||Cloning, Physical Mapping and Expression of Chromosomal Genes Specifying Degradation of the Herbicide 2,4,5-T by 'Pseudomonas cepacia' AC1100.
Sangodkar, U. M. X.;
Chapman, P. J.;
Chakrabarty, A. M.;
||Illinois Univ. at the Medical Center, Chicago. Dept. of Microbiology and Immunology.;Environmental Research Lab., Gulf Breeze, FL.;National Inst. of Environmental Health Sciences, Research Triangle Park, NC.
||EPA-R-812660 ;PHS-ES-04050-3; EPA/600/J-88/372;
Gene expression regulation;
Pseudomonas cepacia AC1100;
T 2-4-5 herbicide;
Nucleic acid hybridization;
Nucleic acid sequence homology
||Most EPA libraries have a fiche copy filed under the call number shown. Check with individual libraries about paper copy.
||A genomic library of total DNA of Pseudomonas cepacia AC1100 was constructed on a broad-host-range cosmid vector pCP13 in E. coli AC80. A 25-kb segment was isolated from the library that complemented a Tn5-generated, 2,4,5-T minus mutant, P. cepacia PT88. A simple colorimetric assay was developed to detect the presence of this active enzyme in intact cells and was used to determine the expression of complementing genes. Subcloning experiments showed that a 4-kb BamHI-PstI fragment and a 290-bp PstI-EcoRI fragment, separated by 1.3-kb, were required for complementation. Both fragments were identified to be chromosomal in origin. Hybridization studies using the subcloned fragments revealed that in addition to a Tn5 insertion, mutant PT88 contained an extensive chromosomal deletion accounting for its 2,4,5-T-phenotype. The cloned fragments did not show homology to plasmid DNAs carrying degradative genes for toluene, naphthalene, and 3-chlorobenzoate. (Copyright 1988 Elsevier Science Publishers B.V. (Biomedical Division).)
||Pub. in Gene 71, n2 p267-277 1988. Sponsored in part by grant EPA-R-812011-01. Sponsored by Environmental Research Lab., Gulf Breeze, FL., and National Inst. of Environmental Health Sciences, Research Triangle Park, NC.
|NTIS Title Notes
||Reprint: Cloning, Physical Mapping and Expression of Chromosomal Genes Specifying Degradation of the Herbicide 2,4,5-T by 'Pseudomonas cepacia' AC1100.
||57F; 57K; 68E
||PC A03/MF A01