accttgatag aattgaaatc atccctgtaa atccaacacg 360
agaagcggaa gargatttag aagcggcgaa gaaagcggtg gcgagcttgt ttac 414
〈210〉 47
〈211〉 142
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 47
Pro Gly Phe Thr Gly Gly Gly Ile Leu Arg Arg Thr Thr Asn Gly Thr
1 5 10 15
Phe Gly Thr Leu Arg Val Thr Val Asn Ser Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Val Arg Val Arg Phe Ala Ser Ser Gly Asn Phe Ser Ile Arg Ile
35 40 45
Leu Arg Gly Asn Thr Ser Ile Ala Tyr Gln Arg Phe Gly Ser Thr Met
50 55 60
Asn Arg Gly Gln Glu Leu Thr Tyr Glu Ser Phe Val Thr Ser Glu Phe
65 70 75 80
Thr Thr Asn Gln Ser Asp Leu Pro Phe Thr Phe Thr Gln Ala Gln Glu
85 90 95
Asn Leu Thr Ile Leu Ala Glu Gly Val Ser Thr Gly Ser Glu Tyr Phe
100 105 110
Ile Asp Arg Ile Glu Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Glu
115 120 125
Glu Asp Leu Glu Ala Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135 140
〈210〉 48
〈211〉 428
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 48
ccaggwttta caggaggggg tatactccga agaacaacta atggcacatt tggaacgtta 60
agagtaacag ttaattcacc attaacacaa agatatcgcg taagagttcg ttttgcttca 120
tcaggaaatt tcagcataag gatactgcgt ggaaatacct ctatagctta tcaaagattt 180
gggagtacaa tgaacagagg acaggaacta acttacgaat catttgtcac aagtgagttc 240
actactaatc agagcgatct gccttttaca tttacacaag ctcaagaaaa tttaacaatc 300
cttgcagaag gtgttagcac cggtagtgaa tattttatag atagaattga aatcatccct 360
gtgaacccgg cacgagaagc agaagaggat ttagaagcag cgaagaaagc ggtggcgagc 420
ttgtttac 428
〈210〉 49
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 49
Pro Gly Phe Ile Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser
1 5 10 15
Leu Gly Val Leu Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr
20 25 30
Arg Ile Arg Val Arg Tyr Ala Ser Thr Thr Asn Ile Arg Leu Ser Val
35 40 45
Asn Gly Ser Phe Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg
50 55 60
Leu Gly Glu Asp Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn
65 70 75 80
Thr Ser Ile Arg Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile
85 90 95
Glu Pro Ser Phe Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe
100 105 110
Ile Pro Val Asn Pro Thr Arg Glu Ala Lys Glu Asp Leu Glu Ala Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 50
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 50
ccaggwttta taggaggagc tctacttcaa aggactgacc atggttcgct tggagtattg 60
agggtccaat ttccacttca cttaagacaa caatatcgta ttagagtccg ttatgcttct 120
acaacaaata ttcgattgag tgtgaatggc agtttcggta ctatttctca aaatctccct 180
agtacaatga gattaggaga ggatttaaga tacggatctt ttgctataag agagtttaat 240
acttctatta gacccactgc aagtccggac caaattcgat tgacaataga accatctttt 300
attagacaag aggtctatgt agatagaatt gagttcattc cagttaatcc gacgcgagag 360
gcgaaagagg atctagaagc agcaaaaaaa gcggtggcga gcttgtttac 410
〈210〉 51
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 51
Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg Thr Gly Val Gly Thr
1 5 10 15
Phe Gly Thr Ile Arg Val Arg Thr Thr Ala Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Ile Arg Phe Arg Phe Ala Ser Thr Thr Asn Leu Phe Ile Gly Ile
35 40 45
Arg Val Gly Asp Arg Gln Val Asn Tyr Phe Asp Phe Gly Arg Thr Met
50 55 60
Asn Arg Gly Asp Glu Leu Arg Tyr Glu Ser Phe Ala Thr Arg Glu Phe
65 70 75 80
Thr Thr Asp Phe Asn Phe Arg Gln Pro Gln Glu Leu Ile Ser Val Phe
85 90 95
Ala Asn Ala Phe Ser Ala Gly Gln Glu Val Tyr Phe Asp Arg Ile Glu
100 105 110
Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Lys Glu Asp Leu Glu Ala
115 120 125
Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 52
〈211〉 412
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 52
ccaggtttta caggagggga tatactccga agaacagggg ttggtacatt tggaacaata 60
agggtaagga ctactgcccc cttaacacaa agatatcgca taagattccg tttcgcttct 120
accacaaatt tgttcattgg tataagagtt ggtgatagac aagtaaatta ttttgacttc 180
ggaagaacaa tgaacagagg agatgaatta aggtacgaat cttttgctac aagggagttt 240
actactgatt ttaattttag acaacctcaa gaattaatct cagtgtttgc aaatgcattt 300
agcgctggtc aagaagttta ttttgataga attgagatta tccccgttaa tcccgcacga 360
gaggcgaaag aggatctaga agcagcaaag aaagcggtgg cgagcttgtt ta 412
〈210〉 53
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 53
Pro Gly Phe Thr Gly Gly Asp Val Ile Arg Arg Thr Asn Thr Gly Gly
1 5 10 15
Phe Gly Ala Ile Arg Val Ser Val Thr Gly Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Ile Arg Phe Arg Tyr Ala Ser Thr Ile Asp Phe Asp Phe Phe Val
35 40 45
Thr Arg Gly Gly Thr Thr Ile Asn Asn Phe Arg Phe Thr Arg Thr Met
50 55 60
Asn Arg Gly Gln Glu Ser Arg Tyr Glu Ser Tyr Arg Thr Val Glu Phe
65 70 75 80
Thr Thr Pro Phe Asn Phe Thr Gln Ser Gln Asp Ile Ile Arg Thr Ser
85 90 95
Ile Gln Gly Leu Ser Gly Asn Gly Glu Val Tyr Leu Asp Arg Ile Glu
100 105 110
Ile Ile Pro Val Asn Pro Thr Arg Glu Ala Glu Glu Asp Xaa Glu Ala
115 120 125
Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 54
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 54
ccaggattta caggaggaga tgtaatccga agaacaaata ctggtggatt cggagcaata 60
agggtgtcgg tcactggacc gctaacacaa cgatatcgca taaggttccg ttatgcttcg 120
acaatagatt ttgatttctt tgtaacacgt ggaggaacta ctataaataa ttttagattt 180
acacgtacaa tgaacagggg acaggaatca agatatgaat cctatcgtac tgtagagttt 240
acaactcctt ttaactttac acaaagtcaa gatataattc gaacatctat ccagggactt 300
agtggaaatg gggaagtata ccttgataga attgaaatca tccctgtaaa tccaacacga 360
gaagcggaag aggatttwga agcggcgaag aaagcggtgg cgagcttgtt tac 413
〈210〉 55
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 55
Pro Gly Phe Ile Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser
1 5 10 15
Leu Gly Val Leu Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr
20 25 30
Arg Ile Arg Val Arg Tyr Ala Ser Thr Thr Asn Ile Arg Leu Ser Val
35 40 45
Asn Gly Ser Phe Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg
50 55 60
Leu Gly Glu Asp Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn
65 70 75 80
Thr Ser Ile Arg Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile
85 90 95
Glu Pro Ser Phe Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe
100 105 110
Ile Pro Val Asn Pro Thr Arg Glu Ala Lys Xaa Asp Leu Xaa Ala Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 56
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 56
ccaggattta taggaggagc tctacttcaa aggactgacc atggttcgct tggagtattg 60
agggtccaat ttccacttca cttaagacaa caatatcgta ttagagtccg ttatgcttct 120
acaacaaata ttcgattgag tgtgaatggc agtttcggta ctatttctca aaatctccct 180
agtacaatga gattaggaga ggatttaaga tacggatctt ttgctataag agagtttaat 240
acttctatta gacccactgc aagtccggac caaattcgat tgacaataga accatctttt 300
attagacaag aggtctatgt agatagaatt gagttcattc cagttaatcc gacgcgagag 360
gcgaaagakg atctabaagc agcaaaaaaa gcggtggcga gcttgtttac 410
〈210〉 57
〈211〉 137
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 57
Pro Gly Phe Thr Gly Gly Asp Val Ile Arg Arg Thr Asn Thr Gly Gly
1 5 10 15
Phe Gly Ala Ile Arg Val Ser Val Thr Gly Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Ile Arg Phe Arg Tyr Ala Ser Thr Ile Asp Phe Asp Phe Phe Val
35 40 45
Thr Arg Gly Gly Thr Thr Ile Asn Asn Phe Arg Phe Thr Arg Thr Met
50 55 60
Asn Arg Gly Gln Glu Ser Arg Tyr Glu Ser Tyr Arg Thr Val Glu Phe
65 70 75 80
Thr Thr Pro Phe Asn Phe Thr Gln Ser Gln Asp Ile Ile Arg Thr Ser
85 90 95
Ile Gln Gly Leu Ser Gly Asn Gly Glu Val Tyr Leu Asp Arg Ile Glu
100 105 110
Ile Ile Pro Val Asn Pro Thr Arg Glu Ala Glu Glu Asp Leu Glu Ala
115 120 125
Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 58
〈211〉 413
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 58
ccaggwttta caggaggaga tgtaatccga agaacaaata ctggtggatt cggagcaata 60
agggtgtcgg tcactggacc gctaacacaa cgatatcgca taaggttccg ttatgcttcg 120
acaatagatt ttgatttctt tgtaacacgt ggaggaacta ctataaataa ttttagattt 180
acacgtacaa tgaacagggg acaggaatca agatatgaat cctatcgtac tgtagagttt 240
acaactcctt ttaactttac acaaagtcaa gatataattc gaacatctat ccagggactt 300
agtggaaatg gggaagtata ccttgataga attgaaatca tccctgtaaa tccaacacga 360
gaagcggaag aggatttaga agcggcgaag aaagcggtgg cgagcttgtt tac 413
〈210〉 59
〈211〉 142
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 59
Pro Gly Phe Xaa Gly Gly Gly Ile Leu Arg Arg Thr Thr Asn Gly Thr
1 5 10 15
Phe Gly Thr Leu Arg Val Thr Val Asn Ser Pro Leu Thr Gln Arg Tyr
20 25 30
Arg Val Arg Val Arg Phe Ala Ser Ser Gly Asn Phe Ser Ile Arg Ile
35 40 45
Leu Arg Gly Asn Thr Ser Ile Ala Tyr Gln Arg Phe Gly Ser Thr Met
50 55 60
Asn Arg Gly Gln Glu Leu Thr Tyr Glu Ser Phe Val Thr Ser Glu Phe
65 70 75 80
Thr Thr Asn Gln Ser Asp Leu Pro Phe Thr Phe Thr Gln Ala Gln Glu
85 90 95
Asn Leu Thr Ile Leu Ala Glu Gly Val Ser Thr Gly Ser Glu Tyr Phe
100 105 110
Ile Asp Arg Ile Glu Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Glu
115 120 125
Glu Asp Leu Glu Ala Ala Lys Lys Ala Val Ala Ser Leu Phe
130 135 140
〈210〉 60
〈211〉 428
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 60
ccaggwttta yaggaggggg tatactccga agaacaacta atggcacatt tggaacgtta 60
agagtaacag ttaattcacc attaacacaa agatatcgcg taagagttcg ttttgcttca 120
tcaggaaatt tcagcataag gatactgcgt ggaaatacct ctatagctta tcaaagattt 180
gggagtacaa tgaacagagg acaggaacta acttacgaat catttgtcac aagtgagttc 240
actactaatc agagcgatct gccttttaca tttacacaag ctcaagaaaa tttaacaatc 300
cttgcagaag gtgttagcac cggtagtgaa tattttatag atagaattga aatcatccct 360
gtgaacccgg cacgagaagc agaagaggat ttagaagcag cgaagaaagc ggtggcgagc 420
ttgtttac 428
〈210〉 61
〈211〉 136
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 61
Pro Gly Phe Ile Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser
1 5 10 15
Leu Gly Val Leu Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr
20 25 30
Arg Ile Arg Val Arg Tyr Ala Ser Thr Thr Asn Ile Arg Leu Ser Val
35 40 45
Asn Gly Ser Phe Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg
50 55 60
Leu Gly Glu Asp Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn
65 70 75 80
Thr Ser Ile Arg Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile
85 90 95
Glu Pro Ser Phe Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe
100 105 110
Ile Pro Val Asn Pro Thr Arg Glu Ala Lys Glu Asp Leu Glu Ala Ala
115 120 125
Lys Lys Ala Val Ala Ser Leu Phe
130 135
〈210〉 62
〈211〉 410
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 62
ccaggtttta taggaggagc tctacttcaa aggactgacc atggttcgct tggagtattg 60
agggtccaat ttccacttca cttaagacaa caatatcgta ttagagtccg ttatgcttct 120
acaacaaata ttcgattgag tgtgaatggc agtttcggta ctatttctca aaatctccct 180
agtacaatga gattaggaga ggatttaaga tacggatctt ttgctataag agagtttaat 240
acttctatta gacccactgc aagtccggac caaattcgat tgacaataga accatctttt 300
attagacaag aggtctatgt agatagaatt gagttcattc cagttaatcc gacgcgagag 360
gcgaaagagg atctagaagc agcaaaaaaa gcggtggcga gcttgtttac 410
〈210〉 63
〈211〉 23
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 63
gttcattggt ataagagttg gtg 23
〈210〉 64
〈211〉 25
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 64
ccactgcaag tccggaccaa attcg 25
〈210〉 65
〈211〉 24
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 65
gaatatattc ccgtcyatct ctgg 24
〈210〉 66
〈211〉 23
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 66
gcacgaatta ctgtagcgat agg 23
〈210〉 67
〈211〉 25
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 67
gctggtaact ttggagatat gcgtg 25
〈210〉 68
〈211〉 23
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 68
gatttctttg taacacgtgg agg 23
〈210〉 69
〈211〉 22
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 69
cactactaat cagagcgatc tg 22
〈210〉 70
〈211〉 1156
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 70
Met Asn Gln Asn Lys His Gly Ile Ile Gly Ala Ser Asn Cys Gly Cys
1 5 10 15
Ala Ser Asp Asp Val Ala Lys Tyr Pro Leu Ala Asn Asn Pro Tyr Ser
20 25 30
Ser Ala Leu Asn Leu Asn Ser Cys Gln Asn Ser Ser Ile Leu Asn Trp
35 40 45
Ile Asn Ile Ile Gly Asp Ala Ala Lys Glu Ala Val Ser Ile Gly Thr
50 55 60
Thr Ile Val Ser Leu Ile Thr Ala Pro Ser Leu Thr Gly Leu Ile Ser
65 70 75 80
Ile Val Tyr Asp Leu Ile Gly Lys Val Leu Gly Gly Ser Ser Gly Gln
85 90 95
Ser Ile Ser Asp Leu Ser Ile Cys Asp Leu Leu Ser Ile Ile Asp Leu
100 105 110
Arg Val Ser Gln Ser Val Leu Asn Asp Gly Ile Ala Asp Phe Asn Gly
115 120 125
Ser Val Leu Leu Tyr Arg Asn Tyr Leu Glu Ala Leu Asp Ser Trp Asn
130 135 140
Lys Asn Pro Asn Ser Ala Ser Ala Glu Glu Leu Arg Thr Arg Phe Arg
145 150 155 160
Ile Ala Asp Ser Glu Phe Asp Arg Ile Leu Thr Arg Gly Ser Leu Thr
165 170 175
Asn Gly Gly Ser Leu Ala Arg Gln Asn Ala Gln Ile Leu Leu Leu Pro
180 185 190
Ser Phe Ala Ser Ala Ala Phe Phe His Leu Leu Leu Leu Arg Asp Ala
195 200 205
Thr Arg Tyr Gly Thr Asn Trp Gly Leu Tyr Asn Ala Thr Pro Phe Ile
210 215 220
Asn Tyr Gln Ser Lys Leu Val Glu Leu Ile Glu Leu Tyr Thr Asp Tyr
225 230 235 240
Cys Val His Trp Tyr Asn Arg Gly Phe Asn Glu Leu Arg Gln Arg Gly
245 250 255
Thr Ser Ala Thr Ala Trp Leu Glu Phe His Arg Tyr Arg Arg Glu Met
260 265 270
Thr Leu Met Val Leu Asp Ile Val Ala Ser Phe Ser Ser Leu Asp Ile
275 280 285
Thr Asn Tyr Pro Ile Glu Thr Asp Phe Gln Leu Ser Arg Val Ile Tyr
290 295 300
Thr Asp Pro Ile Gly Phe Val His Arg Ser Ser Leu Arg Gly Glu Ser
305 310 315 320
Trp Phe Ser Phe Val Asn Arg Ala Asn Phe Ser Asp Leu Glu Asn Ala
325 330 335
Ile Pro Asn Pro Arg Pro Ser Trp Phe Leu Asn Asn Met Ile Ile Ser
340 345 350
Thr Gly Ser Leu Thr Leu Pro Val Ser Pro Ser Thr Asp Arg Ala Arg
355 360 365
Val Trp Tyr Gly Ser Arg Asp Arg Ile Ser Pro Ala Asn Ser Gln Phe
370 375 380
Ile Thr Glu Leu Ile Ser Gly Gln His Thr Thr Ala Thr Gln Thr Ile
385 390 395 400
Leu Gly Arg Asn Ile Phe Arg Val Asp Ser Gln Ala Cys Asn Leu Asn
405 410 415
Asp Thr Thr Tyr Gly Val Asn Arg Ala Val Phe Tyr His Asp Ala Ser
420 425 430
Glu Gly Ser Gln Arg Ser Val Tyr Glu Gly Tyr Ile Arg Thr Thr Gly
435 440 445
Ile Asp Asn Pro Arg Val Gln Asn Ile Asn Thr Tyr Leu Pro Gly Glu
450 455 460
Asn Ser Asp Ile Pro Thr Pro Glu Asp Tyr Thr His Ile Leu Ser Thr
465 470 475 480
Thr Ile Asn Leu Thr Gly Gly Leu Arg Gln Val Ala Ser Asn Arg Arg
485 490 495
Ser Ser Leu Val Met Tyr Gly Trp Thr His Lys Ser Leu Ala Arg Asn
500 505 510
Asn Thr Ile Asn Pro Asp Arg Ile Thr Gln Ile Pro Leu Thr Lys Val
515 520 525
Asp Thr Arg Gly Thr Gly Val Ser Tyr Val Asn Asp Pro Gly Phe Ile
530 535 540
Gly Gly Ala Leu Leu Gln Arg Thr Asp His Gly Ser Leu Gly Val Leu
545 550 555 560
Arg Val Gln Phe Pro Leu His Leu Arg Gln Gln Tyr Arg Ile Arg Val
565 570 575
Arg Tyr Ala Ser Thr Thr Asn Ile Arg Leu Ser Val Asn Gly Ser Phe
580 585 590
Gly Thr Ile Ser Gln Asn Leu Pro Ser Thr Met Arg Leu Gly Glu Asp
595 600 605
Leu Arg Tyr Gly Ser Phe Ala Ile Arg Glu Phe Asn Thr Ser Ile Arg
610 615 620
Pro Thr Ala Ser Pro Asp Gln Ile Arg Leu Thr Ile Glu Pro Ser Phe
625 630 635 640
Ile Arg Gln Glu Val Tyr Val Asp Arg Ile Glu Phe Ile Pro Val Asn
645 650 655
Pro Thr Arg Glu Ala Lys Glu Asp Leu Glu Ala Ala Lys Lys Ala Val
660 665 670
Ala Ser Leu Phe Thr Arg Thr Arg Asp Gly Leu Gln Val Asn Val Lys
675 680 685
Asp Tyr Gln Val Asp Gln Ala Ala Asn Leu Val Ser Cys Leu Ser Asp
690 695 700
Glu Gln Tyr Gly Tyr Asp Lys Lys Met Leu Leu Glu Ala Val Arg Ala
705 710 715 720
Ala Lys Arg Leu Ser Arg Glu Arg Asn Leu Leu Gln Asp Pro Asp Phe
725 730 735
Asn Thr Ile Asn Ser Thr Glu Glu Asn Gly Trp Lys Ala Ser Asn Gly
740 745 750
Val Thr Ile Ser Glu Gly Gly Pro Phe Tyr Lys Gly Arg Ala Ile Gln
755 760 765
Leu Ala Ser Ala Arg Glu Asn Tyr Pro Thr Tyr Ile Tyr Gln Lys Val
770 775 780
Asp Ala Ser Glu Leu Lys Pro Tyr Thr Arg Tyr Arg Leu Asp Gly Phe
785 790 795 800
Val Lys Ser Ser Gln Asp Leu Glu Ile Asp Leu Ile His His His Lys
805 810 815
Val His Leu Val Lys Asn Val Pro Asp Asn Leu Val Ser Asp Thr Tyr
820 825 830
Pro Asp Asp Ser Cys Ser Gly Ile Asn Arg Cys Gln Glu Gln Gln Met
835 840 845
Val Asn Ala Gln Leu Glu Thr Glu His His His Pro Met Asp Cys Cys
850 855 860
Glu Ala Ala Gln Thr His Glu Phe Ser Ser Tyr Ile Asp Thr Gly Asp
865 870 875 880
Leu Asn Ser Ser Val Asp Gln Gly Ile Trp Ala Ile Phe Lys Val Arg
885 890 895
Thr Thr Asp Gly Tyr Ala Thr Leu Gly Asn Leu Glu Leu Val Glu Val
900 905 910
Gly Pro Leu Ser Gly Glu Ser Leu Glu Arg Glu Gln Arg Asp Asn Thr
915 920 925
Lys Trp Ser Ala Glu Leu Gly Arg Lys Arg Ala Glu Thr Asp Arg Val
930 935 940
Tyr Gln Asp Ala Lys Gln Ser Ile Asn His Leu Phe Val Asp Tyr Gln
945 950 955 960
Asp Gln Gln Leu Asn Pro Glu Ile Gly Met Ala Asp Ile Met Asp Ala
965 970 975
Gln Asn Leu Val Ala Ser Ile Ser Asp Val Tyr Ser Asp Ala Val Leu
980 985 990
Gln Ile Pro Gly Ile Asn Tyr Glu Ile Tyr Thr Glu Leu Ser Asn Arg
995 1000 1005
Leu Gln Gln Ala Ser Tyr Leu Tyr Thr Ser Arg Asn Ala Val Gln Asn
1010 1015 1020
Gly Asp Phe Asn Asn Gly Leu Asp Ser Trp Asn Ala Thr Ala Gly Ala
1025 1030 1035 1040
Ser Val Gln Gln Asp Gly Asn Thr His Phe Leu Val Leu Ser His Trp
1045 1050 1055
Asp Ala Gln Val Ser Gln Gln Phe Arg Val Gln Pro Asn Cys Lys Tyr
1060 1065 1070
Val Leu Arg Val Thr Ala Glu Lys Val Gly Gly Gly Asp Gly Tyr Val
1075 1080 1085
Thr Ile Arg Asp Asp Ala His His Thr Glu Thr Leu Thr Phe Asn Ala
1090 1095 1100
Cys Asp Tyr Asp Ile Asn Gly Thr Tyr Val Thr Asp Asn Thr Tyr Leu
1105 1110 1115 1120
Thr Lys Glu Val Val Phe His Pro Glu Thr Gln His Met Trp Val Glu
1125 1130 1135
Val Asn Glu Thr Glu Gly Ala Phe His Ile Asp Ser Ile Glu Phe Val
1140 1145 1150
Glu Thr Glu Lys
1155
〈210〉 71
〈211〉 3471
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 71
atgaatcaaa ataaacacgg aattattggc gcttccaatt gtggttgtgc atctgatgat 60
gttgcgaaat atcctttagc caacaatcca tattcatctg ctttaaattt aaattcttgt 120
caaaatagta gtattctcaa ctggattaac ataataggcg atgcagcaaa agaagcagta 180
tctattggga caaccatagt ctctcttatc acagcacctt ctcttactgg attaatttca 240
atagtatatg accttatagg taaagtacta ggaggtagta gtggacaatc catatcagat 300
ttgtctatat gtgacttatt atctattatt gatttacggg taagtcagag tgttttaaat 360
gatgggattg cagattttaa tggttctgta ctcttataca ggaactattt agaggctctg 420
gatagctgga ataagaatcc taattctgct tctgctgaag aactccgtac tcgttttaga 480
atcgccgact cagaatttga tagaatttta acccgagggt ctttaacgaa tggtggctcg 540
ttagctagac aaaatgccca aatattatta ttaccttctt ttgcgagcgc tgcatttttc 600
catttattac tactaaggga tgctactaga tatggcacta attgggggct atacaatgct 660
acacctttta taaattatca atcaaaacta gtagagctta ttgaactata tactgattat 720
tgcgtacatt ggtataatcg aggtttcaac gaactaagac aacgaggcac tagtgctaca 780
gcttggttag aatttcatag atatcgtaga gagatgacat tgatggtatt agatatagta 840
gcatcatttt caagtcttga tattactaat tacccaatag aaacagattt tcagttgagt 900
agggtcattt atacagatcc aattggtttt gtacatcgta gtagtcttag gggagaaagt 960
tggtttagct ttgttaatag agctaatttc tcagatttag aaaatgcaat acctaatcct 1020
agaccgtctt ggtttttaaa taatatgatt atatctactg gttcacttac attgccggtt 1080
agcccaagta ctgatagagc gagggtatgg tatggaagtc gagatcgaat ttcccctgct 1140
aattcacaat ttattactga actaatctct ggacaacata cgactgctac acaaactatt 1200
ttagggcgaa atatatttag agtagattct caagcttgta atttaaatga taccacatat 1260
ggagtgaata gggcggtatt ttatcatgat gcgagtgaag gttctcaaag atccgtgtac 1320
gaggggtata ttcgaacaac tgggatagat aaccctagag ttcaaaatat taacacttat 1380
ttacctggag aaaattcaga tatcccaact ccagaagact atactcatat attaagcaca 1440
acaataaatt taacaggagg acttagacaa gtagcatcta atcgccgttc atctttagta 1500
atgtatggtt ggacacataa aagtctggct cgtaacaata ccattaatcc agatagaatt 1560
acacagatac cattgacgaa ggttgatacc cgaggcacag gtgtttctta tgtgaatgat 1620
ccaggattta taggaggagc tctacttcaa aggactgacc atggttcgct tggagtattg 1680
agggtccaat ttccacttca cttaagacaa caatatcgta ttagagtccg ttatgcttct 1740
acaacaaata ttcgattgag tgtgaatggc agtttcggta ctatttctca aaatctccct 1800
agtacaatga gattaggaga ggatttaaga tacggatctt ttgctataag agagtttaat 1860
acttctatta gacccactgc aagtccggac caaattcgat tgacaataga accatctttt 1920
attagacaag aggtctatgt agatagaatt gagttcattc cagttaatcc gacgcgagag 1980
gcgaaagagg atctagaagc agcaaaaaaa gcggtggcga gcttgtttac acgcacaagg 2040
gacggattac aagtaaatgt gaaagattat caagtcgatc aagcggcaaa tttagtgtca 2100
tgcttatcag atgaacaata tgggtatgac aaaaagatgt tattggaagc ggtacgtgcg 2160
gcaaaacgac ttagccgaga acgcaactta cttcaggatc cagattttaa tacaatcaat 2220
agtacagaag aaaatggatg gaaagcaagt aacggcgtta ctattagtga gggcgggcca 2280
ttctataaag gccgtgcaat tcagctagca agtgcacgag aaaattaccc aacatacatc 2340
tatcaaaaag tagatgcatc ggagttaaag ccgtatacac gttatagact ggatgggttc 2400
gtgaagagta gtcaagattt agaaattgat ctcattcacc atcataaagt ccatcttgtg 2460
aaaaatgtac cagataattt agtatctgat acttacccag atgattcttg tagtggaatc 2520
aatcgatgtc aggaacaaca gatggtaaat gcgcaactgg aaacagagca tcatcatccg 2580
atggattgct gtgaagcagc tcaaacacat gagttttctt cctatattga tacaggggat 2640
ttaaattcga gtgtagacca gggaatctgg gcgatcttta aagttcgaac aaccgatggt 2700
tatgcgacgt taggaaatct tgaattggta gaggtcggac cgttatcggg tgaatcttta 2760
gaacgtgaac aaagggataa tacaaaatgg agtgcagagc taggaagaaa gcgtgcagaa 2820
acagatcgcg tgtatcaaga tgccaaacaa tccatcaatc atttatttgt ggattatcaa 2880
gatcaacaat taaatccaga aatagggatg gcagatatta tggacgctca aaatcttgtc 2940
gcatcaattt cagatgtata tagcgatgcc gtactgcaaa tccctggaat taactatgag 3000
atttacacag agctgtccaa tcgcttacaa caagcatcgt atctgtatac gtctcgaaat 3060
gcggtgcaaa atggggactt taacaacggg ctagatagct ggaatgcaac agcgggtgca 3120
tcggtacaac aggatggcaa tacgcatttc ttagttcttt ctcattggga tgcacaagtt 3180
tctcaacaat ttagagtgca gccgaattgt aaatatgtat tacgtgtaac agcagagaaa 3240
gtaggcggcg gagacggata cgtgactatc cgggatgatg ctcatcatac agaaacgctt 3300
acatttaatg catgtgatta tgatataaat ggcacgtacg tgactgataa tacgtatcta 3360
acaaaagaag tggtattcca tccggagaca caacacatgt gggtagaggt aaatgaaaca 3420
gaaggtgcat ttcatataga tagtattgaa ttcgttgaaa cagaaaagta a 3471
〈210〉 72
〈211〉 1156
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 72
Met Asn Arg Asn Asn Gln Asn Glu Tyr Glu Ile Ile Asp Ala Pro His
1 5 10 15
Cys Gly Cys Pro Ser Asp Asp Asp Val Arg Tyr Pro Leu Ala Ser Asp
20 25 30
Pro Asn Ala Ala Leu Gln Asn Met Asn Tyr Lys Asp Tyr Leu Gln Met
35 40 45
Thr Asp Glu Asp Tyr Thr Asp Ser Tyr Ile Asn Pro Ser Leu Ser Ile
50 55 60
Ser Gly Arg Asp Ala Val Gln Thr Ala Leu Thr Val Val Gly Arg Ile
65 70 75 80
Leu Gly Ala Leu Gly Val Pro Phe Ser Gly Gln Ile Val Ser Phe Tyr
85 90 95
Gln Phe Leu Leu Asn Thr Leu Trp Pro Val Asn Asp Thr Ala Ile Trp
100 105 110
Glu Ala Phe Met Arg Gln Val Glu Glu Leu Val Asn Gln Gln Ile Thr
115 120 125
Glu Phe Ala Arg Asn Gln Ala Leu Ala Arg Leu Gln Gly Leu Gly Asp
130 135 140
Ser Phe Asn Val Tyr Gln Arg Ser Leu Gln Asn Trp Leu Ala Asp Arg
145 150 155 160
Asn Asp Thr Arg Asn Leu Ser Val Val Arg Ala Gln Phe Ile Ala Leu
165 170 175
Asp Leu Asp Phe Val Asn Ala Ile Pro Leu Phe Ala Val Asn Gly Gln
180 185 190
Gln Val Pro Leu Leu Ser Val Tyr Ala Gln Ala Val Asn Leu His Leu
195 200 205
Leu Leu Leu Lys Asp Ala Ser Leu Phe Gly Glu Gly Trp Gly Phe Thr
210 215 220
Gln Gly Glu Ile Ser Thr Tyr Tyr Asp Arg Gln Leu Glu Leu Thr Ala
225 230 235 240
Lys Tyr Thr Asn Tyr Cys Glu Thr Trp Tyr Asn Thr Gly Leu Asp Arg
245 250 255
Leu Arg Gly Thr Asn Thr Glu Ser Trp Leu Arg Tyr His Gln Phe Arg
260 265 270
Arg Glu Met Thr Leu Val Val Leu Asp Val Val Ala Leu Phe Pro Tyr
275 280 285
Tyr Asp Val Arg Leu Tyr Pro Thr Gly Ser Asn Pro Gln Leu Thr Arg
290 295 300
Glu Val Tyr Thr Asp Pro Ile Val Phe Asn Pro Pro Ala Asn Val Gly
305 310 315 320
Leu Cys Arg Arg Trp Gly Thr Asn Pro Tyr Asn Thr Phe Ser Glu Leu
325 330 335
Glu Asn Ala Phe Ile Arg Pro Pro His Leu Phe Asp Arg Leu Asn Ser
340 345 350
Leu Thr Ile Ser Ser Asn Arg Phe Pro Val Ser Ser Asn Phe Met Asp
355 360 365
Tyr Trp Ser Gly His Thr Leu Arg Arg Ser Tyr Leu Asn Asp Ser Ala
370 375 380
Val Gln Glu Asp Ser Tyr Gly Leu Ile Thr Thr Thr Arg Ala Thr Ile
385 390 395 400
Asn Pro Gly Val Asp Gly Thr Asn Arg Ile Glu Ser Thr Ala Val Asp
405 410 415
Phe Arg Ser Ala Leu Ile Gly Ile Tyr Gly Val Asn Arg Ala Ser Phe
420 425 430
Val Pro Gly Gly Leu Phe Asn Gly Thr Thr Ser Pro Ala Asn Gly Gly
435 440 445
Cys Arg Asp Leu Tyr Asp Thr Asn Asp Glu Leu Pro Pro Asp Glu Ser
450 455 460
Thr Gly Ser Ser Thr His Arg Leu Ser His Val Thr Phe Phe Ser Phe
465 470 475 480
Gln Thr Asn Gln Ala Gly Ser Ile Ala Asn Ala Gly Ser Val Pro Thr
485 490 495
Tyr Val Trp Thr Arg Arg Asp Val Asp Leu Asn Asn Thr Ile Thr Pro
500 505 510
Asn Arg Ile Thr Gln Leu Pro Leu Val Lys Ala Ser Ala Pro Val Ser
515 520 525
Gly Thr Thr Val Leu Lys Gly Pro Gly Phe Thr Gly Gly Gly Ile Leu
530 535 540
Arg Arg Thr Thr Asn Gly Thr Phe Gly Thr Leu Arg Val Thr Val Asn
545 550 555 560
Ser Pro Leu Thr Gln Arg Tyr Arg Val Arg Val Arg Phe Ala Ser Ser
565 570 575
Gly Asn Phe Ser Ile Arg Ile Leu Arg Gly Asn Thr Ser Ile Ala Tyr
580 585 590
Gln Arg Phe Gly Ser Thr Met Asn Arg Gly Gln Glu Leu Thr Tyr Glu
595 600 605
Ser Phe Val Thr Ser Glu Phe Thr Thr Asn Gln Ser Asp Leu Pro Phe
610 615 620
Thr Phe Thr Gln Ala Gln Glu Asn Leu Thr Ile Leu Ala Glu Gly Val
625 630 635 640
Ser Thr Gly Ser Glu Tyr Phe Ile Asp Arg Ile Glu Ile Ile Pro Val
645 650 655
Asn Pro Ala Arg Glu Ala Glu Glu Asp Leu Glu Ala Ala Lys Lys Ala
660 665 670
Val Ala Asn Leu Phe Thr Arg Thr Arg Asp Gly Leu Gln Val Asn Val
675 680 685
Thr Asp Tyr Gln Val Asp Gln Ala Ala Asn Leu Val Ser Cys Leu Ser
690 695 700
Asp Glu Gln Tyr Gly His Asp Lys Lys Met Leu Leu Glu Ala Val Arg
705 710 715 720
Ala Ala Lys Arg Leu Ser Arg Glu Arg Asn Leu Leu Gln Asp Pro Asp
725 730 735
Phe Asn Thr Ile Asn Ser Thr Glu Glu Asn Gly Trp Lys Ala Ser Asn
740 745 750
Gly Val Thr Ile Ser Glu Gly Gly Pro Phe Phe Lys Gly Arg Ala Leu
755 760 765
Gln Leu Ala Ser Ala Arg Glu Asn Tyr Pro Thr Tyr Ile Tyr Gln Lys
770 775 780
Val Asp Ala Ser Val Leu Lys Pro Tyr Thr Arg Tyr Arg Leu Asp Gly
785 790 795 800
Phe Val Lys Ser Ser Gln Asp Leu Glu Ile Asp Leu Ile His His His
805 810 815
Lys Val His Leu Val Lys Asn Val Pro Asp Asn Leu Val Ser Asp Thr
820 825 830
Tyr Ser Asp Gly Ser Cys Ser Gly Ile Asn Arg Cys Asp Glu Gln His
835 840 845
Gln Val Asp Met Gln Leu Asp Ala Glu His His Pro Met Asp Cys Cys
850 855 860
Glu Ala Ala Gln Thr His Glu Phe Ser Ser Tyr Ile Asn Thr Gly Asp
865 870 875 880
Leu Asn Ala Ser Val Asp Gln Gly Ile Trp Val Val Leu Lys Val Arg
885 890 895
Thr Thr Asp Gly Tyr Ala Thr Leu Gly Asn Leu Glu Leu Val Glu Val
900 905 910
Gly Pro Leu Ser Gly Glu Ser Leu Glu Arg Glu Gln Arg Asp Asn Ala
915 920 925
Lys Trp Asn Ala Glu Leu Gly Arg Lys Arg Ala Glu Ile Asp Arg Val
930 935 940
Tyr Leu Ala Ala Lys Gln Ala Ile Asn His Leu Phe Val Asp Tyr Gln
945 950 955 960
Asp Gln Gln Leu Asn Pro Glu Ile Gly Leu Ala Glu Ile Asn Glu Ala
965 970 975
Ser Asn Leu Val Glu Ser Ile Ser Gly Val Tyr Ser Asp Thr Leu Leu
980 985 990
Gln Ile Pro Gly Ile Asn Tyr Glu Ile Tyr Thr Glu Leu Ser Asp Arg
995 1000 1005
Leu Gln Gln Ala Ser Tyr Leu Tyr Thr Ser Arg Asn Ala Val Gln Asn
1010 1015 1020
Gly Asp Phe Asn Ser Gly Leu Asp Ser Trp Asn Thr Thr Met Asp Ala
1025 1030 1035 1040
Ser Val Gln Gln Asp Gly Asn Met His Phe Leu Val Leu Ser His Trp
1045 1050 1055
Asp Ala Gln Val Ser Gln Gln Leu Arg Val Asn Pro Asn Cys Lys Tyr
1060 1065 1070
Val Leu Arg Val Thr Ala Arg Lys Val Gly Gly Gly Asp Gly Tyr Val
1075 1080 1085
Thr Ile Arg Asp Gly Ala His His Gln Glu Thr Leu Thr Phe Asn Ala
1090 1095 1100
Cys Asp Tyr Asp Val Asn Gly Thr Tyr Val Asn Asp Asn Ser Tyr Ile
1105 1110 1115 1120
Thr Glu Glu Val Val Phe Tyr Pro Glu Thr Lys His Met Trp Val Glu
1125 1130 1135
Val Ser Glu Ser Glu Gly Ser Phe Tyr Ile Asp Ser Ile Glu Phe Ile
1140 1145 1150
Glu Thr Gln Glu
1155
〈210〉 73
〈211〉 3471
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 73
atgaatcgaa ataatcaaaa tgaatatgaa attattgatg ccccccattg tgggtgtcca 60
tcagatgacg atgtgaggta tcctttggca agtgacccaa atgcagcgtt acaaaatatg 120
aactataaag attacttaca aatgacagat gaggactaca ctgattctta tataaatcct 180
agtttatcta ttagtggtag agatgcagtt cagactgcgc ttactgttgt tgggagaata 240
ctcggggctt taggtgttcc gttttctgga caaatagtga gtttttatca attcctttta 300
aatacactgt ggccagttaa tgatacagct atatgggaag ctttcatgcg acaggtggag 360
gaacttgtca atcaacaaat aacagaattt gcaagaaatc aggcacttgc aagattgcaa 420
ggattaggag actcttttaa tgtatatcaa cgttcccttc aaaattggtt ggctgatcga 480
aatgatacac gaaatttaag tgttgttcgt gctcaattta tagctttaga ccttgatttt 540
gttaatgcta ttccattgtt tgcagtaaat ggacagcagg ttccattact gtcagtatat 600
gcacaagctg tgaatttaca tttgttatta ttaaaagatg catctctttt tggagaagga 660
tggggattca cacaggggga aatttccaca tattatgacc gtcaattgga actaaccgct 720
aagtacacta attactgtga aacttggtat aatacaggtt tagatcgttt aagaggaaca 780
aatactgaaa gttggttaag atatcatcaa ttccgtagag aaatgacttt agtggtatta 840
gatgttgtgg cgctatttcc atattatgat gtacgacttt atccaacggg atcaaaccca 900
cagcttacac gtgaggtata tacagatccg attgtattta atccaccagc taatgttgga 960
ctttgccgac gttggggtac taatccctat aatacttttt ctgagctcga aaatgccttc 1020
attcgcccac cacatctttt tgataggctg aatagcttaa caatcagcag taatcgattt 1080
ccagtttcat ctaattttat ggattattgg tcaggacata cgttacgccg tagttatctg 1140
aacgattcag cagtacaaga agatagttat ggcctaatta caaccacaag agcaacaatt 1200
aatcctggag ttgatggaac aaaccgcata gagtcaacgg cagtagattt tcgttctgca 1260
ttgataggta tatatggcgt gaatagagct tcttttgtcc caggaggctt gtttaatggt 1320
acgacttctc ctgctaatgg aggatgtaga gatctctatg atacaaatga tgaattacca 1380
ccagatgaaa gtaccggaag ttctacccat agactatctc atgttacctt ttttagtttt 1440
caaactaatc aggctggatc tatagctaat gcaggaagtg tacctactta tgtttggacc 1500
cgtcgtgatg tggaccttaa taatacgatt accccaaata gaattacaca attaccattg 1560
gtaaaggcat ctgcacctgt ttcgggtact acggtcttaa aaggtccagg atttacagga 1620
gggggtatac tccgaagaac aactaatggc acatttggaa cgttaagagt aacagttaat 1680
tcaccattaa cacaaagata tcgcgtaaga gttcgttttg cttcatcagg aaatttcagc 1740
ataaggatac tgcgtggaaa tacctctata gcttatcaaa gatttgggag tacaatgaac 1800
agaggacagg aactaactta cgaatcattt gtcacaagtg agttcactac taatcagagc 1860
gatctgcctt ttacatttac acaagctcaa gaaaatttaa caatccttgc agaaggtgtt 1920
agcaccggta gtgaatattt tatagataga attgaaatca tccctgtgaa cccggcacga 1980
gaagcagaag aggatttaga agcagcgaag aaagcggtgg cgaacttgtt tacacgtaca 2040
agggacggat tacaggtaaa tgtgacagat tatcaagtgg accaagcggc aaatttagtg 2100
tcatgcttat ccgatgaaca atatgggcat gacaaaaaga tgttattgga agcggtaaga 2160
gcggcaaaac gcctcagccg cgaacgcaac ttacttcaag atccagattt taatacaatc 2220
aatagtacag aagagaatgg ctggaaggca agtaacggtg ttactattag cgagggcggt 2280
ccattcttta aaggtcgtgc acttcagtta gcaagcgcaa gagaaaatta tccaacatac 2340
atttatcaaa aagtagatgc atcggtgtta aagccttata cacgctatag actagatgga 2400
tttgtgaaga gtagtcaaga tttagaaatt gatctcatcc accatcataa agtccatctt 2460
gtaaaaaatg taccagataa tttagtatct gatacttact cagatggttc ttgcagcgga 2520
atcaaccgtt gtgatgaaca gcatcaggta gatatgcagc tagatgcgga gcatcatcca 2580
atggattgct gtgaagcggc tcaaacacat gagttttctt cctatattaa tacaggggat 2640
ctaaatgcaa gtgtagatca gggcatttgg gttgtattaa aagttcgaac aacagatggg 2700
tatgcgacgt taggaaatct tgaattggta gaggttgggc cattatcggg tgaatctcta 2760
gaacgggaac aaagagataa tgcgaaatgg aatgcagagc taggaagaaa acgtgcagaa 2820
atagatcgtg tgtatttagc tgcgaaacaa gcaattaatc atctgtttgt agactatcaa 2880
gatcaacaat taaatccaga aattgggcta gcagaaatta atgaagcttc aaatcttgta 2940
gagtcaattt cgggtgtata tagtgataca ctattacaga ttcctgggat taactacgaa 3000
atttacacag agttatccga tcgcttacaa caagcatcgt atctgtatac gtctagaaat 3060
gcggtgcaaa atggagactt taacagtggt ctagatagtt ggaatacaac tatggatgca 3120
tcggttcagc aagatggcaa tatgcatttc ttagttcttt cgcattggga tgcacaagtt 3180
tcccaacaat tgagagtaaa tccgaattgt aagtatgtct tacgtgtgac agcaagaaaa 3240
gtaggaggcg gagatggata cgtcacaatc cgagatggcg ctcatcacca agaaactctt 3300
acatttaatg catgtgacta cgatgtaaat ggtacgtatg tcaatgacaa ttcgtatata 3360
acagaagaag tggtattcta cccagagaca aaacatatgt gggtagaggt gagtgaatcc 3420
gaaggttcat tctatataga cagtattgag tttattgaaa cacaagagta g 3471
〈210〉 74
〈211〉 1150
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 74
Met Asn Arg Asn Asn Pro Asn Glu Tyr Glu Ile Ile Asp Ala Pro Tyr
1 5 10 15
Cys Gly Cys Pro Ser Asp Asp Asp Val Arg Tyr Pro Leu Ala Ser Asp
20 25 30
Pro Asn Ala Ala Phe Gln Asn Met Asn Tyr Lys Glu Tyr Leu Gln Thr
35 40 45
Tyr Asp Gly Asp Tyr Thr Gly Ser Leu Ile Asn Pro Asn Leu Ser Ile
50 55 60
Asn Pro Arg Asp Val Leu Gln Thr Gly Ile Asn Ile Val Gly Arg Ile
65 70 75 80
Leu Gly Phe Leu Gly Val Pro Phe Ala Gly Gln Leu Val Thr Phe Tyr
85 90 95
Thr Phe Leu Leu Asn Gln Leu Trp Pro Thr Asn Asp Asn Ala Val Trp
100 105 110
Glu Ala Phe Met Ala Gln Ile Glu Glu Leu Ile Asp Gln Lys Ile Ser
115 120 125
Ala Gln Val Val Arg Asn Ala Leu Asp Asp Leu Thr Gly Leu His Asp
130 135 140
Tyr Tyr Glu Glu Tyr Leu Ala Ala Leu Glu Glu Trp Leu Glu Arg Pro
145 150 155 160
Asn Gly Ala Arg Ala Asn Leu Val Thr Gln Arg Phe Glu Asn Leu His
165 170 175
Thr Ala Phe Val Thr Arg Met Pro Ser Phe Gly Thr Gly Pro Gly Ser
180 185 190
Gln Arg Asp Ala Val Ala Leu Leu Thr Val Tyr Ala Gln Ala Ala Asn
195 200 205
Leu His Leu Leu Leu Leu Lys Asp Ala Glu Ile Tyr Gly Ala Arg Trp
210 215 220
Gly Leu Gln Gln Gly Gln Ile Asn Leu Tyr Phe Asn Ala Gln Gln Glu
225 230 235 240
Arg Thr Arg Ile Tyr Thr Asn His Cys Val Glu Thr Tyr Asn Arg Gly
245 250 255
Leu Glu Asp Val Arg Gly Thr Asn Thr Glu Ser Trp Leu Asn Tyr His
260 265 270
Arg Phe Arg Arg Glu Met Thr Leu Met Ala Met Asp Leu Val Ala Leu
275 280 285
Phe Pro Phe Tyr Asn Val Arg Gln Tyr Pro Asn Gly Ala Asn Pro Gln
290 295 300
Leu Thr Arg Glu Ile Tyr Thr Asp Pro Ile Val Tyr Asn Pro Pro Ala
305 310 315 320
Asn Gln Gly Ile Cys Arg Arg Trp Gly Asn Asn Pro Tyr Asn Thr Phe
325 330 335
Ser Glu Leu Glu Asn Ala Phe Ile Arg Pro Pro His Leu Phe Glu Arg
340 345 350
Leu Asn Arg Leu Thr Ile Ser Arg Asn Arg Tyr Thr Ala Pro Thr Thr
355 360 365
Asn Ser Phe Leu Asp Tyr Trp Ser Gly His Thr Leu Gln Ser Gln His
370 375 380
Ala Asn Asn Pro Thr Thr Tyr Glu Thr Ser Tyr Gly Gln Ile Thr Ser
385 390 395 400
Asn Thr Arg Leu Phe Asn Thr Thr Asn Gly Ala Arg Ala Ile Asp Ser
405 410 415
Arg Ala Arg Asn Phe Gly Asn Leu Tyr Ala Asn Leu Tyr Gly Val Ser
420 425 430
Ser Leu Asn Ile Phe Pro Thr Gly Val Met Ser Glu Ile Thr Asn Ala
435 440 445
Ala Asn Thr Cys Arg Gln Asp Leu Thr Thr Thr Glu Glu Leu Pro Leu
450 455 460
Glu Asn Asn Asn Phe Asn Leu Leu Ser His Val Thr Phe Leu Arg Phe
465 470 475 480
Asn Thr Thr Gln Gly Gly Pro Leu Ala Thr Leu Gly Phe Val Pro Thr
485 490 495
Tyr Val Trp Thr Arg Glu Asp Val Asp Phe Thr Asn Thr Ile Thr Ala
500 505 510
Asp Arg Ile Thr Gln Leu Pro Trp Val Lys Ala Ser Glu Ile Gly Gly
515 520 525
Gly Thr Thr Val Val Lys Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu
530 535 540
Arg Arg Thr Asp Gly Gly Ala Val Gly Thr Ile Arg Ala Asn Val Asn
545 550 555 560
Ala Pro Leu Thr Gln Gln Tyr Arg Ile Arg Leu Arg Tyr Ala Ser Thr
565 570 575
Thr Ser Phe Val Val Asn Leu Phe Val Asn Asn Ser Ala Ala Gly Phe
580 585 590
Thr Leu Pro Ser Thr Met Ala Gln Asn Gly Ser Leu Thr Tyr Glu Ser
595 600 605
Phe Asn Thr Leu Glu Val Thr His Thr Ile Arg Phe Ser Gln Ser Asp
610 615 620
Thr Thr Leu Arg Leu Asn Ile Phe Pro Ser Ile Ser Gly Gln Glu Val
625 630 635 640
Tyr Val Asp Lys Leu Glu Ile Val Pro Ile Asn Pro Thr Arg Glu Ala
645 650 655
Glu Glu Asp Leu Glu Asp Ala Lys Lys Ala Val Ala Ser Leu Phe Thr
660 665 670
Arg Thr Arg Asp Gly Leu Gln Val Asn Val Thr Asp Tyr Gln Val Asp
675 680 685
Gln Ala Ala Asn Leu Val Ser Cys Leu Ser Asp Glu Gln Tyr Gly His
690 695 700
Asp Lys Lys Met Leu Leu Glu Ala Val Arg Ala Ala Lys Arg Leu Ser
705 710 715 720
Arg Glu Arg Asn Leu Leu Gln Asp Pro Asp Phe Asn Glu Ile Asn Ser
725 730 735
Thr Glu Glu Asn Gly Trp Lys Ala Ser Asn Gly Val Thr Ile Ser Glu
740 745 750
Gly Gly Pro Phe Phe Lys Gly Arg Ala Leu Gln Leu Ala Ser Ala Arg
755 760 765
Glu Asn Tyr Pro Thr Tyr Ile Tyr Gln Lys Val Asp Ala Ser Thr Leu
770 775 780
Lys Pro Tyr Thr Arg Tyr Lys Leu Asp Gly Phe Val Gln Ser Ser Gln
785 790 795 800
Asp Leu Glu Ile Asp Leu Ile His His His Lys Val His Leu Val Lys
805 810 815
Asn Val Pro Asp Asn Leu Val Ser Asp Thr Tyr Ser Asp Gly Ser Cys
820 825 830
Ser Gly Ile Asn Arg Cys Glu Glu Gln His Gln Val Asp Val Gln Leu
835 840 845
Asp Ala Glu Asp His Pro Lys Asp Cys Cys Glu Ala Ala Gln Thr His
850 855 860
Glu Phe Ser Ser Tyr Ile His Thr Gly Asp Leu Asn Ala Ser Val Asp
865 870 875 880
Gln Gly Ile Trp Val Val Leu Gln Val Arg Thr Thr Asp Gly Tyr Ala
885 890 895
Thr Leu Gly Asn Leu Glu Leu Val Glu Val Gly Pro Leu Ser Gly Glu
900 905 910
Ser Leu Glu Arg Glu Gln Arg Asp Asn Ala Lys Trp Asn Glu Glu Val
915 920 925
Gly Arg Lys Arg Ala Glu Thr Asp Arg Ile Tyr Gln Asp Ala Lys Gln
930 935 940
Ala Ile Asn His Leu Phe Val Asp Tyr Gln Asp Gln Gln Leu Ser Pro
945 950 955 960
Glu Val Gly Met Ala Asp Ile Ile Asp Ala Gln Asn Leu Ile Ala Ser
965 970 975
Ile Ser Asp Val Tyr Ser Asp Ala Val Leu Gln Ile Pro Gly Ile Asn
980 985 990
Tyr Glu Met Tyr Thr Glu Leu Ser Asn Arg Leu Gln Gln Ala Ser Tyr
995 1000 1005
Leu Tyr Thr Ser Arg Asn Val Val Gln Asn Gly Asp Phe Asn Ser Gly
1010 1015 1020
Leu Asp Ser Trp Asn Ala Thr Thr Asp Thr Ala Val Gln Gln Asp Gly
1025 1030 1035 1040
Asn Met His Phe Leu Val Leu Ser His Trp Asp Ala Gln Val Ser Gln
1045 1050 1055
Gln Phe Arg Val Gln Pro Asn Cys Lys Tyr Val Leu Arg Val Thr Ala
1060 1065 1070
Lys Lys Val Gly Asn Gly Asp Gly Tyr Val Thr Ile Gln Asp Gly Ala
1075 1080 1085
His His Arg Glu Thr Leu Thr Phe Asn Ala Cys Asp Tyr Asp Val Asn
1090 1095 1100
Gly Thr His Val Asn Asp Asn Ser Tyr Ile Thr Lys Glu Leu Val Phe
1105 1110 1115 1120
Tyr Pro Lys Thr Glu His Met Trp Val Glu Val Ser Glu Thr Glu Gly
1125 1130 1135
Thr Phe Tyr Ile Asp Ser Ile Glu Phe Ile Glu Thr Gln Glu
1140 1145 1150
〈210〉 75
〈211〉 3453
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 75
atgaatcgaa ataatccaaa tgaatatgaa attattgatg ccccctattg tgggtgtccg 60
tcagatgatg atgtgaggta tcctttggca agtgacccaa atgcagcgtt ccaaaatatg 120
aactataaag agtatttaca aacgtatgat ggagactaca caggttctct tatcaatcct 180
aacttatcta ttaatcctag agatgtacta caaacaggta ttaatattgt gggaagaata 240
ctagggtttt taggtgttcc atttgcgggt caactagtta ctttctatac ctttctctta 300
aatcagttgt ggccaactaa tgataatgca gtatgggaag cttttatggc gcaaatagaa 360
gagctaatcg atcaaaaaat atcggcgcaa gtagtaagga atgcactcga tgacttaact 420
ggattacacg attattatga ggagtattta gcagcattag aggagtggct ggaaagaccg 480
aacggagcaa gagctaactt agttacacag aggtttgaaa acctgcatac tgcatttgta 540
actagaatgc caagctttgg tacgggtcct ggtagtcaaa gagatgcggt agcgttgttg 600
acggtatatg cacaagcagc gaatttgcat ttgttattat taaaagatgc agaaatctat 660
ggggcaagat ggggacttca acaagggcaa attaacttat attttaatgc tcaacaagaa 720
cgtactcgaa tttataccaa tcattgcgtg gaaacatata atagaggatt agaagatgta 780
agaggaacaa atacagaaag ttggttaaat taccatcgat tccgtagaga gatgacatta 840
atggcaatgg atttagtggc cctattccca ttctataatg tgcgacaata tccaaatggg 900
gcaaatccac agcttacacg tgaaatatat acagatccaa tcgtatataa tccaccagct 960
aatcagggaa tttgccgacg ttgggggaat aatccgtata atacattttc tgaacttgaa 1020
aatgctttta ttcgcccgcc acatcttttt gaaaggttga acagattaac tatttctaga 1080
aaccgatata cagctccaac aactaatagc ttcctagact attggtcagg tcatacttta 1140
caaagccaac atgcaaataa cccgacgaca tatgaaacta gttacggtca gattacctct 1200
aacacacgtt tattcaatac gactaatgga gcccgtgcaa tagattcaag ggcaagaaat 1260
tttggtaact tatacgctaa tttgtatggc gttagcagct tgaacatttt cccaacaggt 1320
gtgatgagtg aaatcaccaa tgcagctaat acgtgtcggc aagaccttac tacaactgaa 1380
gaactaccac tagagaataa taattttaat cttttatctc atgttacttt cttacgcttc 1440
aatactactc agggtggccc ccttgcaact ctagggtttg tacccacata tgtgtggaca 1500
cgtgaagatg tagattttac gaacacaatt actgcggata gaattacaca actaccatgg 1560
gtaaaggcat ctgaaatagg tgggggtact actgtcgtga aaggtccagg atttacagga 1620
ggggatatac ttcgaagaac ggacggtggt gcagttggaa cgattagagc taatgttaat 1680
gccccattaa cacaacaata tcgtataaga ttacgctatg cttcgacaac aagttttgtt 1740
gttaatttat ttgttaataa tagtgcggct ggctttactt taccgagtac aatggctcaa 1800
aatggttctt taacatacga gtcgtttaat accttagagg taactcatac tattagattt 1860
tcacagtcag atactacact taggttgaat atattcccgt ctatctctgg tcaagaagtg 1920
tatgtagata aacttgaaat cgttccaatt aacccgacac gagaagcgga agaagattta 1980
gaagatgcaa agaaagcggt ggcgagcttg tttacacgta caagggatgg attacaggta 2040
aatgtgacag attaccaagt cgatcaggcg gcaaatttag tgtcgtgctt atcagatgaa 2100
caatatgggc atgataaaaa gatgttattg gaagccgtac gcgcagcaaa acgcctcagc 2160
cgcgaacgca acttacttca agatccagat tttaatgaaa taaatagcac agaagaaaat 2220
ggctggaagg caagtaacgg tgttactatt agcgagggcg gtccattctt taaaggtcgt 2280
gcacttcagt tagcaagcgc acgtgaaaat tacccaacat acatctatca aaaggtagat 2340
gcatcgacgt taaaacctta tacacgatat aaactagatg gatttgtgca aagtagtcaa 2400
gatttagaaa ttgacctcat tcatcatcat aaagtccacc tcgtgaaaaa tgtaccagat 2460
aatttagtat ctgatactta ttctgatggc tcatgtagtg gaattaaccg ttgtgaggaa 2520
caacatcagg tagatgtgca gctagatgcg gaggatcatc caaaggattg ttgtgaagcg 2580
gctcaaacac atgagttttc ttcctatatt catacaggtg atctaaatgc aagtgtagat 2640
caaggcattt gggttgtatt gcaggttcga acaacagatg gttatgcgac gttaggaaat 2700
cttgaattgg tagaggttgg tccattatcg ggtgaatctt tagaacgaga acaaagagat 2760
aatgcgaaat ggaatgaaga ggtaggaaga aagcgtgcag aaacagatcg catatatcaa 2820
gatgcgaaac aagcaattaa ccatctattt gtagactatc aagatcaaca attaagtcca 2880
gaggtaggga tggcggatat tattgatgct caaaatctta tcgcatcaat ttcagatgta 2940
tatagcgatg cagtactgca aatccctggg attaactacg agatgtatac agagttatcc 3000
aatcgattac aacaagcatc gtatctgtat acgtctcgaa atgtcgtgca aaatggggac 3060
tttaacagtg gtttagatag ttggaatgca acaactgata cagctgttca gcaggatggc 3120
aatatgcatt tcttagttct ttcccattgg gatgcacaag tttctcaaca atttagagta 3180
cagccgaatt gtaaatatgt gttacgtgtg acagcgaaga aagtagggaa cggagatgga 3240
tatgttacga tccaagatgg cgctcatcac cgagaaacac tgacattcaa tgcatgtgac 3300
tacgatgtaa atggtacgca tgtaaatgat aattcgtata ttacaaaaga attggtgttc 3360
tatccaaaga cggaacatat gtgggtagag gtaagtgaaa cagaaggtac cttctatata 3420
gacagcattg agttcattga aacacaagag tag 3453
〈210〉 76
〈211〉 1134
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 76
Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu
1 5 10 15
Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Gly Asn Val Arg
20 25 30
Thr Gly Leu Gln Thr Gly Ile Asp Ile Val Ala Val Val Val Gly Ala
35 40 45
Leu Gly Gly Pro Val Gly Gly Ile Leu Thr Gly Phe Leu Ser Thr Leu
50 55 60
Phe Gly Phe Leu Trp Pro Ser Asn Asp Gln Ala Val Trp Glu Ala Phe
65 70 75 80
Ile Glu Gln Met Glu Glu Leu Ile Glu Gln Arg Ile Ser Asp Gln Val
85 90 95
Val Arg Thr Ala Leu Asp Asp Leu Thr Gly Ile Gln Asn Tyr Tyr Asn
100 105 110
Gln Tyr Leu Ile Ala Leu Lys Glu Trp Glu Glu Arg Pro Asn Gly Val
115 120 125
Arg Ala Asn Leu Val Leu Gln Arg Phe Glu Ile Leu His Ala Leu Phe
130 135 140
Val Ser Ser Met Pro Ser Phe Gly Ser Gly Pro Gly Ser Gln Arg Phe
145 150 155 160
Gln Ala Gln Leu Leu Val Val Tyr Ala Gln Ala Ala Asn Leu His Leu
165 170 175
Leu Leu Leu Ala Asp Ala Glu Lys Tyr Gly Ala Arg Trp Gly Leu Arg
180 185 190
Glu Ser Gln Ile Gly Asn Leu Tyr Phe Asn Glu Leu Gln Thr Arg Thr
195 200 205
Arg Asp Tyr Thr Asn His Cys Val Asn Ala Tyr Asn Asn Gly Leu Ala
210 215 220
Gly Leu Arg Gly Thr Ser Ala Glu Ser Trp Leu Lys Tyr His Gln Phe
225 230 235 240
Arg Arg Glu Ala Thr Leu Met Ala Met Asp Leu Ile Ala Leu Phe Pro
245 250 255
Tyr Tyr Asn Thr Arg Arg Tyr Pro Ile Ala Val Asn Pro Gln Leu Thr
260 265 270
Arg Glu Val Tyr Thr Asp Pro Leu Gly Val Pro Ser Glu Glu Ser Ser
275 280 285
Leu Phe Pro Glu Leu Arg Cys Leu Arg Trp Gln Glu Thr Ser Ala Met
290 295 300
Thr Phe Ser Asn Leu Glu Asn Ala Ile Ile Ser Ser Pro His Leu Phe
305 310 315 320
Asp Thr Ile Asn Asn Leu Met Ile Tyr Thr Gly Ser Phe Ser Val His
325 330 335
Leu Thr Asn Gln Leu Ile Glu Gly Trp Ile Gly His Ser Val Thr Ser
340 345 350
Ser Leu Leu Ala Ser Gly Pro Thr Thr Val Leu Arg Arg Asn Tyr Gly
355 360 365
Ser Thr Thr Ser Ile Val Asn Tyr Phe Ser Phe Asn Asp Arg Asp Val
370 375 380
Tyr Gln Ile Asn Thr Arg Ser His Thr Gly Leu Gly Phe Gln Asn Ala
385 390 395 400
Pro Leu Phe Gly Ile Thr Arg Ala Gln Phe Tyr Pro Gly Gly Thr Tyr
405 410 415
Ser Val Thr Gln Arg Asn Ala Leu Thr Cys Glu Gln Asn Tyr Asn Ser
420 425 430
Ile Asp Glu Leu Pro Ser Leu Asp Pro Asn Glu Pro Ile Ser Arg Ser
435 440 445
Tyr Ser His Arg Leu Ser His Ile Thr Ser Tyr Leu His Arg Val Leu
450 455 460
Thr Ile Asp Gly Ile Asn Ile Tyr Ser Gly Asn Leu Pro Thr Tyr Val
465 470 475 480
Trp Thr His Arg Asp Val Asp Leu Thr Asn Thr Ile Thr Ala Asp Arg
485 490 495
Ile Thr Gln Leu Pro Leu Val Lys Ser Phe Glu Ile Pro Ala Gly Thr
500 505 510
Thr Val Val Arg Gly Pro Gly Phe Thr Gly Gly Asp Ile Leu Arg Arg
515 520 525
Thr Gly Val Gly Thr Phe Gly Thr Ile Arg Val Arg Thr Thr Ala Pro
530 535 540
Leu Thr Gln Arg Tyr Arg Ile Arg Phe Arg Phe Ala Ser Thr Thr Asn
545 550 555 560
Leu Phe Ile Gly Ile Arg Val Gly Asp Arg Gln Val Asn Tyr Phe Asp
565 570 575
Phe Gly Arg Thr Met Asn Arg Gly Asp Glu Leu Arg Tyr Glu Ser Phe
580 585 590
Ala Thr Arg Glu Phe Thr Thr Asp Phe Asn Phe Arg Gln Pro Gln Glu
595 600 605
Leu Ile Ser Val Phe Ala Asn Ala Phe Ser Ala Gly Gln Glu Val Tyr
610 615 620
Phe Asp Arg Ile Glu Ile Ile Pro Val Asn Pro Ala Arg Glu Ala Lys
625 630 635 640
Glu Asp Leu Glu Ala Ala Lys Lys Ala Val Ala Ser Leu Phe Thr Arg
645 650 655
Thr Arg Asp Gly Leu Gln Val Asn Val Lys Asp Tyr Gln Val Asp Gln
660 665 670
Ala Ala Asn Leu Val Ser Cys Leu Ser Asp Glu Gln Tyr Gly Tyr Asp
675 680 685
Lys Lys Met Leu Leu Glu Ala Val Arg Ala Ala Lys Arg Leu Ser Arg
690 695 700
Glu Arg Asn Leu Leu Gln Asp Pro Asp Phe Asn Thr Ile Asn Ser Thr
705 710 715 720
Glu Glu Asn Gly Trp Lys Ala Ser Asn Gly Val Thr Ile Ser Glu Gly
725 730 735
Gly Pro Phe Tyr Lys Gly Arg Ala Leu Gln Leu Ala Ser Ala Arg Glu
740 745 750
Asn Tyr Pro Thr Tyr Ile Tyr Gln Lys Val Asp Ala Ser Glu Leu Lys
755 760 765
Pro Tyr Thr Arg Tyr Arg Ser Asp Gly Phe Val Lys Ser Ser Gln Asp
770 775 780
Leu Glu Ile Asp Leu Ile His His His Lys Val His Leu Val Lys Asn
785 790 795 800
Val Pro Asp Asn Leu Val Ser Asp Thr Tyr Pro Asp Asp Ser Cys Ser
805 810 815
Gly Ile Asn Arg Cys Gln Glu Gln Gln Met Val Asn Ala Gln Leu Glu
820 825 830
Thr Glu His His His Pro Met Asp Cys Cys Glu Ala Ala Gln Thr His
835 840 845
Glu Phe Ser Ser Tyr Ile Asp Thr Gly Asp Leu Asn Ser Ser Val Asp
850 855 860
Gln Gly Ile Trp Ala Ile Phe Lys Val Arg Thr Thr Asp Gly Tyr Ala
865 870 875 880
Thr Leu Gly Asn Leu Glu Leu Val Glu Val Gly Pro Leu Ser Gly Glu
885 890 895
Ser Leu Glu Arg Glu Gln Arg Asp Asn Thr Lys Trp Ser Ala Glu Leu
900 905 910
Gly Arg Lys Arg Ala Glu Thr Asp Arg Val Tyr Gln Asp Ala Lys Gln
915 920 925
Ser Ile Asn His Leu Phe Val Asp Tyr Gln Asp Gln Gln Leu Asn Pro
930 935 940
Glu Ile Gly Met Ala Asp Ile Met Asp Ala Gln Asn Leu Val Ala Ser
945 950 955 960
Ile Ser Asp Val Tyr Ser Asp Ala Val Leu Gln Ile Pro Gly Ile Asn
965 970 975
Tyr Glu Ile Tyr Thr Glu Leu Ser Asn Arg Leu Gln Gln Ala Ser Tyr
980 985 990
Leu Tyr Thr Ser Arg Asn Ala Val Gln Asn Gly Asp Phe Asn Asn Gly
995 1000 1005
Leu Asp Ser Trp Asn Ala Thr Ala Gly Ala Ser Val Gln Gln Asp Gly
1010 1015 1020
Asn Thr His Phe Leu Val Leu Ser His Trp Asp Ala Gln Val Ser Gln
1025 1030 1035 1040
Gln Phe Arg Val Gln Pro Asn Cys Lys Tyr Val Leu Arg Val Thr Ala
1045 1050 1055
Glu Lys Val Gly Gly Gly Asp Gly Tyr Val Thr Ile Arg Asp Gly Ala
1060 1065 1070
His His Thr Glu Thr Leu Thr Phe Asn Ala Cys Asp Tyr Asp Ile Asn
1075 1080 1085
Gly Thr Tyr Val Thr Asp Asn Thr Tyr Leu Thr Lys Glu Val Ile Phe
1090 1095 1100
Tyr Ser His Thr Glu His Met Trp Val Glu Val Asn Glu Thr Glu Gly
1105 1110 1115 1120
Ala Phe His Ile Asp Ser Ile Glu Phe Val Glu Thr Glu Lys
1125 1130
〈210〉 77
〈211〉 3411
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 77
atggataaca atccgaacat caatgaatgc attccttata attgtttaag taaccctgaa 60
gtagaagtat taggtggaga aagaggaaat gttagaactg gactacaaac tggaattgat 120
attgttgcag tagtagtagg tgctttaggt ggaccagttg gtggcatact cactggtttt 180
ctttctactc tttttggttt tctttggcca tctaatgatc aagcagtatg ggaagctttt 240
atagaacaaa tggaagaact gattgaacaa aggatatcag atcaagtagt aaggactgca 300
ctcgatgact taactggaat tcaaaattat tataatcaat atctaatagc attaaaggaa 360
tgggaggaaa gaccaaacgg cgtaagagca aacttagttt tgcaaagatt tgaaatcttg 420
cacgcgctat ttgtaagtag tatgccaagt tttggtagtg gccctggaag tcaaaggttt 480
caggcacaat tgttggttgt ttatgcgcaa gcagcaaatc ttcatttact attattagct 540
gatgctgaaa agtatggggc aagatgggga ctccgtgaat cccagatagg aaatttatat 600
tttaatgaac tacaaactcg tactcgagat tacaccaacc attgtgtaaa cgcgtataat 660
aacgggttag ccgggttacg aggaacgagc gctgaaagtt ggttaaagta ccatcaattc 720
cgcagagaag caaccttaat ggcaatggat ttgatagctt tatttccata ttataacacc 780
cggcgatatc caatcgcagt aaatcctcag cttacacgtg aggtatatac agatccatta 840
ggcgttcctt ctgaagaatc aagtttattt ccagaattga gatgcttaag atggcaagag 900
acttctgcca tgactttttc aaatttggaa aatgcaataa tttcgtcacc acatctattt 960
gacacaataa acaatttaat gatttatacc ggttcctttt ccgttcacct aaccaatcaa 1020
ttaattgaag ggtggattgg acattctgta actagtagtt tgttggccag tggaccaaca 1080
acagtactga gaagaaatta cggtagcacg acatctattg taaactattt tagttttaat 1140
gatcgtgatg tttatcagat taatacgaga tcacatactg ggttgggatt ccagaacgca 1200
cctttatttg gaatcactag agctcaattt tacccaggtg ggacttattc agtaactcaa 1260
cgaaatgcat taacatgtga acaaaattat aattcaattg atgagttacc gagcctagac 1320
ccaaatgaac ctatcagtag aagttatagt catagattat ctcatattac ctcctatttg 1380
catcgtgtat tgactattga tggtattaat atatattcag gaaatctccc tacttatgta 1440
tggacccatc gcgatgtgga ccttacaaac acgattaccg cagatagaat tacacaacta 1500
ccattggtaa agtcatttga aatacctgcg ggtactactg tcgtaagagg accaggtttt 1560
acaggagggg atatactccg aagaacaggg gttggtacat ttggaacaat aagggtaagg 1620
actactgccc ccttaacaca aagatatcgc ataagattcc gtttcgcttc taccacaaat 1680
ttgttcattg gtataagagt tggtgataga caagtaaatt attttgactt cggaagaaca 1740
atgaacagag gagatgaatt aaggtacgaa tcttttgcta caagggagtt tactactgat 1800
tttaatttta gacaacctca agaattaatc tcagtgtttg caaatgcatt tagcgctggt 1860
caagaagttt attttgatag aattgagatt atccccgtta atcccgcacg agaggcgaaa 1920
gaggatctag aagcagcaaa gaaagcggtg gcgagcttgt ttacacgcac aagggacgga 1980
ttacaagtaa atgtgaaaga ttatcaagtc gatcaagcgg caaatttagt gtcatgctta 2040
tcagatgaac aatatgggta tgacaaaaag atgttattgg aagcggtacg cgcggcaaaa 2100
cgcctcagcc gagaacgtaa cttacttcag gatccagatt ttaatacaat caatagtaca 2160
gaagaaaatg gatggaaagc aagtaacggc gttactatta gtgagggcgg tccattctat 2220
aaaggccgtg cacttcagct agcaagtgca cgagaaaatt atccaacata catttatcaa 2280
aaagtagatg catcggagtt aaaaccttat acacgttata gatcagatgg gttcgtgaag 2340
agtagtcaag atttagaaat tgatctcatt caccatcata aagtccatct tgtgaaaaat 2400
gtaccagata atttagtatc tgatacttac ccagatgatt cttgtagtgg aatcaatcga 2460
tgtcaggaac aacagatggt aaatgcgcaa ctggaaacag agcatcatca tccgatggat 2520
tgctgtgaag cagctcaaac acatgagttt tcttcctata ttgatacagg ggatttaaat 2580
tcgagtgtag accagggaat ctgggcgatc tttaaagttc gaacaaccga tggttatgcg 2640
acgttaggaa atcttgaatt ggtagaggtc ggaccgttat cgggtgaatc tttagaacgt 2700
gaacaaaggg ataatacaaa atggagtgca gagctaggaa gaaagcgtgc agaaacagat 2760
cgcgtgtatc aagatgccaa acaatccatc aatcatttat ttgtggatta tcaagatcaa 2820
caattaaatc cagaaatagg gatggcagat attatggacg ctcaaaatct tgtcgcatca 2880
atttcagatg tatatagcga tgccgtactg caaatccctg gaattaacta tgagatttac 2940
acagagctgt ccaatcgctt acaacaagca tcgtatctgt atacgtctcg aaatgcggtg 3000
caaaatgggg actttaacaa cgggctagat agctggaatg caacagcggg tgcatcggta 3060
caacaggatg gcaatacgca tttcttagtt ctttctcatt gggatgcaca agtttctcaa 3120
caatttagag tgcagccgaa ttgtaaatat gtattacgtg taacagcaga gaaagtaggc 3180
ggcggagacg gatacgtgac tatccgggat ggtgctcatc atacagaaac gcttacattt 3240
aatgcatgtg attatgatat aaatggcacg tacgtgactg ataatacgta tctaacaaaa 3300
gaagtgatat tctattcaca tacagaacac atgtgggtag aggtaaatga aacagaaggt 3360
gcatttcata tagatagtat tgaattcgtt gaaacagaaa agtaaggtac c 3411
〈210〉 78
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 78
Met Asn Lys Asn Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asp Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Asp Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu Arg Val Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Ser Ser Lys Val Lys Lys Asp Gly Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Thr Lys Glu Asn Val Lys Ala Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly His Ala Leu Ile Gly Phe Glu Ile Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Met Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Glu Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Glu Asp Tyr Gln Thr Ile Asn Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr Asn Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 79
〈211〉 2370
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 79
atgaacaaga ataatactaa attaagcaca agagccttac caagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaacatgat ttttaaaacg 120
gatacaggtg gtgatctaac cctagacgaa attttaaaga atcagcagtt actaaatgat 180
atttctggta aattggatgg ggtgaatgga agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatatta aaaattgcaa atgaacaaaa tcaagtttta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcgggtata tctacctaaa 360
attacctcta tgttgagtga tgtaatgaaa caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agagatttct gataagttgg atattattaa tgtaaatgta 480
cttattaact ctacacttac tgaaattaca cctgcgtatc aaaggattaa atatgtgaac 540
gaaaaatttg aggaattaac ttttgctaca gaaactagtt caaaagtaaa aaaggatggc 600
tctcctgcag atattcttga tgagttaact gagttaactg aactagcgaa aagtgtaaca 660
aaaaatgatg tggatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcatcgg aattaattac taaagaaaat 780
gtgaaagcaa gtggcagtga ggtcggaaat gtttataact tcttaattgt attaacagct 840
ctgcaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatcctcc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gacatgcatt gattgggttt 1080
gaaattagta atgattcaat tacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagtcg ataaggattc cttatcggaa gttatttatg gtgatatgga taaattattg 1200
tgcccagatc aatctgaaca aatctattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatgggg tgtatatgcc gttaggtgtc 1440
atcagtgaaa catttttgac tccgattaat gggtttggcc tccaagctga tgaaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aacagactta 1560
agcaataaag aaactaaatt gattgtcccg ccaagtggtt ttattagcaa tattgtagag 1620
aacgggtcca tagaagagga caatttagag ccgtggaaag caaataataa gaatgcgtat 1680
gtagatcata caggcggagt gaatggaact aaagctttat atgttcataa ggacggagga 1740
atttcacaat ttattggaga taagttaaaa ccgaaaactg agtatgtaat ccaatatact 1800
gttaaaggaa aaccttctat tcatttaaaa gatgaaaata ctggatatat tcattatgaa 1860
gatacaaata ataatttaga agattatcaa actattaata aacgttttac tacaggaact 1920
gatttaaagg gagtgtattt aattttaaaa agtcaaaatg gagatgaagc ttggggagat 1980
aactttatta ttttggaaat tagtccttct gaaaagttat taagtccaga attaattaat 2040
acaaataatt ggacgagtac gggatcaact aatattagcg gtaatacact cactctttat 2100
cagggaggac gagggattct aaaacaaaac cttcaattag atagtttttc aacttataga 2160
gtgtattttt ctgtgtccgg agatgctaat gtaaggatta gaaattctag ggaagtgtta 2220
tttgaaaaaa gatatatgag cggtgctaaa gatgtttctg aaatgttcac tacaaaattt 2280
gagaaagata acttttatat agagctttct caagggaata atttatatgg tggtcctatt 2340
gtacattttt acgatgtctc tattaagtaa 2370
〈210〉 80
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 80
Met Asn Lys Asp Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asp Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Asp Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Glu Val Asn Asn Lys Leu Glu Ala Ile Ser Thr
100 105 110
Ile Phe Arg Val Tyr Leu Pro Lys Asn Thr Ser Arg Gly Gly Gly Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Met Glu Asn Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Val Lys Trp Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Ser Ser Lys Val Lys Lys Asp Gly Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Thr Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly His Ala Leu Ile Gly Phe Glu Ile Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Met Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Glu Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Xaa Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Glu Asp Tyr Gln Thr Ile Asn Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr Asn Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 81
〈211〉 2375
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 81
atgaacaagg ataatactaa attaagcaca agagccttac caagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaacatgat ttttaaaacg 120
gatacaggtg gtgatctaac cctagacgaa attttaaaga atcagcagtt actaaatgat 180
atttctggta aattggatgg ggtgaatgga agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatatta aaaattgcaa atgaacaaaa tcaagtttta 300
aatgaggtta ataacaaact cgaggcgata agtacgattt ttcgggtata tttacctaaa 360
aatacctcta gggggggggg ggtaatgaaa caaaattatg cgctaagtct gcaaatggaa 420
aacttgagta aacaattaca agagatttct gttaagtggg atattattaa tgtaaatgta 480
cttattaact ctacacttac cgaaattaca cctgcgtatc aaaggattaa atatgtgaac 540
gaaaaatttg aggaattaac ttttgctaca gaaactagtt caaaagtaaa aaaggatggc 600
tctcccgcag atattcttga tgagttaact gagttaactg aactagcgaa aagtgtaaca 660
aaaaatgatg tggatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcatcgg aattaattac taaagaaaat 780
gtgaaaacaa gtggcagtga ggtcggaaat gtttataact tcttaattgt attaacagct 840
ctgcaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatcctcc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gacatgcatt gattgggttt 1080
gaaattagta atgattcaat tacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagtcg ataaggattc cttatcggaa gttatttatg gtgatatgga taaattattg 1200
tgcccagatc aatctgaaca aatctattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatgggg tgtatatgcc gttaggtgtc 1440
atcagtgaaa catttttgac tccgattaat gggtttggcc tccaagctga tgaaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aaccgactta 1560
agcaataaag aaactaaatt gatcgtcccg ccaagtggtt ttattagcsa tattgtagag 1620
aacgggtcca tagaagagga caatttagag ccgtggaaag caaataataa gaatgcgtat 1680
gtagatcata caggcggagt gaatggaact aaagctttat atgttcataa ggacggagga 1740
atttcacaat ttattggaga taagttaaaa ccgaaaactg agtatgtaat ccaatatact 1800
gttaaaggaa aaccttctat tcatttaaaa gatgaaaata ctggatatat tcattatgaa 1860
gatacaaata ataatttaga agattatcaa actattaata aacgttttac tacaggaact 1920
gatttaaagg gagtgtattt aattttaaaa agtcaaaatg gagatgaagc ttggggagat 1980
aactttatta ttttggaaat tagtccttct gaaaagttat taagtccaga attaattaat 2040
acaaataatt ggacgagtac gggatcaact aatattagcg gtaatacact cactctttat 2100
cagggaggac gagggattct aaaacaaaac cttcaattag atagtttttc aacttataga 2160
gtgtattttt ctgtgtccgg agatgctaat gtaaggatta gaaattctag ggaagtgtta 2220
tttgaaaaaa gatatatgag cggtgctaaa gatgtttctg aaatgttcac tacaaaattt 2280
gagaaagata acttttatat agagctttct caagggaata atttatatgg tggtcctatt 2340
gttcattttt acgatgtctc tattaagtaa cccaa 2375
〈210〉 82
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 82
Met Asn Lys Asn Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asn Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Glu Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu His Ile Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Thr Leu Lys Val Lys Lys Asp Ser Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Ala Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly Tyr Ala Leu Val Gly Phe Glu Met Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Thr Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Gly Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Leu Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Phe Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Lys Asp Tyr Gln Thr Ile Thr Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr His Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Asn
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 83
〈211〉 2375
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 83
atgaacaaga ataatactaa attaagcaca agagccttac caagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaatatgat ttttaaaacg 120
gatacaggtg gtaatctaac cttagatgaa atcctaaaga atcagcagtt actaaatgag 180
atttctggta aattggatgg ggtaaatggg agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatctta aaaattgcaa atgaacagaa tcaagtctta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcatatata tctacctaaa 360
attacatcta tgttaagtga tgtaatgaag caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agaaatttct gataaattag atattattaa cgtaaatgtt 480
cttattaact ctacacttac tgaaattaca cctgcatatc aacggattaa atatgtgaat 540
gaaaaatttg aagaattaac ttttgctaca gaaaccactt taaaagtaaa aaaggatagc 600
tcgcctgctg atattcttga tgagttaact gaattaactg aactagcgaa aagtgttaca 660
aaaaatgacg tggatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcttcag aattaattgc taaagaaaat 780
gtgaaaacaa gtggcagtga agtaggaaat gtttataact tcttaattgt attaacagct 840
ctacaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatccttc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gatatgcatt ggttgggttt 1080
gaaatgagca atgattcaat cacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagttg ataaggattc cttatcggaa gttatttatg gtgatacgga taaattattg 1200
tgtccagatc aatctgaaca aatatattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatggag tgtatatgcc attaggtgtc 1440
atcagtgaaa catttttgac tccgataaat gggtttggcc tccaagctga tggaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aacagactta 1560
agcaataaag aaactaaatt gatcgtcctg ccaagtggtt ttattagcaa tattgtagag 1620
aacgggtcca tagaagagga caatttagag ccgtggaaag caaataataa gaatgcgtat 1680
gtagatcata caggcggagt gaatggaact aaagctttat atgttcataa ggacggagga 1740
ttttcacaat ttattggaga taagttaaaa ccgaaaactg agtatgtaat ccaatatact 1800
gttaaaggaa aaccttctat tcatttaaaa gatgaaaata ctggatatat tcattatgaa 1860
gatacaaata ataatttaaa agattatcaa actattacta aacgttttac tacaggaact 1920
gatttaaagg gagtgtattt aattttaaaa agtcaaaatg gagatgaagc ttggggagat 1980
aactttatta ttttggaaat tagtccttct gaaaagttat taagtccaga attaattaat 2040
acaaataatt ggacgagtac gggatcaact catattagcg gtaatacact cactctttat 2100
cagggaggac gaggaattct aaaacaaaac cttcaattag atagtttttc aacttataga 2160
gtgtattttt ctgtgtccgg agatgctaat gtaaggatta gaaattctag ggaagtgtta 2220
tttgaaaaaa gatatatgag cggtgctaaa gatgtttctg aaatgttcac tacaaaattt 2280
gagaaagata acttttatat agagctttct caagggaata atttatatgg tggtcctatt 2340
gtacatttta acgatgtctc tattaagtaa cccaa 2375
〈210〉 84
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 84
Met Asn Lys Asn Asn Thr Lys Leu Ser Ala Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asn Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Glu Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu His Ile Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Ser Ser Lys Val Lys Lys Asp Ser Pro Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Ala Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly Tyr Ala Leu Val Gly Phe Glu Met Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Thr Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Gly Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Phe Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Lys Asp Tyr Gln Thr Ile Thr Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr His Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Gly Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 85
〈211〉 2375
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 85
atgaacaaga ataatactaa attaagcgca agggccctac cgagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaatatgat ttttaaaacg 120
gatacaggtg gtaatctaac cttagatgaa atcctaaaga atcagcagtt actaaatgag 180
atttctggta aattggatgg ggtaaatggg agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatctta aaaattgcaa atgaacagaa tcaagtctta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcatatata tctacctaaa 360
attacatcta tgttaagtga tgtaatgaaa caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agaaatttct gataaattag atattattaa cgtaaatgtc 480
cttattaact ctacacttac tgaaattaca cctgcatatc aacggattaa atatgtgaat 540
gaaaaatttg aagaattaac ttttgctaca gaaactagtt caaaagtaaa aaaggatagc 600
ccccctgctg atattcttga tgagttaact gaattaactg aactagcgaa aagtgtaaca 660
aaaaatgacg tggatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcttcag aattaattgc taaagaaaat 780
gtgaaaacaa gtggcagtga agtaggaaat gtttataatt tcttaattgt attaacagct 840
ctacaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatccttc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gatatgcatt ggttggtttt 1080
gaaatgagca atgattcaat cacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagttg ataaggattc cttatcggag gttatttatg gtgatacgga taaattattg 1200
tgtccagatc aatctgaaca aatatattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatggag tgtatatgcc attaggtgtc 1440
atcagtgaaa catttttgac tccgataaat gggtttggcc tccaagctga tggaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aacagactta 1560
agcaataaag aaactaaatt gatcgtcccg ccaagtggtt ttattagcaa tattgtagag 1620
aacgggtcca tagaagagga caatttagag ccgtggaaag caaataataa gaatgcgtat 1680
gtagatcata caggcggagt gaatggaact aaagctttat atgttcataa ggacggagga 1740
ttttcacaat ttattggaga taagttaaaa ccgaaaactg agtatgtaat ccaatatact 1800
gttaaaggaa aaccttctat tcatttaaaa gatgaaaata ctggatatat tcattatgaa 1860
gatacaaata ataatttaaa agattatcaa actattacta aacgttttac tacaggaact 1920
gatttaaagg gagtgtattt aattttaaaa agtcaaaatg gagatgaagc ttggggagat 1980
aactttatta ttttggaaat tagtccttct gaaaagttat taagtccaga attaattaat 2040
acaaataatt ggacgagtac gggatcaact catattagcg gtaatacact cactctttat 2100
cagggaggac gaggaattct aaaacaaaac cttcaattag atagtttttc aacttataga 2160
gtgtattttt ctgtgtccgg agatgctaat gtaaggatta gaaattctag ggaagtgtta 2220
tttgaaaaag gatatatgag cggtgctaaa gatgtttctg aaatgttcac tacaaaattt 2280
gagaaagata acttttatat agagctttct caagggaata atttatatgg tggtcctatt 2340
gtacattttt acgatgtctc tattaagtaa ccaag 2375
〈210〉 86
〈211〉 759
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 86
Met Asn Lys Asn Asn Thr Lys Leu Ser Ala Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asn Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Glu Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu Arg Ile Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Asn Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Xaa Ser Lys Val Lys Lys Asp Gly Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Ile Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Ile Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Xaa Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly Tyr Ala Leu Val Gly Phe Glu Met Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Thr Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Glu Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser His
530 535 540
Arg Arg Gly Gln Phe Arg Ala Val Glu Ser Lys Glu Cys Val Cys Arg
545 550 555 560
Ser Tyr Arg Arg Ser Glu Trp Asn Ser Phe Ile Cys Ser Gly Arg Arg
565 570 575
Asn Phe Thr Ile Tyr Trp Arg Val Lys Thr Glu Asn Val Cys Asn Pro
580 585 590
Ile Tyr Cys Arg Lys Thr Phe Tyr Ser Phe Lys Arg Lys Tyr Trp Ile
595 600 605
Tyr Ser Leu Arg Tyr Lys Phe Lys Arg Leu Ser Asn Tyr Tyr Thr Phe
610 615 620
Tyr Tyr Arg Asn Phe Lys Gly Ser Val Phe Asn Phe Lys Lys Ser Lys
625 630 635 640
Trp Arg Ser Leu Gly Arg Leu Tyr Tyr Phe Gly Asn Ser Phe Lys Val
645 650 655
Ile Lys Ser Arg Ile Asn Tyr Lys Leu Asp Glu Tyr Gly Ile Asn Ser
660 665 670
Tyr Arg Tyr Thr His Ser Leu Ser Gly Arg Thr Arg Asn Ser Lys Thr
675 680 685
Lys Pro Ser Ile Arg Phe Phe Asn Leu Ser Val Phe Phe Cys Val Arg
690 695 700
Arg Cys Cys Lys Asp Lys Phe Gly Ser Val Ile Lys Lys Ile Tyr Glu
705 710 715 720
Arg Cys Arg Cys Phe Asn Val His Tyr Lys Ile Glu Arg Leu Leu Tyr
725 730 735
Arg Ala Phe Ser Arg Glu Phe Ile Trp Trp Ser Tyr Cys Thr Phe Leu
740 745 750
Arg Cys Leu Tyr Val Thr Gln
755
〈210〉 87
〈211〉 2376
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 87
atgaacaaga ataatactaa attaagcgca agagccctac cgagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaatatgat ttttaaaacg 120
gatacaggtg gtaatctaac cttagatgaa atcctaaaga atcagcagtt actaaatgag 180
atttctggta aattggatgg ggtaaatggg agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatctta aaaattgcaa atgaacaaaa tcaagtctta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcggatata tctacctaaa 360
attacatcta tgttaagtga tgtaatgaac caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agaaatttct gataaattgg atattattaa tgtaaatgta 480
cttattaact ctacacttac tgaaattaca cctgcgtatc aaaggattaa atatgtgaac 540
gaaaaatttg aggaattaac ttttgctaca gaaactaktt caaaagtaaa aaaggatggc 600
tctcctgcag atattcttga tgagttaact gagttaactg aactagcgaa aagtgtaaca 660
aaaaatgatg tggatggttt tgaaatttac cttaatacat tccacgatgt aatggtagga 720
aataatttaa tcgggcgttc agctttaaaa actgcatcgg aattaattas taaagaaaat 780
gtgaaaacaa gtggcagtga ggtaggaaat gtttataact tcttaattgt attaacagct 840
ctacaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatccttc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gatatgcatt ggttggtttt 1080
gaaatgagca atgattcaat cacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagttg ataaggattc cttatcggag gttatttatg gtgatacgga taaattattg 1200
tgtccagatc aatctgaaca aatatattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatggag tgtatatgcc gttaggtgtc 1440
atcagtgaaa catttttgac tccgattaat gggtttggcc tccaagctga tgaaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aacagactta 1560
agcaataaag aaactaaatt gatcgtcccg ccaagtggtt ttattagcaa tattgtagag 1620
aacgggtccc atagaagagg acaatttaga gccgtggaaa gcaaataata agaatgcgta 1680
tgtagatcat acaggcggag tgaatggaac taaagcttta tatgttcata aggacggagg 1740
aatttcacaa tttattggag ataagttaaa accgaaaact gagtatgtaa tccaatatac 1800
tgttaaagga aaaccttcta ttcatttaaa agatgaaaat actggatata ttcattatga 1860
agatacaaat aataatttaa aagattatca aactattact aaacgtttta ctacaggaac 1920
tgatttaaag ggagtgtatt taattttaaa aagtcaaaat ggagatgaag cttggggaga 1980
taactttatt attttggaaa ttagtccttc tgaaaagtta ttaagtccag aattaattaa 2040
tacaaataat tggacgagta cgggatcaac tcatattagc ggtaatacac tcactcttta 2100
tcagggagga cgaggaattc taaaacaaaa ccttcaatta gatagttttt caacttatag 2160
agtgtatttt tctgtgtccg gagatgctaa tgtaaggatt agaaattcta gggaagtgtt 2220
atttgaaaaa agatatatga gcggtgctaa agatgtttct gaaatgttca ctacaaaatt 2280
tgagaaagat aacttttata tagagctttc tcaagggaat aatttatatg gtggtcctat 2340
tgtacatttt tacgatgtct ctattaagta acccaa 2376
〈210〉 88
〈211〉 511
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 88
Tyr Leu Ser Lys Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile
1 5 10 15
Asn Val Asn Val Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala
20 25 30
Tyr Gln Arg Ile Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe
35 40 45
Ala Thr Glu Thr Thr Leu Lys Val Lys Lys Asp Ser Ser Pro Ala Asp
50 55 60
Ile Leu Asp Glu Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr
65 70 75 80
Lys Asn Asp Val Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp
85 90 95
Val Met Val Gly Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala
100 105 110
Ser Glu Leu Ile Ala Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val
115 120 125
Gly Asn Val Tyr Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys
130 135 140
Ala Phe Leu Thr Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp
145 150 155 160
Ile Asp Tyr Thr Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu
165 170 175
Glu Phe Arg Val Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn
180 185 190
Pro Asn Tyr Ala Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile
195 200 205
Val Glu Ala Lys Pro Gly Tyr Ala Leu Val Gly Phe Glu Met Ser Asn
210 215 220
Asp Ser Ile Thr Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn
225 230 235 240
Tyr Gln Val Asp Lys Asp Pro Leu Ser Glu Val Ile Tyr Gly Asp Thr
245 250 255
Asp Lys Leu Leu Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn
260 265 270
Asn Ile Val Phe Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr
275 280 285
Lys Lys Met Lys Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp
290 295 300
Ser Ser Thr Gly Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser
305 310 315 320
Glu Ala Glu Tyr Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met
325 330 335
Pro Leu Gly Val Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe
340 345 350
Gly Leu Gln Ala Asp Gly Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys
355 360 365
Ser Tyr Leu Arg Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu
370 375 380
Thr Lys Leu Ile Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu
385 390 395 400
Asn Gly Ser Ile Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn
405 410 415
Lys Asn Ala Tyr Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala
420 425 430
Leu Tyr Val His Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys
435 440 445
Leu Lys Pro Lys Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys
450 455 460
Pro Ser Ile His Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu
465 470 475 480
Asp Thr Asn Asn Asn Leu Lys Asp Tyr Gln Thr Ile Thr Lys Arg Phe
485 490 495
Thr Thr Gly Thr Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser
500 505 510
〈210〉 89
〈211〉 1533
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 89
tacttaagta aacaattgca agaaatttct gataaattag atattattaa cgtaaatgtt 60
cttattaact ctacacttac tgaaattaca cctgcatatc aacggattaa atatgtgaat 120
gaaaaatttg aagaattaac ttttgctaca gaaaccactt taaaagtaaa aaaggatagc 180
tcgcctgctg atattcttga tgagttaact gaattaactg aactagcgaa aagtgttaca 240
aaaaatgacg ttgatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 300
aataatttat tcgggcgttc agctttaaaa actgcttcag aattaattgc taaagaaaat 360
gtgaaaacaa gtggcagtga agtaggaaat gtttataatt tcttaattgt attaacagct 420
ctacaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 480
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 540
aacatcctyc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 600
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gatatgcatt ggttggtttt 660
gaaatgagca atgattcaat cacagtatta aaagtatatg aggctaagct aaaacaaaat 720
tatcaagttg ataaggatcc cttatcggag gttatttatg gtgatacgga taaattattg 780
tgtccagatc aatctgaaca aatatattat acaaataaca tagtatttcc aaatgaatat 840
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 900
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 960
gaagcggagt atagaacgtt aagtgctaat gatgatggag tgtatatgcc attaggtgtc 1020
atcagtgaaa catttttgac tccgattaat gggtttggcc tccaagctga tggaaattca 1080
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aacagactta 1140
agcaataaag aaactaaatt gatcgtcccg ccaagtggtt ttattagcaa tattgtagag 1200
aacgggtcca tagaagagga caatttagag ccgtggaaag caaataataa gaatgcgtat 1260
gtagatcata caggcggagt gaatggaact aaagctttat atgttcataa ggacggagga 1320
atttcacaat ttattggaga taagttaaaa ccgaaaactg agtatgtaat ccaatatact 1380
gttaaaggaa aaccttctat tcatttaaaa gatgaaaata ctggatatat tcattatgaa 1440
gatacaaata ataatttaaa agattatcaa actattacta aacgttttac tacaggaact 1500
gatttaaagg gagtgtattt aattttaaaa agt 1533
〈210〉 90
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 90
Met Asn Lys Asn Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asp Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Asp Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asp Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu Arg Val Tyr Leu Pro Lys Ile Thr Xaa Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Ser Ser Lys Val Lys Lys Asp Gly Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Thr Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly His Ala Leu Val Gly Phe Glu Ile Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Met Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Pro Gln Ala
485 490 495
Asp Glu Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Lys Leu Leu Leu Ala Thr Asp Phe Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Leu Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Xaa Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Gly Lys Ala Asn Asn Arg Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Glu Asp Tyr Gln Thr Ile Thr Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr Asn Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Ile Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Asn Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 91
〈211〉 2367
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 91
atgaacaaga ataatactaa attaagcaca agagccttac caagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaacatgat ttttaaaacg 120
gatacaggtg gtgatctaac cctagacgaa attttaaaga atcagcagtt actaaatgat 180
atttctggta aattggatgg ggtgaatgga agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa agaaatatta aaaattgcaa atgaacaaaa tcaagtttta 300
aatgatgttg ataacaaact cgatgcgata aatacgatgc ttcgggtata tctacctaaa 360
attaccctat gttgagtgat gtaatgaaac aaaattatgc gctaagtctg caaatagaat 420
acttaagtaa acaattgcaa gagatttctg ataagttgga tattattaat gtaaatgtac 480
ttattaactc tacacttact gaaattacac ctgcgtatca aaggattaaa tatgtgaacg 540
aaaaatttga ggaattaact tttgctacag aaactagttc aaaagtaaaa aaggatggct 600
ctcctgcaga tattcttgat gagttaactg agttaactga actagcgaaa agtgtaacaa 660
aaaatgatgt ggatggtttt gaattttacc ttaatacatt ccacgatgta atggtaggaa 720
ataatttatt cgggcgttca gctttaaaaa ctgcatcgga attaattact aaagaaaatg 780
tgaaaacaag tggcagtgag gtcggaaatg tttataactt cttaattgta ttaacagctc 840
tgcaagcaaa agcttttctt actttaacaa catgccgaaa attattaggc ttagcagata 900
ttgattatac ttctattatg aatgaacatt taaataagga aaaagaggaa tttagagtaa 960
acatcctccc tacactttct aatacttttt ctaatcctaa ttatgcaaaa gttaaaggaa 1020
gtgatgaaga tgcaaagatg attgtggaag ctaaaccagg acatgcattg gttgggtttg 1080
aaattagtaa tgattcaatt acagtattaa aagtatatga ggctaagcta aaacaaaatt 1140
atcaagttga taaggattcc ttatcggaag ttatttatgg tgatatggat aaattattgt 1200
gcccagatca atctgaacaa atctattata caaataacat agtatttcca aatgaatatg 1260
taattactaa aattgatttt actaaaaaaa tgaaaacttt aagatatgag gtaacagcga 1320
atttttatga ttcttctaca ggagaaattg acttaaataa gaaaaaagta gaatcaagtg 1380
aagcggagta tagaacgtta agtgctaatg atgatggagt gtatatgccg ttaggtgtca 1440
tcagtgaaac atttttgact ccgattaatg ggtttggccc ccaagctgat gaaaattcaa 1500
gattaattac tttaacatgt aaatcatatt taagaaaact actgctagca acagacttta 1560
gcaataaaga aactaaattg atcctcccgc caagtggttt tattagcaat attgtagaaa 1620
cgggtccata gaagaggaca atttagagcc ggggaaagca aataatagga atgcgtatgt 1680
agatcataca ggcggagtga atggaactaa agctttatat gttcataagg acggaggaat 1740
ttcacaattt attggagata agttaaaacc gaaaactgag tatgtaatcc aatatactgt 1800
taaaggaaaa ccttctattc atttaaaaga tgaaaatact ggatatattc attatgaaga 1860
tacaaataat aatttagaag attatcaaac tattactaaa cgttttacta caggaactga 1920
tttaaaggga gtgtatttaa ttttaaaaag tcaaaatgga gatgaagctt ggggagataa 1980
ctttattatt ttggaaatta gtccttctga aaagttatta agtccagaat taattaatac 2040
aaataattgg acgagtacgg gatcaactaa tattagcggt aatacactca ctctttatca 2100
gggaggacga ggaattctaa aacaaaacct tcaattagat agtttttcaa cttatagagt 2160
gtatttttct gtgtccggag atgctaatgt aaggattaga aattctaggg aagtgttatt 2220
tgaaaaaaga tatatgagcg gtgctaaaga tgtttctgaa attttcacta caaaatttga 2280
gaaagataac ttttatatag agctttctca agggaataat ttaaatggtg gccctattgt 2340
acatttttac gatgtctcta ttaagta 2367
〈210〉 92
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 92
Met Asn Lys Asn Asn Thr Lys Leu Ser Ala Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asn Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Glu Ile Ser Gly Lys
50 55 60
Leu Gly Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu His Ile Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Thr Leu Lys Val Lys Lys Asp Ser Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Ala Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly Tyr Ala Leu Val Gly Phe Glu Met Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Thr Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Gly Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Lys Asp Tyr Gln Thr Ile Thr Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr His Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 93
〈211〉 2369
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 93
atgaacaaga ataatactaa attaagcgca agggccctac cgagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaatatgat ttttaaaacg 120
gatacaggtg gtaatctaac cttagatgaa atcctaaaga atcagcagtt actaaatgag 180
atttctggta aattgggggg ggtaaatggg agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatctta aaaattgcaa atgaacaaat caagtcttaa 300
atgatgttaa taacaaactc gatgcgataa atacgatgct tcatatatat ctacctaaaa 360
ttacatctat gttaagtgat gtaatgaagc aaaattatgc gctaagtctg caaatagaat 420
acttaagtaa acaattgcaa gaaatttctg ataaattaga tattattaac gtaaatgttc 480
ttattaactc tacacttact gaaattacac ctgcatatca acggattaaa tatgtgaatg 540
aaaaatttga agaattaact tttgctacag aaaccacttt aaaagtaaaa aaggatagct 600
cgcctgctga tattcttgat gagttaactg aattaactga actagcgaaa agtgttacaa 660
aaaatgacgt tgatggtttt gaattttacc ttaatacatt ccacgatgta atggtaggaa 720
ataatttatt cgggcgttca gctttaaaaa ctgcttcaga attaattgct aaagaaaatg 780
tgaaaacaag tggcagtgaa gtaggaaatg tttataattt cttaattgta ttaacagctc 840
tacaagcaaa agcttttctt actttaacaa catgccgaaa attattaggc ttagcagata 900
ttgattatac ttctattatg aatgaacatt taaataagga aaaagaggaa tttagagtaa 960
acatccttcc tacactttct aatacttttt ctaatcctaa ttatgcaaaa gttaaaggaa 1020
gtgatgaaga tgcaaagatg attgtggaag ctaaaccagg atatgcattg gttggttttg 1080
aaatgagcaa tgattcaatc acagtattaa aagtatatga ggctaagcta aaacaaaatt 1140
atcaagttga taaggattcc ttatcggagg ttatttatgg tgatacggat aaattattgt 1200
gtccagatca atctgaacaa atatattata caaataacat agtatttcca aatgaatatg 1260
taattactaa aattgatttc actaaaaaaa tgaaaacttt aagatatgag gtaacagcga 1320
atttttatga ttcttctaca ggagaaattg acttaaataa gaaaaaagta gaatcaagtg 1380
aagcggagta tagaacgtta agtgctaatg atgatggagt gtatatgcca ttaggtgtca 1440
tcagtgaaac atttttgact ccgataaatg ggtttggcct ccaagctgat ggaaattcaa 1500
gattaattac tttaacatgt aaatcatatt taagagaact actgctagca acagacttaa 1560
gcaataaaga aactaaattg attgtcccgc caagtggttt tattagcaat attgtagaga 1620
acgggtccat agaagaggac aatttagagc cgtggaaagc aaataataag aatgcgtatg 1680
tagatcatac aggcggagtg aatggaacta aagctttata tgttcataag gacggaggaa 1740
tttcacaatt tattggagat aagttaaaac cgaaaactga gtatgtaatc caatatactg 1800
ttaaaggaaa accttctatt catttaaaag atgaaaatac tggatatatt cattatgaag 1860
atacaaataa taatttaaaa gattatcaaa ctattactaa acgttttact acaggaactg 1920
atttaaaggg agtgtattta attttaaaaa gtcaaaatgg agatgaagct tggggagata 1980
actttattat tttggaaatt agtccttctg aaaagttatt aagtccagaa ttaattaata 2040
caaataattg gacgagtacg ggatcaactc atattagcgg taatacactc actctttatc 2100
agggaggacg aggaattcta aaacaaaacc ttcaattaga tagtttttca acttatagag 2160
tgtatttttc tgtgtccgga gatgctaatg taaggattag aaattctagg gaagtgttat 2220
ttgaaaaaag atatatgagc ggtgctaaag atgtttctga aatgttcact acaaaatttg 2280
agaaagataa cttttatata gagctttctc aagggaataa tttatatggt ggtcctattg 2340
tacattttta cgatgtctct attaagtaa 2369
〈210〉 94
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 94
Met Asn Lys Asn Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asp Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Asp Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu Arg Val Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Ser Ser Lys Val Lys Lys Asp Gly Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Ala Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Thr Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly His Ala Leu Ile Gly Phe Glu Ile Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Met Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Glu Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Glu Asp Tyr Gln Thr Ile Asn Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr Asn Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 95
〈211〉 2370
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 95
ttgaacaaga ataatactaa attaagcaca agagccttac caagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaacatgat ttttaaaacg 120
gatacaggtg gtgatctaac cctagacgaa attttaaaga atcagcagtt actaaatgat 180
atttctggta aattggatgg ggtgaatgga agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatatta aaaattgcaa atgaacaaaa tcaagtttta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcgggtata tctacctaaa 360
attacctcta tgttgagtga tgtaatgaaa caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agagatttct gataagttgg atattattaa tgtaaatgta 480
cttattaact ctacacttac tgaaattaca cctgcgtatc aaaggattaa atatgtgaac 540
gaaaaatttg aggaattaac ttttgctaca gaaactagtt caaaagtaaa aaaggatggc 600
tctcctgcag atattcttga tgagttagct gagttaactg aactagcgaa aagtgtaaca 660
aaaaatgatg tggatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcatcgg aattaattac taaagaaaat 780
gtgaaaacaa gtggcagtga ggtcggaaat gtttataact tcttaattgt attaacagct 840
ctgcaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatcctcc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gacatgcatt gattgggttt 1080
gaaattagta atgattcaat tacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagtcg ataaggattc cttatcggaa gttatttatg gtgatatgga taaattattg 1200
tgcccagatc aatctgaaca aatctattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatgggg tgtatatgcc gttaggtgtc 1440
atcagtgaaa catttttgac tccgattaat gggtttggcc tccaagctga tgaaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aacagactta 1560
agcaataaag aaactaaatt gattgtcccg ccaagtggtt ttattagcaa tattgtagag 1620
aacgggtcca tagaagagga caatttagag ccgtggaaag caaataataa gaatgcgtat 1680
gtagatcata caggcggagt gaatggaact aaagctttat atgttcataa ggacggagga 1740
atttcacaat ttattggaga taagttaaaa ccgaaaactg agtatgtaat ccaatatact 1800
gttaaaggaa aaccttctat tcatttaaaa gatgaaaata ctggatatat tcattatgaa 1860
gatacaaata ataatttaga agattatcaa actattaata aacgttttac tacaggaact 1920
gatttaaagg gagtgtattt aattttaaaa agtcaaaatg gagatgaagc ttggggagat 1980
aactttatta ttttggaaat tagtccttct gaaaagttat taagtccaga attaattaat 2040
acaaataatt ggacgagtac gggatcaact aatattagcg gtaatacact cactctttat 2100
cagggaggac gagggattct aaaacaaaac cttcaattag atagtttttc aacttataga 2160
gtgtattttt ctgtgtccgg agatgctaat gtaaggatta gaaattctag ggaagtgtta 2220
tttgaaaaaa gatatatgag cggtgctaaa gatgtttctg aaatgttcac tacaaaattt 2280
gagaaagata acttttatat agagctttct caagggaata atttatatgg tggtcctatt 2340
gtacattttt acgatgtctc tattaagtaa 2370
〈210〉 96
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 96
Met Asn Lys Asn Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asp Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Asp Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu Arg Val Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Ser Ser Lys Val Lys Lys Asp Gly Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Thr Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly His Ala Leu Ile Gly Phe Glu Ile Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Met Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Asn Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Glu Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Glu Asp Tyr Gln Thr Ile Asn Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr Asn Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 97
〈211〉 2374
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 97
atgaacaaga ataatactaa attaagcaca agagccttac caagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaacatgat ttttaaaacg 120
gatacaggtg gtgatctaac cctagacgaa attttaaaga atcagcagtt actaaatgat 180
atttctggta aattggatgg ggtgaatgga agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatatta aaaattgcaa atgaacaaaa tcaagtttta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcgggtata tctacctaaa 360
attacctcta tgttgagtga tgtaatgaaa caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agagatttct gataagttgg atattattaa tgtaaatgta 480
cttattaact ctacacttac tgaaattaca cctgcgtatc aaaggattaa atatgtgaac 540
gaaaaatttg aggaattaac ttttgctaca gaaactagtt caaaagtaaa aaaggatggc 600
tctcctgcag atattcttga tgagttaact gagttaactg aactagcgaa aagtgtaaca 660
aaaaatgatg tggatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcatcgg aattaattac taaagaaaat 780
gtgaaaacaa gtggcagtga ggtcggaaat gtttataact tcttaattgt attaacagct 840
ctgcaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatcctcc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gacatgcatt gattgggttt 1080
gaaattagta atgattcaat tacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagtcg ataaggattc cttatcggaa gttatttatg gtgatatgga taaattattg 1200
tgcccagatc aatctgaaca aatctattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaacgt cgaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatgggg tgtatatgcc gttaggtgtc 1440
atcagtgaaa catttttgac tccgattaat gggtttggcc tccaagctga tgaaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagagaac tactgctagc aacagactta 1560
agcaataaag aaactaaatt gatgtcccgc caagtggttt tattagcaat attgtagaga 1620
acgggtccat agaagaggac aatttagagc cgtggaaagc aaataataag aatgcgtatg 1680
tagatcatac aggcggagtg aatggaacta aagctttata tgttcataag gacggaggaa 1740
tttcacaatt tattggagat aagttaaaac cgaaaactga gtatgtaatc caatatactg 1800
ttaaaggaaa accttctatt catttaaaag atgaaaatac tggatatatt cattatgaag 1860
atacaaataa taatttagaa gattatcaaa ctattaataa acgttttact acaggaactg 1920
atttaaaggg agtgtattta attttaaaaa gtcaaaatgg agatgaagct tggggagata 1980
actttattat tttggaaatt agtccttctg aaaagttatt aagtccagaa ttaattaata 2040
caaataattg gacgagtacg ggatcaacta atattagcgg taatacactc actctttatc 2100
agggaggacg agggattcta aaacaaaacc ttcaattaga tagtttttca acttatagag 2160
tgtatttttc tgtgtccgga gatgctaatg taaggattag aaattctagg gaagtgttat 2220
ttgaaaaaag atatatgagc ggtgctaaag atgtttctga aatgttcact acaaaatttg 2280
agaaagataa cttttatata gagctttctc aagggaataa tttatatggt ggtcctattg 2340
tacattttta cgatgtctct attaagtaac ccaa 2374
〈210〉 98
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 98
Met Asn Lys Asn Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asn Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Glu Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu His Ile Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Xaa Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Thr Leu Lys Val Lys Lys Asp Ser Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Ala Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Xaa Lys Leu Leu Gly Leu Ala Asn Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly Tyr Ala Leu Val Gly Phe Glu Met Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Thr Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Xaa Ile Xaa Gly Phe Gly Leu Gln Ala
485 490 495
Asp Gly Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Phe Ser Gln Phe Ile Gly Asp Xaa Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Xaa Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Lys Asp Tyr Gln Thr Ile Thr Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr His Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 99
〈211〉 2366
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 99
atgaacaaga ataatactaa attaagcaca agagccttac cgagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaatatgat ttttaaaacg 120
gatacaggtg gtaatctaac cttagatgaa atcctaaaga atcagcagtt actaaatgag 180
atttctggta aattggatgg ggtaaatggg agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatctta aaaattgcaa atgaacagaa tcaagtctta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcatatata tctacctaaa 360
attacatcta tgttaagtga tgtaatgaag caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca gaatttctga taaattagat attattaacg taaatgttct 480
tattaactct acacttactg aaattacacc tgcatatcaa cggattaaat atgtgaagaa 540
aaatttgaag aattaacttt tgctacagaa accactttaa aagtaaaaaa ggatagctcg 600
cctgctgata ttcttgatga gttaactgaa ttaactgaac tagcgaaaag tgttacaaaa 660
aatgacgttg atggttttga attttacctt aatacattcc acgatgtaat ggtaggaaat 720
aatttattcg ggcgttcagc tttaaaaact gcttcagaat taattgctaa agaaaatgtg 780
aaaacaagtg gcagtgaagt aggaaatgtt tataatttct taattgtatt aacagctcta 840
caagcaaaag cttttcttac tttaacaaca tgccaaaatt attaggctta gcaaatattg 900
attatacttc tattatgaat gaacatttaa ataaggaaaa agaggaattt agagtaaaca 960
tccttcctac actttctaat actttttcta atcctaatta tgcaaaagtt aaaggaagtg 1020
atgaagatgc aaagatgatt gtggaagcta aaccaggata tgcattggtt ggttttgaaa 1080
tgagcaatga ttcaatcaca gtattaaaag tatatgaggc taagctaaaa caaaattatc 1140
aagttgataa ggattcctta tcggaggtta tttatggtga tacggataaa ttattgtgtc 1200
cagatcaatc tgaacaaata tattatacaa ataacatagt atttccaaat gaatatgtaa 1260
ttactaaaat tgatttcact aaaaaaatga aaactttaag atatgaggta acagcgaatt 1320
tttatgattc ttctacagga gaaattgact taaataagaa aaaagtagaa tcaagtgaag 1380
cggagtatag aacgttaagt gctaatgatg atggagtgta tatgccatta ggtgtcatca 1440
gtgaaacatt tttgactcga ttatgggttt ggcctccaag ctgatggaaa ttcaagatta 1500
attactttaa catgtaaatc atatttaaga gaactactgc tagcaacaga cttaagcaat 1560
aaagaaacta aattgattgt cccccaagtg gttttattag caatattgta gagaacgggt 1620
ccatagaaga ggacaattta gagccgtgga aagcaaataa taagaatgcg tatgtagatc 1680
atacaggcgg agtgaatgga actaaagctt tatatgttca taaggacgga ggattttcac 1740
aatttattgg agataattaa aaccgaaaac tgagtattaa tccaatatac tgttaaagga 1800
aaaccttcta ttcatttaaa agatgaaaat actggatata ttcattatga agatacaaat 1860
aataatttaa aagattatca aactattact aaacgtttta ctacaggaac tgatttaaag 1920
ggagtgtatt taattttaaa aagtcaaaat ggagatgaag cttggggaga taactttatt 1980
attttggaaa ttagtccttc tgaaaagtta ttaagtccag aattaattaa tacaaataat 2040
tggacgagta cgggatcaac tcatattagc ggtaatacac tcactcttta tcagggagga 2100
cgaggaattc taaaacaaaa ccttcaatta gatagttttt caacttatag agtgtatttt 2160
tctgtgtccg gagatgctaa tgtaaggatt agaaattcta gggaagtgtt atttgaaaaa 2220
agatatatga gcggtgctaa agatgtttct gaaatgttca ctacaaaatt tgagaaagat 2280
aacttttata tagagctttc tcaagggaat aatttatatg gtggtcctat tgtacatttt 2340
tacgatgtct ctattaagta acccaa 2366
〈210〉 100
〈211〉 789
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 100
Met Asn Lys Asn Asn Thr Lys Leu Ser Thr Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asp Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Asp Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Asn Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu Arg Val Tyr Leu Pro Lys Ile Thr Phe Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Ser Ser Lys Val Lys Lys Asp Gly Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Thr Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly His Ala Leu Ile Gly Phe Glu Ile Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Met Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Glu Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Glu Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Xaa Asn Xaa Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Asn Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Lys Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Xaa Xaa Tyr Gln Thr Ile Asn Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Xaa Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Xaa Leu Ile Asn Thr Xaa Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr Asn Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Xaa Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Xaa Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Ser Ile Lys
785
〈210〉 101
〈211〉 2362
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 101
atgaacaaga ataatactaa attaagcaca agagccttac caagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaacatgat ttttaaaacg 120
gatacaggtg gtgatctaac cctagacgaa attttaaaga atcagcagtt actaaatgat 180
atttctggta aattggatgg ggtgaatgga agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatatta aaaattgcaa atgaacaaaa tcaagtttta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcgggtata tctacctaaa 360
attaccttta tgttgagtga tgtaatgaaa caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agagatttct gataagttgg atattattaa tgtaaatgta 480
cttattaact ctacacttac tgaaattaca cctgcgtatc aaaggattaa atatgtgaac 540
gaaaaatttg aggaattaac ttttgctaca gaaactagtt caaaagtaaa aaaggatggc 600
tctcctgcag atattcttga tgagttaact gagttaactg aactagcgaa aagtgtaaca 660
aaaaatgatg tggatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcatcgg aattaattac taaagaaaat 780
gtgaaaacaa gtggcagtga ggtcggaaat gtttataact tcttaattgt attaacagct 840
ctgcaagcaa aagcttttct tactttaaca acatgccgaa aattattagg gttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatcctcc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gacatgcatt gattgggttt 1080
gaaattagta atgattcaat tacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagtcg ataaggattc cttatcggaa gttatttatg gtgatatgga taaattattg 1200
tgcccagatc aatctgaaca aatctattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatgggg tgtatatgcc gttaggtgtc 1440
atcagtgaaa catttttgac tccgattaat gggtttggct ccaagctgat gaaaattcaa 1500
gattaattac tttaacatgt aaatcatatt taagagaact actgctagca acagacttaa 1560
gcaataaaga aactaaattg atcgtcccgc caagtggttt tattagcaat attgtagaga 1620
acgggtccat agaagaggac aatttagagc cctggaaagc aataatagaa tgcgtatgta 1680
gatcatacag gcggagtgaa tggaactaaa gctttatatg ttcataagga cggaggaatt 1740
tcacaattta ttggagataa gttaaaaccg aaaactgagt atgtaatcca atatactgtt 1800
aaaggaaaac cttctattca tttaaaagat gaaaatactg gatatattca ttatgaagat 1860
acaaataata atttaaatta tcaaactatt aataaacgtt ttactacagg aactgattta 1920
aagggagtgt atttaatttt aaaaagtcaa aatggaatga agcttgggga gataacttta 1980
ttattttgga aattagtcct tctgaaaagt tattaagtcc aaattaatta atacaataat 2040
tggacagtac gggatcaact aatattagcg gtaatacact cactctttat cagggaggac 2100
gagggattct aaaacaaaac cttcaattag atagttttca acttatagag tgtatttttc 2160
tgtgtccgga gatgctaatg taaggattag aaattctagg gaagtgttat ttgaaaaaag 2220
atatatgagc ggtgctaaaa tgtttctgaa atgttcacac aaaatttgag aaagataact 2280
tttatataga gctttctcaa gggaataatt tatatggtgg tcctattgta catttttacg 2340
atgtctctat taagtaaccc aa 2362
〈210〉 102
〈211〉 790
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 102
Met His Glu Asn Asn Thr Lys Leu Ser Ala Arg Ala Leu Pro Ser Phe
1 5 10 15
Ile Asp Tyr Phe Asn Gly Ile Tyr Gly Phe Ala Thr Gly Ile Lys Asp
20 25 30
Ile Met Asn Met Ile Phe Lys Thr Asp Thr Gly Gly Asn Leu Thr Leu
35 40 45
Asp Glu Ile Leu Lys Asn Gln Gln Leu Leu Asn Glu Ile Ser Gly Lys
50 55 60
Leu Asp Gly Val Asn Gly Ser Leu Asn Asp Leu Ile Ala Gln Gly Asn
65 70 75 80
Leu Asn Thr Glu Leu Ser Lys Glu Ile Leu Lys Ile Ala Asn Glu Gln
85 90 95
Ser Gln Val Leu Asn Asp Val Asn Asn Lys Leu Asp Ala Ile Asn Thr
100 105 110
Met Leu His Ile Tyr Leu Pro Lys Ile Thr Ser Met Leu Ser Asp Val
115 120 125
Met Lys Gln Asn Tyr Ala Leu Ser Leu Gln Ile Glu Tyr Leu Ser Lys
130 135 140
Gln Leu Gln Glu Ile Ser Asp Lys Leu Asp Ile Ile Asn Val Asn Val
145 150 155 160
Leu Ile Asn Ser Thr Leu Thr Glu Ile Thr Pro Ala Tyr Gln Arg Ile
165 170 175
Lys Tyr Val Asn Glu Lys Phe Glu Glu Leu Thr Phe Ala Thr Glu Thr
180 185 190
Thr Leu Lys Val Lys Lys Asp Xaa Ser Pro Ala Asp Ile Leu Asp Glu
195 200 205
Leu Thr Glu Leu Thr Glu Leu Ala Lys Ser Val Thr Lys Asn Asp Val
210 215 220
Asp Gly Phe Glu Phe Tyr Leu Asn Thr Phe His Asp Val Met Val Gly
225 230 235 240
Asn Asn Leu Phe Gly Arg Ser Ala Leu Lys Thr Ala Ser Glu Leu Ile
245 250 255
Ala Lys Glu Asn Val Lys Thr Ser Gly Ser Glu Val Gly Asn Val Tyr
260 265 270
Asn Phe Leu Ile Val Leu Thr Ala Leu Gln Ala Lys Ala Phe Leu Thr
275 280 285
Leu Thr Thr Cys Arg Lys Leu Leu Gly Leu Ala Asp Ile Asp Tyr Thr
290 295 300
Ser Ile Met Asn Glu His Leu Asn Lys Glu Lys Glu Glu Phe Arg Val
305 310 315 320
Asn Ile Leu Pro Thr Leu Ser Asn Thr Phe Ser Asn Pro Asn Tyr Ala
325 330 335
Lys Val Lys Gly Ser Asp Glu Asp Ala Lys Met Ile Val Glu Ala Lys
340 345 350
Pro Gly Tyr Ala Leu Val Gly Phe Glu Met Ser Asn Asp Ser Ile Thr
355 360 365
Val Leu Lys Val Tyr Glu Ala Lys Leu Lys Gln Asn Tyr Gln Val Asp
370 375 380
Lys Asp Ser Leu Ser Glu Val Ile Tyr Gly Asp Thr Asp Lys Leu Leu
385 390 395 400
Cys Pro Asp Gln Ser Glu Gln Ile Tyr Tyr Thr Asn Asn Ile Val Phe
405 410 415
Pro Asn Glu Tyr Val Ile Thr Lys Ile Asp Phe Thr Lys Lys Met Lys
420 425 430
Thr Leu Arg Tyr Glu Val Thr Ala Asn Phe Tyr Asp Ser Ser Thr Gly
435 440 445
Glu Ile Asp Leu Asn Lys Lys Lys Val Glu Ser Ser Glu Ala Glu Tyr
450 455 460
Arg Thr Leu Ser Ala Asn Asp Asp Gly Val Tyr Met Pro Leu Gly Val
465 470 475 480
Ile Ser Glu Thr Phe Leu Thr Pro Ile Asn Gly Phe Gly Leu Gln Ala
485 490 495
Asp Gly Asn Ser Arg Leu Ile Thr Leu Thr Cys Lys Ser Tyr Leu Arg
500 505 510
Lys Leu Leu Leu Ala Thr Asp Leu Ser Asn Lys Glu Thr Lys Leu Ile
515 520 525
Val Pro Pro Ser Gly Phe Ile Ser Asn Ile Val Glu Asn Gly Ser Ile
530 535 540
Glu Glu Asp Asn Leu Glu Pro Trp Lys Ala Asn Asn Lys Asn Ala Tyr
545 550 555 560
Val Asp His Thr Gly Gly Val Lys Gly Thr Lys Ala Leu Tyr Val His
565 570 575
Lys Asp Gly Gly Ile Ser Gln Phe Ile Gly Asp Xaa Leu Lys Pro Lys
580 585 590
Thr Glu Tyr Val Ile Gln Tyr Thr Val Lys Gly Lys Pro Ser Ile His
595 600 605
Leu Lys Asp Glu Asn Thr Gly Tyr Ile His Tyr Glu Asp Thr Asn Asn
610 615 620
Asn Leu Lys Asp Tyr Gln Thr Ile Thr Lys Arg Phe Thr Thr Gly Thr
625 630 635 640
Asp Leu Lys Gly Val Tyr Leu Ile Leu Lys Ser Gln Asn Gly Asp Glu
645 650 655
Ala Trp Gly Asp Asn Phe Ile Ile Leu Glu Ile Ser Pro Ser Glu Lys
660 665 670
Leu Leu Ser Pro Glu Leu Ile Asn Thr Asn Asn Trp Thr Ser Thr Gly
675 680 685
Ser Thr His Ile Ser Gly Asn Thr Leu Thr Leu Tyr Gln Gly Gly Arg
690 695 700
Gly Ile Leu Lys Gln Asn Leu Gln Leu Asp Ser Phe Ser Thr Tyr Arg
705 710 715 720
Val Tyr Phe Ser Val Ser Gly Asp Ala Asn Val Arg Ile Arg Asn Ser
725 730 735
Arg Glu Val Leu Phe Glu Lys Arg Tyr Met Ser Gly Ala Lys Asp Val
740 745 750
Ser Glu Met Phe Thr Thr Lys Phe Glu Lys Asp Asn Phe Tyr Ile Glu
755 760 765
Leu Ser Gln Gly Asn Asn Leu Tyr Gly Gly Pro Ile Val His Phe Tyr
770 775 780
Asp Val Xaa Ile Lys Pro
785 790
〈210〉 103
〈211〉 2375
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 103
atgcacgaga ataatactaa attaagcgca agggccttac cgagttttat tgattatttt 60
aatggcattt atggatttgc cactggtatc aaagacatta tgaatatgat ttttaaaacg 120
gatacaggtg gtaatctaac cttagatgaa atcctaaaga atcagcagtt actaaatgag 180
atttctggta aattggatgg ggtaaatggg agcttaaatg atcttatcgc acagggaaac 240
ttaaatacag aattatctaa ggaaatctta aaaattgcaa atgaacagag tcaagtttta 300
aatgatgtta ataacaaact cgatgcgata aatacgatgc ttcatatata tctacctaaa 360
attacatcta tgttaagtga tgtaatgaag caaaattatg cgctaagtct gcaaatagaa 420
tacttaagta aacaattgca agaaatttct gataaattag atattattaa cgtaaatgtt 480
cttattaact ctacacttac tgaaattaca cctgcatatc aacggattaa atatgtgaat 540
gaaaaatttg aagaattaac ttttgctaca gaaaccactt taaaagtaaa aaaggatrac 600
tcgcctgctg atattcttga tgaattaact gaattaactg aactagcgaa aagtgttaca 660
aaaaatgacg ttgatggttt tgaattttac cttaatacat tccacgatgt aatggtagga 720
aataatttat tcgggcgttc agctttaaaa actgcttcag aattaattgc taaagaaaat 780
gtgaaaacaa gtggcagtga agtaggaaat gtttataatt tcttaattgt attaacagct 840
ctacaagcaa aagcttttct tactttaaca acatgccgaa aattattagg cttagcagat 900
attgattata cttctattat gaatgaacat ttaaataagg aaaaagagga atttagagta 960
aacatccttc ctacactttc taatactttt tctaatccta attatgcaaa agttaaagga 1020
agtgatgaag atgcaaagat gattgtggaa gctaaaccag gatatgcatt ggttggtttt 1080
gaaatgagca atgattcaat cacagtatta aaagtatatg aggctaagct aaaacaaaat 1140
tatcaagttg ataaggattc cttatcggag gttatttatg gtgatacgga taaattattg 1200
tgtccagatc aatctgaaca aatatattat acaaataaca tagtatttcc aaatgaatat 1260
gtaattacta aaattgattt cactaaaaaa atgaaaactt taagatatga ggtaacagcg 1320
aatttttatg attcttctac aggagaaatt gacttaaata agaaaaaagt agaatcaagt 1380
gaagcggagt atagaacgtt aagtgctaat gatgatggag tgtatatgcc attaggtgtc 1440
atcagtgaaa catttttgac tccgataaat gggtttggcc tccaagctga tggaaattca 1500
agattaatta ctttaacatg taaatcatat ttaagaaaac tactgctagc aacagactta 1560
agcaataaag aaactaaatt gatcgtcccg ccaagtggtt ttattagcaa tattgtagag 1620
aacgggtcca tagaagagga caatttagag ccgtggaaag caaataataa gaatgcgtat 1680
gtagatcata caggcggagt gaaaggaact aaagctttat atgttcataa ggacggagga 1740
atttcacaat ttattggaga taakttaaaa ccgaaaactg agtatgtaat ccaatatact 1800
gttaaaggaa aaccttctat tcatttaaaa gatgaaaata ctggatatat tcattatgaa 1860
gatacaaata ataatttaaa agattatcaa actattacta aacgttttac tacaggaact 1920
gatttaaagg gagtgtattt aattttaaaa agtcaaaatg gagatgaagc ttggggagat 1980
aactttatta ttttggaaat tagtccttct gaaaagttat taagtccaga attaattaat 2040
acaaataatt ggacgagtac gggatcaact catattagcg gtaatacact cactctttat 2100
cagggaggac gaggaattct aaaacaaaac cttcaattag atagtttttc aacttataga 2160
gtgtattttt ctgtgtccgg agatgctaat gtaaggatta gaaattctag ggaagtgtta 2220
tttgaaaaaa gatatatgag cggtgctaaa gatgtttctg aaatgttcac tacaaaattt 2280
gagaaagata acttttatat agagctttct caagggaata atttatatgg tggtcctatt 2340
gtgcattttt acgatgtcyc tattaagtaa cccaa 2375
〈210〉 104
〈211〉 554
〈212〉 PRT
〈213〉 Bacillus thuringiensis
〈400〉 104
Thr Leu His Leu Leu Lys Leu His Leu Arg Ile Lys Gly Leu Asn Met
1 5 10 15
Thr Lys Asn Leu Arg Asn Leu Leu Leu Xaa Xaa Leu Xaa Gln Lys Lys
20 25 30
Arg Met Ala Leu Leu Gln Ile Phe Xaa Met Ser Leu Ser Xaa Asn Arg
35 40 45
Lys Val Gln Lys Met Met Trp Met Val Leu Asn Phe Thr Leu Ile His
50 55 60
Ser Thr Met Xaa Glu Ile Ile Tyr Ser Gly Val Gln Leu Lys Leu Xaa
65 70 75 80
Arg Asn Leu Leu Lys Lys Met Lys Gln Val Ala Val Xaa Xaa Glu Met
85 90 95
Phe Ile Xaa Ser Leu Tyr Gln Leu Xaa Lys Gln Lys Leu Phe Leu Leu
100 105 110
Gln His Ala Glu Asn Tyr Xaa Gln Ile Leu Ile Ile Leu Leu Leu Met
115 120 125
Asn Ile Ile Arg Lys Lys Arg Asn Leu Glu Thr Ser Xaa Leu His Phe
130 135 140
Leu Ile Leu Phe Leu Ile Leu Ile Met Gln Lys Leu Lys Glu Val Met
145 150 155 160
Lys Met Gln Arg Leu Trp Lys Leu Asn Gln Asp Met His Trp Leu Val
165 170 175
Leu Lys Ala Met Ile Gln Ser Gln Tyr Lys Tyr Met Arg Leu Ser Asn
180 185 190
Lys Ile Ile Lys Leu Ile Arg Ile Pro Tyr Arg Arg Leu Phe Met Val
195 200 205
Ile Arg Ile Asn Tyr Cys Val Gln Ile Asn Leu Asn Lys Tyr Ile Ile
210 215 220
Gln Ile Thr Tyr Phe Gln Met Asn Met Leu Leu Lys Leu Ile Ser Leu
225 230 235 240
Lys Lys Lys Leu Asp Met Arg Gln Arg Ile Phe Met Ile Leu Leu Gln
245 250 255
Glu Lys Leu Thr Ile Arg Lys Lys Asn Gln Val Lys Arg Ser Ile Glu
260 265 270
Arg Val Leu Met Met Met Xaa Cys Ile Cys His Val Ser Ser Val Lys
275 280 285
His Phe Leu Arg Met Gly Leu Ala Ser Lys Leu Arg Gln Ile Gln Asp
290 295 300
Leu Leu His Val Asn His Ile Glu Asn Tyr Cys Gln Gln Thr Ala Ile
305 310 315 320
Arg Lys Leu Asn Ser Ser Arg Gln Val Phe Tyr Gln Tyr Cys Arg Glu
325 330 335
Arg Val Leu Arg Arg Gly Gln Phe Arg Ala Val Glu Ser Lys Glu Cys
340 345 350
Val Cys Arg Ser Tyr Arg Arg Ser Glu Trp Asn Ser Phe Ile Cys Ser
355 360 365
Gly Arg Arg Asn Phe Thr Ile Tyr Trp Arg Val Lys Thr Glu Asn Val
370 375 380
Cys Asn Pro Ile Tyr Cys Arg Lys Thr Phe Tyr Ser Phe Lys Arg Lys
385 390 395 400
Tyr Trp Ile Tyr Ser Leu Arg Tyr Lys Phe Lys Arg Leu Ser Asn Tyr
405 410 415
Tyr Thr Phe Tyr Tyr Arg Asn Phe Lys Gly Ser Val Phe Asn Phe Lys
420 425 430
Lys Ser Lys Trp Arg Ser Leu Gly Arg Leu Tyr Tyr Phe Gly Asn Ser
435 440 445
Phe Lys Val Ile Lys Ser Arg Ile Asn Tyr Lys Leu Asp Glu Tyr Gly
450 455 460
Ile Asn Ser Tyr Arg Tyr Thr His Ser Leu Ser Gly Arg Thr Arg Asn
465 470 475 480
Ser Lys Thr Lys Pro Ser Ile Arg Phe Phe Asn Leu Ser Val Phe Phe
485 490 495
Cys Val Arg Arg Cys Cys Lys Asp Lys Phe Gly Ser Val Ile Lys Lys
500 505 510
Ile Tyr Glu Arg Cys Arg Cys Phe Asn Val His Tyr Lys Ile Glu Arg
515 520 525
Leu Leu Tyr Arg Ala Phe Ser Arg Glu Phe Ile Trp Trp Ser Tyr Cys
530 535 540
Thr Phe Leu Arg Cys Leu Tyr Val Thr Gln
545 550
〈210〉 105
〈211〉 1888
〈212〉 DNA
〈213〉 Bacillus thuringiensis
〈400〉 105
actctacact tactgaaatt acacctgcgt atcaaaggat taaatatgtg aacgaaaaat 60
ttgaggaatt aacttttgct acrgamacta kttcaaaagt aaaaamggat ggctctccts 120
cagatattct kgatgagtta actgagttaa cwgaactagc gaaaagtgta acaaaaaatg 180
atgtggatgg ttttraattt taccttaata cattccacga tgtaakggta ggaaataatt 240
tattcgggcg ttcagcttta aaaactgcwt cggaattaat trctaaagaa aatgtgaaaa 300
caagtggcag tgargtmgga aatgtttata ayttcttaat tgtattaaca gctctrcaag 360
caaaagcttt tcttacttta acaacatgcc gaaaattatt aggsttagca gatattgatt 420
atacttctat tatgaatgaa catttaaata aggaaaaaga ggaatttaga gtaaacatcc 480
tycctacact ttctaatact ttttctaatc ctaattatgc aaaagttaaa ggaagtgatg 540
aagatgcaaa gatgattgtg gaagctaaac caggatatgc attggttggt tttgaaatga 600
gcaatgattc aatcacagta ttaaaagtat atgaggctaa gctaaaacaa aattatcaag 660
ttgataagga ttccttatcg gaggttattt atggtgatac ggataaatta ttgtgtccag 720
atcaatctga acaaatatat tatacaaata acatagtatt tccaaatgaa tatgtaatta 780
ctaaaattga tttcactaaa aaaatgaaaa ctttaagata tgaggtaaca gcgaattttt 840
atgattcttc tacaggagaa attgacttaa ataagaaaaa agtagaatca agtgaagcgg 900
agtatagaac gttaagtgct aatgatgatg grgtgtatat gccattaggt gtcatcagtg 960
aaacattttt gactccgata aatgggtttg gcctccaagc tgaggcaaat tcaagattaa 1020
ttactttaac atgtaaatca tatttaagag aactactgct agcaacagac ttaagcaatw 1080
aggaaactaa attgatcttc ccgccaagtg ttttattagc aatattgtag agaacgggtc 1140
cttagaagag gacaatttag agccgtggaa agcaaataat aagaatgcgt atgtagatca 1200
tacaggcgga gtgaatggaa ctaaagcttt atatgttcat aaggacggag gaatttcaca 1260
atttattgga gataagttaa aaccgaaaac tgagtatgta atccaatata ctgttaaagg 1320
aaaaccttct attcatttaa aagatgaaaa tactggatat attcattatg aagatacaaa 1380
taataattta aaagattatc aaactattac taaacgtttt actacaggaa ctgatttaaa 1440
gggagtgtat ttaattttaa aaagtcaaaa tggagatgaa gcttggggag ataactttat 1500
tattttggaa attagtcctt ctgaaaagtt attaagtcca gaattaatta atacaaataa 1560
ttggacgagt acgggatcaa ctcatattag cggtaataca ctcactcttt atcagggagg 1620
acgaggaatt ctaaaacaaa accttcaatt agatagtttt tcaacttata gagtgtattt 1680
ttctgtgtcc ggagatgcta atgtaaggat tagaaattct agggaagtgt tatttgaaaa 1740
aagatatatg agcggtgcta aagatgtttc tgaaatgttc actacaaaat ttgagaaaga 1800
taacttttat atagagcttt ctcaagggaa taatttatat ggtggtccta ttgtacattt 1860
ttacgatgtc tctattaagt aacccaaa 1888
Claims (2)
- 유럽 옥수수 좀벌레[European corn borer; 오스트리니아 누빌라리스(Ostrinia nubilalis)]의 박멸 방법으로서, 상기 방법이 상기 해충과 살충 유효량의 바실러스 츄린겐시스(Bacillus thuringensis) 독소를 접촉시키는 것을 포함하며, 여기서 상기 독소는(a) 상기 독소는 서열 70, 서열 72, 서열 74, 서열 76, 서열 78, 서열 80, 서열 82, 서열 84, 서열 86, 서열 88, 서열 90, 서열 92, 서열 94, 서열 96, 서열 98, 서열 100, 서열 102 및 서열 104로 구성된 그룹으로부터 선택된 서열과 약 75% 이상의 상동성을 갖는 아미노산 서열을 포함하는 독소 ;(b) 상기 독소는 서열 70, 서열 72, 서열 74, 서열 76, 서열 78, 서열 80, 서열 82, 서열 84, 서열 86, 서열 88, 서열 90, 서열 92, 서열 94, 서열 96, 서열 98, 서열 100, 서열 102 및 서열 104로 구성된 그룹으로부터 선택된 아미노산 서열을 코드화하는 뉴클레오티드 서열과 혼성화되는 뉴클레오티드에 의해서 코드화되는 아미노산 서열을 포함하는 독소 ; 및(c) 상기 독소는 서열 70, 서열 72, 서열 74, 서열 76, 서열 78, 서열 80, 서열 82, 서열 84, 서열 86, 서열 88, 서열 90, 서열 92, 서열 94, 서열 96, 서열 98, 서열 100, 서열 102 및 서열 104로 구성된 그룹으로부터 선택된 독소에 대한 항체와 면역 반응을 하는 독소로 구성된 그룹으로부터 선택되는 것을 특징으로 하는 방법.
- 제 1항에 있어서, 상기 독소가 서열 74에 나타낸 아미노산 서열, 또는 이의 살충성 절편(pesticidal fragment)을 갖는 방법.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US9/002,285 | 1997-12-31 | ||
| US09/002,285 US6369213B1 (en) | 1996-07-01 | 1997-12-31 | Toxins active against pests |
| PCT/US1998/026585 WO1999033991A2 (en) | 1997-12-31 | 1998-12-15 | TOXINS ACTIVE AGAINST $i(OSTRINIA NUBILALIS) |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20010033398A true KR20010033398A (ko) | 2001-04-25 |
Family
ID=21700070
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020007006867A KR20010033398A (ko) | 1997-12-31 | 1998-12-15 | 오스트리니아 누빌라리스에 활성인 독소 |
Country Status (9)
| Country | Link |
|---|---|
| US (2) | US6369213B1 (ko) |
| EP (1) | EP1042481A2 (ko) |
| JP (1) | JP2002500166A (ko) |
| KR (1) | KR20010033398A (ko) |
| AR (1) | AR014233A1 (ko) |
| AU (1) | AU1826499A (ko) |
| BR (1) | BR9814567A (ko) |
| CA (1) | CA2315106A1 (ko) |
| WO (1) | WO1999033991A2 (ko) |
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| FI981809A0 (fi) * | 1998-08-24 | 1998-08-24 | Viktor Kuvshinov | Muunnetut synteettiset DNA-sekvenssit, jotka koodaavat cry-geeniin perustuvaa proteiinia |
| CN1390259A (zh) | 1999-09-15 | 2003-01-08 | 孟山都技术有限公司 | 对鳞翅目昆虫有活性的苏云金芽孢杆菌δ内毒素组合物及其使用方法 |
| DE60041505D1 (de) * | 1999-12-28 | 2009-03-19 | Bayer Bioscience Nv | Inzektizide proteinen von bacillus thuringiensis |
| AR035799A1 (es) | 2001-03-30 | 2004-07-14 | Syngenta Participations Ag | Toxinas insecticidas aisladas de bacillus thuringiensis y sus usos. |
| JP2006500908A (ja) * | 2002-03-06 | 2006-01-12 | シンジェンタ パーティシペーションズ アクチェンゲゼルシャフト | 新規なＶｉｐ３トキシン及び使用方法 |
| WO2003080656A1 (en) * | 2002-03-22 | 2003-10-02 | Bayer Bioscience N.V. | Novel bacillus thuringiensis insecticidal proteins |
| DE60332711D1 (de) * | 2002-03-22 | 2010-07-08 | Bayer Bioscience Nv | Neue insektizide proteine von bazillus thuringiensis |
| US8088478B2 (en) * | 2005-06-21 | 2012-01-03 | Weyerhaeuser Nr Company | Barrier material |
| CN110734919A (zh) | 2008-07-02 | 2020-01-31 | 阿森尼克斯公司 | Axmi-il5、axmi-113、axmi-005、axmi-163和axmi-184∶vip3a杀虫蛋白及其使用方法 |
| CA2825049A1 (en) * | 2011-01-24 | 2012-08-02 | Pioneer Hi-Bred International, Inc. | Novel bacillus thuringiensis genes with lepidopteran activity |
| EA031448B1 (ru) * | 2012-02-16 | 2019-01-31 | Зингента Партисипейшнс Аг | Сконструированные пестицидные белки |
| CN104651377A (zh) * | 2013-11-25 | 2015-05-27 | 浙江大学 | 新型的杀虫蛋白 |
| EP3230456B1 (en) * | 2014-12-12 | 2024-05-22 | Syngenta Participations AG | Compositions and methods for controlling plant pests |
| US11129906B1 (en) | 2016-12-07 | 2021-09-28 | David Gordon Bermudes | Chimeric protein toxins for expression by therapeutic bacteria |
| UY39585A (es) * | 2020-12-23 | 2022-07-29 | Monsanto Technology Llc | Proteínas que exhiben actividad inhibidora de insectos frente a plagas con importancia agrícola de plantas de cultivo y semillas |
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-
1997
- 1997-12-31 US US09/002,285 patent/US6369213B1/en not_active Expired - Lifetime
-
1998
- 1998-12-15 AU AU18264/99A patent/AU1826499A/en not_active Abandoned
- 1998-12-15 EP EP98963186A patent/EP1042481A2/en not_active Withdrawn
- 1998-12-15 JP JP2000526647A patent/JP2002500166A/ja active Pending
- 1998-12-15 BR BR9814567-3A patent/BR9814567A/pt not_active IP Right Cessation
- 1998-12-15 CA CA002315106A patent/CA2315106A1/en not_active Abandoned
- 1998-12-15 KR KR1020007006867A patent/KR20010033398A/ko not_active Application Discontinuation
- 1998-12-15 WO PCT/US1998/026585 patent/WO1999033991A2/en not_active Application Discontinuation
- 1998-12-30 AR ARP980106768A patent/AR014233A1/es not_active Application Discontinuation
-
2002
- 2002-03-15 US US10/099,285 patent/US6752992B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| US20030105319A1 (en) | 2003-06-05 |
| US6752992B2 (en) | 2004-06-22 |
| AU1826499A (en) | 1999-07-19 |
| WO1999033991A2 (en) | 1999-07-08 |
| JP2002500166A (ja) | 2002-01-08 |
| US6369213B1 (en) | 2002-04-09 |
| CA2315106A1 (en) | 1999-07-08 |
| AR014233A1 (es) | 2001-02-07 |
| WO1999033991A3 (en) | 1999-09-10 |
| BR9814567A (pt) | 2000-10-10 |
| EP1042481A2 (en) | 2000-10-11 |
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