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Main Title Cloning, Physical Mapping and Expression of Chromosomal Genes Specifying Degradation of the Herbicide 2,4,5-T by 'Pseudomonas cepacia' AC1100.
Author Sangodkar, U. M. X. ; Chapman, P. J. ; Chakrabarty, A. M. ;
CORP Author 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.
Publisher c1988
Year Published 1988
Report Number EPA-R-812660 ;PHS-ES-04050-3; EPA/600/J-88/372;
Stock Number PB90-100520
Additional Subjects Herbicides ; Pseudomonas ; Biodeterioration ; Deoxyribonucleic acids ; Naphthalene ; Toulene ; Reprints ; Molecular cloning ; Gene expression regulation ; Chromosome mapping ; Pseudomonas cepacia AC1100 ; T 2-4-5 herbicide ; Chlorobenzoates ; Nucleic acid hybridization ; Nucleic acid sequence homology
Library Call Number Additional Info Location Last
NTIS  PB90-100520 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 13p
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).)