Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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The γ-Benzenehexachloride Degradation Using Transgenic Tobacco Plant

담배 형질전환 식물체를 이용한 γ-Benzenehexachloride의 분해

  • Lee, Jeong-Kyung (Faculty of Life Resources, Catholic University of Daegu) ;
  • Park, Soon-Ki (Division of Plant Bioscience, Kyungpook National University) ;
  • Chung, Il-Kyung (Faculty of Life Resources, Catholic University of Daegu)
  • 이정경 (대구가톨릭대학교 자연대학 생명자원학부) ;
  • 박순기 (경북대학교 농과대학 농학과) ;
  • 정일경 (대구가톨릭대학교 자연대학 생명자원학부)
  • Published : 2003.03.01
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Abstract

LinA gene involving in the ${\gamma}$-benzenehexachloride degradation have been cloned from Sphingmonas paucimobilis UT26. This linA gene which catalyzes the first dechlorination step of ${\gamma}$-benzenehexachloride is known to play a key role in the ${\gamma}$-benzenehexachloride degradation pathway in UT26. In this study, the linA gene was designed to clean-up the ${\gamma}$-benzenehexachloride and its derivatives contaminated in soil, water and air using transgenic tobacco plants. The linA transgene was introduced into the chromosome of tobacco using leaf-disk transformation approach as revealed by Southern blot analysis. In addition, mRNA and protein produced by linA gene was expressed at a high level in the leaf tissue as demonstrated by both northern blot analysis and Western bolt analysis with polyclonal antibody against S. paucimobilis UT26. in vitro analysis using GC-MS showed that transgenic tobacco plant produced the linA protein which effectively degraded ${\gamma}$-benzenehexachloride into ${\gamma}$- pentachlorocyclohexene and 1,2,4-trichlobenzene compounds which are less toxic.

본 연구는 난분해성 농약의 자연분해를 유도하기 위하여 ${\gamma}$-BHC를 탄소원으로 이용하여 호기적으로 자화하는 미생물 (Sphingomonas paucimobilis)에서 분리된 탈염소화효소 중 ${\gamma}$-BHC분해의 첫단계에 관여하는 linA유전자를 담배에 도입하여 BHC에 활성을 나타내는 형질전환 시물체를 만들었다. LinA 유전자를 함유한 pJK 108 백터를 제작하여. Agrobacterium를 이용한 leaf disk transformation 방법으로 담배에 형질전환시켰으며, northern분석 및 Southern분석을 통해 형질전환 개체에서 linA유전자의 도입과 linA유전자 유래의 mRNA가 전사됨을 확인하였다. Western 분석에서 linA단백질의 발현을 확인하였고, gas chromatography를 이용하여 담배 형질전환 개체에서 BHC가 ${\gamma}$-PCCH와 1,2.4-TCB로 분해되는 것을 확인하였다.

Keywords

References

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