Research BriefPlasmodium falciparum:A Simple, Rapid Method for Detecting Parasite Clones in Microtiter Plates
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Cited by (47)
Plasmodium falciparum LipB mutants display altered redox and carbon metabolism in asexual stages and cannot complete sporogony in Anopheles mosquitoes
2021, International Journal for ParasitologyCitation Excerpt :Resistant parasites were then treated with 1.0 µM 5-Fluorocytosine (in DMSO) to remove single crossover-integrated plasmid and episomal forms, both of which contain the cdup negative selection marker (Maier et al., 2006). PTET-BSD-Cre (a kind gift from Alan Cowman, Walter and Eliza Hall Institute, Australia) was transfected into recombinant NF54 parasites and cultured in 2.5 µM blasticidin S-hydrochloride for 7 days, after which single-cell cloning was performed (Goodyer and Taraschi, 1997). Gametocytes were induced from PfΔlipB and wild-type (WT) NF54 parasite lines by multiple rounds of sub-culturing for 14–18 days with nutrient deprivation, as described (Ponnudurai et al., 1986).
Perforin-like protein PPLP4 is crucial for mosquito midgut infection by Plasmodium falciparum
2015, Molecular and Biochemical ParasitologyCitation Excerpt :After plasmid integration was confirmed, a mixed culture with >3% ring stages was diluted and transferred to a 96-well-plate. After three weeks of cultivation, clones were identified via Malstat assay [20] and subsequent diagnostic PCR was performed to confirm vector integration and the absence of the WT pplp4 gene locus. Two clonal lines, PPLP4(−) 1C10 and PPLP4(−) 2D6, were isolated.
A primaquine-chloroquine hybrid with dual activity against Plasmodium liver and blood stages
2013, International Journal of Medical MicrobiologyCitation Excerpt :IC50 values represent the means of four values of two independent assays carried out in duplicate. Parasite viability was determined by the Malstat assay (Goodyer and Taraschi, 1997). Synchronized ring stages of the 3D7 or Dd2 strains of P. falciparum were plated at 1% parasitemia and 5% hematocrit in RPMI 1640 medium with Albumax II (5 g/L) in 96-well microtiter plates.
Probing the multifactorial basis of Plasmodium falciparum quinine resistance: Evidence for a strain-specific contribution of the sodium-proton exchanger PfNHE
2009, Molecular and Biochemical ParasitologyPlasmodium falciparum purine nucleoside phosphorylase is critical for viability of malaria parasites
2008, Journal of Biological ChemistryCitation Excerpt :In short, plasmid-transfected parasites were selected by the addition of 2.5 nm blasticidin (InvivoGen, San Diego, CA) to the culture medium, starting 48 h post-transfection. Parasite clones were obtained by two rounds of limiting dilution using 500 μm hypoxanthine in the culture media, and identified using the MALSTAT assay reagent specific for P. falciparum lactate dehydrogenase (15, 16). Nucleic Acid Analysis—Genomic DNA of P. falciparum was purified as previously described (17).
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