Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Construction of New P4-Derived Vector Plasmid Containing Tetracyclin Resistance Marker for the Bacteriophage P2-P4 System

박테리오파아지 P2-P4 시스템을 위한 tetracyclin resistance marker 함유 P4 유도체 벡터 플라스미드 조성

  • Kim, Kyoung-Jin (Department of Applied Molecular Biology, Division of Applied Biology, Sunmoon University)
  • 김경진 (선문대학교 응용생물과학부 응용분자생물학전공)
  • Published : 2003.06.01
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Abstract

To develop vector plasmid for the bacteriophage P2-P4 system which is a useful experimental tool for the study of viral capsid assembly, we constructed a new P4-derived vector plasmid starting from P4 ash8 sid71 With recombinant DNA technology, a portion of P4 genome was deleted and tetracyclin resistance gene (terR) was introduced into P4 genome to give P4 selectivity. Resulting P4 ash8(sid71) terR was 12.09 kb long and could be converted to a viable bacteriophage with P2 infection. The burst size of induced bacteriophage form of P4 ash8(sid71) terR was determined. The CsCl buoyant equilibrium density gradient experiment of new P4 derivative suggested the upper limit of packaging capacity in P2-size head.

바이러스 조립 과정 기작 연구를 위한 좋은 재료인 박데리오파아지 P2-P4시스템에 이용될 벡터 플라스미드를 개발하기 위하여, P4 ash8 sid71을 출발 물질로 삼아 새로운 P4 유도체 벡터를 조성하였다. 유전자 재조합 기법을 써서 쉽게 선택 가능한 tetracyclin내성 유전자(tetR)를 도입하고 플라스미드P 4의 크기를 조절하였다. 이를 통해 얻어진 P4 ash8(sid7l) tetR은 12.09 kb의 크기를 가지며, 필요할 때 P2로 induction하면 생물학적 활성을 가지는 박데리오파아지로 전환 가능하였다. 전환된 파아지의 burst size를 결정하고, CsCi 부양균등밀도 편차실험을 수행하였다. 균등밀도 실험 분포도에서 P2크기 파아지 머리의 packaging 상한을 추정할 수 있었다.

Keywords

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