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. 2008 Feb 29;4(2):e1000008. doi: 10.1371/journal.ppat.1000008

Figure 5. Generation of PbACα- parasite lines.

Figure 5

(A) RNA from WT P. berghei sporozoites was reverse transcribed into cDNA and used as template to amplify ACα. Water was used as negative control (Neg) and wild type P. berghei genomic DNA (gDNA) as positive control. (B) Schematic representation of the ACα locus and the replacement vector. Correct integration of the construct results in the disrupted ACα gene as shown. Arrows indicate the position of the primers used for PCR in C. (C) Disruption of ACα was shown by PCR (left) and by Southern analysis (right). PCR on DNA of WT transfected population (before cloning) and PbACα- clones (C1 and C2) results in the amplification of two 0.7-kb WT fragments and a 0.8 and a 0.9-kb disrupted fragments when using the primers indicated in (B). Genomic Southern blot hybridization of WT and the PbACα- C1. The probe used for hybridization is represented in B. Integration of the targeting plasmid causes reduction in size of a 1.6-kb fragment in WT parasites to a 1.0-kb fragment in the PbACα- parasites. Similar results were found for PbACα- C2.