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Actinobacillus actinomycetemcomitans의 cytolethal distending toxin subunit CdtA 유전자 클로닝과 단백질 발현

Cloning and protein expression of Actinobacillus actinomycetemcomitans cytolethal distending toxin subunit CdtA

정동근|유소현|김형섭|고선영
대한치주과학회 (The Korean Academy Of Periodontology)|2007.08|Journal of Periodontal & Implant Science|제37권 제5호|pp. 339-351

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참고문헌

키워드

Actinobacillus actinomycetemcomitanscloningCytolethal distending toxin
주제분류치의학

초록

Cytolethal distending toxin(CDT)은 세포 주기 중 G2에서 M 기로의 전환을 막아 세포의 증식을 억제할 수 있는 세균 단백 독소의 일종이다. 구강 미생물 중 유일하게 Actinobacillus　actinomycetemcomitans (A. actinomycetemcomitans)만이 이 CDT를 생성 할 수 있는 것으로 알려져 있다. A. actinomycetemcomitans는 localized aggressive periodontitis (LAP)의 원인균으로 여겨지며 비 운동성의 그람 음성 구간균이고 37℃, 5% CO2 하에 성장이 왕성하다. A. actinomycetemcomitans 의 CDT는 3개의 인접한 유전자인 cdtA, cdtB, cdtC 에 의해 형성 되며 각각의 유전자에 대한 단백질의 기능은 아직 완전히 밝혀지지 않았다. 현재까지 연구에 의하면 cdtA는 CDT의 세포부착과 관련이 있는 것으로 여겨지며 이 유전자의 기능 이상 시 CDT 의 독성 효과가 현저히 감소한다고 알려져 있다. 따라서 본 연구는 A. actinomycetemcomitans 의 cdtA 유전자를 클로닝, 단백질 발현하여 향후 치주질환의 발병 과정에서 CdtA의 역할을 규명하고 질환의 예방 및 치료법에 도움을 주고자 하였다. A. actinomycetemcomitans Y4 균주를 cdtA 유전자 클로닝을 위해 사용하였다. A. actinomycetemcomitans 의 genomic DNA는 genomic DNA 추출 kit를 사용하여 분리하고 cdtA에 특이적인 primer를 이용하여 PCR을 통해 cdtA 유전자를 증폭하였다. 증폭된 cdtA 유전자를 T-vector에 클로닝 하였으며, 클로닝 된 cdtA 유전자는 단백질 발현을 위해 pRSET A vector에 서브클로닝 한 후 발현 균주인 BL21(DE3)를 이용하여 발현시켰다. 발현 후 Ni-NTA AP conjugate를 이용한 Western blot을 통해 pRSET-CDTA를 확인하였다.

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관련 논문AI추천

Cloning and protein expression of Aggregatibacter actinomycetemcomitans cytolethal distending toxin C김형섭, 박소영, 이은선, 이은숙대한치주과학회 · Journal of Periodontal & Implant Science2008 · 제38권 제2호Co-expression of CdtA and CdtC subunits of cytolethal distending toxin from Aggregatibacter actinomycetemcomitans김형섭, 이경열, Seung-Jae Lee대한치주과학회 · Journal of Periodontal & Implant Science2009 · 제39권 제2호Production of the polyclonal subunit C protein antibody against Aggregatibacter actinomycetemcomitans cytolethal distending toxin고선영, 김형섭, 박소영, 유소현, 이수정대한치주과학회 · Journal of Periodontal & Implant Science2008 · 제38권 제2호Purification and refolding of the recombinant subunit B protein of the Aggregatibacter actinomycetemcomitans cytolethal distending toxin고선영, 권진희, 김형섭, 서성찬, 전용선대한치주과학회 · Journal of Periodontal & Implant Science2008 · 제38권 제2호치면세균막내의 Fusobacterium nucleatum과 Actinobacillus actinomycetemcomitans의 동정을 위한 세균배양법 및 Multiplex PCR법의 비교김화숙, 임선아한국치위생과학회 · 치위생과학회지2009 · 제9권 제2호Actinobacillus actinomycetemcomitans 내독소가？백서 치주조직에 미치는 영향김영준, 김종철, 최득철대한치주과학회 · Journal of Periodontal & Implant Science2007 · 제37권 제5호

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