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. 2008 Sep 22;7:183. doi: 10.1186/1475-2875-7-183

Figure 2.

Figure 2

Integration of PbCON into P. chabaudi ssu-rrna. Schematic representation of P. chabaudi ssu-rrna locus on contig827 (A). Schematic representation of linearized plasmid PbCON containing the Tgdhfr-ts cassette, conferring pyrimethamine resistance and gfp, flanked by P. berghei d-ssu-rrna target sequences for integration (B). Schematic representation of the contig827 ssu-rrna locus in PcGFPCON clone 2.3, following integration of the PbCON plasmid (C). D: tgdhfr is present in PcGFPCON (lane 1) but not wt (lane 2) DNA; amplification with L190/L191. Circular PbGFPCON plasmid is absent in PcGFPCON (lane 3); circular PbGFPCON (lane 4); amplification with PlF2/Pl5'R. Verification of the 5' integration site; Pc5'F/Pl5'R amplify a product in PcGFPCON (lane 5) but not in wt (lane 6) DNA. Verification of the 3' integration site; Pl3'F/Pc3'R amplify a product in PcGFPCON (lane 7) but not wt (lane 8) DNA. E: Tgdhfr is present in the genome of PcGFPCON but not wt parasites. wt (lanes 1 and 2) and PcGFPCON (lanes 3 and 4) genomic DNA digested with NheI (lanes 1 and 3) or HindIII (lanes 2 and 4). Integration of PcGFP into the P. chabaudi ssu-rrna locus on contig827 results in restriction enzyme digestion products of 9.4 and 15.4 kb following NheI and HindIII digestion respectively. N, NheI; H, HindIII.