Molecular cloning of gene xylS of the TOL plasmid: evidence for positive regulation of the xylDEGF operon by xylS.
The xylDEGF operon and the regulatory gene xylS of the TOL plasmid found in Pseudomonas putida mt-2 were cloned onto Escherichia coli vector plasmids. A 9.5-kilobase fragment, derived from the TOL segment of pTN2 deoxyribonucleic acid, carried the xyl genes D, E, G, and F, which encode toluate oxygenase, catechol 2,3-oxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, and 2-hydroxymuconic semialdehyde hydrolase, respectively. The enzymes were noninducible unless a 3-kilobase PstI fragment, derived also from the TOL segment, was provided in either cis or trans. The PstI fragment appeared to contain the regulatory gene xylS, which produced a positive regulator. The regulator was activated by m-toluate or benzoate, but not by m-xylene or m-methylbenzyl alcohol. the map positions of xylG and xylF were also determined.
D002264:Carboxylic Acids / Q000378:metabolism*; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005809:Genes, Regulator; D009876:Operon; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics* / Q000378:metabolism
D002264:Carboxylic Acids; D004262:DNA Restriction Enzymes