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Fig. 1 | Malaria Journal

Fig. 1

From: Plasmodium Rab5b is secreted to the cytoplasmic face of the tubovesicular network in infected red blood cells together with N-acylated adenylate kinase 2

Fig. 1

Complementation of Plasmodium berghei N-acylated Rab5b with conventional Rab5 and Rab5b from other apicomplexan parasites. a Schematic representation of PbRab5b genomic-locus replacement strategy. The targeting construct, consisting of the 5′ and 3′ regions (two black boxes) and PbRab5b open reading frame (ORF) (orange) fused to mAG (dark green), P. berghei DHFR 3′ untranslated region (PbDT) (blue), and the selectable marker TgDHFR expression cassette (white), was integrated into the PbRab5b genomic locus (orange) by double-crossover homologous recombination. The positions of the four PCR primers used to confirm the plasmid integration (right panel) are indicated. b Growth curve of the wild-type parasites (blue) and transgenic parasites for which the PbRab5b open reading frame replaced with PbRab5b-mAG (red). Bars standard deviation (n = 5). c Multiple amino acid sequence alignment of Plasmodium Rab5 isoforms. The GTP-binding consensus sequences and lipid modification sites are indicated red and blue boxes, respectively. The effector regions are shown as grey, black or magenta boxes. The conserved glutamine residue which was mutated to create a constitutively active mutant is shown in a red arrow. d Complementation test of the PbRab5b knockout. The coding region of PbRab5b was replaced with each gene-of-interest (GOI). The result of the complementation analysis is shown at the right; +, complemented or −, uncomplemented (n = 3)

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