Genetica

, Volume 98, Issue 1, pp 33–41 | Cite as

Excision of the piggyBac transposable element in vitro is a precise event that is enhanced by the expression of its encoded transposase

  • Teresa A. Elick
  • Christopher A. Bauser
  • M. J. Fraser
Article

Abstract

The piggyBac Lepidopteran transposable element moves from the cellular genome into infecting baculovirus genomes during passage of the virus in cultured TN-368 cells. We have constructed genetically tagged piggyBac elements that permit analysis of excision when transiently introduced on plasmids into the piggyBac-deficient Spodoptera frugiperda IPLB-SF21AE cell line. Precise excision of the element from these plasmids occurs at a higher frequency in the presence of a helper plasmid that presumably supplies the piggyBac transposase. The results suggest that the piggyBac transposon encodes a protein that functions to facilitate not only insertion, but precise excision as well. This is the first demonstration of piggyBac mobility from plasmid sources in uninfected Lepidopteran cells.

Key words

piggyBac Lepidopteran transposable element 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ariza, R.R., T. Roldan-Arjona, C. Hera & C. Pueyo, 1993. A method for selection of forward mutations in a supF gene carried by shuttle vector plasmids. Carcinogenesis 14: 303–305.Google Scholar
  2. Atkinson, P.W., W.D. Warren & D.A. O'Brochta, 1993. The hobo transposable element of Drosophila can be cross-mobilized in houseflies and excises like the Ac element of maize. Proc. Nat. Acad. Sci. USA 90: 9693–9697.Google Scholar
  3. Beames, B. & M.D. Summers, 1988. Comparisons of host cell DNA insertions and altered transcription at the site of insertions in few polyhedra baculovirus mutants. Virology 162: 206–220.Google Scholar
  4. Beames, B. & M.D. Summers, 1990. Sequence comparison of cellular and viral copies of host cell DNA insertions found in Autographa californica nuclear polyhedrosis virus. Virology 174: 354–363.Google Scholar
  5. Berg, D.E. & M.M. Howe, 1989. Mobile DNA, American Society for Microbiology.Google Scholar
  6. Carstens, E.B., 1987. Identification and nucleotide sequence of the regions of Autographa californica nuclear polyhedrosis virus genome carrying insertion elements derived from Spodoptera frugiperda. Virology 161: 8–17.Google Scholar
  7. Cary, L.C., M. Goebel, B.G. Corsaro, H.G. Wang, E. Rosen & M.J. Fraser, 1989. Transposon mutagenesis of Baculoviruses: Analysis of Trichoplusia ni transposon IFP2 insertions within the FP-Locus of nuclear polyhedrosis viruses. Virology 172: 156–169.Google Scholar
  8. Coen, E.S. & R. Carpenter, 1988. A semi-dominant allele, niv-525, works in trans to inhibit expression of its wild-type homologue in Antirrhinum majus. J. Eur. Mol. Bio. Org. 7: 877–883.Google Scholar
  9. Corsaro, B.G. & M.J. Fraser, 1989. Transfection of Lepidopteran insect cells with Baculovirus DNA. J. Tiss. Cul. Meth. 12: 7–12.Google Scholar
  10. Elick, T.A., C.A. Bauser, N.M. Principe & M.J. Fraser, 1996. PCR analysis of insertion site specificity, transcription, and structural uniformity of the Lepidopteran transposable element IFP2 in the TN-368 cell genome. Genetica 97: 127–139.Google Scholar
  11. Engels, W.R., D.M. Johnson-Schlitz, W.B. Eggleston & J. Sveg, 1990. High-frequency P element loss in Drosophila is homolog dependent. Cell 62: 515–525.Google Scholar
  12. Finnegan, D.J., 1990. Transposable elements and DNA transposition in eukaryotes. Curr. Opin. Cell Bio. 2: 471–477.Google Scholar
  13. Fraser, M.J., J.S. Brusca, G.E. Smith & M.D. Summers, 1985. Transposon-mediated mutagenesis of a baculovirus. Virology 145: 356–361.Google Scholar
  14. Fraser, M.J., L. Cary, K. Boonvisudhi & H.H. Wang, 1995. Assay for movement of the Lepidopteran transposon IFP2 in insect cells using a baculovirus genome as a target DNA. Virology 211: 397–407.Google Scholar
  15. Fraser, M.J., T. Ciszczon, T. Elick & C. Bauser, 1995. Precise excision of TTAA-specific Lepidopteran transposons TFP3 and IFP2 from the baculovirus genome in cell lines from two species of Lepidoptera. J. Insect Mol. Bio., in press.Google Scholar
  16. Fraser, M.J., G.E. Smith & M.D. Summers, 1983. Acquisition of host cell DNA sequences by baculoviruses: relationship between host DNA insertions and FP mutants of Autographa californica and Galleria mellonella nuclear polyhedrosis viruses. J. Virol. 47: 287–300.Google Scholar
  17. Graham, F.L. & A.J.Van der Eb, 1973. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52: 456–467.Google Scholar
  18. Handler, A.M. & S.P. Gomez, 1995. The hobo transposable element has transposase-dependent and -independent excision activity in drosophilid species. Mol. Gen. Genet. 247: 399–408.Google Scholar
  19. Hirt, B., 1967. Selective extraction of polyoma DNA from infected mouse cell cultures. J. Mol. Bio. 26: 365–369.Google Scholar
  20. Lidholm, D.A., A.R. Lohe & D.L. Hartl, 1993. The transposable element mariner mediates germline transformation in Drosophila melanogaster. Genetics 134: 859–868.Google Scholar
  21. Moerman, D.G., J.E. Kiff & R.H. Waterston, 1991. Germline excision of the transposable element Tc1 in C. elegans. Nuc. Acids Res. 19: 5669–5672.Google Scholar
  22. O'Brochta, D.A., S.P. Gomez & A.M. Handler, 1991. P element excision in Drosophila melanogaster and related drosophilids. Mol. Gen. Genet. 225: 387–394.Google Scholar
  23. O'Brochta, D.A. & A.M. Handler, 1993. Prospects and possibilities for gene transfer techniques in insects, pp. 451–488 in Molecular Approaches to Fundamental and Applied Entomology, edited by J. Oakeshott and M.J. Whitten. Springer-Verlag, New York.Google Scholar
  24. O'Brochta, D.A., W.D. Warren, K.J. Saville & P.W. Atkinson, 1994. Interplasmid transposition of Drosophila hobo elements in non-drosophilid insects. Mol. Gen. Genet 244: 9–14.Google Scholar
  25. Oellig, C., B. Happ, T. Mueller & W. Doerfler, 1987. Overlapping sets of viral RNAs reflect the array of polypeptides in the EcoRI J and N fragments (map positions 81.2 to 85.0) of the Autographa californica nuclear polyhedrosis virus. J. Virol. 61: 3048–3057.Google Scholar
  26. Rio, D.C., F.A. Laski & G.M. Rubin, 1986. Identification and immunochemical analysis of biologically active Drosophila P element transposase. Cell 44: 21–32.Google Scholar
  27. Robertson, H.M., 1995. The Tc1-mariner superfamily of transposons in animals. J. Insect Phys. 41: 99–105.Google Scholar
  28. Rubin, G.M. & A.C. Spradling, 1982. Genetic transformation of Drosophila with transposable element vectors. Science 218: 348–353.Google Scholar
  29. Sambrook, J., E.F. Fritsch & T. Maniatis, 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Press.Google Scholar
  30. Sanger, F., S. Nickien & A.R. Coulson, 1977. DNA sequencing with chain-terminating inhibitors. Proc. Nat. Acad. Sci. USA 74: 5463–5467.Google Scholar
  31. Saedler, H. & P. Nevers, 1985. Transposition in plants: a molecular model. J. Eur. Mol. Bio. Org. 4: 585–590.Google Scholar
  32. Schetter, C., C. Oellig & W. Doerffler, 1990. An insertion of insect cell DNA in the 81-map-unit segment of Autographa californica nuclear polyhedrosis virus. J. Virol. 64: 1844–1850.Google Scholar
  33. Summers, M.D. & G.E. Smith, 1987. A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin.Google Scholar
  34. Takasu-Ishikawa, E., M. Yoshihara & Y. Hotta, 1992. Extra sequences found at P element excision sites in Drosophila melanogaster. Mol. Gen. Genet. 232: 17–23.Google Scholar
  35. Vaugha, J.L., R.H. Goodwin, G. Thompkins & P. McCawley, 1977. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera: Noctuidae). In Vitro 13: 213–217.Google Scholar
  36. van Leunen, H.G.A.M., S.D. Colloms & R.H.A. Plasterk, 1994. The mechanism of transposition of Tc3 in C. elegans. Cell 79: 293–301.Google Scholar
  37. Wang, H.H. & M.J. Fraser, 1992. TTAA serves as the target site for TFP3 Lepidopteran transposon insertions in both nuclear polyhedrosis virus and Trichoplusia ni genomes. J. Insect Mol. Bio. 1: 1–7.Google Scholar
  38. Wang, H.H., M.J. Fraser & L.C. Cary, 1989. Transposon mutagenesis of baculoviruses: analysis of TFP3 lepidopteran insertions at the FP locus of nuclear polyhedrosis viruses. Gene 81: 97–108.Google Scholar
  39. Warren, W.D., P.W. Atkinson & D.A. O'Brochta, 1994. The Hermes transposable element from the house fly, Musca domestica, is a short inverted repeat-type element of the hobo, Ac, and Tum3 (hAT) element family. Genet. Res. 64: 87–97.Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Teresa A. Elick
    • 1
  • Christopher A. Bauser
    • 1
  • M. J. Fraser
    • 1
  1. 1.Department of Biological SciencesUniversity of Notre DameNotre DameUSA

Personalised recommendations