Protocol

Malaria

Volume 923 of the series Methods in Molecular Biology pp 99-125

Date:

Transfection of Rodent Malaria Parasites

  • Nisha PhilipAffiliated withDivision of Infection and Immunity Glasgow Biomedical Research Centre, Faculty of Biomedical Life Sciences and Wellcome Trust Center for Molecular Parasitology, University of Glasgow
  • , Rachael OrrAffiliated withDivision of Infection and Immunity Glasgow Biomedical Research Centre, Faculty of Biomedical Life Sciences and Wellcome Trust Center for Molecular Parasitology, University of Glasgow
  • , Andrew P. WatersAffiliated withDivision of Infection and Immunity Faculty of Biomedical Life Sciences and Wellcome Trust, Center for Molecular Parasitology Glasgow Biomedical Research Centre University of Glasgow Email author 

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Abstract

Gene manipulation is an invaluable tool to investigate and understand the biology of an organism. Although this technology has been applied to both the human and rodent malarial parasites (RMP), Plasmodium berghei in particular offers a more robust system due to a higher and more efficient transformation rate. Here, we describe a comprehensive transfection and selection protocol using P. berghei including a variant negative selection protocol administering 5-fluorocytosine to the animals in drinking water. Additionally, we discuss and assess the latest advances in gene manipulation technologies developed in RMP to gain a better understanding of Plasmodium biology.

Key words

Plasmodium berghei Transfection Reverse genetics Forward genetics Episomes Selectable markers Promoter swap Artificial chromosome