title: Tn5 tagging of the phenol-degrading gene on the chromosome of Pseudomonas putida.
abstract: Transposon mutagenesis was performed by the method of conjugational transfer in order to identify and characterize genes encoding enzymes involved in the pathway of phenol utilization as a carbon source. Escherichia coli, which carries the Tn5-132, Was mated with Pseudomonas putida SM25 as a host. We selected a mutant that could not utilize phenol as a carbon source. Chromosomal integration of the transposon was confirmed by Southern analysis, successfully tagging the gene related to a phenol-utilizing pathway. By cell-free enzyme and genetic complementation assays, the inactivated enzyme through the mutation of the corresponding gene was identified as the catB gene, which encodes a cis,cis-muconate lactonizing enzyme.
authors.affiliation: Department of Biology, Sookmyung Women's University, Seoul, Korea.
authors.forename: E M
authors.initials: EM
authors.lastname: Han
authors.forename: Y H
authors.initials: YH
authors.lastname: Jung
authors.forename: H Y
authors.initials: HY
authors.lastname: On
authors.forename: M S
authors.initials: MS
authors.lastname: Lee
authors.forename: Y K
authors.initials: YK
authors.lastname: Yang
authors.forename: Y
authors.initials: Y
authors.lastname: Kim
authors.forename: C K
authors.initials: CK
authors.lastname: Kim
authors.forename: K S
authors.initials: KS
authors.lastname: Lee
authors.forename: K H
authors.initials: KH
authors.lastname: Min
mesh_terms: D001673:Biodegradation, Environmental; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003227:Conjugation, Genetic; D004251:DNA Transposable Elements; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D019753:Intramolecular Lyases; D007535:Isomerases / Q000235:genetics / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009154:Mutation; D019800:Phenol; D010636:Phenols / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism*; D016324:Sequence Tagged Sites
chemical_list: D004251:DNA Transposable Elements; D010636:Phenols; D019800:Phenol; D007535:Isomerases; D019753:Intramolecular Lyases; C021811:muconate cycloisomerase