Skip to main content
SpringerLink
Log in

Tn611 Transposon Mutagenesis in Mycobacterium smegmatis Using a Temperature-Sensitive Delivery System

  • Protocol
Mycobacteria Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 101))

  • 1669 Accesses

Abstract

Mobile generic elements are discrete DNA fragments encoding their own transposltlon functions, which enable them to move from one site in the DNA to another, independently of host-recombination functions (such as RecA). Recombination by transposition occurs without homology between the sequences of the transposon and the target sequence (1). Moreover, most of these elements do not exhibit a strong specificity for the target-site sequence, thus allowing these elements to be widely used for mutagenesis in bacteria (2).

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Galas, D. J and Chandler, M (1989) Bacterial insertion sequences, in Mobile DNA (Berg, D E and Howe, M M., eds), American Society for Microbiology, Washington, DC, pp 109–162.

    Google Scholar 

  2. Berg, D. E (1989) Transposon Tn5, in Mobile DNA, (Berg, D. E and Howe, MM., eds.), American Society for Microbiology, Washington, DC, pp 185–210.

    Google Scholar 

  3. Craig, N L. and Kleckner, N (1988) Transposition and site-specific recombination, in Escherichia coli and Salmonella typhimurium Cellular and Molecular Biology,(Neidhart, F E A, eds.), American Society for Microbiology, Washington, DC, pp. 1054–1070.

    Google Scholar 

  4. Murphy, E (1989) Transposable elements in Gram-positive bacteria, in Mobile DNA (Berg, D. E and Howe, M. M, eds), American Society for Microbiology, Washington DC, pp. 269–288

    Google Scholar 

  5. McAdam, R A, Guilhot, C, and Gicquel, B. (1994) Transposition in Mycobacteria, in Tuberculosis (Bloom, B. R, eds), American Society for Microbiology, Washington, D C, pp. 199–216

    Google Scholar 

  6. Green, E P, Tizard, M. L V, Moss, M T, Thomson, J., Winterbourne, D J, McFadden, J. J., and Hermon-Taylor, J (1989) Sequence and characteristics of IS900, an insertion element identified in a human Crohn’s disease isolate of Mycobacterium paratuberculosls. Nucleic Acids Res 17, 9063–9073

    Article  PubMed  CAS  Google Scholar 

  7. Thierry, D., Cave, M D, Ersenach, K. D, Crawford, J. T., Bates, J H., Gicquel, B, and Guesdon, J. L (1990) IS61I0, an IS-like element of Mycobacterium tuberculosis complex Nucleic Acids Res. 18, 188.

    Article  PubMed  CAS  Google Scholar 

  8. Guilhot, C, Gicquel, B, Davies, J., and Martin, C. (1992) Isolation and analysis of IS6120, a new insertion sequence from Mycobacterium smegmatis. Mol Microbiol 6, 107–113

    Article  PubMed  CAS  Google Scholar 

  9. Cirillo, J D., Barletta, R G, Bloom, B R, and Jacobs, W R. J (1991) A novel transposon trap for mycobacteria isolation and characterization of IS1096 J Bacteriol 173, 7772–7780

    CAS  Google Scholar 

  10. Garcia, M J, Guilhot, C., Lathigra, R, Mendez, C, Domenech, P., Moreno, C, Gicquel, B, and Martin, C. (1994) Insertion sequence IS1137, a new IS3 family element from Mycobacterium smegmatis. Microbiology 140, 2821–2828

    Article  PubMed  CAS  Google Scholar 

  11. Martin, C, Timm, J., Rauzier, J., GÓmez-Lus, R., Davies, J., and Gicquel, B (1990) Transposition of an antibiotic resistance element in mycobacteria Nature 345, 739–743.

    Article  PubMed  CAS  Google Scholar 

  12. Guilhot, C., Otal, I., Rompaey, I. V., Martin, C., and Gicquel, B (1994) Efficient transposition in mycobacteria: construction of Mycobacterium smegmatis insertional mutant libraries J Bacteriol 176, 535–539

    PubMed  CAS  Google Scholar 

  13. Guilhot, C, Gicquel, B, and Martin, C (1992) Temperature-sensitive mutants of the Mycobacterium plasmid pAL5000. FEMS Microbiol Lett. 98, 181–186

    Article  CAS  Google Scholar 

  14. Sambrook, J., Fritsch, E. F, and Maniatis, T (1989) Molecular Cloning A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  15. Gavigan, J, Guilhot, C, Gicquel, B, and Martin, C (1995) Use of conjugative and thermosensitive cloning vectors for transposon delivery to Mycobacterium smegmatis. FEMS Microbiol Lett 121, 35–39

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departamento de Microbiologia, Medicina, Preventiva y Salud Publica, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain

    Esther Pérez & Isabel Otal

  2. Department of Microbiology, University College, Cork, Ireland

    Julie-Ann Gavigan

  3. Unité de Génétique Mycobacterienne, Department de Bacteriologie et Mycologie, Institut Pasteur, Paris, France

    Christophe Guilhot & Brigitte Giquel

  4. Departmento de Microbiologia, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain

    Vladimir Pelicic & Carlos Martin

Authors
  1. Esther Pérez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Julie-Ann Gavigan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Isabel Otal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christophe Guilhot
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vladimir Pelicic
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Brigitte Giquel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Carlos Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. London School of Hygiene and Tropical Medicine, London, UK

    Tanya Parish  & Neil G. Stoker  & 

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Pérez, E. et al. (1998). Tn611 Transposon Mutagenesis in Mycobacterium smegmatis Using a Temperature-Sensitive Delivery System. In: Parish, T., Stoker, N.G. (eds) Mycobacteria Protocols. Methods in Molecular Biology™, vol 101. Humana Press. https://doi.org/10.1385/0-89603-471-2:187

Download citation

  • DOI: https://doi.org/10.1385/0-89603-471-2:187

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-471-6

  • Online ISBN: 978-1-59259-576-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation