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Fig. 1 | BMC Medicine

Fig. 1

From: A novel approach to identifying patterns of human invasion-inhibitory antibodies guides the design of malaria vaccines incorporating polymorphic antigens

Fig. 1

Generation and phenotypic analysis of P. falciparum lines genetically engineered to express different AMA1 alleles. a Plasmid design and integration: HB3, XIE and Pf2006 codon-optimised AMA1 alleles were transfected into W2Mef parental parasites. The single crossover event for allelic replacement of the W2Mef wild-type (WT) AMA1 with the codon-optimised AMA1 alleles (AMA1co) is illustrated. b Southern blots: genomic DNA from parental W2Mef (W2Mef WT) and genetically engineered parasite lines (W2-W2, W2-3D7, W2-FVO, W2-HB3, W2-XIE and W2-2006) were digested with restriction enzymes as indicated. Expected sizes for WT, non-integrated plasmid and for integrated codon-optimised AMA1 alleles are shown in kilobases (kb). c Differential invasion inhibition of W2Mef and genetically engineered parasite lines in the presence of W2Mef AMA1 rabbit anti-serum (1:10 dilution) in a two-cycle growth inhibition assay (GIA). Columns represent the mean percentage growth inhibition (relative to a non-inhibitory control) achieved in two separate assays tested in triplicate wells. Antibody inhibition was significantly lower for W2-3D7, W2-HB3, W2-FVO, W2-2006 and W2-XIE than for W2mef parental or W2-W2 (P < 0.05 by paired t test). d Antibody-mediated invasion inhibition of wild-type or genetically engineered P. falciparum expressing the same AMA1 allele was compared in GIAs. Total inhibition is expressed as the mean of two separate assays performed in triplicate. n = 6 for each group listed. Error bars indicate + standard error of the mean (SEM). Statistical analysis was by paired t test

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