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Characterization of the bacteriophage P4 sid+ derivative overcoming P2sir-associated helper inefficiency through DNA conformational adaptation

DNA 형태 적응을 거쳐 P2sir-관련 도움파지 비효율성을 극복하는 박테리오파지 P4 sid+ 유도체 정성 연구

  • Kim, Kyoung-Jin (Department of BT-Convergent Pharmaceutical Engineering, College of Health Sciences, Sunmoon University)
  • 김경진 (선문대학교 건강보건대학 BT-융합제약공학과)
  • Received : 2016.02.25
  • Accepted : 2016.03.21
  • Published : 2016.03.31

Abstract

A certain size of DNA (28-29 kb long) to be packaged into P2-size head and the mutation in sid gene of bacteriophage P4 are the major factors to overcome "P2 sir-associated helper inefficiency". To clarify whether the presence of sid mutation is essential to overcome "P2 sir-associated helper inefficiency" or not, we tested the P4 derivative, P4 delRI::kmr, which is $sid^+$ and whose genome size supposed to be 28.5 kb long in the case of being packaged into $P2_{sir3}$-sized large head. As P4 delRI::kmr showed the low EOP with P2 sir3 lysogen, P4 delRI::kmr phage stock was prepared in P2 sir3 lysogen host to increase the EOP with P2 sir3 lysogen. Through this process, P4 delRI::kmr had been adapted for P2 sir3 lysogen. With a CsCl buoyant equilibrium density gradient experiment and gel electrophoresis of the isolated DNA, it was evident that the adaptation of P4 delRI::kmr for P2 sir3 lysogen was caused by the conformational change of DNA to be packaged into large head. The burst size determination experiments with P4 delRI::kmr phage stock adapted for P2 sir3 lysogen and normal P4 delRI::kmr phage stock showed that not the sid mutation but the size of DNA to be packaged (28-29 kb long) was essential to overcome "P2 sir-associated helper inefficiency".

Keywords

adaptation;bacteriophage P2-P4;DNA conformation;sid mutation;sir mutation

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