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


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.


  1. Curreny protocols in molecular biology Ausubel,F.M.;R.Brent;R.E.Kingston;D.D.Moore;J.G.Seidmen;J.A.Smith;K.Struhl
  2. FEMS Microbiol. Lett. v.81 Cloning and expression of the Arthrobacter globiformis fcbAgene encoding dehalogenase (4-chlorobenzoate-4-hydroxylase) in Escherichia coli Tsoi,T.V.;G.M.Zaitsev;E.G.Plotnikova;I.A.Kosheleva;A.M.Boronin
  3. Microbiol. Rev. v.56 Microbial reductive dehalogenation Mohn,W.W.;J.M.Tiedje
  4. Kor. J. Microbiol. v.30 Divergence of the cbp genes in 4CB catabolizing bacteria Yun,D.J.;J.J.Han;C.K.Kim;Y.S.Kim
  5. J. Bacteriol. v.168 Cloning of Pseudomonas sp. strain CBS3 genes specifying dehalogenation of 4-chlorobenzoate Savard,D.;L.Peloquin;M.Sylverstre
  6. Kor. J. Microbiol. v.25 Isolation and characterization of bacteria degrading chlorinated aromatic hydrocarbons Kim,C.K.;J.W.Kim;Y.C.Kim
  7. FEMS Microbiol. Rev. v.103 Microbial breakdown of halogenated aromatic pesticides and related compounds Haggblom,M.M.;
  8. Kor. J. Microbiol. v.32 Cloning of dechlorination genes specifying biodegradation of toxic 4-chlorobiphenyl Kim,C.K.;J.C.Chae;J.J.Han
  9. Kor. J. Microbiol. v.32 Cloning and expression of pcbCD genes in Escherichia coli from Pseudomonas sp. DJ-12 Kim,C.K.;T.K.Sung;J.H.Nam;Y.C.Kim;J.K.Lee
  10. Molecular cloning(2nd ed.) Sambrook,J.;E.F.Fritsch;T.Maniatis
  11. Appl. Environ. Microbiol. v.53 Construction of chlorobenzene-utilizing recombinants by progenitive manifestation of a rare event Krockel,L.;D.D.Focht
  12. J. Bacteriol. v.175 Characterization of a Flavobacterium glutathion S-transferase gene involved in reductive dechlorination Orser,C.S.;J.Dutton;C.Lange;P.Jablonski;L.Xun;M.Hargis
  13. Appl. Environ. Microbiol. v.58 Clonig and sequence of genes for dehalogenation of 4-chlorobenzoate from Arthrobacter sp. strain SU Schmitz,A.;K.H.Gartemann;J.Fiedler;E.Grund;R.Eichenlaub
  14. Gene. v.144 The biphenyl/polychlorinated biphenyl-degradation locus(bph) of Pseudomonas sp. LB400 encodes four additional metabolic enzymes Hofer,B.;S.Backhaus;K.N.Timmis
  15. Ann. Rev. Microbiol. v.42 Microbial degradation of haloaromatics Reineke,W.