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Figure 4 | Malaria Journal

Figure 4

From: Characterization and tissue-specific expression patterns of the Plasmodium chabaudi cir multigene family

Figure 4

RT-PCR with subfamily-specific primers. (A) The cir gene structure and the location of the subfamily-specific primer pairs for cir subfamily 1 and 2. The f1up/f1lo (subfamily 1) and f2/f2lo (subfamily 2) primers are indicated as black arrows. Most cir genes consist of 3 exons (blue rectangles) with about 15 bp, 800-840 bp (highly polymorphic with highly conserved motifs), and 80-90 bp coding sequence, respectively, and two introns (100-150 bp) in conserved positions. (B) RT-PCR products of cir subfamily 1 (f1) and 2 (f2). As an example for all RT-PCR products used in the following RFLP, the RT-PCR products obtained from blood of a female NMRI mouse infected with 100 pRBCs (parasitaemia about 30%) are shown. Expected product sizes were about 600 bp. An additional smaller but much weaker RT-PCR product (approx. 300 bp) appeared in most of the amplifications which was revealed by sequencing as a further putative cir gene splice variant. These smaller cir products were excluded from all following RFLPs and only the expected RT-PCR products of about 600 bp were purified and digested. For size determination a 100 bp ladder was used.

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