Isolation of Magnetospirillum magneticum AMB-1 Mutants Defective in Bacterial Magnetic Particle Synthesis by Transposon Mutagenesis

  • Aris Tri Wahyudi
  • Haruko Takeyama
  • Tadashi Matsunaga
Part of the ABAB Symposium book series (ABAB)


Nonmagnetic mutants of Magnetospirillum magneticum AMB-1 were recovered following mini-Tn5 transposon mutagenesis. Transconjugants with kanamycin resistance were obtained at a frequency of 2.7 × 10−7 per recipient. Of 3327 transconjugants, 62 were defective for bacterial magnetic particle (BMP) synthesis. The frequency of independent transposition events for nonmagnetic mutants was about 1.4% in transconjugants. Further analy- sis of DNA sequences flanking transposon by inverted polymerase chain reaction allowed isolation of at least 10 genes or DNA sequences involved in BMP synthesis in M. magneticum AMB-1.

Index Entries

Transposon mutagenesis mini-Tn5 Magnetospirillum magneticum AMB-1 inverse polymerase chain reaction bacterial magnetic particles sequence analysis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Schuler, D. and Frankel, R. B. (1999), Appl. Microbiol. Biotechnol. 52, 464–473.CrossRefGoogle Scholar
  2. 2.
    Blackemore, R. P., Maratea, D., and Wolfe, R. S. (1979), J. Bacteriol. 140, 720–729.Google Scholar
  3. 3.
    Matsunaga, T., Sakaguchi, T., and Tadokoro, F. (1991), Appl. Microbiol. Biotechnol. 35, 651–655.CrossRefGoogle Scholar
  4. 4.
    Matsunaga, T., Nakamura, C., Burgess, J. G., and Sode, K. (1992), J. Bacteriol. 174, 2748–2753.Google Scholar
  5. 5.
    DeBruijn, F. J. and Lupski, J. R. (1984), Gene 27, 131–149.CrossRefGoogle Scholar
  6. 6.
    Simon, R., Preifer, U., and Puhler, A. (1983), Bio/Technology 1, 784–791.CrossRefGoogle Scholar
  7. 7.
    Berg, D. E. (1989), in Mobile DNA, Berg, D. E and Howe, M. M., eds., American Society for Microbiology, Washington, DC, pp. 185–210.Google Scholar
  8. 8.
    Herrero, M., deLorenzo, V., and Timmis, K. N. (1990), J. Bacteriol. 172, 6557–6567.Google Scholar
  9. 9.
    Holtwick, R., Meinhardt, F., and Keweloh, H. (1997), Appl. Environ. Microbiol. 63, 4292–4297.Google Scholar
  10. 10.
    Jiang, Z. Y., Rushing, B. G., Bai, Y., Gest, H., and Bauer, C. E. (1998), J. Bacteriol. 180, 1248–1255.Google Scholar
  11. 11.
    DeLorenzo, V., Herrero, M., Martinko, J. M., and Parker, J. (1990), J. Bacteriol. 172, 6568–6572.Google Scholar
  12. 12.
    Martin, V. J. J. and Mohn, W. W. (1999), J. Microbiol. Methods 35, 163–166.CrossRefGoogle Scholar
  13. 13.
    Nakamura, C., Burgess, J. G., Sode, K., and Matsunaga, T. (1995), J. Biol. Chetn. 270, 28,392–28,396.Google Scholar
  14. 14.
    Wilson, K. (1994), in Current Protocols in Molecular Biology, vol. 1, Ausubel, F. M., Brent, R., and Kingston, R. E., eds., John Wiley & Sons, New York, pp. 2.4.1–2.4.5.Google Scholar
  15. 15.
    Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989), Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, New York.Google Scholar
  16. 16.
    Klarskov, K., Van Driessche, G., Barkers, K., Dumortier, C., Meyer, T. E., Tollin, G., Cusanovich, M. A., and Beeumen, J. J. V. (1998), Biochemistry 37, 5995–6002.CrossRefGoogle Scholar
  17. 17.
    Wilson, R., Ainscough, R., Anderso, K., et al. (1994), Nature 368, 32–38.CrossRefGoogle Scholar
  18. 18.
    Freiberg, C., Fellay, R., Bairoch, A., Broughton, W. J., Rosenthal, A., and Parret, X. (1997), Nature 387, 394–401.CrossRefGoogle Scholar
  19. 19.
    Kaneko, T., Sato, S., Kotani, H., et al. (1996), DNA Res. 3, 109–136.CrossRefGoogle Scholar
  20. 20.
    Tietze, E. (1998), Plasmid 39, 165–168.CrossRefGoogle Scholar
  21. 21.
    Sherman, D. H., Malpartida, F., Bibb, M. J., Kieser, H. M., and Hopwood, D. A. (1989), EMBO J. 8, 2717–2725Google Scholar
  22. 22.
    Ramakrishnan, G., Zhao, J. L., and Newton, A. (1994), J. Bacteriol. 176, 7587–7600.Google Scholar
  23. 23.
    Deckart, G. and Warren, P. V. (1998), Nature 392, 353–358.CrossRefGoogle Scholar
  24. 24.
    Davison, J., Brunei, F., Phanopoulos, A., Prozzi, D., and Terpstra, P. (1992), Gene 114, 19–24.CrossRefGoogle Scholar
  25. 25.
    Ooi, J., Yatnik, V., Immanuel, D., Gordon, M., Moskow, J. J., Buchberg, A. M., and Margolis, B. (1995), Oncogene 10, 1621–1630.Google Scholar

Copyright information

© Springer Science+Business Media New York 2001

Authors and Affiliations

  • Aris Tri Wahyudi
    • 1
  • Haruko Takeyama
    • 1
  • Tadashi Matsunaga
    • 1
  1. 1.Department of BiotechnologyTokyo University of Agriculture and TechnologyKoganei, TokyoJapan

Personalised recommendations