Summary: A 309 bp DNA fragment from was amplified by PCR using primers derived from the consensus TATA-box binding protein amino acid sequence. The amplified fragment was used to isolate cDNA and genomic DNA clones containing an ORF encoding the complete TATA-box binding protein ( 702 bp, 234 aa, molecular mass 26 kDa). The EhTBP functional domain showed 55% sequence identity to that of 54% to and 37% to TBPs. In Southern blot experiments we detected a single band, which was transcribed as a 1.3 kb mRNA containing a 420 nt 5′ untranslated region. However, the probe hybridized with the 0.8 and 1.5 Mb chromosomes, suggesting that this sequence is diploid. PCR assays showed two signals in 95% of trophozoites, one located in the nucleus and another in EhkO, the novel DNA-containing organelle recently reported. The recombinant TATA-box binding protein was expressed in Antibodies against it recognized two proteins of 26 and 29 kDa in nuclear extracts. Confocal microscopy immunofluorescence analysis located the protein in both the nucleus and EhkO.


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