Chromosomal Mapping and Cloning of the Lipase Gene of Pseudomonas aeruginosa

• Published in: Journal of general microbiology Vol. 134; no. 2; pp. 433 - 440
• Main Authors: WOHLFARTH, SUSANNE, WINKLER, ULRICH K
• Format: Journal Article
• Language: English
• Published: London Soc General Microbiol 01.02.1988; New York, NY Cambridge University Press
• Subjects: Bacterial Outer Membrane Proteins - analysis; Bacteriology; Biological and medical sciences; Chromosome Mapping; Cloning, Molecular; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genetic Linkage; Genetics; Lipase - genetics; Microbiology; Mutation; Phenotype; Pseudomonas aeruginosa; Pseudomonas aeruginosa - enzymology; Pseudomonas aeruginosa - genetics; Pseudomonadales; Genetic mapping; Enzyme; Genetic variant; Triacylglycerol lipase; Chromosome; Extracellular; Virus; Characterization; Gene; Bacteria; Pseudomonadaceae; Bactériophage; Isolation; Pseudomonas aeruginosa; Transduction; Localization
• ISSN: 0022-1287; 1350-0872; 1465-2080
• DOI: 10.1099/00221287-134-2-433

Ruhr-Universität Bochum, Lehrstuhl für Biologie der Mikroorganismen, D-4630 Bochum, FRG ABSTRACT SUMMARY: Various mutants ( lip ) of Pseudomonas aeruginosa PAO 2302 that lacked extracellular lipase activity were isolated. They were selected on a calcium-triolein agar. The phenotypic characteristics of two of these mutants suggested that they were defective in the gene coding for lipase: both lip mutants produced no lipase in liquid- and on solid medium. They were non-pleiotropic with regard to various other exoproducts. None of the mutants released any putatively cell-bound lipase after treatment of cells with Triton X-100 or alginate. The electrophoretic protein- and LPS-profiles of outer membranes derived from lip mutants and the parental strain were identical. The lip locus was mapped on the chromosome of P. aeruginosa PAO 1 by FP5- and R68.45-mediated crossings and by transduction with phage G101. The lip locus was cotransduced with pyrF only (60%) indicating a map position at about 57 min. The lipase gene was cloned on a 3.1 kb Sal I fragment using vector pKT248. The newly constructed plasmid was able to complement the lipase deficiency of the two lip mutants of P. aeruginosa.