Gene expression constructs defining elements of PfoTPT essential for targeting to the outer membrane of the apicoplast. TMDs are represented by numbered boxes and are joined by loops. All PfoTPT constructs are episomally expressed under the PfCRT promoter and tagged with triple HA at the C-terminus detected with anti-HA and secondary antibody conjugated to FITC (green) in parasites within erythrocytes. Co-localisation of the apicoplast using antisera against apicoplast stromal marker, ACP, is shown in red. Nuclei are stained with Hoechst (blue), and transmitted light images of the parasites within their host red blood cell are shown on the right. Scale bars = 2μm. A. Full length, synthetic PfoTPT colocalises

with ACP. B. Point mutation of tyrosine residue at position 10 (✪) in the synthetic PfoTPT (Y10A) abrogates targeting to the apicoplast showing no co-localisation with the apicoplast marker. C. Removal of TMD 10 (TMD1to9) abrogates targeting to the apicoplast showing no co-localisation with the apicoplast marker. D. Removal of TMDs 9 and 10 (TMD1to8) abrogates targeting to the apicoplast showing no co-localisation with the apicoplast marker. marker.

Lim L, Sayers CP, Goodman CD, McFadden GI. Targeting of a Transporter to the Outer Apicoplast Membrane in the Human Malaria Parasite Plasmodium falciparum. PLoS One. 2016 11(7):e0159603.