Development of Selectable Vector Plasmid in Bacteriophage P2-P4 System and Its Stability

박테리오파지 P2-P4 시스템을 위한 벡터 플라스미드 개발과 안정성

  • Kim, Kyoung-Jin (Department of Microbiology, College of Natural Sciences, Sunmoon University)
  • 김경진 (선문대학교 자연과학대학 미생물학과)
  • Received : 1998.07.16
  • Accepted : 1998.09.25
  • Published : 1998.12.01

Abstract

While bacteriophage P2-P4 system is very useful experimental tool for the study of viral capsid assembly, there is no useful plasmid vector for the DNA manipulation in bacteriophage P2-P4 system. In this study, a new vector plasmid, P4 ash8 (sid71) kmr, was constructed by swapping the non-essential region of P4 DNA for kanamycin resistance(kmr) gene cassette of plasmid pUC4-K. P4 ash8 sid71 was starting material for the construction, since it tends to be maintained as a plasmid in the absence of the helper phage. The total size of this chimera was designed to be packaged into P4 or P2 size heads with induction by P2 infection. The conversion of plasmid P4 ash8 (sid71) kmr to bacteriophage was proved by burst size determination experiment and CsCl buoyant equilibrium density gradient experiment. Integrase destructed P4 derivative, P4 ash8 sid71 kmr intS, was able to be constructed easily by in vitro DNA manipulation of P4 ash8 sid71 kmr. The plasmid stability experiment with P4 ash8 sid71 kmr if/tS showed that the integrase of P4 affects the stable maintenance of plasmid P4 state.

Keywords

bacteriophage P2-P4;Escherichia coli;integrase;phagemid;plasmid