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Fig. 1 | Parasites & Vectors

Fig. 1

From: The Entamoeba histolytica TBP and TRF1 transcription factors are GAAC-box binding proteins, which display differential gene expression under different stress stimuli and during the interaction with mammalian cells

Fig. 1

EhTBP and EhTRF1 have DNA-binding activity to different TATA variants in vitro. Recombinant EhTBP and EhTRF1 were expressed in E. coli BL21(DE3) pLysS, purified by IMAC and analyzed by electrophoresis as described in Methods. a SDS-PAGE (12%) gel analysis of extracts from bacteria transformed with pRSET A (Lane 2), pcEhtrf1 (Lanes 3 and 4), pEhtbp (Lanes 6 and 7). Protein expression was induced with 1 mM IPTG for rEhTRF1 and rEhTBP (Lanes 4, and 7, respectively). rEhTRF1 and rEhTBP purified proteins (arrows) are shown in Lanes 5 and 8, respectively. b Western blot of the gel shown in a using anti-His tag monoclonal antibodies. EMSA assay to determine the DNA-binding activity of purified rEhTRF1 and rEhTBP using different probes: (c) TATTTAAA(1); (d) TAgTgAAA(2); (e) TATgTAAA(5); (f) gAgTTAAA(6); (g) TAcTcAAA(7); (h) cAcTcAAA(8). Abbreviations: UC, unspecific competitor; SC, specific competitor

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