Fig. 1

Plasmid constructs used to evaluate Plasmodium promoter sequences in P. knowlesi. Expression of hdhfr from the plasmids confers resistance to WR99210 and a fLuc or NanoLuc reporter. The pD-pfcam-Luc plasmid has the fLuc cassette driven by the P. falciparum calmodulin 5′ UTR (pfcam 5′) and the hdhfr cassette driven by the P. chabaudi dts 5′ UTR (pcdts 5′). pD-pvcam-Luc, pD-pvcrt-Luc and pD-pvhsp70-Luc have the 5′ UTR sequences of P. vivax calmodulin (pvcam 5′), chloroquine resistance transporter (pvcrt 5′) and heat shock protein 70 (pvhsp70 5′) driving fLuc expression, respectively, replacing the pfcam promoter from the pD-pfcam-Luc. Plasmids pvhsp70D-pfcam-Luc and pkef1D-pfcam-Luc have the 5′ UTR sequences of P. vivax heat shock protein 70 (pvhsp70 5′) and P. knowlesi elongation factor 1 α (pkef1-α 5′), respectively, replacing the pcdts promoter from the pD-pfcam-Luc to drive the expression of hdhfr. The pvcen-pvhsp70-D-NanoLuc plasmid includes P. vivax centromeric sequence repeats from chromosome 11 (Pv centromeric repeats) and has hdhfr-NanoLuc fusion expression driven by the P. vivax heat shock protein 70 5′ UTR (pvhsp70 5′). The arrows indicate directions of transcription