Identification of Pseudomonas alcaligenes chromosomal DNA in the plasmid DNA of the chlorobenzene-degrading recombinant Pseudomonas putida strain CB1-9.
The recombinant Pseudomonas putida strain CB1-9, which acquired the ability to grow on chlorobenzenes, contains a 33-kilobase (kb) plasmid (pKFL3) which lacked homology to an indigenous 15-kb plasmid (pKFL1) in Pseudomonas alcaligenes C-0 parent but was homologous to a 55-kb plasmid (pKFL2) from the P. putida R5-3 parent. Chromosomal DNA of P. alcaligenes C-0 hybridized to probes prepared from pKFL3 but not to probes prepared from pKFL2. A single clone from a genomic library of P. alcaligenes C-0 hybridized to EcoRI-digested pKFL3. Southern blot hybridization with the insert DNA from that clone identified homology with specific restriction enzyme fragments in pKFL3. The ability of the recombinant to utilize 3-chlorobenzoate, chlorobenzene, and 1,4-dichlorobenzene as well as its loss of utilization of xylenes and methylbenzoates appears to be associated with the transfer and integration of chromosomal DNA from P. alcaligenes into a Tol-like plasmid of P. putida R5-3.
Carney
B F
BF
Department of Plant Pathology, University of California, Riverside 92521-0122.
Kröckel
L
L
Leary
J V
JV
Focht
D D
DD
D015139:Blotting, Southern; D002722:Chlorobenzenes / Q000378:metabolism*; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015183:Restriction Mapping
D002722:Chlorobenzenes; D015342:DNA Probes; D004269:DNA, Bacterial