Gene Activation by T-DNA Tagging

  • Klaus Fritze
  • Richard Walden
Part of the Methods in Molecular Biology™ book series (MIMB, volume 44)


Agrobacterium-mediated transformation of plants has not only become a useful tool for the introduction of chimeric genes into plants, but it has also recently been utilized in gene tagging (for reviews, see refs. 1, 2, 3). T-DNA tagging has proven a powerful genetic approach for the isolation of novel plant genes. Compared with other approaches to gene isolation (for a review, see ref. 4), mutagenesis by insertion of defined DNA sequences allows the rapid cloning and characterization of the mutated locus. Furthermore, T-DNA is an ideal gene tag, as it has the potential to insert preferentially into potentially transcribed regions of the genome (5).


Naphthalene Acetic Acid Protoplast Culture Tobacco Protoplast Ampicillin Resistance Gene Benchtop Centrifuge 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Walden, R., Hayashi, H., and Schell, J. (1991) T-DNA as a gene tag. Plant J. 3, 281–288.Google Scholar
  2. 2.
    Koncz, C., Nemeth, K., Redei, G., and Schell, J. (1993) T-DNA insertional mutagenesis in Arabidopsis. Plant Mol. Biol. 20, 963–976.CrossRefGoogle Scholar
  3. 3.
    Coomber, S. and Feldmann, K. A. (1993) Gene tagging in transgenic plants, in Transgenic Plants, vol. 1 (Kung, S. and Wu, R., eds.), Academic, New York, pp. 225–240.Google Scholar
  4. 4.
    Gibson, S. and Somerville, C. (1993) Isolating plant genes. TibTech 11, 306–313.Google Scholar
  5. 5.
    Koncz, C., Martini, N., Mayerhofer, R., Koncz-Kalmann, Zs., Korber, H., Redei, G. P., and Schell, J. (1989) High frequency T-DNA mediated gene tagging in plants. Proc. Natl. Acad. Sci. USA 86, 8467–8471.PubMedCrossRefGoogle Scholar
  6. 6.
    Feldmann, K. A. (1991) T-DNA insertion mutagenesis in Arabidopsis. mutational spectrum. Plant J. 1, 71–82.CrossRefGoogle Scholar
  7. 7.
    Teeri, T. H., Herrera-Estrella, L., DePicker, A., Van Montagu, M., and Palva, E. T. (1986) Identification of plant promoters in situ by T-DNA mediated transcriptional fusions of the NPT II gene. EMBO J. 5, 1755–1760.PubMedGoogle Scholar
  8. 8.
    Kerbundit, S., De Greve, H., Deboeck, F., Van Montagu, M., and Hernalsteens, J.-P. (1991) In vivo random β-glucuronidase gene fusions in Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 88, 5212–5216.CrossRefGoogle Scholar
  9. 9.
    Goldsborough, A. and Bevan, M. (1990) New patterns of gene activity in plants detected using an Agobacterium vector. Plant Mol. Biol. 16, 263–269.CrossRefGoogle Scholar
  10. 10.
    Topping J. F., Wei, W., and Lindsay, K. (1991) Functional tagging of regulatory elements in the plant genome. Development 112, 1009–1019.PubMedGoogle Scholar
  11. 11.
    Hayashi, H., Czaja, I., Lubenow, H., Schell, J., and Walden, R. (1992) Activation of a plant gene by T-DNA tagging: auxin independent growth in vitro. Science 258, 1350–1353.PubMedCrossRefGoogle Scholar
  12. 12.
    Maliga, P., Sz.-Breznovitis, A., and Marton, L. (1973) Streptomycin resistant plants from callus culture of haploid tobacco. Nature 244, 623–625.Google Scholar
  13. 13.
    Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15, 473–497.CrossRefGoogle Scholar
  14. 14.
    Koncz, C. and Schell, J. (1986) The promoter of the TL-DNA gene 5 controls the tissue-specific expression of chimeric genes carried by a novel type of Agrobacterium binary vector. Mol. Gen. Genet. 213, 285–290.Google Scholar
  15. 15.
    Van Larebeke, N., Genetello, C., Hernalsteens, J.-P., DePicker, A., Zaenen, I., Messens, E., et al (1977) Transfer of Ti-plasmids between Agrobacterium strains by mobilisation with the conjugative plasmid RP4. Mol. Gen. Genet. 152, 119–124.CrossRefGoogle Scholar
  16. 16.
    Draper, J., Scott, R., Armitage, P., and Walden, R. (1988) Plant Genetic Transformation and Gene Expression: A Laboratory Manual. Blackwell, Oxford, UK.Google Scholar
  17. 17.
    Sambrook, J., Fritsch, E. F., and Mamaus, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
  18. 18.
    Dellaporta, S. L., Wood, J., and Hicks, J. B. (1983) A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1, 19–21.CrossRefGoogle Scholar
  19. 19.
    Odell, J. T., Nagy, F., and Chua, N. H. (1985) Identification of sequences required for activity of the Cauliflower mosaic virus 35 S promoter. Nature 313, 810–812.PubMedCrossRefGoogle Scholar
  20. 20.
    Marsh, J. L., Erfle, M., and Wykes, E. J. (1984) The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene 32, 481–485.PubMedCrossRefGoogle Scholar
  21. 21.
    Nagy, J. I. and Maliga, P. (1976) Callus induction and plant regeneration from mesophyll protoplasts of Nicotiana sylvestris. Z. Pfanzenphysiol. 78, 453–455.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1995

Authors and Affiliations

  • Klaus Fritze
    • 1
  • Richard Walden
    • 1
  1. 1.Max Planck Institut fur ZuchtungsforschungKolnGermany

Personalised recommendations