미생물학회지 (Korean Journal of Microbiology)

  • 제49권1호
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  • Pages.99-104
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  • 2013
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  • 0440-2413(pISSN)
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  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
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박테리오파지 P2의 cos 지역을 함유하는 박테리오파지 P4 유도체인 P4 sid71 cosP2의 In vitro 조성과 정성 연구

In Vitro Construction and Characterization of the Bacteriophage P4 Derivative, P4 sid71 cosP2, Containing the Bacteriophage P2 cos Region

  • 김경진 (선문대학교 건강보건대학 의생명과학과)
  • Kim, Kyoung-Jin (Department of Biomedical Sciences, College of Health Sciences, Sun Moon University)
  • 투고 : 2013.02.13
  • 심사 : 2013.03.08
  • 발행 : 2013.03.31
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초록

박테리오파지 P2 sir 변이체는 그것의 위성파지인 박테리오 파지 P4를 위해 비효율적인 도움파지로 알려졌다. 이러한 현상을 "P2 sir-관련 도움파지 비효율성"이라고 부르고 있으며, P2와 P4의 cos 지역에서의 DNA 염기배열 순서 차이가 이러한 현상을 나타나게 한다고 생각되었다. 이를 검증하기 위해, 여러 단계의 in vitro DNA 조작을 거쳐 P2의 cos 지역을 함유하는 유도체 P4인 P4 sid71 cosP2를 조성하였다. 일단계 생장 실험을 통해 그것의 후손 방출량을 결정하였다. 그 결과는, P4 sid71 cosP2에서 P4의 cos 지역을 P2의 cos 지역으로 대체한 것이 "P2 sir-관련 도움파지 비효율성"을 극복한 것으로 나타났다. P2 야생형과 P2 sir 돌연변이형을 도움파지로 삼아 준비한 P4 sid71 cosP2 파지 농축액들을 CsCl 부양 균등밀도 편차실험으로 분석하였다. 그 결과, 양 경우 모두 3개의 P4 유전체를 가진 파지 입자가 우세한 것으로 나타났다.

Bacteriophage P2 sir mutants are inefficient helpers for their satellite bacteriophage P4. The term, "P2 sir-associated helper inefficiency" has been used to define this phenomenon and it has been suggested that the DNA sequence difference between the cos region of P2 and that of P4 is responsible. To test this hypothesis, P4 derivative phage, P4 sid71 cosP2, containing the cos region of P2 and sid71 allele was constructed through several in vitro DNA manipulation steps. Its burst size was determined using a one-step growth experiment. The results showed that the substitution of the cos region of P2 for the cos region of P4 in P4 sid71 cosP2 overcame "P2 sir-associated helper inefficiency". P4 sid71 cosP2 stock phages prepared with P2 wild type helper and P2 sir helper were analyzed using a CsCl buoyant equilibrium density gradient experiment. The results revealed that the phage particles containing three copies of the P4 genome were the predominant particles in both cases.

키워드

참고문헌

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피인용 문헌

  1. Construction and characterization of the bacteriophage P4 derivatives whose genome size suitable for packaging into a P2sir3-sized head vol.51, pp.1, 2015, https://doi.org/10.7845/kjm.2015.5009