Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Expression and Purification of Bacteriophage Lambda Integrase by Fusion Protein System

단백질 융합 시스템을 이용한 Bacteriophage Lambda Integrase의 발현 및 정제

  • 이나영 (연세대학교 공과대학 식품생물공학과 및 생물산업소재연구센터) ;
  • 유승구 (연세대학교 공과대학 식품생물공학과 및 생물산업소재연구센터)
  • Published : 1995.12.01
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Abstract

The lambda Integrase (Int) carries out site-specific recombination between the two partner DNA sequences, attachment P (attP) and attachment B (attB). In order to study the recombination mechanism, a large quantity of pure integrase is required. Then, we constructed an int gene inserted recombinant plasmid (pNYL3) by using the pQE31 HIS-Tag vector, and produced the fusion protein, 6xHIS-Int from the E. coli TG1 strain carrying the pNYL3 plasmid. The recombinant protein produced was purified by phosphocellulose and Ni$^{++}$-NTA affinity column chromatographies. The result of the in vitro recombination assay using the standard reaction mixture containing 6xHIS-Int and partially purified integration host factor (IHF) showed that the 6xHIS-Int tagged recombination Integrase had the full recombination activity.

Keywords

References

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