Molecular cloning of the xylL-xylE region from the P. putida TOL plasmid, pDK1.
A 5.2 kilobase EcoRI restriction fragment from the Pseudomonas putida HS1 TOL plasmid pDK1, encoding a portion of the lower toluene degradation pathway, was cloned into the E. coli plasmid pBR325. A detailed map of the restriction endonuclease sites was constructed and the nucleotide sequence of three contiguous XhoI fragments, with a combined total length of approximately 3.9 kilobases, has been investigated. This region was determined to contain a total of four separate open reading frames, each preceded by an identical putative ribosome-binding site (nucleotide sequence of 5'-GAGGTG-3'). These open reading frames have been tentatively identified as encoding the lower pathway enzymes catechol 2,3-dioxygenase (C23O) and 1,2-dihydroxycyclohexa-3,5-diene carboxylate dehydrogenase (DHCDH) and a subunit of the toluate 1,2-dioxygenase complex (TO).
Department of Biology, University of North Texas, Denton 76203.
J A
JA
Voss
H
H
Khedairy
R F
RF
Baker
R C
RC
Benjamin
D050557:Catechol 2,3-Dioxygenase; D003001:Cloning, Molecular; D049308:Dioxygenases; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics*; D015183:Restriction Mapping; D014050:Toluene / Q000378:metabolism; D014162:Transfection / Q000235:genetics
D014050:Toluene; D010088:Oxidoreductases; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase; C023172:toluene dioxygenase; C025312:3,5-cyclohexadiene-1,2-diol-1-carboxylate dehydrogenase