Skip to main content

A Method to Introduce an Internal Tag Sequence into a Salmonella Chromosomal Gene

  • Protocol
  • First Online:
Salmonella

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1225))

Abstract

Epitope tags are short peptide sequences that are particularly useful for the characterization of proteins against which no antibody has been developed. Influenza hemagglutinin (HA) tag is one of the most widely used epitope tags as several valuable monoclonal and polyclonal antibodies that can be used in various techniques are commercially available. Therefore, adding a HA tag to a protein of interest is quite helpful to get rapid and cost less information regarding its localization, its expression or its biological function. In this chapter, we describe a process, derived from the Datsenko and Wanner procedure, which allows the introduction of an internal 2HA tag sequence into a chromosomal gene of the bacterial pathogen Salmonella.

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

Access this chapter

eBook
USD 39.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 54.99
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. Ochman H, Groisman EA (1996) Distribution of pathogenicity islands in Salmonella spp. Infect Immun 64:5410–5412

    PubMed  CAS  PubMed Central  Google Scholar 

  2. Hensel M, Shea JE, Gleeson C et al (1995) Simultaneous identification of bacterial virulence genes by negative selection. Science 269:400–403

    Article  PubMed  CAS  Google Scholar 

  3. Figueira R, Holden DW (2012) Functions of the Salmonella pathogenicity island 2 (SPI-2) type III secretion system effectors. Microbiology 158:1147–1161. doi:10.1099/mic.0.058115-0

    Article  PubMed  CAS  Google Scholar 

  4. Stein MA, Leung KY, Zwick M et al (1996) Identification of a Salmonella virulence gene required for formation of filamentous structures containing lysosomal membrane glycoproteins within epithelial cells. Mol Microbiol 20:151–164. doi:10.1111/j.1365-2958.1996.tb02497.x

    Article  PubMed  CAS  Google Scholar 

  5. Brumell JH, Goosney DL, Finlay BB (2002) SifA, a type III secreted effector of Salmonella typhimurium, directs Salmonella-induced filament (Sif) formation along microtubules. Traffic 3:407–415

    Article  PubMed  CAS  Google Scholar 

  6. Zhang L, Hernan R, Brizzard B (2001) Multiple tandem epitope tagging for enhanced detection of protein expressed in mammalian cells. Mol Biotechnol 19:313–321. doi:10.1385/MB:19:3:313

    Article  PubMed  CAS  Google Scholar 

  7. Henry T, Couillault C, Rockenfeller P et al (2006) The Salmonella effector protein PipB2 is a linker for kinesin-1. Proc Natl Acad Sci U S A 103:13497–13502. doi:10.1073/pnas.0605443103

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  8. Freeman JA, Ohl ME, Miller SI (2003) The Salmonella enterica serovar typhimurium translocated effectors SseJ and SifB are targeted to the Salmonella-containing vacuole. Infect Immun 71:418–427

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  9. Schroeder N, Henry T, de Chastellier C et al (2010) The virulence protein SopD2 regulates membrane dynamics of Salmonella-containing vacuoles. PLoS Pathog 6:e1001002. doi:10.1371/journal.ppat.1001002

    Article  PubMed  PubMed Central  Google Scholar 

  10. Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645. doi:10.1073/pnas.120163297

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  11. Brown NF, Szeto J, Jiang X et al (2006) Mutational analysis of Salmonella translocated effector members SifA and SopD2 reveals domains implicated in translocation, subcellular localization and function. Microbiology 152:2323–2343. doi:10.1099/mic.0.28995-0

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

W.Z. was supported by the China Scholarship Council (CSC Grant). The authors thank Aude-Agnès Blanchoin for her excellent technical assistance.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Stéphane Méresse .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Zhao, W., Méresse, S. (2015). A Method to Introduce an Internal Tag Sequence into a Salmonella Chromosomal Gene. In: Schatten, H., Eisenstark, A. (eds) Salmonella. Methods in Molecular Biology, vol 1225. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1625-2_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1625-2_5

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1624-5

  • Online ISBN: 978-1-4939-1625-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics