Protocol

Parasite Genomics Protocols

Volume 270 of the series Methods in Molecular Biology™ pp 263-276

Transfection of the Human Malaria Parasite Plasmodium falciparum

  • Brendan S. CrabbAffiliated withDivision of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research
  • , Melanie RugAffiliated withDivision of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research
  • , Tim-Wolf GilbergerAffiliated withDivision of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research
  • , Jennifer K. ThompsonAffiliated withDivision of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research
  • , Tony TrigliaAffiliated withDivision of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research
  • , Alexander G. MaierAffiliated withDivision of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research
  • , Alan F. CowmanAffiliated withDivision of Infection and Immunity, The Walter and Eliza Hall Institute of Medical Research

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Abstract

Methods to transiently and stably transfect blood stages of the human malaria parasite Plasmodium falciparum have been developed and adapted for gene-knockout, allelic replacement, and transgene expression in this organism. These methods are detailed in this chapter, as are approaches used to monitor transfectants during the selection process. The different plasmid vectors that are currently used for gene targeting and transgene expression (including green fluorescent protein expression) are also described.

Key Words

Allelic replacement DHFR GFP knockout malaria negative selection Plasmodium falciparum Rep20 selectable marker thymidine kinase transfection transgene expression