Figure 4
Generation and validation of Δpfaop knockout parasites. (a) Schematic summary of the knockout strategy by double crossover using the plasmid pL7-PFAOP. Expected product sizes from analytical PCR reactions 1–3 are highlighted. The two external primers should anneal to chromosome 7, whereas the two internal primers should anneal to the gene that encodes the selection marker human dihydrofolate reductase (hDHFR). (b) Following the genetic manipulation of the wild type strain 3D7 (3D7 WT) and isolation of three clonal cell lines (Δaop#1-3), products from PCR reactions 1-3 were analysed by agarose gel electrophoresis. Marker (M) and expected product sizes are indicated on the left and right side, respectively. (c) Western blot analysis of parasite cell lines Δaop#1-3. Protein extracts from 107 parasites were loaded per lane. Hsp70 was decorated as a control.
