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Pseudomonas putida NCIB 9869 carries a transmissible plasmid pRA500 of approximately 500 kb which encodes the degradation of 3,5-xylenol via a gentisate pathway. Several mutant strains which were unable to utilize 3,5-xylenol were isolated and these strains either carried deleted derivatives of pRA500 or lacked plasmid DNA. Biochemical and restriction endonuclease analysis of the wild-type and mutant strains showed that the structural and/or regulatory genes for 3,5-xylenol metabolism were encoded within a 130–140 kb region of pRA500 and that, with the exception of the first enzyme of the pathway, 3,5-xylenol methylhydroxylase, all the enzymes were encoded within a 50–70 kb segment of that region. pRA500 also encoded for resistance to inorganic mercuric ions; the genes for this phenotype were located separately from those for the degradation of 3,5-xylenol.
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