RP4::Mu3A-mediated in vivo cloning and transfer of a chlorobiphenyl catabolic pathway.
Chromosomal DNA fragments encoding the ability to utilize biphenyl as sole carbon source (Bph+) were mobilized by means of plasmid RP4::Mu3A from strain JB1 (tentatively identified as Burkholderia sp.) to Alcaligenes eutrophus CH34 at a frequency of 10(-3) per transferred plasmid. The mobilized DNA integrated into the recipient chromosome or was recovered as catabolic prime plasmids. Three Bph+ prime plasmids were transferred from A. eutrophus to Escherichia coli and back to A. eutrophus without modification of the phenotype. The transferred Bph+ DNA segments allowed metabolism of biphenyl, 2-, 3- and 4-chlorobiphenyl, and diphenylmethane. Genes involved in biphenyl degradation were identified on the prime plasmids by DNA-DNA hybridization and by gene cloning. Bph+ prime plasmids were transferred to Burkholderia cepacia, Pseudomonas aeruginosa, Comamonas testosteroni and A. eutrophus and the catabolic genes were expressed in those hosts. Transfer of the plasmid to the 3-chlorobenzoate-degrading bacterium Pseudomonas sp. B13 allowed the recipient to mineralize 3-chlorobiphenyl. Other catabolic prime plasmids were obtained from JB1 by selection on m-hydroxybenzoate and tyrosine as carbon sources. 16S rRNA sequence data demonstrated that the in vivo transfer of bph was achieved between bacteria belonging to two different branches of the beta-Proteobacteria.
Springael
D
D
Environmental Technology, Flemish Institute for Technological Research (VITO), Mol, Belgium.
van Thor
J
J
Goorissen
H
H
Ryngaert
A
A
De Baere
R
R
Van Hauwe
P
P
Commandeur
L C
LC
Parsons
J R
JR
De Wachter
R
R
Mergeay
M
M
D000421:Alcaligenes / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001673:Biodegradation, Environmental; D005075:Biological Evolution; D001713:Biphenyl Compounds / Q000378:metabolism*; D019117:Burkholderia / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012988:Soil Microbiology
D001713:Biphenyl Compounds; D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; C034280:3-chlorobiphenyl