KR20060096515A - 외표면 단백질들, 그들의 유전자들 및 그들의 사용 - Google Patents
외표면 단백질들, 그들의 유전자들 및 그들의 사용 Download PDFInfo
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- KR20060096515A KR20060096515A KR1020067017195A KR20067017195A KR20060096515A KR 20060096515 A KR20060096515 A KR 20060096515A KR 1020067017195 A KR1020067017195 A KR 1020067017195A KR 20067017195 A KR20067017195 A KR 20067017195A KR 20060096515 A KR20060096515 A KR 20060096515A
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- South Korea
- Prior art keywords
- ala
- gly
- leu
- val
- glu
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Abstract
본 발명에 따르면, 일련의 유전자들이 B군 스트렙토코커스에서 동정되고, 그것들의 생성물들은 유기체의 외표면에 위치될 것이다. 이 유전자들 또는 그것들의 기능성 단편들은 예를들면, 미생물 감염에 대한 환자의 면역화를 위한 백신 등의 치료제 제조에 유용할 것이다.
스트렙토코커스, 요로 감염, 병소 감염
Description
본 발명은 외표면 단백질들 및 그들의 유전자들의 확인(동정) 및 그들의 사용에 관한 것이다. 더욱 구체적으로는, 면역화용 치료법 및 약품 선별에서의 그들의 사용에 관한 것이다.
Streptococcus agalactiae로도 알려져 있는, B군 스트렙토코커스 (GBS)는 다양한 상태의 원인제이다. 특히, GBS는 다음을 일으킨다:
초기
발증
신생아 감염(Early onset neonatal infection)
이 감염은 보통 자궁내에서 시작되며 유아의 심한 패혈증 및 폐염을 일으키며, 이것은 치료하지 않으면 치명적이며 치료한다고 해도 10 내지 20%의 사망율을 갖는다.
말기
발증
신생아 감염(Late onset neonatal infection)
이 감염은 탄생 직후부터 약 3개월까지의 기간에 발생한다. 이것은 폐혈증을 일으키며, 환자의 약 90%는 합병증으로 수막염이 발생한다. 골수염, 패혈성관절염, 농양 및 내안구염을 포함한 기타의 병소(focal) 감염이 또한 발생한다.
성인 감염
이 감염은 일반적으로 점점 더 증가하는 것으로 생각되며, 방금 아기를 출산한 산모, 노인층 및 면역저항성 감약자들에게 가장 빈번하게 일어난다. 이 감염은 패혈증 및 골수염, 패혈성관절염, 농양 및 내안구염을 포함한 병소감염으로 특징된다.
요로 감염
GBS는 요로감염의 원인이며 임신 중의 모든 감염의 약 10%의 원인이 된다.
수의적 감염
GBS는 젖소의 만성 유선염을 일으킨다. 이것은 우유 생산을 감소시키고 따라서 경제적으로 상당히 중요한 것이다.
GBS 감염들은 항생물질로 치료될 수 있다. 그러나, 면역화가 바람직하다. 그러므로, 치료학적으로-효과적인 백신에 사용될 수 있는 면역원을 개발하는 것이 필요하다.
본 발명은 GBS, 및 관련 유기체의 일련의 유전자들의 동정에 기초하며, 그것들의 생성물들은 유기체의 외표면과 연관될 것이고, 따라서 면역치료법의 표적으로 유용할 것이다.
본 발명의 한 면에 따르면, 펩티드는 B군 스트렙토코커스로부터 얻을 수 있는, MS4, MS10, MS11, MS14 및 MS16으로 확인된 유전자중 어느 것, 또는 그들의 상동체(homologue) 또는 기능성 단편을 포함하는 오페론에 의해 코드화된다. 이와 같 은 펩티드는 예를 들면, 단리되었을 때 치료용으로 적당하다.
본 발명에서 "기능성 단편"이라는 용어는 전체 유전자 또는 펩티드의 활성을 보유하는 유전자 또는 펩티드의 일부를 정의하기 위해 사용된다. 예를 들면, 펩티드의 기능성 단편은 백신 또는 항체 제조에 있어서 유용한, 항원성 결정기로서 사용될 수 있다.
유전자 단편은 활성 펩티드를 코드화하는데 사용할 수 있다. 선택적으로, 유전자 단편은, 치료효과를 발휘하기 위해 생체내의 야생형 유전자를 표적화하는 유전자 요법에 유용할 수 있다.
본 발명에 따른 펩티드는 본 명세서에서 서열 ID 번호 2, 4, 6, 8, 10 및 12로 확인된 아미노산 서열 또는 그들의 기능성 단편 중 어느 것을 포함할 수 있다.
세포외 또는 세포 표면 위치 때문에, 본 발명의 펩티드는 GBS에 대한 치료학적으로-효과적인 백신 제조에 대한 적당한 후보일 것이다. "치료학적으로-효과적" 이란 용어는 백신의 예방효과를 포함하는 의도이다. 예를 들면, 백신은 본 발명에 따른 펩티드, 또는 감염 치료를 위한 그것의 발현 수단을 포함할 수 있다.
백신은 GBS에 의한 감염에 대해 산모와 신생아를 보호하기 위해 임신 전 또는 임신 중 여성에게 투여될 수 있다.
본 발명의 다른 면에 따르면, 펩티드들 또는 유전자들은 유력한 항미생물제의 선별 또는 독성(virulence)의 검출에 사용될 수 있다.
본 발명의 또 다른 면은 본 발명에서 동정된 생성물들을 B군 스트렙토코커스 균주에 의한 감염과 관련된 상태의 치료 또는 예방에 사용하는 것이다.
비록 그 단백질이 환자의 치료에의 사용이 기재되었지만, 본 발명의 생성물들의 수의적 사용 또한 본 발명의 범위 내에서 고려된다. 특히, 펩티드 또는 백신은 특히 젖소의 만성 유선염 치료에 사용될 수 있다.
본 발명은 B군 스트렙토코커스 균주 M732에 관하여 기재한다. 그러나, 모든 GBS 균주들 및 많은 기타 세균성 균주들은 M732의 펩티드와 아미노산 서열 상동성를 갖는 관련 펩티드 또는 단백질을 포함할 수 있다. 펩티드를 함유할 수 있는 유기체는, S. pneumoniae, S. pyogenes, S. suis, S. milleri, C 및 G군 Streptococci 및 Enterococci를 포함하지만, 이것으로 한정되는 것은 아니다. 이들 각각에 대한 백신은 GBS에 대해 기재한 것과 동일한 방법으로 개발될 수 있다.
바람직하게는, 백신제조에 유용한 펩티드들은 본 발명에서 확인된 펩티드들과 40%를 넘는 서열 상사성(相似性)을 갖는 것이다. 더 바람직하게는, 펩티드는 60 %를 넘는 서열 상사성을 갖는다. 가장 바람직하게는, 펩티드는 80%를 넘는 서열 유사성, 예를 들면 95%의 상사성을 갖는다.
본 발명에 따른 유전자를 특성화함으로써, 기타 미생물에 상동성을 수립하는데 그 유전자 서열을 사용하는 것이 가능하다. 이 방법으로, 기타 미생물이 유사한 외표면 생성물들을 갖는가를 결정하는 것이 가능하다. 서열 상동성은 예를 들면 EMBL 또는 Genbank과 같은 존재하는 데이터베이스에서 조사함으로써 수립될 수 있다.
본 발명에 따른 펩티드들 또는 단백질들은 공지의 방법으로 정제되고 단리될 수 있다. 특히, 유전자 서열을 확인함으로써, 적당한 숙주에 유전자를 발현시키는 재조합 기술을 사용하는 것이 가능할 것이다. 활성 단편들 및 상동체들은 동정될 수 있고 그리고 치료법에 유용할 것이다. 예를 들면, 펩티드들 또는 그들의 활성 단편들은 면역 반응을 얻기 위해, 백신 중 항원성 결정기로서 사용될 수 있다. 이들은 또한 수동면역, 또는 진단적 응용을 위한 항체들의 제조에 사용될 수 있다. 적당한 항체는 모노클로날 항체들, 또는 단일 사슬 fv 단편을 포함한 그들의 단편을 포함한다. 항체의 제조방법은 당업자들에게는 명백할 것이다.
감쇠된(attenuated) 미생물을 기초한 백신의 제조는 당업자들에게 알려져 있다. 백신 조성물들은 필요 또는 요구에 따라, 예를 들면 명반과 같은 적당한 담체 또는 보조제와 제제화될 수 있고 그리고 B군 스트렙토코키(streptococci) 또는 기타 관련 미생물에 대한 효과적인 면역화를 제공하기 위한 치료법에 사용될 수 있다. 백신 제제의 제조는 당업자에게는 명백할 것이다.
더욱 일반적으로, 그리고 당업자들에게 잘 알려진 바와 같이, 본 발명의 활성 성분의 적당한 양은 치료 용도로서 적당한 담체 또는 부형제에 따라, 및 투여 경로에 따라 선택될 수 있다. 이들 인자는 치료될 상태의 성질/심각성, 대상자의 유형 또는 건강등과 같은 알려진 기준들에 따라 선택 또는 결정될 수 있다.
본 발명의 생성물들은 다음과 같이 동정되었다:
토드-헤위트(Todd-Hewitt) 브로스를 GBS로 접종했고 그리고 37℃에서 하룻밤성장시켰다. 세포들을 원심분리로 수확하고 그리고 인산염 완충된 염수(PBS)로 세척하였다. 세포들을 프로테아제 저해제들 (1mM PMSF, 10μM 인돌 아세트산, 10mM 1,10-페난트롤린, 1μM 펩스타틴 A) 및 뮤타노리신을 마이크로리터당 4 유니트의 최종 농도로 함유하는 삼투압 완충액 (20%(W/V) 슈크로스, 20mM 트리스-HCl pH 7.0, 10mM MgCl2)에 재현탁하였다. 이것을 37℃에서 2시간동안 인큐베이트(진동하면서) 했다.
세포들 및 파편들을 고속 원심분리로 일차 제거하고 그리고 나서 1시간 동안 초원심분리하였다. 결과적으로 생성되는 세포벽 단백질들을 함유하는 상징액을 한외여과 장치를 사용하여 압력하에 농축하였다 (10,000 분자량 컷-오프).
시료를 초 고질(high quality) 수에 대해 투석하고 그리고 동결건조하였다. 로딩 완충액에 재현탁한 후, 단백질들을 예비 2-차원-겔 전기영동으로 분리하였다. 전기영동에 이어서, 개개의 스팟(spot)을 연구를 위해 선택하였다. 스팟을 겔내에서 트립신 소화시켰다. 결과적으로 생성되는 펩티드들을 겔로부터 추출해 내고 그리고 Microbore RP-HPLC를 사용하여 정제하였다. 분획들을 매45초 마다 모으고 그리고 UV 흡수의 영역들에 부합하는 이들의 부분을 플라이트 질량 분광분석기(Flight Mass Spectrometry)의 지연된 추출-매트리스 조력된 레이저 탈착-시간 (DE-MALDI-TOF-MS)으로 분석하였다. 그리고 나서 공(blank) 조제물에서 관찰되지 않은 펩티드들을 Nanospray-MS/Ms를 사용하여 서열화하였다.
이 펩티드 서열 정보를 사용하여, 확인된 펩티드 서열들 간에 놓여있는 DNA 절편을 증폭하는 중합효소 연쇄 반응(PCR)에 사용될 축퇴(degenerate) 올리고뉴클레오티드들을 설계하였다.
PCR 증폭은 여러개의 폴리뉴클레오티드 단편들의 생산을 가져왔고, 이들 각각을 제조자의 프로토콜에 따라서 pCR 2.1-TOPO 벡터 (Invitrogen BV, Netherlands)로 클론하였다.
각각의 플라스미드 내의 DNA 단편을 서열화로 확인하였고, 그리고 나서 하기와 같이 전체-길이 유전자 서열을 얻기 위하여 사용하였다.
확인된 DNA 단편을 사용하여, 게놈성 DNA 서열화를 위한 올리고뉴클레오티드 프라이머들을 설계하였다. 이들 프라이머를 얻은 서열로부터 "외부" 방향으로 서열화되도록 설계하였다. 일단 서열을 판독하면, 얻은 서열을 유전자의 5' 및 3' 말단에 도달했는지를 보기 위해 체크하였다. 이들 특성의 존재는 상동 서열들에 대해, 5' 말단의 경우, 샤인-달가르노 공통 서열(Shine-Dalgarno consensus sequence)에 의해 선행되는 AUG 개시코돈의 존재, 및 3' 말단의 경우, 번역 종결(중지) 코돈의 존재에 대해 체크함으로써 확인하였다.
전체-길이 유전자를 확인한 후, GBS 게놈성 DNA로부터 전체-길이 생성물을 증폭하기 위해 프라이머를 설계하였다. 사용된 프라이머들은 생성물이 사용된 락토코키(Lactococcal) 발현 시스템으로 연이은 클로닝을 허용하도록 제한 효소 인식부위들(5' 말단에서 NcoI 및 3' 말단에서 Eco0109I)을 포함한다.
PCR은 상기 프라이머를 사용하여 실시하였고 그리고 생성물들을 pCR 2.1 클로닝 벡터(In Vitrogen)로 클론하였다. 클론된 단편의 존재의 확인에 이어서, DNA를 제한 효소 NcoI 및 Eco0109I를 사용하여 절개하였다.
이 단편이 삽입된 벡터는 pNZ8048의 변형된 형이었다 (Kuipers, O.P. et al.(1998) J. Biotech 64:15-21). 복제의 락토코커스 기원, 클로람페니콜 내성 마커, 유도성 니신 프로모터 및 다중클로닝 부위를 장착하는 이 벡터는, NcoI 부위를 갖는 5-최말단의 측면에 접하고 다중클로닝 부위(Eco0109I 부위 포함)의 중앙에서 분할된 두개의 10X His 태그, 및 His 태그의 3' 말단의 종결(중지) 코돈으로 다중클로닝 부위를 대치함에 의해 변경되었다.
관심있는 유전자는 10x His 태그가 코딩 영역에 대해 3' 위치에 있도록 삽입되었다. 재조합 플라즈미드를 L. lactis (균주 NZ9000 - Kuipers, O.P. et al. (1998)Supra)로 형질전환 후, 액체 배양물 400ml를 설정하고 그리고 이 배양물에 니신을 첨가하여 단백질의 번역을 유도하였다. 2시간 동안 배양시킨 후, 세포들을 수확하고 비드 비팅(bead beating)으로 용해시켰다. 생성된 용해물은 원심분리하여 청정시키고 금속 친화성 칼럼(Talon, Clonetech)을 통과시켰다. 결합된 단백질들을 이미다졸로 용출시키기 전에 칼럼을 되풀이해서 세척하였다.
His-태그된 재조합 단백질을 함유하는 분획을 확인하기 위해, 각 분획 유래의 분취량을 웨스턴 블롯되고 항-His 항체들로 프로브된 SDS-PAGE로 분석하였다.
얻어진 재조합 단백질을 면역화 전에 수확된 예비-면역 혈청을 갖는, 뉴질랜드 흰 토끼를 면역시키는데 사용하였다. 부스터 후, 토끼들을 희생시키고 혈청을 모았다. 이 혈청을 웨스턴 블롯, ELISA 및 동물 보호 모델에 사용하였다.
동물 연구로부터 얻은 혈청을 사용하여 면역흡착 연구를 실시하였다.
B군 스트렙토코커스를 8시간 동안 토드 헤위트 브로스(THB)에서 성장시키고, 수확하고 그리고 PBS 5ml에 재현탁하였다. 이것의 50㎕ 분취량을 96웰 플레이트의 웰(Nunc Immuno-Sorb)을 코팅하는데 사용하였다. 이것을 세균이 플레이트상에 흡착되도록 하기 위해 4℃에서 밤새 방치하였다. 플레이트를 PBS로 두번 세척하고, 그 다음 37℃에서 1시간 동안 PBS 중의 3% BSA로 차단시켰다. 다시 플레이트를 세척하였다. 혈청의 일련의 10배 희석액 PBS에서 제조하고 그리고 이 희석액 50㎕를, 이중으로, 플레이트의 웰들에 첨가하였다. 플레이트를 덮고 1 시간동안 37℃에서 인큐베이트하였다. 플레이트를 세척하고, 그 다음 1:5000의 농도로 항-토끼 알칼리 포스파타제 컨쥬게이트된 2차 항체 50 ㎕를 각 웰에 참가하였다. 37℃에서 1 시간동안 인큐베이트한 후, 플레이트를 다시 세척하였다. 기질(PNPP) 50㎕을 각 웰에 첨가하고, 그리고 30분 동안 반응을 진행시킨 후, 405nm에서 흡광도를 기록하였다.
또한, 동물 보호 연구를 면역화된 토끼에 대한 보호 효과를 시험하기 위해 수행하였다.
GBS M732를 THB에서 정-대수기(mid-log phase)에 다다를 때까지- 약 5 시간 성장시켰다. 계수 챔버에서 세포의 수를 세고, 세균을 면역전 또는 시험 혈청에 2x 107 세균/ml의 농도로 희석하였다. 이것의 50㎕를 생후 0-1 일의 마우스들에 복강 내 경로를 통해 주입하였다. 마우스들을 48 시간에 걸쳐 생존을 관찰하였다.
이하의 실시예들은 본 발명을 설명한다.
실시예
실시예
1
첫번째 플라스미드는 MS4로 명명하였다. 클론된 DNA 단편을 서열화하고 그리 고 뉴클레오티드 및 추론된 아미노산 서열 (SEQ ID NOS. 1 및 2)을 단백질 데이터베이스를 검색하기 위해 사용하였다.
GBS MS4 유전자 생성물에 상동체들은 Clostridium perfingens, Haemophilus influenzae, Neisseria flavescens 및 Thermatoga maritima에서 확인될 수 있다. 모든 경우에서, 상동체들은 오르니틴 카바모일트랜스퍼라제 (OCT)의 유전자들이다. 진핵계에서, 이 효소는 요소 사이클에서 두번째 단계, 카르보밀 포스페이트를 요구하는 반응인 오르니틴을 시트룰린으로의 전환을 촉매한다. 원핵계에서, ODC는 산 손상으로부터 세균을 보호하는 시스템인 아르기닌 디아미나제 활성에 관련된 세개의 효소중 하나이다. 특히, ODC는 시트룰린을 오르니틴 및 카르보밀 포스페이트 (진핵에서의 그것의 반대 역할)로의 전환을 책임진다 (Casiano-Colon, A and Marquis, R. E. 1988. Appl. Envirion. Microbiol. 54: 1318-1324, Cunin, R. et al. 1986. Microbiol. Rev. 50:314-352).
동물 보호 연구를 하기에 기재된 바와 같이 수행하였다. 그 결과는 표 1과 같다:
표 1
처 리 (치료) | # 퍼프(pups) | # 생존 퍼프 (시간) |
PBS 예비-면역 시험(테스트) | 15 41 41 | 24 48 6 0 18 1 33 14 |
실시예
2
두번째 플라스미드를 MS11로 명명하였다. 뉴클레오티드 및 추론된 아미노산 서열 (SEQ ID NOS. 3 및 4)을 단백질 데이터베이스를 검색하기 위해 사용하였다.
GBS MS11 유전자 생성물에 상동체들은 Lactobacillus delbrueckii, Thermatoga maritima , Clostridium acetobulylicum, Bacillus megaterium, Triticum aestivium 및 Synechocystis PCC6803에서 확인될 수 있다.
모든 경우에서, 상동체들은 단백질 포스포글리세레이트 키나제 (PGK)의 유전자들이다. PGK는 글리세르알데히드-3-포스페이트를 포스포에놀피루베이트로의 전환에 관련된 해당과정에서 주요한 효소이다. 특히, 그것은 이전 반응에서의 포스페이트를 방출하는, 글리세레이트-1,3-디포스페이트와 3-포스포-글리세레이트 간의 반응의 촉매에 관여된다.
실시예
3
세번째 플라스미드는 pMS16으로 명명하였다. 5' 및 3' 클론된 DNA 단편들을 서열화하고 그리고 각각에 대한 뉴클레오티드 및 추론된 아미노산 서열들을 5' 단편에 대해서는 SEQ ID NOS. 5 및 6으로 그리고 3' 단편에 대해서는 SEQ ID NOS. 7 및 8로서 도시하였다.
GBS MS16 유전자 생성물에 상동체들을 Bacillus stearothermophilus, Bacillus subtilis 및 Mycoplasma genitalium에서 확인할 수 있다.
모든 경우에서, 상동체들은 단백질 글루코스-6-포스페이트 이소메라제 (GPI)의 유전자들이다.
효소 글루코스-6-포스페이트 이소메라제는 해당과정 (G6p에서 F6p로) 및 글루코스 생성과정 (F6p에서 G6P로)에서 글루코스-6-포스페이트와 프룩토스-6-포스페이트 간의 반응을 촉매한다. gpi 유전자에서 돌연변이들은 유기체들에 퓨린 유사체 민감성을 부여하는 것으로 보여졌다.
실시예
4
네번째 플라스미드는 pMS14로 명명하였다. 클론된 DNA 단편을 서열화하고 그리고 뉴클레오티드 및 추론된 아미노산 서열 (SEQ ID NOS. 9 및 10)을 단백질 데이터베이스를 검색하기 위해 사용하였다.
GBS MS14 유전자 생성물에 상동체들은 Bacillus subtilis, Bacillus stearothermophilus, Mus musculus, Bos taurus 및 Zea mays에서 확인될 수 있다. 모든 경우에서, 상동체들은 단백질 퓨린 뉴클레오시드 포스파타제 (RNP)의 유전자들이다. 이 효소의 기능은 오르소포스페이트의 존재하에서 뉴클레오시드들 구아노신 또는 이노신을 그들 각각의 염기 및 당-1-포스페이트 분자들로 분해하는 것이다.
실시예
5
다섯번째 플라스미드를 pMS10으로 명명하였다. 클론된 DNA 단편을 서열화하였다. 뉴클레오티드 및 추론된 아미노산 서열 (SEQ ID NOS. 11 및 12)을 단백질 데이터베이스를 검색하기 위해 사용하였다.
GBS MS10 유전자 생성물에 상동체들을 Streptococcus mutans, Nicotiana plumb, Pisum sativum 및 Zea mays에서 확인될 수 있다. 모든 경우에서, 상동체들은 단백질 비인산화, NADP-의존 글리세르알데히드-3-포스페이트 디히드로게나제 (NPGAP-3-DH)의 유전자들이다. NPGAP-3-DH는 S. mutans에서 생합성 반응들을 위한 NADPH를 생산하는 (해당과정의 요구를 만족시키는 NAD-특이적 GAP-3-DH에 반대로 서) 중요한 수단으로서 보고되어 왔다 (Boyd, D.A., Cvitkovitch, D. G. and Hamilton, I. R 1995 J. Bacteriol. 177: 2622-2727).
본 발명에 따른 외표면 단백질들 및 그들의 유전자들은 면역화용 치료법 및 약품 선별에 유용하다. 또한, 본 발명의 펩티드들 또는 유전자들은 유력한 항미생물제의 선별 또는 독성(virulence)의 검출에 사용될 수 있다. 본 발명에서 동정된 생성물들을 B군 스트렙토코커스 균주에 의한 감염과 관련된 상태의 치료 또는 예방에 유용하다. 또한 본 발명의 생성물들의 수의적 사용 또한 가능하며, 특히, 펩티드 또는 백신은 특히 젖소의 만성 유선염 치료에 사용될 수 있다.
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1 5 10 15
Ser Arg Glu Glu Phe Glu Tyr Leu Ile Asp Phe Ser Ala His Leu Lys
20 25 30
Asp Leu Lys Lys Arg Gly Val Pro His His Tyr Leu Glu Gly Lys Asn
35 40 45
Ile Ala Leu Leu Phe Glu Lys Thr Ser Thr Arg Thr Arg Ala Ala Phe
50 55 60
Thr Thr Ala Ala Ile Asp Leu Gly Ala His Pro Glu Tyr Leu Gly Ala
65 70 75 80
Asn Asp Ile Gln Leu Gly Lys Lys Glu Ser Thr Glu Asp Thr Ala Lys
85 90 95
Val Leu Gly Arg Met Phe Asp Gly Ile Glu Phe Arg Gly Phe Ser Gln
100 105 110
Arg Met Val Glu Glu Leu Ala Glu Phe Ser Gly Val Pro Val Trp Asn
115 120 125
Gly Leu Thr Asp Glu Trp His Pro Thr Gln Met Leu Ala Asp Tyr Leu
130 135 140
Thr Ile Lys Glu Asn Phe Gly Lys Leu Glu Gly Ile Thr Leu Val Tyr
145 150 155 160
Cys Gly Asp Gly Arg Asn Asn Val Ala Asn Ser Leu Leu Val Ala Gly
165 170 175
Thr Leu Met Gly Val Asn Val His Ile Phe Ser Pro Lys Glu Leu Phe
180 185 190
Xaa Ala Glu Glu Ile Val Lys Leu Ala Glu Gly Tyr Ala Lys Glu Ser
195 200 205
Gly Ala His Val Leu Val Thr Asp Asn Val Asp Glu Ala Val Lys Gly
210 215 220
Ala Asp Val Phe Tyr Thr Asp Val Trp Val Ser Met Gly Glu Glu Asp
225 230 235 240
Lys Phe Lys Glu Arg Val Glu Leu Leu Gln Pro Tyr Gln Val Asn Met
245 250 255
Glu Leu Ile Lys Lys Ala Asn Asn Asp Asn Leu Ile Phe Leu His Cys
260 265 270
Leu Pro Ala Phe His Asp Thr Asn Thr Val Tyr Gly Lys Asp Val Ala
275 280 285
Glu Lys Phe Gly Val Lys Glu Met Glu Val Thr Asp Glu Val Phe Arg
290 295 300
Ser Lys Tyr Ala Arg His Phe Asp Gln Ala Glu Asn Arg Met His Thr
305 310 315 320
Ile Lys Ala Val Met Ala Ala Thr Leu Gly Asn Leu Phe Ile Pro Lys
325 330 335
Val
<210> 3
<211> 1197
<212> DNA
<213> group B streptococcus
<220>
<221> CDS
<222> (1)..(1197)
<400> 3
atg gct aaa ttg act gtt aaa gac gtt gat ttg aag gta aaa aaa gtc 48
Met Ala Lys Leu Thr Val Lys Asp Val Asp Leu Lys Val Lys Lys Val
1 5 10 15
ctc gtt cgt gtt gac ttt aat gtg cct ttg aaa gac ggc gtt atc act 96
Leu Val Arg Val Asp Phe Asn Val Pro Leu Lys Asp Gly Val Ile Thr
20 25 30
aac gac aac cgt atc act gcg gct ctt cca aca atc aag tat atc atc 144
Asn Asp Asn Arg Ile Thr Ala Ala Leu Pro Thr Ile Lys Tyr Ile Ile
35 40 45
gaa caa ggt ggt cgt gct atc ctc ttc tct cac ctt gga cgt gtt aaa 192
Glu Gln Gly Gly Arg Ala Ile Leu Phe Ser His Leu Gly Arg Val Lys
50 55 60
gaa gaa gct gac aaa gaa gga aaa tca ctt gca ccg gta gct gct gat 240
Glu Glu Ala Asp Lys Glu Gly Lys Ser Leu Ala Pro Val Ala Ala Asp
65 70 75 80
tta gct gct aaa ctt ggt caa gat gtt gta ttc cca ggt gtt act cgt 288
Leu Ala Ala Lys Leu Gly Gln Asp Val Val Phe Pro Gly Val Thr Arg
85 90 95
ggt gca aaa tta gaa gaa gca atc aat gct ttg gaa gat gga caa gtt 336
Gly Ala Lys Leu Glu Glu Ala Ile Asn Ala Leu Glu Asp Gly Gln Val
100 105 110
ctt ttg gtt gaa aac act cgt ttt gaa gat gtt gac ggt aag aaa gaa 384
Leu Leu Val Glu Asn Thr Arg Phe Glu Asp Val Asp Gly Lys Lys Glu
115 120 125
tct aag aat gac gaa gaa ctt ggt aaa tac tgg gct tca ctt gga gat 432
Ser Lys Asn Asp Glu Glu Leu Gly Lys Tyr Trp Ala Ser Leu Gly Asp
130 135 140
gga atc ttc gtt aac gat gca ttt ggt aca gca cac cgt gct cat gca 480
Gly Ile Phe Val Asn Asp Ala Phe Gly Thr Ala His Arg Ala His Ala
145 150 155 160
tca aac gta ggt att tca gca aac gtt gaa aaa gct gta gct ggt ttc 528
Ser Asn Val Gly Ile Ser Ala Asn Val Glu Lys Ala Val Ala Gly Phe
165 170 175
ctt ctt gaa aac gaa att gct tac atc caa gaa gca gtt gaa act cca 576
Leu Leu Glu Asn Glu Ile Ala Tyr Ile Gln Glu Ala Val Glu Thr Pro
180 185 190
gaa cgc cca ttc gta gct att ctt ggt ggc tca aaa gtt tct gat aag 624
Glu Arg Pro Phe Val Ala Ile Leu Gly Gly Ser Lys Val Ser Asp Lys
195 200 205
att ggt gtt atc gaa aac ctt ctt gaa aaa gct gat aaa gtt ctt atc 672
Ile Gly Val Ile Glu Asn Leu Leu Glu Lys Ala Asp Lys Val Leu Ile
210 215 220
ggt ggt ggt atg act tac aca ttc tac aaa gct caa ggt atc gaa atc 720
Gly Gly Gly Met Thr Tyr Thr Phe Tyr Lys Ala Gln Gly Ile Glu Ile
225 230 235 240
ggt aac tca ctt gta gaa gaa gac aaa ttg gat gtt gct aaa gac ctc 768
Gly Asn Ser Leu Val Glu Glu Asp Lys Leu Asp Val Ala Lys Asp Leu
245 250 255
ctt gaa aaa tca aac ggt aaa ttg atc ttg cca gtt gac tca aaa gaa 816
Leu Glu Lys Ser Asn Gly Lys Leu Ile Leu Pro Val Asp Ser Lys Glu
260 265 270
gca aac gca ttt gct ggt tat act gaa gtt cgc gac act gaa ggt gaa 864
Ala Asn Ala Phe Ala Gly Tyr Thr Glu Val Arg Asp Thr Glu Gly Glu
275 280 285
gca gtt tca gaa ggg ttc ctt ggt ctt gac atc ggt cct aaa tca atc 912
Ala Val Ser Glu Gly Phe Leu Gly Leu Asp Ile Gly Pro Lys Ser Ile
290 295 300
gct aaa ttt gat gaa gca ctt act ggt gct aaa aca gtt gta tgg aac 960
Ala Lys Phe Asp Glu Ala Leu Thr Gly Ala Lys Thr Val Val Trp Asn
305 310 315 320
gga cct atg ggt gtc ttt gaa aac cct gac ttc caa gct ggt aca atc 1008
Gly Pro Met Gly Val Phe Glu Asn Pro Asp Phe Gln Ala Gly Thr Ile
325 330 335
ggt gta atg gac gct atc gtt aaa caa cca ggc gtt aaa tca atc atc 1056
Gly Val Met Asp Ala Ile Val Lys Gln Pro Gly Val Lys Ser Ile Ile
340 345 350
ggt ggt ggt gat tca gca gca gct gct atc aac ctt ggt cgt gct gac 1104
Gly Gly Gly Asp Ser Ala Ala Ala Ala Ile Asn Leu Gly Arg Ala Asp
355 360 365
aaa ttc tca tgg atc tct act ggt ggt gga gca agc atg gaa ttg ctc 1152
Lys Phe Ser Trp Ile Ser Thr Gly Gly Gly Ala Ser Met Glu Leu Leu
370 375 380
gaa ggt aaa gta tta cca ggt ttg gca gca ttg act gaa aaa taa 1197
Glu Gly Lys Val Leu Pro Gly Leu Ala Ala Leu Thr Glu Lys
385 390 395
<210> 4
<211> 398
<212> PRT
<213> group B streptococcus
<400> 4
Met Ala Lys Leu Thr Val Lys Asp Val Asp Leu Lys Val Lys Lys Val
1 5 10 15
Leu Val Arg Val Asp Phe Asn Val Pro Leu Lys Asp Gly Val Ile Thr
20 25 30
Asn Asp Asn Arg Ile Thr Ala Ala Leu Pro Thr Ile Lys Tyr Ile Ile
35 40 45
Glu Gln Gly Gly Arg Ala Ile Leu Phe Ser His Leu Gly Arg Val Lys
50 55 60
Glu Glu Ala Asp Lys Glu Gly Lys Ser Leu Ala Pro Val Ala Ala Asp
65 70 75 80
Leu Ala Ala Lys Leu Gly Gln Asp Val Val Phe Pro Gly Val Thr Arg
85 90 95
Gly Ala Lys Leu Glu Glu Ala Ile Asn Ala Leu Glu Asp Gly Gln Val
100 105 110
Leu Leu Val Glu Asn Thr Arg Phe Glu Asp Val Asp Gly Lys Lys Glu
115 120 125
Ser Lys Asn Asp Glu Glu Leu Gly Lys Tyr Trp Ala Ser Leu Gly Asp
130 135 140
Gly Ile Phe Val Asn Asp Ala Phe Gly Thr Ala His Arg Ala His Ala
145 150 155 160
Ser Asn Val Gly Ile Ser Ala Asn Val Glu Lys Ala Val Ala Gly Phe
165 170 175
Leu Leu Glu Asn Glu Ile Ala Tyr Ile Gln Glu Ala Val Glu Thr Pro
180 185 190
Glu Arg Pro Phe Val Ala Ile Leu Gly Gly Ser Lys Val Ser Asp Lys
195 200 205
Ile Gly Val Ile Glu Asn Leu Leu Glu Lys Ala Asp Lys Val Leu Ile
210 215 220
Gly Gly Gly Met Thr Tyr Thr Phe Tyr Lys Ala Gln Gly Ile Glu Ile
225 230 235 240
Gly Asn Ser Leu Val Glu Glu Asp Lys Leu Asp Val Ala Lys Asp Leu
245 250 255
Leu Glu Lys Ser Asn Gly Lys Leu Ile Leu Pro Val Asp Ser Lys Glu
260 265 270
Ala Asn Ala Phe Ala Gly Tyr Thr Glu Val Arg Asp Thr Glu Gly Glu
275 280 285
Ala Val Ser Glu Gly Phe Leu Gly Leu Asp Ile Gly Pro Lys Ser Ile
290 295 300
Ala Lys Phe Asp Glu Ala Leu Thr Gly Ala Lys Thr Val Val Trp Asn
305 310 315 320
Gly Pro Met Gly Val Phe Glu Asn Pro Asp Phe Gln Ala Gly Thr Ile
325 330 335
Gly Val Met Asp Ala Ile Val Lys Gln Pro Gly Val Lys Ser Ile Ile
340 345 350
Gly Gly Gly Asp Ser Ala Ala Ala Ala Ile Asn Leu Gly Arg Ala Asp
355 360 365
Lys Phe Ser Trp Ile Ser Thr Gly Gly Gly Ala Ser Met Glu Leu Leu
370 375 380
Glu Gly Lys Val Leu Pro Gly Leu Ala Ala Leu Thr Glu Lys
385 390 395
<210> 5
<211> 516
<212> DNA
<213> group B streptococcus
<220>
<221> CDS
<222> (1)..(516)
<400> 5
atg aca cat att aca ttt gac tta ttc aaa gtc ttg ggt caa ttt gta 48
Met Thr His Ile Thr Phe Asp Leu Phe Lys Val Leu Gly Gln Phe Val
1 5 10 15
ggc gaa cac gag tta gac tac cta cca cca caa gta agt gca gca gat 96
Gly Glu His Glu Leu Asp Tyr Leu Pro Pro Gln Val Ser Ala Ala Asp
20 25 30
gct ttc ctt cgt caa ggg act ggt cct gga tca gat ttt ctc gga tgg 144
Ala Phe Leu Arg Gln Gly Thr Gly Pro Gly Ser Asp Phe Leu Gly Trp
35 40 45
atg gaa cct cca gaa aac tat gac aaa gaa gaa ttt tct cgc att caa 192
Met Glu Pro Pro Glu Asn Tyr Asp Lys Glu Glu Phe Ser Arg Ile Gln
50 55 60
aaa gcc gct gaa aag att aaa tca gat agc gaa gta ctc gtg gtt att 240
Lys Ala Ala Glu Lys Ile Lys Ser Asp Ser Glu Val Leu Val Val Ile
65 70 75 80
ggt att ggt ggt tcg tac ctt ggt gca aaa gca gca att gac ttt ttg 288
Gly Ile Gly Gly Ser Tyr Leu Gly Ala Lys Ala Ala Ile Asp Phe Leu
85 90 95
aat aat cat ttt gct aat ttg caa acc gca gaa gaa cgt aaa gcg cct 336
Asn Asn His Phe Ala Asn Leu Gln Thr Ala Glu Glu Arg Lys Ala Pro
100 105 110
cag att ctt tat gct gga aat tct att tca tct act tac ctt gcc gat 384
Gln Ile Leu Tyr Ala Gly Asn Ser Ile Ser Ser Thr Tyr Leu Ala Asp
115 120 125
tta gtt gaa tac gtc caa gat aaa gaa ttc tca gta aat gtc att tca 432
Leu Val Glu Tyr Val Gln Asp Lys Glu Phe Ser Val Asn Val Ile Ser
130 135 140
aaa tca ggt aca aca act gaa cca gcg att gct ttc cgt gta ttt aaa 480
Lys Ser Gly Thr Thr Thr Glu Pro Ala Ile Ala Phe Arg Val Phe Lys
145 150 155 160
gaa ctt cta gtt aaa aag tac cgg tca aga aga agc 516
Glu Leu Leu Val Lys Lys Tyr Arg Ser Arg Arg Ser
165 170
<210> 6
<211> 172
<212> PRT
<213> group B streptococcus
<400> 6
Met Thr His Ile Thr Phe Asp Leu Phe Lys Val Leu Gly Gln Phe Val
1 5 10 15
Gly Glu His Glu Leu Asp Tyr Leu Pro Pro Gln Val Ser Ala Ala Asp
20 25 30
Ala Phe Leu Arg Gln Gly Thr Gly Pro Gly Ser Asp Phe Leu Gly Trp
35 40 45
Met Glu Pro Pro Glu Asn Tyr Asp Lys Glu Glu Phe Ser Arg Ile Gln
50 55 60
Lys Ala Ala Glu Lys Ile Lys Ser Asp Ser Glu Val Leu Val Val Ile
65 70 75 80
Gly Ile Gly Gly Ser Tyr Leu Gly Ala Lys Ala Ala Ile Asp Phe Leu
85 90 95
Asn Asn His Phe Ala Asn Leu Gln Thr Ala Glu Glu Arg Lys Ala Pro
100 105 110
Gln Ile Leu Tyr Ala Gly Asn Ser Ile Ser Ser Thr Tyr Leu Ala Asp
115 120 125
Leu Val Glu Tyr Val Gln Asp Lys Glu Phe Ser Val Asn Val Ile Ser
130 135 140
Lys Ser Gly Thr Thr Thr Glu Pro Ala Ile Ala Phe Arg Val Phe Lys
145 150 155 160
Glu Leu Leu Val Lys Lys Tyr Arg Ser Arg Arg Ser
165 170
<210> 7
<211> 318
<212> DNA
<213> group B streptococcus
<220>
<221> CDS
<222> (1)..(318)
<400> 7
att aac cga aga ttt aga tgg tct tgg tta tct tca aga aaa gat gta 48
Ile Asn Arg Arg Phe Arg Trp Ser Trp Leu Ser Ser Arg Lys Asp Val
1 5 10 15
gat ttt gtt aat aaa aaa gca aca gat ggt gtg ctt ctt gct cat aca 96
Asp Phe Val Asn Lys Lys Ala Thr Asp Gly Val Leu Leu Ala His Thr
20 25 30
gat ggt ggg gtt cca aat atg ttt gta acg ctt cct aca caa gac gct 144
Asp Gly Gly Val Pro Asn Met Phe Val Thr Leu Pro Thr Gln Asp Ala
35 40 45
tac act ctt ggt tac act att tac ttc ttt gag tta gca att ggc ctt 192
Tyr Thr Leu Gly Tyr Thr Ile Tyr Phe Phe Glu Leu Ala Ile Gly Leu
50 55 60
tca ggt tat ctt aac tca gta aat cca ttt gat caa ccg ggg gta gaa 240
Ser Gly Tyr Leu Asn Ser Val Asn Pro Phe Asp Gln Pro Gly Val Glu
65 70 75 80
gca tat aaa cgt aat atg ttc gca ttt ggt aaa cct gga ttc gaa gag 288
Ala Tyr Lys Arg Asn Met Phe Ala Phe Gly Lys Pro Gly Phe Glu Glu
85 90 95
ctt agc gct gaa ttg aat gca cgt ctt taa 318
Leu Ser Ala Glu Leu Asn Ala Arg Leu
100 105
<210> 8
<211> 105
<212> PRT
<213> group B streptococcus
<400> 8
Ile Asn Arg Arg Phe Arg Trp Ser Trp Leu Ser Ser Arg Lys Asp Val
1 5 10 15
Asp Phe Val Asn Lys Lys Ala Thr Asp Gly Val Leu Leu Ala His Thr
20 25 30
Asp Gly Gly Val Pro Asn Met Phe Val Thr Leu Pro Thr Gln Asp Ala
35 40 45
Tyr Thr Leu Gly Tyr Thr Ile Tyr Phe Phe Glu Leu Ala Ile Gly Leu
50 55 60
Ser Gly Tyr Leu Asn Ser Val Asn Pro Phe Asp Gln Pro Gly Val Glu
65 70 75 80
Ala Tyr Lys Arg Asn Met Phe Ala Phe Gly Lys Pro Gly Phe Glu Glu
85 90 95
Leu Ser Ala Glu Leu Asn Ala Arg Leu
100 105
<210> 9
<211> 804
<212> DNA
<213> group B streptococcus
<220>
<221> CDS
<222> (1)..(804)
<400> 9
atg aca tta tta gaa aaa att aat gag act aga gac ttt ttg caa gca 48
Met Thr Leu Leu Glu Lys Ile Asn Glu Thr Arg Asp Phe Leu Gln Ala
1 5 10 15
aaa ggc gtc aca gca cca gaa ttt ggy ctt att tta ggc tct ggt tta 96
Lys Gly Val Thr Ala Pro Glu Phe Xaa Leu Ile Leu Gly Ser Gly Leu
20 25 30
gga gaa ttg gct gaa gaa atc gaa aat cct att gtt gtg gat tat gca 144
Gly Glu Leu Ala Glu Glu Ile Glu Asn Pro Ile Val Val Asp Tyr Ala
35 40 45
gac atc ccm aat tgg gga cag tca aca gta gtt ggt cat gct gga aaa 192
Asp Ile Xaa Asn Trp Gly Gln Ser Thr Val Val Gly His Ala Gly Lys
50 55 60
ttt agt gta tgg gat tta tca ggc cgt aag gta tta gcg ctt caa ggt 240
Phe Ser Val Trp Asp Leu Ser Gly Arg Lys Val Leu Ala Leu Gln Gly
65 70 75 80
cgt ttt cat ttt tay gaa ggw aat aca atg gaa gtc gtt act ttc cca 288
Arg Phe His Phe Tyr Glu Xaa Asn Thr Met Glu Val Val Thr Phe Pro
85 90 95
gta cgt atc atg aga gca ttg gct tgc cac agt gtg ctt gtg act aat 336
Val Arg Ile Met Arg Ala Leu Ala Cys His Ser Val Leu Val Thr Asn
100 105 110
gca gcg ggt ggg att gga tac gga cca gga act tta atg ctg atc aaa 384
Ala Ala Gly Gly Ile Gly Tyr Gly Pro Gly Thr Leu Met Leu Ile Lys
115 120 125
gac cac atc aat atg att ggg act aac cct ctc ata ggt gag aac ctt 432
Asp His Ile Asn Met Ile Gly Thr Asn Pro Leu Ile Gly Glu Asn Leu
130 135 140
gaa gaa ttt gga cca cgt ttc cca gac atg tcg gat gct tay aca gca 480
Glu Glu Phe Gly Pro Arg Phe Pro Asp Met Ser Asp Ala Tyr Thr Ala
145 150 155 160
aca tat cga caa aaa gct cac caa att gct gaa aac gat atc aaa ctc 528
Thr Tyr Arg Gln Lys Ala His Gln Ile Ala Glu Asn Asp Ile Lys Leu
165 170 175
gaa gaa ggt gtg tac ttg ggt gta tca gga ccc act tat gaa aca cct 576
Glu Glu Gly Val Tyr Leu Gly Val Ser Gly Pro Thr Tyr Glu Thr Pro
180 185 190
gca gaa att cgt gca ttc caa aca atg ggc gca caa gcg gta ggt atg 624
Ala Glu Ile Arg Ala Phe Gln Thr Met Gly Ala Gln Ala Val Gly Met
195 200 205
tcc acg gtt cca gag gtg atc gtt gca gct cac tca ggg ctt aaa gtg 672
Ser Thr Val Pro Glu Val Ile Val Ala Ala His Ser Gly Leu Lys Val
210 215 220
tta gga att tca gca att act aac ctt gcc gct ggc ttc caa tca gag 720
Leu Gly Ile Ser Ala Ile Thr Asn Leu Ala Ala Gly Phe Gln Ser Glu
225 230 235 240
ctc aat cat gag gag gtc gtt gaa gtt act cag cgt att aaa gaa gat 768
Leu Asn His Glu Glu Val Val Glu Val Thr Gln Arg Ile Lys Glu Asp
245 250 255
ttc aag gga tta ggt aaa tca tta gtt gct gaa ctc 804
Phe Lys Gly Leu Gly Lys Ser Leu Val Ala Glu Leu
260 265
<210> 10
<211> 268
<212> PRT
<213> group B streptococcus
<400> 10
Met Thr Leu Leu Glu Lys Ile Asn Glu Thr Arg Asp Phe Leu Gln Ala
1 5 10 15
Lys Gly Val Thr Ala Pro Glu Phe Xaa Leu Ile Leu Gly Ser Gly Leu
20 25 30
Gly Glu Leu Ala Glu Glu Ile Glu Asn Pro Ile Val Val Asp Tyr Ala
35 40 45
Asp Ile Xaa Asn Trp Gly Gln Ser Thr Val Val Gly His Ala Gly Lys
50 55 60
Phe Ser Val Trp Asp Leu Ser Gly Arg Lys Val Leu Ala Leu Gln Gly
65 70 75 80
Arg Phe His Phe Tyr Glu Xaa Asn Thr Met Glu Val Val Thr Phe Pro
85 90 95
Val Arg Ile Met Arg Ala Leu Ala Cys His Ser Val Leu Val Thr Asn
100 105 110
Ala Ala Gly Gly Ile Gly Tyr Gly Pro Gly Thr Leu Met Leu Ile Lys
115 120 125
Asp His Ile Asn Met Ile Gly Thr Asn Pro Leu Ile Gly Glu Asn Leu
130 135 140
Glu Glu Phe Gly Pro Arg Phe Pro Asp Met Ser Asp Ala Tyr Thr Ala
145 150 155 160
Thr Tyr Arg Gln Lys Ala His Gln Ile Ala Glu Asn Asp Ile Lys Leu
165 170 175
Glu Glu Gly Val Tyr Leu Gly Val Ser Gly Pro Thr Tyr Glu Thr Pro
180 185 190
Ala Glu Ile Arg Ala Phe Gln Thr Met Gly Ala Gln Ala Val Gly Met
195 200 205
Ser Thr Val Pro Glu Val Ile Val Ala Ala His Ser Gly Leu Lys Val
210 215 220
Leu Gly Ile Ser Ala Ile Thr Asn Leu Ala Ala Gly Phe Gln Ser Glu
225 230 235 240
Leu Asn His Glu Glu Val Val Glu Val Thr Gln Arg Ile Lys Glu Asp
245 250 255
Phe Lys Gly Leu Gly Lys Ser Leu Val Ala Glu Leu
260 265
<210> 11
<211> 1428
<212> DNA
<213> group B streptococcus
<220>
<221> CDS
<222> (1)..(1428)
<400> 11
ttg aca aaa gaa tat caa aat tat gtc aat ggc gaa tgg aaa tca tct 48
Leu Thr Lys Glu Tyr Gln Asn Tyr Val Asn Gly Glu Trp Lys Ser Ser
1 5 10 15
gtt aat cag att gag att ttg tca cca att gat gat tct tca ttg gga 96
Val Asn Gln Ile Glu Ile Leu Ser Pro Ile Asp Asp Ser Ser Leu Gly
20 25 30
ttc gtg cca gcg atg act cga gaa gaa gtt gat cat gct atg aaa gcg 144
Phe Val Pro Ala Met Thr Arg Glu Glu Val Asp His Ala Met Lys Ala
35 40 45
ggt cgt gag gct tta cca gct tgg gct gct tta aca gta tat gaa cgt 192
Gly Arg Glu Ala Leu Pro Ala Trp Ala Ala Leu Thr Val Tyr Glu Arg
50 55 60
gca caa tac ctt cat aaa gcc gca gac att att gaa cgt gat aaa gaa 240
Ala Gln Tyr Leu His Lys Ala Ala Asp Ile Ile Glu Arg Asp Lys Glu
65 70 75 80
gaa att gct act gtt tta gca aaa gaa att tct aaa gct tac aat gct 288
Glu Ile Ala Thr Val Leu Ala Lys Glu Ile Ser Lys Ala Tyr Asn Ala
85 90 95
tca gta act gag gtt gta agg aca gct gat ctt att cgt tat gca gca 336
Ser Val Thr Glu Val Val Arg Thr Ala Asp Leu Ile Arg Tyr Ala Ala
100 105 110
gaa gaa gga att cgt tta tca act tca gct gac gaa ggt gga aaa atg 384
Glu Glu Gly Ile Arg Leu Ser Thr Ser Ala Asp Glu Gly Gly Lys Met
115 120 125
gat gct tca aca ggt cat aag ttg gct gtt att cgt cgt caa cca gta 432
Asp Ala Ser Thr Gly His Lys Leu Ala Val Ile Arg Arg Gln Pro Val
130 135 140
ggt atc gtt tta gca atc gca cct tat aat tac cct gtt aac ctc tca 480
Gly Ile Val Leu Ala Ile Ala Pro Tyr Asn Tyr Pro Val Asn Leu Ser
145 150 155 160
gga tca aaa att gcg cca gct cta att ggt gga aac gtt gtg atg ttt 528
Gly Ser Lys Ile Ala Pro Ala Leu Ile Gly Gly Asn Val Val Met Phe
165 170 175
aaa cca cca aca caa ggt tca gtc tca gga ctt gtt tta gca aaa gct 576
Lys Pro Pro Thr Gln Gly Ser Val Ser Gly Leu Val Leu Ala Lys Ala
180 185 190
ttt gca gaa gca ggt ctt cca gca ggt gtc ttt aat act att aca gga 624
Phe Ala Glu Ala Gly Leu Pro Ala Gly Val Phe Asn Thr Ile Thr Gly
195 200 205
cgc ggt tct gag att gga gat tac att gtt gag cat gaa gaa gtt aat 672
Arg Gly Ser Glu Ile Gly Asp Tyr Ile Val Glu His Glu Glu Val Asn
210 215 220
ttt att aac ttt aca gga tca acg cca gtt gga caa cgt att ggt aag 720
Phe Ile Asn Phe Thr Gly Ser Thr Pro Val Gly Gln Arg Ile Gly Lys
225 230 235 240
ttg gca gga atg cgt cca att atg ctt gag ttg ggc ggt aag gat gca 768
Leu Ala Gly Met Arg Pro Ile Met Leu Glu Leu Gly Gly Lys Asp Ala
245 250 255
ggt atc gtc tta gct gat gct gac ctt gat aac gct gct aaa caa atc 816
Gly Ile Val Leu Ala Asp Ala Asp Leu Asp Asn Ala Ala Lys Gln Ile
260 265 270
gtt gca ggt gct tat gat tac tct gga caa cgc tgt acg gca att aag 864
Val Ala Gly Ala Tyr Asp Tyr Ser Gly Gln Arg Cys Thr Ala Ile Lys
275 280 285
cgt gtg ctt gtc gtt gaa gaa gtt gcw gat gaa ttg gca gaa aaa ata 912
Arg Val Leu Val Val Glu Glu Val Xaa Asp Glu Leu Ala Glu Lys Ile
290 295 300
tct gaa aat gta gca aaa tta tca gta ggt gat cca ttt gat aat gca 960
Ser Glu Asn Val Ala Lys Leu Ser Val Gly Asp Pro Phe Asp Asn Ala
305 310 315 320
acg gtg aca ccg gtt att gat gat aat tca gct gac ttt att gaa agc 1008
Thr Val Thr Pro Val Ile Asp Asp Asn Ser Ala Asp Phe Ile Glu Ser
325 330 335
tta gta gta gat gca cgt caa aaa ggt gcg aaa gaa ttg aat gaa ttt 1056
Leu Val Val Asp Ala Arg Gln Lys Gly Ala Lys Glu Leu Asn Glu Phe
340 345 350
aaa cgt gat ggt cgt cta tta act cca gga ttg ttt gat cat gtt act 1104
Lys Arg Asp Gly Arg Leu Leu Thr Pro Gly Leu Phe Asp His Val Thr
355 360 365
tta gat atg aaa cta gct tgg gaa gag cct ttt gga cca att ctc cca 1152
Leu Asp Met Lys Leu Ala Trp Glu Glu Pro Phe Gly Pro Ile Leu Pro
370 375 380
att att cgt gtc aag gat gca gaa gaa gct gtt gct att gcc aac aaa 1200
Ile Ile Arg Val Lys Asp Ala Glu Glu Ala Val Ala Ile Ala Asn Lys
385 390 395 400
tct gat ttt gga tta caa tca tca gtc ttt aca cgt gat ttc caa aaa 1248
Ser Asp Phe Gly Leu Gln Ser Ser Val Phe Thr Arg Asp Phe Gln Lys
405 410 415
gca ttt gat ata gca aat aaa ctt gaa gtt ggt aca gtt cac att aac 1296
Ala Phe Asp Ile Ala Asn Lys Leu Glu Val Gly Thr Val His Ile Asn
420 425 430
aat aag act gga cgt ggt ccw gat aat ttc cca ttc tta gga ctc aaa 1344
Asn Lys Thr Gly Arg Gly Xaa Asp Asn Phe Pro Phe Leu Gly Leu Lys
435 440 445
gga tct ggt gca ggt gtt caa ggt atc aga tat tca att gaa gca atg 1392
Gly Ser Gly Ala Gly Val Gln Gly Ile Arg Tyr Ser Ile Glu Ala Met
450 455 460
aca aat gta aaa tcg att gtt ctc gat atg aaa tag 1428
Thr Asn Val Lys Ser Ile Val Leu Asp Met Lys
465 470 475
<210> 12
<211> 475
<212> PRT
<213> group B streptococcus
<400> 12
Leu Thr Lys Glu Tyr Gln Asn Tyr Val Asn Gly Glu Trp Lys Ser Ser
1 5 10 15
Val Asn Gln Ile Glu Ile Leu Ser Pro Ile Asp Asp Ser Ser Leu Gly
20 25 30
Phe Val Pro Ala Met Thr Arg Glu Glu Val Asp His Ala Met Lys Ala
35 40 45
Gly Arg Glu Ala Leu Pro Ala Trp Ala Ala Leu Thr Val Tyr Glu Arg
50 55 60
Ala Gln Tyr Leu His Lys Ala Ala Asp Ile Ile Glu Arg Asp Lys Glu
65 70 75 80
Glu Ile Ala Thr Val Leu Ala Lys Glu Ile Ser Lys Ala Tyr Asn Ala
85 90 95
Ser Val Thr Glu Val Val Arg Thr Ala Asp Leu Ile Arg Tyr Ala Ala
100 105 110
Glu Glu Gly Ile Arg Leu Ser Thr Ser Ala Asp Glu Gly Gly Lys Met
115 120 125
Asp Ala Ser Thr Gly His Lys Leu Ala Val Ile Arg Arg Gln Pro Val
130 135 140
Gly Ile Val Leu Ala Ile Ala Pro Tyr Asn Tyr Pro Val Asn Leu Ser
145 150 155 160
Gly Ser Lys Ile Ala Pro Ala Leu Ile Gly Gly Asn Val Val Met Phe
165 170 175
Lys Pro Pro Thr Gln Gly Ser Val Ser Gly Leu Val Leu Ala Lys Ala
180 185 190
Phe Ala Glu Ala Gly Leu Pro Ala Gly Val Phe Asn Thr Ile Thr Gly
195 200 205
Arg Gly Ser Glu Ile Gly Asp Tyr Ile Val Glu His Glu Glu Val Asn
210 215 220
Phe Ile Asn Phe Thr Gly Ser Thr Pro Val Gly Gln Arg Ile Gly Lys
225 230 235 240
Leu Ala Gly Met Arg Pro Ile Met Leu Glu Leu Gly Gly Lys Asp Ala
245 250 255
Gly Ile Val Leu Ala Asp Ala Asp Leu Asp Asn Ala Ala Lys Gln Ile
260 265 270
Val Ala Gly Ala Tyr Asp Tyr Ser Gly Gln Arg Cys Thr Ala Ile Lys
275 280 285
Arg Val Leu Val Val Glu Glu Val Xaa Asp Glu Leu Ala Glu Lys Ile
290 295 300
Ser Glu Asn Val Ala Lys Leu Ser Val Gly Asp Pro Phe Asp Asn Ala
305 310 315 320
Thr Val Thr Pro Val Ile Asp Asp Asn Ser Ala Asp Phe Ile Glu Ser
325 330 335
Leu Val Val Asp Ala Arg Gln Lys Gly Ala Lys Glu Leu Asn Glu Phe
340 345 350
Lys Arg Asp Gly Arg Leu Leu Thr Pro Gly Leu Phe Asp His Val Thr
355 360 365
Leu Asp Met Lys Leu Ala Trp Glu Glu Pro Phe Gly Pro Ile Leu Pro
370 375 380
Ile Ile Arg Val Lys Asp Ala Glu Glu Ala Val Ala Ile Ala Asn Lys
385 390 395 400
Ser Asp Phe Gly Leu Gln Ser Ser Val Phe Thr Arg Asp Phe Gln Lys
405 410 415
Ala Phe Asp Ile Ala Asn Lys Leu Glu Val Gly Thr Val His Ile Asn
420 425 430
Asn Lys Thr Gly Arg Gly Xaa Asp Asn Phe Pro Phe Leu Gly Leu Lys
435 440 445
Gly Ser Gly Ala Gly Val Gln Gly Ile Arg Tyr Ser Ile Glu Ala Met
450 455 460
Thr Asn Val Lys Ser Ile Val Leu Asp Met Lys
465 470 475
Claims (11)
- B군 스트렙토코커스로부터 얻을 수 있는 SEQ ID NOS. 2, 4, 6, 8 또는 10에 나타낸 아미노산 서열, 또는 그것의 상동체, 또는 전체-길이의 펩티드에 대한 면역반응을 유도할 수 있는 능력을 보유하는 그것의 기능성 단편을 포함하는, 치료학적 사용을 위한 펩티드.
- 제 1항에 있어서, 상기 상동체는 아미노산 수준 (level)으로 80% 초과의 상사성 (similarity)을 갖는 것인 펩티드.
- 치료학적 사용을 위한, 제 1항에 따른 펩티드를 코드화하는 폴리뉴클레오티드.
- 제 1항에 따른 펩티드를 포함하는 백신.
- 제 1항 또는 제 3항에 따른 펩티드 또는 폴리뉴클레오티드를 사용하는 것을 특징으로 하는 유력한 항미생물 약제의 선별방법.
- 제 1항 또는 제 3항에 따른 펩티드 또는 폴리뉴클레오티드를 포함하는, 세균성 감염과 연관된 질환의 치료 또는 예방용 의약.
- 제 6항에 있어서, 상기 감염이 B군 스트렙토코커스 감염인 것을 특징으로 하는 의약.
- 제 6항에 있어서, 상기 감염이 병소 감염인 것을 특징으로 하는 의약.
- 제 6항에 있어서, 상기 감염은 요로 감염인 것을 특징으로 하는 의약.
- SEQ ID NOS. 2, 4, 6, 8 또는 10에 나타낸 아미노산 서열에 특이적으로 결합하는 항체.
- 제 1항에 따른 펩티드를 사용하는 것을 특징으로 하는 항체의 생성방법.
Applications Claiming Priority (12)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GBGB9828346.8A GB9828346D0 (en) | 1998-12-22 | 1998-12-22 | Protein and compositions containing it |
GB9828346.8 | 1998-12-22 | ||
GBGB9901234.6A GB9901234D0 (en) | 1999-01-20 | 1999-01-20 | Protein and compositions containing it |
GBGB9901233.8A GB9901233D0 (en) | 1999-01-20 | 1999-01-20 | Protein and compositions containing it |
GB9901234.6 | 1999-01-20 | ||
GB9901233.8 | 1999-01-20 | ||
GBGB9908321.4A GB9908321D0 (en) | 1999-04-12 | 1999-04-12 | Purine nucleoside phosphatase and compositions containing it |
GB9908321.4 | 1999-04-12 | ||
GB9912036.2 | 1999-05-24 | ||
GBGB9912036.2A GB9912036D0 (en) | 1999-05-24 | 1999-05-24 | Glucose-6-phosphate isomerase and compositions containing it |
GB9922596.3 | 1999-09-23 | ||
GBGB9922596.3A GB9922596D0 (en) | 1999-09-23 | 1999-09-23 | Protein and compositions containing it |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
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KR1020017007910A Division KR100818815B1 (ko) | 1998-12-22 | 1999-12-22 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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KR1020070008368A Division KR20070027665A (ko) | 1998-12-22 | 2007-01-26 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
Publications (2)
Publication Number | Publication Date |
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KR20060096515A true KR20060096515A (ko) | 2006-09-11 |
KR100866170B1 KR100866170B1 (ko) | 2008-10-30 |
Family
ID=27547330
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
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KR1020017007910A KR100818815B1 (ko) | 1998-12-22 | 1999-12-22 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
KR1020067017195A KR100866170B1 (ko) | 1998-12-22 | 1999-12-22 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
KR1020070008368A KR20070027665A (ko) | 1998-12-22 | 2007-01-26 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
KR1020080041439A KR20080052527A (ko) | 1998-12-22 | 2008-05-02 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
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KR1020017007910A KR100818815B1 (ko) | 1998-12-22 | 1999-12-22 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
Family Applications After (2)
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KR1020070008368A KR20070027665A (ko) | 1998-12-22 | 2007-01-26 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
KR1020080041439A KR20080052527A (ko) | 1998-12-22 | 2008-05-02 | 외표면 단백질들, 그들의 유전자들 및 그들의 사용 |
Country Status (24)
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US (3) | US7217415B1 (ko) |
EP (2) | EP1574579A3 (ko) |
JP (1) | JP2002533065A (ko) |
KR (4) | KR100818815B1 (ko) |
CN (3) | CN1733800A (ko) |
AP (1) | AP2006003504A0 (ko) |
AT (1) | ATE297995T1 (ko) |
AU (1) | AU758722B2 (ko) |
BR (1) | BR9916473A (ko) |
CA (1) | CA2354843A1 (ko) |
CZ (1) | CZ20012174A3 (ko) |
DE (1) | DE69925866T2 (ko) |
DK (1) | DK1140994T3 (ko) |
EA (1) | EA009885B1 (ko) |
ES (1) | ES2241352T3 (ko) |
HK (1) | HK1039339A1 (ko) |
HU (1) | HUP0201022A3 (ko) |
NO (1) | NO20013101L (ko) |
NZ (4) | NZ525538A (ko) |
OA (1) | OA12425A (ko) |
PL (1) | PL351027A1 (ko) |
PT (1) | PT1140994E (ko) |
SI (1) | SI1140994T1 (ko) |
WO (1) | WO2000037490A2 (ko) |
Families Citing this family (6)
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HUP0201022A3 (en) * | 1998-12-22 | 2004-07-28 | Microscience Ltd Wokingham | Outer surface proteins, their genes, and their use |
ES2281977T3 (es) * | 1998-12-22 | 2007-10-01 | Microscience Limited | Proteinas de estreptococo del grupo b y su utilizacion. |
US6890539B2 (en) * | 1998-12-22 | 2005-05-10 | Microscience, Ltd. | Genes and proteins, and their use |
WO2008152448A2 (en) * | 2006-12-21 | 2008-12-18 | Emergent Product Development Uk Limited | Streptococcus proteins, and their use in vaccination |
CN103060284A (zh) * | 2012-10-31 | 2013-04-24 | 内蒙古民族大学 | 一种无乳链球菌pgk亚单位重组蛋白及其制备方法 |
CN110669711A (zh) * | 2019-08-09 | 2020-01-10 | 中国水产科学研究院珠江水产研究所 | 基于pgk基因的重组乳酸乳球菌和无乳链球菌病疫苗 |
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DE3678702D1 (de) * | 1985-12-28 | 1991-05-16 | Ohgen Research Lab Ltd | Antitumorprotein-gen von streptococcus pyogenes su, plasmide, die das gen enthalten, transformantenzellen, die die plasmide beherbergen, und verfahren zur herstellung des antitumorproteins. |
US5328996A (en) * | 1989-03-29 | 1994-07-12 | University Of Florida Research Foundation, Inc. | Bacterial plasmin receptors as fibrinolytic agents |
WO1993014198A1 (en) * | 1992-01-08 | 1993-07-22 | The Rockefeller University | Multifunctional surface protein of streptococci |
IL107458A0 (en) * | 1992-11-02 | 1994-02-27 | Gen Hospital Corp | Conjugate vaccine against group b streptococcus |
US6015889A (en) * | 1993-03-19 | 2000-01-18 | Gunnar Lindahl | Protein rib, a cell surface protein that confers immunity to many strains of the group B streptococcus: process for purification of the protein, reagent kit and pharmaceutical composition |
EP0656014B1 (en) * | 1993-03-19 | 2003-05-14 | LINDAHL, Gunnar | Protein rib, a cell surface protein that confers immunity to many strains of the group b streptococcus; process for purification of the protein, reagent kit and pharmaceutical composition |
US6346392B1 (en) * | 1996-11-27 | 2002-02-12 | Smithkline Beecham Corporation | Polynucleotides encoding a novel glutamine transport ATP-binding protein |
EP1770164B1 (en) * | 1996-10-31 | 2010-09-01 | Human Genome Sciences, Inc. | Streptococcus pneumoniae antigens and vaccines |
US7196164B2 (en) * | 1997-07-08 | 2007-03-27 | Human Genome Sciences, Inc. | Secreted protein HHTLF25 |
US6800744B1 (en) * | 1997-07-02 | 2004-10-05 | Genome Therapeutics Corporation | Nucleic acid and amino acid sequences relating to Streptococcus pneumoniae for diagnostics and therapeutics |
US7081530B1 (en) * | 1997-07-02 | 2006-07-25 | sanofi pasteur limitée | Nucleic acid and amino acid sequences relating to Streptococcus pneumoniae for diagnostics and therapeutics |
US6380370B1 (en) * | 1997-08-14 | 2002-04-30 | Genome Therapeutics Corporation | Nucleic acid and amino acid sequences relating to Staphylococcus epidermidis for diagnostics and therapeutics |
US6165763A (en) * | 1997-10-30 | 2000-12-26 | Smithkline Beecham Corporation | Ornithine carbamoyltransferase |
US6248329B1 (en) * | 1998-06-01 | 2001-06-19 | Ramaswamy Chandrashekar | Parasitic helminth cuticlin nucleic acid molecules and uses thereof |
ES2281977T3 (es) * | 1998-12-22 | 2007-10-01 | Microscience Limited | Proteinas de estreptococo del grupo b y su utilizacion. |
US6890539B2 (en) * | 1998-12-22 | 2005-05-10 | Microscience, Ltd. | Genes and proteins, and their use |
HUP0201022A3 (en) * | 1998-12-22 | 2004-07-28 | Microscience Ltd Wokingham | Outer surface proteins, their genes, and their use |
US6605709B1 (en) * | 1999-04-09 | 2003-08-12 | Genome Therapeutics Corporation | Nucleic acid and amino acid sequences relating to Proteus mirabilis for diagnostics and therapeutics |
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