Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Expression of the lux Genes in Escherichia coli for the Basis of Development of Biosensor

바이오센서 개발을 위한 발광 유전자의 대장균에서의 발현 시스템 분석

  • Cho, Mi-Mi (Department of Biochemistry, Chungnam National University) ;
  • Kim, Young-Doo (Department of Biochemistry, Chungnam National University) ;
  • Kang, Kyung-Sook (Department of Biochemistry, Chungnam National University) ;
  • Kim, Sook-Kyung (Center for Bioanalysis, Korea Research Institute of Standards and Science) ;
  • Yang, In-Chul (Center for Bioanalysis, Korea Research Institute of Standards and Science) ;
  • Park, Sang-Ryoul (Center for Bioanalysis, Korea Research Institute of Standards and Science) ;
  • Lee, Chan-Yong (Department of Biochemistry, Chungnam National University)
  • 조미미 (충남대학교 생화학과) ;
  • 김영두 (충남대학교 생화학과) ;
  • 강경숙 (충남대학교 생화학과) ;
  • 김숙경 (한국표준과학연구원 바이오임상표준센터) ;
  • 양인철 (한국표준과학연구원 바이오임상표준센터) ;
  • 박상열 (한국표준과학연구원 바이오임상표준센터) ;
  • 이찬용 (충남대학교 생화학과)
  • Received : 2009.12.07
  • Accepted : 2009.12.16
  • Published : 2009.12.31
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Abstract

To provide the basis of biosensor based on the lux genes from bioluminescent bacteria of Photobacterium leiognathi and Vibrio harveyi, we test the expression of lux genes in several strains of Escherichia coli. The expression of the recombinant plasmid of PlXba.pT7-3, containing all lux genes requiring for light emission without adding substrate, in E. coli 43R was so strong to see the blue-green light in single colony as well as in the alginate immobilized cell. In addition, the light intensity was decreased by adding heavy metal ion such as cadmium and zinc ions. These result raise the possibility that a biosensor can be developed using the lux genes system.

발광 세균 Photobacterium과 Vibrio의 lux 유전자가 삽입된 재조합 플라스미드를 유전자 전이시킨 발광표현형 대장균이 내는 빛의 세기를 조사하였다. 여러 대장균 균주에 형질전환 시켰는 바, 빛을 내는데 관여하는 효소들을 코드하는 유전자를 모두 포함하는 Photobacterium leiognathi lux 오페론이 삽입된 재조합플라스미드(PlXba.pT7-3)가 형질전환된 대장균 43R (Escherichia coli 43R) 균주에서는 발광세기가 다른 균주에 비해 1,000배 이상 되었으며, Vibrio harveyi luxA 유전자와 luxB 유전자를 융합시킨 유전자가 삽입된 재조합플라스미드(VhluxAB.pT7-5)가 형질전환 된 대장균 43R (E. coli 43R)에서는 기질인 fatty aldehyde를 가하면 단일 콜로니에서도 빛을 볼 수 있게 되는 대장균 형질전환체를 얻었다. 또한 이들이 중금속에 노출되었을 때 생물발광이 감소하였으며, 이들 발광 대장균 형질 전환체가 담긴 고정화된 세포에서도 빛을 내는 것을 관측함으로써 이들 실험 결과들이 lux 유전자를 활용한 바이오센서 시스템 개발의 기반 기술이 될 가능성을 보였다.

Keywords

References

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