Fig. 4

Deletion of the NcACBP gene did not affect parasite morphology, virulence or apicoplast biogenesis. a Schematic illustration of the NcACBP knockout. b-i Identified the knockout plasmids by PCR; each lane represents a different clone. b-ii The genomic PCR identification of the ΔNcACBP strain. The position of the primers was shown in the pattern diagram. The numbers #1-#8 represent different clones. c Quantitative RT-PCR was used to analyze the transcription levels of the NcACBP gene in the ΔNcACBP clones and in the wild-type. d Plaque assay comparing the growth of ΔNcACBP clones and wild-type parasites. The growth ability of parasites was evaluated by the number of plaques (d-ii) and the plaque sizes (d-iii). e Intracellular parasite replication of ΔNcACBP was compared with wild-type. Data were compiled from three independent assays, and 100 total PVs of each strain were counted in each assay. f Mouse survival after infection with ΔNcACBP or Nc-1. BALB/c mouse (n = 5) were injected i.p. with 5 × 10⁶ parasites. The data were representative of three experiments with similar outcomes. g Detection of apicoplasts in the Nc-1 and ΔNcACBP strains. Apicoplasts were stained with mouse anti-NcENR antibodies. Scale-bars: 2.5 μm