Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Transformation of the rodent malaria parasite Plasmodium chabaudi


The rodent malaria parasite Plasmodium chabaudi chabaudi shares many features with human malaria species, including P. falciparum, and is the in vivo model of choice for many aspects of malaria research in the mammalian host, from sequestration of parasitized erythrocytes, to antigenic variation and host immunity and immunopathology. This protocol describes an optimized method for the transformation of mature blood-stage P.c. chabaudi and a description of a vector that targets efficient, single crossover integration into the P.c. chabaudi genome. Transformed lines are reproducibly generated and selected within 14–20 d, and show stable long-term protein expression even in the absence of drug selection. This protocol, therefore, provides the scientific community with a robust and reproducible method to generate transformed P.c. chabaudi parasites expressing fluorescent, bioluminescent and model antigens that can be used in vivo to dissect many of the fundamental principles of malaria infection.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type



Prices may be subject to local taxes which are calculated during checkout

Figure 1: Overview of the generation of transformed P.c. chabaudi.
Figure 2
Figure 3: Analysis of mCherry expression in transformed P.c. chabaudi.
Figure 4: An example of vector design and genotypic analysis of transformed P.c. chabaudi parasites.


  1. Langhorne, J., Quin, S.J. & Sanni, L.A. Mouse models of blood-stage malaria infections: immune responses and cytokines involved in protection and pathology. Chem. Immunol. 80, 204–228 (2002).

    Article  CAS  PubMed  Google Scholar 

  2. Stevenson, M.M. & Riley, E.M. Innate immunity to malaria. Nat. Rev. Immunol. 4, 169–180 (2004).

    Article  CAS  PubMed  Google Scholar 

  3. Reece, S.E., Duncan, A.B., West, S.A. & Read, A.F. Host cell preference and variable transmission strategies in malaria parasites. Proc. Biol. Sci. 272, 511–517 (2005).

    Article  PubMed  PubMed Central  Google Scholar 

  4. de Roode, J.C. et al. Virulence and competitive ability in genetically diverse malaria infections. Proc. Natl. Acad. Sci. USA 102, 7624–7628 (2005).

    Article  CAS  PubMed  Google Scholar 

  5. Reece, S.E. & Thompson, J. Transformation of the rodent malaria parasite Plasmodium chabaudi and generation of a stable fluorescent line PcGFPCON. Malar. J. 7, 183 (2008).

    Article  PubMed  PubMed Central  Google Scholar 

  6. Janse, C.J., Ramesar, J. & Waters, A.P. High-efficiency transfection and drug selection of genetically transformed blood stages of the rodent malaria parasite Plasmodium berghei. Nat. Protoc. 1, 346–356 (2006).

    Article  CAS  PubMed  Google Scholar 

  7. Sponaas, A.M. et al. Migrating monocytes recruited to the spleen play an important role in control of blood stage malaria. Blood 114, 5522–5531 (2009).

    Article  CAS  PubMed  Google Scholar 

  8. Belyaev, N.N. et al. Induction of an IL7-R(+)c-Kit(hi) myelolymphoid progenitor critically dependent on IFN-gamma signaling during acute malaria. Nat. Immunol. 11, 477–485 (2010).

    Article  CAS  PubMed  Google Scholar 

  9. Franke-Fayard, B. et al. A Plasmodium berghei reference line that constitutively expresses GFP at a high level throughout the complete life cycle. Mol. Biochem. Parasitol. 137, 23–33 (2004).

    Article  CAS  Google Scholar 

  10. Menard, R. & Janse, C. Gene targeting in malaria parasites. Methods 13, 148–157 (1997).

    Article  CAS  PubMed  Google Scholar 

Download references


We thank B. Franke-Fayard (Leiden University Medical Centre, The Netherlands) for the kind gift of plasmid pL0017. This work was supported by the Medical Research Council (MRC; reference U117584248) and the Wellcome Trust (048684). P.J.S. is the recipient of a fellowship from the Leverhulme Trust, and J. Lawton is funded by an MRC PhD studentship.

Author information

Authors and Affiliations



P.J.S., D.C., W.J., J. Langhorne and J.T. designed the experiments and wrote the manuscript. P.J.S., D.C., W.J. and J. Lawton performed the parasite purification, transfection optimization and parasite visualization experiments. J.T. designed and generated transfection plasmids.

Corresponding authors

Correspondence to Jean Langhorne or Joanne Thompson.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Spence, P., Cunningham, D., Jarra, W. et al. Transformation of the rodent malaria parasite Plasmodium chabaudi. Nat Protoc 6, 553–561 (2011).

Download citation

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing