Advantages and Limitations of Using Spm as a Transposon Tag

  • Karen C. Cone
  • Robert J. Schmidt
  • Benjamin Burr
  • Frances A. Burr
Part of the Basic Life Sciences book series (BLSC, volume 47)


Transposon tagging has become the method of choice for isolating genes whose products are in low abundance. We have recently used the transposable element Spm to tag and clone maize regulatory loci. Our choice of Spm was dictated by several factors: The frequency of transposition of Spm is high enough to obtain detectable transposition events, into loci affecting kernel traits, in populations of <106 seed. Although the copy number of Spm is high in the maize genome, insertions into the gene of interest can be distinguished from other Spm copies by digesting DNAs from segregating populations with methyl-sensitive restriction enzymes, and hybridizing with Spm-specific probes. Since all members of the Spm family thus far examined share DNA homology, hybridization with appropriate probes allows detection of insertions of both autonomous and defective elements. Thus, if a mutable allele can be shown to be under Spm control, one can be reasonably confident of successfully cloning that allele.


Transposable Element Mutable Allele Transposition Event Mapping Restriction Fragment Length Polymorphism Sensitive Restriction Enzyme 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Alleman, M., and M. Freeling (1986) The Mu transposable elements of maize: Evidence for transposition and copy number regulation during development. Genetics 112:107–119.PubMedGoogle Scholar
  2. 2.
    Chandler, V.L., and V. Walbot (1986) DNA modification of a maize transposable element correlates with loss of activity. Proc. Natl. Acad. Sci., USA 83:1767–1771.PubMedCrossRefGoogle Scholar
  3. 3.
    Chen, C.-H., K. Oishi, B. Kloeckner-Gruissem, and M. Freeling (1987) Organ-specific expression of maize Adh1 is altered after a Mu transposon insertion. Genetics 116:469–477.PubMedGoogle Scholar
  4. 4.
    Cone, K.C., F.A. Burr and B. Burr (1986) Molecular analysis of the maize anthocyanin regulatory locus C1. Proc. Natl. Acad. Sci., USA 83:9631–9635.PubMedCrossRefGoogle Scholar
  5. 5.
    Dellaporta, S.L., and P.S. Chomet (1985) The activation of maize controlling elements. In Plant Gene Research: Genetic Flux in Plants, B. Hohn and E. Dennis, eds. Springer, New York, pp. 170–217.Google Scholar
  6. 6.
    Evola, S.V., F.A. Burr, and B. Burr (1986) The suitability of restriction fragment length polymorphisms as genetic markers in maize. Theor. Appl. Genet. 71:765–771.CrossRefGoogle Scholar
  7. 7.
    Fedoroff, N., S. Wessler, and M. Shure (1983) Isolation of the transposable maize controlling elements Ac and Ds. Cell 35:235–242.PubMedCrossRefGoogle Scholar
  8. 8.
    Greenblatt, I.M. (1984) A chromosome replication pattern deduced from pericarp phenotypes resulting from movements of the transposable element, Modulator, in maize. Genetics 108:471–485.PubMedGoogle Scholar
  9. 9.
    Helentjaris, T., D.F. Weber, and S. Wright (1986) Use of monosomics to map cloned DNA fragments in maize. Proc. Natl. Acad. Sci., USA 83:6035–6039.PubMedCrossRefGoogle Scholar
  10. 10.
    Leder, P., D. Tiemeier, and L. Enquist (1977) EK2 derivatives of bacteriophage lambda useful in the cloning of DNA from higher organisms: The λgtWES system. Science 196:175–177.PubMedCrossRefGoogle Scholar
  11. 11.
    Marotta, R., G. Ponziani, M. Motto, H. Hartings, A. Gierl, N. Di Fonzo, C. Soave, A. Bianchi, and F. Salamini (1986) Genetic instability at the shrunken and waxy loci in the o2-m(r)-Bg strain of maize. Maydica 31:131–151.Google Scholar
  12. 12.
    McClintock, B. (1956) Controlling elements and the gene. Cold Spring Harbor Symposia on Quantitative Biology 21:197–216.PubMedCrossRefGoogle Scholar
  13. 13.
    McClintock, B. (1962) Topographical relations between elements of control systems in maize. Carnegie Institution of Washington Yearbook 61:448–461.Google Scholar
  14. 14.
    Motto, M., R. Marotta, N. Di Fonzo, C. Soave, and F. Salamini (1986) Ds-induced alleles at the opague-2 locus of maize. Genetics 112:121–133.PubMedGoogle Scholar
  15. 15.
    Nelson, O.E., and A.S. Klein (1984) Characterization of an Spm-controlled bronze-mutable allele in maize. Genetics 106:769–779.PubMedGoogle Scholar
  16. 16.
    Pereira, A., H. Cuypers, A. Gierl, Zs. Schwarz-Sommer, and H. Saedler (1986) Molecular analysis of the En/Spm transposable element system of Zea mays. EMBO J. 5:835–841.PubMedGoogle Scholar
  17. 17.
    Peterson, P.A. (1970) The En mutable system in maize. III. Transposition associated with mutational events. Theor. Appl. Genet. 40:367–377.CrossRefGoogle Scholar
  18. 18.
    Peterson, P.A. (1978) Controlling elements: The induction of mutability at the A2 and C loci in maize. In Maize Breeding and Genetics, D.B. Waiden, ed. John Wiley and Sons, New York, pp. 601–631.Google Scholar
  19. 19.
    Robertson, D.S. (1980) The timing of Mu activity in maize. Genetics 94:969–978.PubMedGoogle Scholar
  20. 20.
    Robertson, D.S. (1985) Differential activity of the maize mutator Muat different loci and in different cell lineages. Molec. Gen. Genet. 200:9–13.CrossRefGoogle Scholar
  21. 21.
    Schmidt, R.J., F.A. Burr, and B. Burr (1987) Transposon tagging and molecular analysis of the maize regulatory locus opague-2. Science 238:960–963.PubMedCrossRefGoogle Scholar
  22. 22.
    Shepherd, N.S. (1987) Transposable elements and gene-tagging. In Plant Molecular Biology: A Practical Approach, C.H. Shaw, ed. IRL Press, London (in press).Google Scholar
  23. 23.
    van Schaik, N.W., and R.A. Brink (1959) Transpositions of Modulator, A component of the variegated pericarp allele in maize. Genetics 44:725–738.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Karen C. Cone
    • 1
  • Robert J. Schmidt
    • 1
  • Benjamin Burr
    • 1
  • Frances A. Burr
    • 1
  1. 1.Biology DepartmentBrookhaven National LaboratoryUptonUSA

Personalised recommendations