Selection by flow-sorting of genetically transformed, GFP-expressing blood stages of the rodent malaria parasite, Plasmodium berghei
- 6 July 2006
- journal article
- research article
- Published by Springer Nature in Nature Protocols
- Vol. 1 (2), 614-623
- https://doi.org/10.1038/nprot.2006.88
Abstract
This protocol describes a methodology for the genetic transformation of the rodent malaria parasite Plasmodium berghei and the subsequent selection of transformed parasites expressing green fluorescent protein (GFP) by flow-sorting. It provides methods for: transfection of the schizont stage with DNA constructs that contain gfp as the selectable marker; selection of fluorescent mutants by flow-sorting; and injection of flow-sorted, GFP-expressing parasites into mice and the subsequent collection of transformed parasites. The use of two different promoters for the expression of GFP is described; these two promoters require slightly different procedures for the selection of mutants. The protocol enables the collection of transformed parasites within 10–12 days after transfection. The genetic modification of P. berghei is widely used to investigate gene function in Plasmodium sp. The application of flow-sorting to the selection of transformed parasites increases the possibilities of parasite mutagenesis, by effectively expanding the range of selectable markers.Keywords
This publication has 11 references indexed in Scilit:
- Genetic Manipulation ofPlasmodium falciparumPublished by American Society for Microbiology ,2014
- High-efficiency transfection and drug selection of genetically transformed blood stages of the rodent malaria parasite Plasmodium bergheiNature Protocols, 2006
- In vivo imaging enters parasitologyTrends in Parasitology, 2006
- High efficiency transfection of Plasmodium berghei facilitates novel selection proceduresMolecular and Biochemical Parasitology, 2006
- In vivo imaging of malaria parasites — recent advances and future directionsCurrent Opinion in Microbiology, 2005
- Murine malaria parasite sequestration: CD36 is the major receptor, but cerebral pathology is unlinked to sequestrationProceedings of the National Academy of Sciences, 2005
- The Development of Genetic Tools for Dissecting the Biology of Malaria ParasitesAnnual Review of Microbiology, 2000
- Transfection of Malaria ParasitesMethods, 1997
- Gene Targeting in Malaria ParasitesMethods, 1997
- Host diet in experimental rodent malaria: a variable which can compromise experimental design and interpretationParasitology, 1989