Transfer and Random Integration of Tn5 in Azospirillum

  • M. Vanstockem
  • K. Michiels
  • J. Vanderleyden
  • A. Van Gool
Conference paper

Summary

A method is described for the isolation of at random Tn5 insertion mutants of Azospirillum spp. For this purpose a set of plasmid vectors of Tn5, that are suicidal in Rhizobium and Agrobacterium species, were introduced in A. brasilense Sp7 and A. lipoferum SpBrl7 by conjugation. The plasmids pSUP2021 and pGS9 are suicidal in these Azospirillum strains as shown by hybridization experiments of digested total DNA of twenty bona fide Kmr ex-conjugants with 32P labeled ColEl:: Tn5. Kanamycm-resistant colonies were detected at a frequency of 10−8 per recipient cell.

Keywords

Azospirillum transposon mutagenesis suicide plasmids Southern hybridization 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Döbereiner, J. and Day, J.M. (1976). In: Symposium on Nitrogen Fixation. Edited by W.E. Newton and C.J. Nyman. Washington State University Press, Pullman, pp. 518–538.Google Scholar
  2. 2.
    Elmerich, C. (1984). BIO/TECHNOLOGY 967–978.Google Scholar
  3. 3.
    Singh, M. (1983). In: Klingmüller, W. (ed.). Azospirillum Genetics Physiology Ecology. EXS 48 pp 47–55, Birkhauser, Basel.CrossRefGoogle Scholar
  4. 4.
    Elmerich, C. and Franche, C. (1982). In: Klingmüller, W. (ed.). Azospirillum Genetics Physiology Ecology EXS 42 pp 44–52, Birkhaiiser, Basel.Google Scholar
  5. 5.
    Simon, R., Priefer, U. and Pühler, A. (1983). BIO/TECHNOLOGY, nov., 184–190.Google Scholar
  6. 6.
    Selvaraj, G. and Iyer, V.N. (1983). J. Bacteriol. 156, 1292–1300.PubMedGoogle Scholar
  7. 7.
    Wood, W.B. (1966). J. Mol. Biol. 16, 118–133.PubMedCrossRefGoogle Scholar
  8. 8.
    Franche, C. and Elmerich, C. (1981). Annales de microbiologie 132, n°l, 3–18.Google Scholar
  9. 9.
    In “Cours de microbiologie générale. Etude de la fixation de l’azote chez diverses bacteries”. Elmerich, Institut Pasteur (1983).Google Scholar
  10. 10.
    Kado, C.I. and Liu, S.T. (1981). J. Bacteriol. 145, 1365–1373.PubMedGoogle Scholar
  11. 11.
    EMBO Course on “The use of Ti plasmids as cloning vectors for genetic engineering in plant”. 4–23 August 1982. Lab. genetics, Rijksuniversiteit Gent, Belgium.Google Scholar
  12. 12.
    Silhavy, T.J., Berman, M.L. and Enquist, L.W. (1984). In: “Experiments with gene fusions”. CSH Laboratory 1984.Google Scholar
  13. 13.
    Shaw, K.J. and Berg, C.M. (1979). Genetics 92, 741–747.PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1985

Authors and Affiliations

  • M. Vanstockem
    • 1
  • K. Michiels
    • 1
  • J. Vanderleyden
    • 1
  • A. Van Gool
    • 1
  1. 1.F.A. Janssens Laboratory of GeneticsCatholic University of LeuvenHeverleeBelgium

Personalised recommendations