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Fig. 1 | Malaria Journal

Fig. 1

From: Identification of Plasmodium falciparum specific translation inhibitors from the MMV Malaria Box using a high throughput in vitro translation screen

Fig. 1

Development of a luciferase based in vitro translation assay in Plasmodium falciparum. a Workflow for preparing lysate for in vitro translation assay. b Plasmid construct to generate mRNA transcripts containing a P. falciparum specific 5′ and 3′ UTR with a firefly luciferase open reading frame. Maxipreps of the plasmid were digested with PvuII and BamHI to create the templates for T7 transcription to make a final mRNA with the P. falciparum UTRs and firefly luciferase. c Lysates were incubated in the presence of a 10 × magnesium-containing translation buffer and luciferase mRNAs for a time course of 30 min to 120 min followed by the addition of luciferin reagent to assay for luciferase activity. d Lysates were incubated with DMSO control only or with 5 μM cycloheximide before or after the 120-min incubation followed by assaying for luciferase output

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Malaria Journal

ISSN: 1475-2875

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