Skip to main content

Transgenesis in Drosophila melanogaster

  • Protocol
  • First Online:
Transgenesis Techniques

Part of the book series: Methods in Molecular Biology ((MIMB,volume 561))

Summary

Transgenesis in Drosophila melanogaster relies upon direct microinjection of embryos and subsequent crossing of surviving adults. The necessity of crossing single flies to screen for transgenic events limits the range of useful transgenesis techniques to those that have a very high frequency of integration, so that about 1 in 10 to 1 in 100 surviving adult flies carry a transgene. Until recently, only random P-element transgenesis fulfilled these criteria. However, recent advances have brought homologous recombination and site-directed integration up to and beyond this level of efficiency. For all transgenesis techniques in Drosophila melanogaster, microinjection of embryos is the central procedure. This chapter gives a detailed protocol for microinjection, and aims to enable the reader to use it for both site-directed integration and for P-element transgenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Venken, K.J. and Bellen, H.J. (2005) Emerging technologies for gene manipulation in Drosophila melanogaster. Nat Rev Genet, 6, 167–178

    Article  PubMed  CAS  Google Scholar 

  2. Venken, K.J. and Bellen, H.J. (2007) Transgenesis upgrades for Drosophila melanogaster. Development, 134, 3571–3584

    Article  PubMed  CAS  Google Scholar 

  3. Rubin, G.M. and Spradling, A.C. (1982) Genetic transformation of Drosophila with transposable element vectors. Science, 218, 348–353

    Article  PubMed  CAS  Google Scholar 

  4. Gong, W.J. and Golic, K.G. (2003) Ends-out, or replacement, gene targeting in Drosophila. Proc Natl Acad Sci U S A, 100, 2556–2561

    Article  PubMed  CAS  Google Scholar 

  5. Xie, H.B. and Golic, K.G. (2004) Gene deletions by ends-in targeting in Drosophila melanogaster. Genetics, 168, 1477–1489

    Article  PubMed  CAS  Google Scholar 

  6. Groth, A.C., Fish, M., Nusse, R. and Calos, M.P. (2004) Construction of transgenic Drosophila by using the site-specific integrase from phage phiC31. Genetics, 166, 1775–1782

    Article  PubMed  CAS  Google Scholar 

  7. Bischof, J., Maeda, R.K., Hediger, M., Karch, F. and Basler, K. (2007) An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases. Proc Natl Acad Sci U S A, 104, 3312–3317

    Article  PubMed  CAS  Google Scholar 

  8. Bateman, J.R., Lee, A.M. and Wu, C.T. (2006) Site-specific transformation of Drosophila via phiC31 integrase-mediated cassette exchange. Genetics, 173, 769–777

    Article  PubMed  CAS  Google Scholar 

  9. Fish, M.P., Groth, A.C., Calos, M.P. and Nusse, R. (2007) Creating transgenic Drosophila by microinjecting the site-specific phiC31 integrase mRNA and a transgene-containing donor plasmid. Nat Protoc, 2, 2325–2331

    Article  PubMed  CAS  Google Scholar 

  10. Venken, K.J., He, Y., Hoskins, R.A. and Bellen, H.J. (2006) P[acman]: a BAC transgenic platform for targeted insertion of large DNA fragments in D. melanogaster. Science, 314, 1747–1751

    Article  PubMed  CAS  Google Scholar 

  11. Ni, J.Q., Markstein, M., Binari, R., Pfeiffer, B., Liu, L.P., Villalta, C., Booker, M., Perkins, L. and Perrimon, N. (2008) Vector and parameters for targeted transgenic RNA interference in Drosophila melanogaster. Nat Methods, 5, 49–51

    Article  PubMed  CAS  Google Scholar 

  12. O’Connor, M.J. and Chia, W. (2002) Gene transfer in Drosophila. Methods Mol Biol, 180, 27–36

    PubMed  Google Scholar 

  13. Voie, A.M. and Cohen, S. (1998) Germ-Line Transformation of Drosophila melanogaster. Cell Biology: A Laboratory Handbook, Academic Press, New York, vol. 3, pp. 510–518

    Google Scholar 

Download references

Acknowledgements

I wish to thank Sara Farina Lopez and Sylvia Oppel for sharing technical expertise, and Sheetal Bhalerao for sharing unpublished data and for helpful discussions during preparation of this chapter.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Leonie Ringrose .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Ringrose, L. (2009). Transgenesis in Drosophila melanogaster . In: Cartwright, E. (eds) Transgenesis Techniques. Methods in Molecular Biology, vol 561. Humana Press. https://doi.org/10.1007/978-1-60327-019-9_1

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-019-9_1

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-018-2

  • Online ISBN: 978-1-60327-019-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics