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Malaria pp 307-319 | Cite as

Expressing Full-Length Functional PfEMP1 Proteins in the HEK293 Expression System

  • Anand Srivastava
  • Yves Durocher
  • Benoît GamainEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 923)

Abstract

Due to the A/T-richness of the genome of Plasmodium falciparum, expressing P. falciparum proteins in heterologous expression systems is challenging. In addition, many P. falciparum proteins have high cysteine content and high molecular weight, which further complicates expression of these proteins in heterologous systems. The high molecular weight Plasmodium falciparum Erythrocyte Membrane Protein 1 (PfEMP1) adhesins expressed on the surface of the infected erythrocytes are among the most difficult proteins to express. Cost reduction in synthetic gene synthesis, as well as improved eukaryotic expression systems, now makes it possible to express such proteins. In this chapter, we describe the construction, production, purification, and functional assessment of the full-length extracellular region of the var2CSA PfEMP1 protein involved in pregnancy-associated malaria (PAM), using a human embryonic kidney (HEK) expression system.

Key words

Plasmodium falciparum Malaria PfEMP1 var var2CSA HEK293 expression system 

Notes

Acknowledgments

The research leading to these results has received funding from the European Community’s Seventh Framework Programme Grant ([FP7/2007-2013]) under Grant agreement 201222. A.S. is supported by a grant from the Fondation pour la Recherche Médicale (FRM) (SPF20101220957).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Anand Srivastava
    • 1
  • Yves Durocher
    • 2
  • Benoît Gamain
    • 1
    Email author
  1. 1.Institut National de Transfusion SanguineUniversité Paris DiderotParisFrance
  2. 2.Biotechnology Research Institute, National Research Council CanadaMontrealCanada

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