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Main Title Multiple Replicons Constituting the Genome of 'Pseudomonas cepacia' 17616.
Author Cheng, H. P. ; Lessie, T. G. ;
CORP Author Environmental Research Lab., Gulf Breeze, FL. Office of Research and Development. ;Massachusetts Univ., Amherst. Dept. of Microbiology.;Department of Energy, Washington, DC.
Publisher c1994
Year Published 1994
Report Number EPA/600/J-94/446; DE-FG02-ER20051;
Stock Number PB95-112132
Additional Subjects Replicon ; Pseudomonas cepacia ; Restriction mapping ; Southern blotting ; Plasmids ; Ribosomal RNA ; Carbohydrates ; Bacterial genes ; Pulsed-field gel electrophoresis ; Reprints ;
Library Call Number Additional Info Location Last
NTIS  PB95-112132 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 11p
Macrorestriction fragment analysis of DNA from Pseudomonas cepacia 17616, in conjunction with Southern hybridization experiments using junction fragments containing rare restriction enzyme sites as probes, indicated that this bacterium contains three large circular replicons of 3.4, 2.5, and 0.9 megabases (Mb). Inclusion of the 170-kb cryptic plasmid present in this strain gave an overall estimate of genome size of 7 Mb. Other Southern hybridization experiments indicated that the three large replicons contained rRNA genes as well as insertion sequence elements identified previously in this strain. The distribution of SwaI, PacI, and PmeI sites on the three replicons was determined. A derivative of Tn5-751 carrying a SwaI site was used to inactivate and map genes on the 2.5- and 3.4-Mb replicons. Mutants were isolated in which the 2.5- and 0.9-Mb replicons had been reduced in size to 1.8 and 0.65 Mb, respectively. The loss of DNA from the 2.5-Mb replicon was associated with lysine auxotrophy. Beta-lactamase deficiency, and failure to utilize ribitol and trehalose as carbon and energy sources. DNA fragments corresponding in size to randomly linearized forms of the different replicons were detected in unrestricted DNA by pulsed-field gel electrophoresis. The results provide a framework for further genetic analysis of strain 17616 and for evaluation of the genomic complexities of other P. cepacia isolates. (Copyright (c) 1994, American Society for Microbiology.)