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