Fig. 3.

The single copy LmPPEF and TbPPEF genes express RNA constitutively through the parasite life cycles. (A) L. major genomic DNA blotting and hybridisation with a LmPPEF probe detects single bands following digestion with XhoI (X), HindIII (H), EcoRI (E), BamHI (B). Kb marker sizes are shown. (B) Trypanosoma brucei genomic DNA blotting and hybridisation with a TbPPEF probe detects single bands following digestion with BglII (Bg), EcoRI (E), HindIII (H), KpnI (K), XhoI (X). Ci, expression of LmPPEF detected by RT-PCR. Amplification of constitutively expressed NMT was used to monitor equal amounts of cDNA in the reactions. cDNAs from L. major procyclics (P), metacyclics (M), amastigotes (A), no cDNA template (0). Cii, LmPPEF RNA expression profile confirmed by blotting and hybridisation of total L. major RNA with the LmPPEF probe used in (A), using NMT hybridisation as a loading control. Di, Expression of TbPPEF detected by RT-PCR. Amplification of constitutively expressed Rab4 was used to monitor equal amounts of cDNA in the reactions. cDNAs from T. brucei procyclic (PCF) and bloodstream (BSF) forms. Dii, TbPPEF RNA expression profile confirmed by blotting and hybridisation of total T. brucei RNA with the TbPPEF probe used in (A), using β-tubulin hybridisation as a loading control.