Transfection of malaria parasites.
The stable genetic transformation of three phylogenetically diverse species of Plasmodium, the parasitic etiological agent of malaria, is now possible. The parasite is haploid throughout the vast majority of its life cycle. Therefore with the single selectable marker activity and protocols currently available, it is possible not only to express introduced transgenes but also to study the effects of site-specific homologous recombination such as gene knockout. Transgene expression will allow the detailed study of many aspects of the cellular biology of malaria parasites, for example, the mechanisms underlying drug resistance and protein trafficking. We describe here the methods for propagation of the two animal models (Plasmodium berghei and Plasmodium knowlesi) and for transfection of these two species and the human parasite, Plasmodium falciparum. Examples of transgene expression are given.
Waters
A P
AP
Department of Parasitology, University of Leiden, Leiden, 2300 RC, The Netherlands.
Thomas
A W
AW
van Dijk
M R
MR
Janse
C J
CJ
D000818:Animals; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D008288:Malaria / Q000469:parasitology; D016296:Mutagenesis; D010961:Plasmodium / Q000235:genetics* / Q000502:physiology; D010962:Plasmodium berghei / Q000235:genetics; D010963:Plasmodium falciparum / Q000235:genetics; D016790:Plasmodium knowlesi / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis; D013940:Thymidylate Synthase / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*; D014170:Transformation, Genetic
D016054:DNA, Protozoan; D011994:Recombinant Proteins; D013762:Tetrahydrofolate Dehydrogenase; D013940:Thymidylate Synthase