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Main Title Characterization of 'Pseudomonas putida' Mutants Unable to Catabolize Benzoate: Cloning and Characterization of 'Pseudomonas' Genes Involved in Benzoate Catabolism and Isolation of a Chromosomal DNA Fragment Able to Substitute for xylS in Activation of the TOL Lower-Pathway Promoter.
Author Jeffrey, W. H. ; Cuskey, S. M. ; Chapman, P. J. ; Resnick, S. ; Olsen, R. H. ;
CORP Author Environmental Research Lab., Gulf Breeze, FL. ;Technical Resources, Inc., Gulf Breeze, FL. ;Michigan Univ., Ann Arbor. Dept. of Microbiology and Immunology.
Publisher 1992
Year Published 1992
Report Number EPA/600/J-92/381;
Stock Number PB93-121135
Additional Subjects Pseudomonas putida ; Mutations ; Deoxyribonucleic acid ; Benzoic acid ; Promotor regions(Genetics) ; Molecular cloning ; Bacterial chromosomes ; Plasmids ; Restriction mapping ; Southern blotting ; Nucleic acid sequence homology ; Reprints ; Benzote dioxygenases ; Toluene dioxygenases
Library Call Number Additional Info Location Last
NTIS  PB93-121135 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 13p
Previous work suggested that (benR) can substitute for the TOL plasmid-encoded xylS regulatory gene which promotes gene expression from the OP2 region of the lower pathway operon. Accordingly, structural and regulatory gene mutations were distinguished by the ability of benzoate-grown mutant strains to induce expression from OP2 without xylS using the TOL plasmid xylE gene (encoding catechol 2,3-dioxygenase) as reporter. A cloned 12 kb BamHI chromosomal DNA fragment from the P. aeruginosa PA01 chromosome complemented all mutations as shown by restoration of growth on benzoate minimal medium. Southern hybridizations demonstrated that DNA encoding the benzoate dioxygenase structural genes showed homology to DNA encoding toluate dioxygenase from the TOL plasmid pWWO, but benR did not show homology to xylS. Evolutionary relationships between regulatory systems of chromosomal and plasmid encoded genes for catabolism of benzoate and related compounds are suggested.