Gene-specific transposon mutagenesis of the biphenyl/polychlorinated biphenyl-degradation-controlling bph operon in soil bacteria.
A transposon, Tn5-B21, was gene-specifically inserted into the chromosomal biphenyl/polychlorinated biphenyl-catabolic operon (bph operon) of soil bacteria. The cloned bphA, bphB and bphC genes of Pseudomonas pseudoalcaligenes KF707, coding for conversion of biphenyl into a ring meta-cleavage product (2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid), carried random insertions of Tn5-B21. The mutagenized bphABC DNA, carried by a suicide plasmid, was introduced back into the parent strain KF707, resulting in the appearance of gene-specific transposon mutants by double crossover homologous recombination: the bphA::Tn5-B21 mutant did not attack 4-chlorobiphenyl, the bphB::Tn5-B21 mutant accumulated dihydrodiol, and the bphC::Tn5-B21 mutant produced dihydroxy compound. Gene-specific transposon mutants of the bph operon were also obtained for some other biphenyl-utilizing strains which possess bph operons nearly identical to that of KF707.
Furukawa
K
K
Department of Agricultural Chemistry, Kyushu University, Fukuoka, Japan.
Hayashida
S
S
Taira
K
K
D001713:Biphenyl Compounds / Q000378:metabolism*; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D005798:Genes, Bacterial; D015394:Molecular Structure; D016254:Mutagenesis, Insertional; D009876:Operon; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics*; D015183:Restriction Mapping; D012988:Soil Microbiology
D001713:Biphenyl Compounds; D004251:DNA Transposable Elements; C010574:biphenyl; C006983:4-chlorobiphenyl