Cloning Genes Involved in Aniline Degradation from Delftia acidovorans.

Delftia acidovorans로부터 Aniline 분해관련 유전자의 분리

  • 김현주 (한국화학연구원 생물화학부) ;
  • 김성은 (충남대학교 농생물학과) ;
  • 김정건 (서울대학교 응용생물화학부) ;
  • 김진철 (한국화학연구원 생물화학부) ;
  • 최경자 (한국화학연구원 생물화학부) ;
  • 김흥태 (충북대학교 농생물학과) ;
  • 황인규 (서울대학교 응용생물화학부) ;
  • 김홍기 (충남대학교 농생물학과) ;
  • 조광연 (한국화학연구원 생물화학부)
  • Published : 2003.03.01

Abstract

Delftia acidovorans 51-A isolated from river water degrades aniline. In order to clone genes involved in aniline degradation, transposon Tn5-B20 was inserted into the strain 51-A to generate a mutant strain 10-4-2 that cannot utilize aniline as a carbon source. The mutant strain was not an auxotroph but could not degrade aniline. Southern hybridization analysis indicated that the transposon was inserted into the mutant bacterial DNA as a single copy. Flanking DNA fragment of Tn5-B2O insertion was cloned and sequenced. DNA sequence analysis revealed three ORFs encoding TdnQ, TdnT, and TdnA 1 that arc responsible for catechol formation from aniline through oxidative deamination. The analysis also confirmed that Tn5-B2O was inserted at the immediate downstream of tdnA1. The result suggests that the transposon insertion behind tdirA1 disrupted the pathway of the catechol formation from aniline, resulting in the mutant phenotype, which cannot degrade aniline. A large plasmid over 100-kb in size was detected from D. acidovorans 51-A and Southern hybridization analysis with Tn5-B2O probe showed that the transposon was inserted on the plasmid named pTDN51. Our results indicated that the tdn genes on pTDN51 of D. acidovorans 51-A are involved in aniline degradation.

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