FIG. 1.

(A) Map of the UBP2-UBP1 region of chromosome 11. Distances in kilobases are shown above the map, and the gene identity numbers are above the open reading frames. The locations of two array clones, 12I09 and 25I16, are indicated, and the positions deduced for the transcripts are beneath the map with corresponding predicted sizes. The dark bars below the map and above the putative transcripts represent EST sequences from the EMBL database. Three hypothetical open reading frames shown as open boxes downstream of TbUBP1 and TbUBP2 have repetitive AT-rich sequences, are not conserved between trypanosomatids, and are unlikely to encode proteins. (B) TbUBP2 and TbUBP1 mRNA identification. Two independent Northern blots are shown. Total RNA was prepared from either bloodstream-form (BS) or procyclic-form (PC) trypanosomes containing an integrated plasmid designed for tetracycline-inducible expression of a UBP2 stem-loop. After 24 h in the presence of tetracycline (+) the level of mRNA encoding TbUBP1 and TbUBP2 is reduced through RNAi (downward arrowheads). The first blot was hybridized with clone 12I09, stripped, and then rehybridized with a TbUBP2 coding region probe, which hybridizes with both TbUBP1 and TbUBP2 mRNA. The second blot was hybridized with clone 25I16, followed by TbUBP2. Marker sizes are in kilobases. Ethidium bromide-stained rRNA is shown beneath the blots.