Identification of 4-Chlorobenzoate Dechlorinating Bacteria by Simple Plate Assay

평판배지법에 의한 4-chlorobenzoate 탈염소화 세균의 검색

  • Chae, Jong-Chan ;
  • Kim, Chi-Kyung ;
  • Min, Kyung-Hee ;
  • Park, Yong-Keun
  • 채종찬 (충북대학교 미생물학과) ;
  • 김치경 (충북대학교 미생물학과) ;
  • 민경희 (서울대학교 분자미생물학 연구센터) ;
  • 박용근 (서울대학교 분자미생물학 연구센터)
  • Published : 1995.02.01

Abstract

The gene responsible for dechlorination of 4-chlorobenzoate (4CBA) was cloned in E. coli XL1-Blue from Pseudomonas sp. DJ-12. The cloned cell of E. coli Cjl had the hybrid pBluescript SK(+) plasmid, into which about 9.5 kb genomic DNA fragment of PseudOmonas sp. DJ-12 was inserted. The subclone of pCJlOl was constructed by inserting the 3.4 kb EcoRI-HindIII fragment of pCJl into the vector. Those cloned cells could be simply selected by halo formation around the colonies which was the precipitate of AgCl produced by reaction of AgNO$_{3}$ and chloride ion liberated by bacterial dechlorination of 4CBA- Such a plate assay method was standardized by the procedure that the colonies grown for 2 days on the Cl$^{-}$-free plate medium containing 1 mM 4CBA were flooded with 0.1 M AgNO$_{3}$ solution.

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