1887

Abstract

Tn mutagenesis of different fluorescent pseudomonads was achieved by conjugational transfer of the suicide vector pSUP 10141. Pyoverdine negative (Pvd) mutants were detected by the absence of fluorescence on King's B medium and by their inability to grow in the presence of the iron chelator EDDHA [ethylenediamine di(hydroxyphenylacetic acid)]. In ATCC 17400 and three rhizosphere isolates (one and two ), the percentage of Pvdmutants ranged between 0 and 0.54%. In a rhizosphere isolate, this percentage was higher (4%). In these mutants both of the Tn antibiotic resistances (Km and Tc) were stable and the transposon could be detected by hybridization. In Pvdmutants of ATCC 17400, the transposon was found to be inserted twice in the chromosome while single insertions were detected in the DNA of other, randomly tested mutants. In PAO1, where 13.1% of the mutants were Pvd, both antibiotic resistances were rapidly lost and accordingly no transposon insertion could be detected by hybridization. However, the Pvdphenotype was generally stable in these mutants. The plasmid pNK862 containing a mini-Tn transposon was introduced by electroporation into PAO1 and Kmmutants were recovered, 89% of which were Pvdand confirmed to be by PCR amplification of the lipoprotein gene. The mini-Tninsertions were also found to be unstable in PAO1.

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1992-07-01
2021-07-31
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References

  1. Bagdasarian M. M., Amann E., Lurz R., RÜckert B., Bagdasarian M. 1983; Activity of the hybrid trp–lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled expression vectors. Gene 26 273 282
    [Google Scholar]
  2. Berg D. E. 1977; Insertion and excision of the transposable kanamycin resistance determinant Tn5. In DNA Insertion Elements, Plasmids and Episomes pp. 205 212 Edited by Bukhari A. I., Shapiro J. A., Adhya S. L. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  3. Berg D. E., Berg C. M. 1983; The prokaryotic transposable element Tn5. Biol. Technology 1 417 435
    [Google Scholar]
  4. Berg D. E., Schmandt M. A., Lowe J. B. 1983; Specificity of transposon Tn5 insertion. Genetics 105 813 828
    [Google Scholar]
  5. Cornelis P., Bouia A., Belarbi A., Guyonvarch A., Kammerer B., Hannaert V., Hubert J. C. 1989; Cloning and analysis of the gene for the major outer membrane lipoprotein from Pseudomonas aeruginosa. Molecular Microbiology 3 421 428
    [Google Scholar]
  6. Egner C., Berg D. E. 1981; Excision of transposon Tn5 is dependent on the inverted repeats but not on the transposase function of Tn5. Proceedings of the National Academy of Sciences of the United States of America 78 459 463
    [Google Scholar]
  7. Frey J., Bagdasarian M., Feiss D., Franklin F. C., Deshusses J. 1983; Stable cosmid vectors that enable the introduction of cloned fragments into a wide range of Gram-negative bacteria. Gene 24 299 308
    [Google Scholar]
  8. Goldberg J. B., Won J., &Ohman D. E. 1990; Precise excision and instability of the transposon Tn5 in Pseudomonas aeruginosa. Journal of General Microbiology 136 789 796
    [Google Scholar]
  9. Höfte M., Seong K. Y., Jurkevitch E., Verstraete W. 1991; Pyoverdin production by the plant-growth-beneficial Pseudomonas strain 7NSK2: ecological significance in soil. Plant and Soil 130 249 257
    [Google Scholar]
  10. Hohnadel D., Meyer J. M. 1988; Specificity of pyoverdine-mediated iron uptake among fluorescent Pseudomonas strains. Journal of Bacteriology 170 4865 4873
    [Google Scholar]
  11. Hohnadel D., Haas D., Meyer J. M. 1986; Mapping of mutations affecting pyoverdine production in Pseudomonas aeruginosa. FEMS Microbiology Letters 36 195 199
    [Google Scholar]
  12. Holloway B. W., Dharmsthiti S., Johnson C., Kearney A., Krishnapillai V., Morgan A. F., Ratnaningsih E., Saffery R., Sinclair M., Strom D., Zhang C. 1990; Chromosome organization in Pseudomonas aeruginosa and Pseudomonas putida. In Pseudomonas: Biotransformation, Pathogenesis, and Evolving Biotechnology pp. 269 278 Edited by Silver S., Chakrabarty A. M., Iglewski B., Kaplan S. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  13. Kleckner N. 1981; Transposable elements in prokaryotes. Annual Review of Genetics 15 341 404
    [Google Scholar]
  14. Kreike C. M., de Koning J. R. A., Krem F. A. 1990; Nonradioactive detection of single-copy DNA–DNA hybrids. Plant Molecular Biology Reports 8 172 179
    [Google Scholar]
  15. Krishnapillai V., Royle P., Lehrer J. 1981; Insertions of the transposon Tn1 into the Pseudomonas aeruginosa chromosome. Genetics 97 495 511
    [Google Scholar]
  16. Leong J. 1986; Siderophores: their biochemistry and possible role in the biocontrol of plant pathogens. Annual Review of Phytopathology 24 187 209
    [Google Scholar]
  17. Lodge J. K., Weston-Hafer K., Berg D. E. 1988; Transposon Tn5 target specificity: preference for insertion at G/C pairs. Genetics 120 645 650
    [Google Scholar]
  18. Lodge J. K., Berg D. E. 1990; Mutations that affect Tn5 insertion into the major Tn5 hotspot in pBR322: importance of local DNA supercoiling. Journal of Bacteriology 172 5956 5960
    [Google Scholar]
  19. Lodge J. K., Weston-Hafer K., Berg D. E. 1991; Tn5 insertion specificity is not influenced by IS50 end sequences in target DNA. Molecular and General Genetics 228 312 315
    [Google Scholar]
  20. Lundblad V., Taylor A. F., Smith G. R., Kleckner N. 1984; Unusual alleles of RecB and RecC stimulate excision of inverted repeat transposons Tn10 and Tn5. Proceedings of the National Academy of Sciences of the United States of America 81 824 828
    [Google Scholar]
  21. Marugg J. D., van Spanje M., Hoeksta W. P., Schippers B., Weisbeek P. 1985; Isolation and analysis of genes involved in siderophore biosynthesis in plant-growth-stimulating Pseudomonas putida WCS358. Journal of Bacteriology 164 563 570
    [Google Scholar]
  22. Moores J. C., Magazin M., Ditta G. S., Leong J. 1984; Cloning of genes involved in the biosynthesis of pseudobactin, a high-affinity iron transport agent of a plant growth-promoting Pseudomonas strain. Journal of Bacteriology 157 53 58
    [Google Scholar]
  23. O’Hoy K., Krishnapillai V. 1987; Recalibration of the Pseudomonas aeruginosa strain PAO chromosome map in time units using high-frequency-of-recombination donors. Genetics 115 611 618
    [Google Scholar]
  24. O’Sullivan D. J., O’Gara F. 1990; Iron regulation of ferric iron uptake in a fluorescent pseudomonad: cloning of a regulatory gene. Molecular Plant–Microbe Interactions 3 86 93
    [Google Scholar]
  25. Römling U., Grothues D., Bautsch W., TÜmmler B. 1989; A physical genome map of Pseudomonas aeruginosa. EMBO Journal 8 4081 4089
    [Google Scholar]
  26. Simon R., Priefer U., Pühler A. 1983; A broad host range mobilization system for in υiυo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Bio/Technology 1 784 791
    [Google Scholar]
  27. Smith A. W., Iglewsky B. H. 1989; Transformation of Pseudomonas aeruginosa by electroporation. Nucleic Acids Research 17 10509
    [Google Scholar]
  28. Stapleton M. J., Jagger K. S., Warren R. I. 1984; Transposon mutagenesis of Pseudomonas aeruginosa exoprotease genes. Journal of Bacteriology 157 7 12
    [Google Scholar]
  29. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. 1984; New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene 32 369 379
    [Google Scholar]
  30. Wendenbaum S., Demange P., Dell A., Meyer J. M., Abdallah M. A. 1983; The structure of pyoverdine Pa, the siderophore of Pseudomonas aeruginosa. Tetrahedron Letters 24 4877 4880
    [Google Scholar]
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