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Complementation of Plasmodium berghei TRAP knockout parasites using human dihydrofolate reductase gene as a selectable marker
journal contribution
posted on 2001-03-01, 00:00 authored by A A Sultan, V Thathy, Tania De Koning-WardTania De Koning-Ward, V NussenzweigPreviously we have used the Plasmodium dihydrofolate reductase thymidylate synthase (DHFR-TS) selectable marker to generate Plasmodium berghei TRAP null mutant parasites. These TRAP null mutants do not glide and they showed a great reduction in their ability to infect mosquito salivary glands and the hepatocytes of the vertebrate host. Thus far, complementation of these knockout parasites was not possible due to the lack of additional selectable markers. Recently, a new selectable marker, based on the human dihydrofolate reductase (hDHFR) gene, has been developed which confers resistance to the antifolate drug WR99210. This drug has been found to be highly active against pyrimethamine-sensitive and -resistant strains of P. berghei. In this study, we have used the hDHFR gene as a second selectable marker for the complementation of P. berghei TRAP null mutant parasites. Restoration of the TRAP null mutant parasites to the wild-type phenotype was achieved in this study via autonomously replicating episomes bearing a wild-type copy of the TRAP gene. This is the first report of complementation of a mutant phenotype in malaria parasites.
History
Journal
Molecular & biochemical parasitologyVolume
113Issue
1Pagination
151 - 156Publisher
ElsevierLocation
Amsterdam, The NetherlandsPublisher DOI
ISSN
0166-6851Language
engPublication classification
C1.1 Refereed article in a scholarly journalCopyright notice
2001, Elsevier ScienceUsage metrics
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No categories selectedKeywords
AnimalsFolic Acid AntagonistsGenetic MarkersHumansMutationPlasmodium bergheiProtozoan ProteinsSensitivity and SpecificityTetrahydrofolate DehydrogenaseTransfectionThrombospondin-related anonymous protein (TRAP)Human dihydrofolate reductase (hDHFR)genetic complementationScience & TechnologyLife Sciences & BiomedicineBiochemistry & Molecular BiologyParasitologyPlamodium bergheiMALARIA PARASITESTRANSFORMATIONSPOROZOITESMOSQUITOCTRPFALCIPARUMDISRUPTIONINFECTIONMOTILITYPROTEIN
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